
1
UI  - 21025
AU  - Lipmann F
TI  - Coupling between pyruvic acid dehydrogenation and adenylic acid phosphorylation.
MH  - ACID
MH  - coupling
MH  - Phosphorylation
RP  - NOT IN FILE
SO  - Nature 1939  ;143():281-283

2
UI  - 21026
AU  - Lipmann F
TI  - Metabolic generation and utilization of phosphate bond energy
MH  - Oxidative Phosphorylation
RP  - NOT IN FILE
SO  - Advances Enzymol 1941  ;I():99-105

3
UI  - 9894
AU  - Williams RJP
MH  - Enzymes
MH  - local coupling
MH  - chemiosmotic theory
T2  - The Enzymes
A2  - Myrback BaL
PB  - New York: Academic Press
RP  - NOT IN FILE
SO  -  1959  ;2(1):

4
UI  - 8325
AU  - Mitchell P
TI  - Coupling of photophosphorylation to electron and hydrogen transfer by a chemiosmotic type of mechanism
MH  - electron
MH  - Hydrogen
MH  - mechanism
MH  - Photophosphorylation
MH  - transport
RP  - IN FILE
SO  - Nature 1961  ;191():144-148

5
UI  - 8423
AU  - Williams RJP
TI  - Possible functions of chains of catalysts
MH  - local coupling
MH  - chemiosmotic theory
RP  - IN FILE
SO  - J Theor Biol 1961  ;1():1-13

6
UI  - 19852
AU  - Boyer PD
AU  - Slater EC
TI  - Labelling rates and detection of intermediates in mitochondrial phosphorylations and other sequential reactions
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - England
MH  - Phosphorus
MH  - Phosphorylation
RP  - NOT IN FILE
NT  - UI - 66156299LA - engRN - 0 (Phosphorus Isotopes)RN - 56-65-5 (Adenosine Triphosphate)RN - 7723-14-0 (Phosphorus)PT - Journal ArticleDA - 19660930IS - 0028-0836SB - IMCY - ENGLANDJC - NSC
UR  - PM:5885856
SO  - Nature 1965 Jul 24 ;207(995):409-412

7
UI  - 1055
AU  - Mitchell P
AU  - Moyle J
TI  - Stoichiometry of proton translocation through the respiratory chain and adenosine triphosphatase systems of rat liver mitochondria
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - Animal
MH  - enzymology
MH  - Hydroxides
MH  - Hydroxybutyrates
MH  - In Vitro
MH  - Liver
MH  - metabolism
MH  - Mitochondria
MH  - pharmacology
MH  - Potassium
MH  - Potassium Chloride
MH  - Rats
MH  - Succinates
RP  - NOT IN FILE
SO  - Nature 1965 Oct 9 ;208(6):147-151

8
UI  - 1054
AU  - Mitchell P
AU  - Moyle J
TI  - Evidence discriminating between the chemical and the chemiosmotic mechanisms of electron transport phosphorylation
MH  - Animal
MH  - Cattle
MH  - Electron Transport
MH  - Liver
MH  - Mitochondria
MH  - Myocardium
MH  - Oxidative Phosphorylation
MH  - Phosphorylation
MH  - Rats
RP  - NOT IN FILE
SO  - Nature 1965 Dec 18 ;208(16):1205-1206

9
UI  - 671
AU  - Penefsky HS
AU  - Warner RC
TI  - Partial resolution of the enzymes catalyzing oxidative phosphorylation. VI. Studies on the mechanism of cold inactivation of mitochondrial adenosine triphosphatase
RP  - NOT IN FILE
NT  - UI - 66047457LA - engRN - 0 (Nitrates)RN - 7447-40-7 (Potassium Chloride)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19660212IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:4221250
SO  - J Biol Chem 1965 Dec ;240(12):4694-4702

10
UI  - 19851
AU  - Bieber LL
AU  - Boyer PD
TI  - 32P-labeling of mitochondrial protein and lipid fractions and their relation to oxidative phosphorylation
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - Glycosides
MH  - Oligomycins
MH  - Oxidative Phosphorylation
MH  - Phospholipids
MH  - Phosphorus
MH  - Phosphorylation
MH  - Proteins
RP  - NOT IN FILE
NT  - UI - 67058417LA - engRN - 0 (Glycosides)RN - 0 (Oligomycins)RN - 0 (Phospholipids)RN - 0 (Phosphorus Isotopes)RN - 0 (Proteins)RN - 56-65-5 (Adenosine Triphosphate)PT - Journal ArticleDA - 19670306IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:5954803
SO  - J Biol Chem 1966 Nov 25 ;241(22):5375-5383

11
UI  - 19850
AU  - Boyer PD
AU  - Bieber LL
AU  - Mitchell RA
AU  - Szabolcsi G
TI  - The apparent independence of the phosphorylation and water formation reactions from the oxidation reactions of oxidative phosphorylation
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - Antimycin A
MH  - Chelating Agents
MH  - Cyanides
MH  - Oxidative Phosphorylation
MH  - Phosphorylation
MH  - Rotenone
MH  - Water
RP  - NOT IN FILE
NT  - UI - 67058418LA - engRN - 0 (Chelating Agents)RN - 0 (Cyanides)RN - 56-65-5 (Adenosine Triphosphate)RN - 57-43-2 (Amobarbital)RN - 642-15-9 (Antimycin A)RN - 7732-18-5 (Water)RN - 83-79-4 (Rotenone)PT - Journal ArticleDA - 19670306IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:5954804
SO  - J Biol Chem 1966 Nov 25 ;241(22):5384-5390

12
UI  - 8083
AU  - Mitchell P
TI  - Chemiosmotic coupling in oxidative and photosynthetic phosphorylation
MH  - chemiosmotic theory
MH  - Phosphorylation
RP  - NOT IN FILE
SO  - Physiol Rev 1966  ;41():445-502

13
UI  - 1005
AU  - Mitchell P
TI  - Chemiosmotic coupling in oxidative and photosynthetic phosphorylation
MH  - Phosphorus
MH  - Phosphorylation
RP  - NOT IN FILE
NT  - UI - 66171111LA - engRN - 7723-14-0 (Phosphorus)PT - Journal ArticlePT - ReviewDA - 19661018IS - 0006-3231SB - IMCY - ENGLANDJC - A40
UR  - PM:5329743
SO  - Biol Rev Camb Philos Soc 1966 Aug ;41(3):445-502

14
UI  - 1053
AU  - Mitchell P
TI  - Chemiosmotic coupling in oxidative and photosynthetic phosphorylation
MH  - Electron Transport
MH  - metabolism
MH  - Oxidative Phosphorylation
MH  - Phosphorus
MH  - Phosphorylation
RP  - NOT IN FILE
SO  - Biol Rev Camb Philos Soc 1966 Aug ;41(3):445-502

15
UI  - 564
AU  - Pedersen TA
AU  - Kirk M
AU  - Bassham JA
TI  - Inhibition of photophosphorylation and photosynthetic carbon cycle reactions by fatty acids and esters
RP  - NOT IN FILE
NT  - UI - 66162177LA - engRN - 0 (Antimetabolites)RN - 0 (Fatty Acids)RN - 0 (Heptoses)RN - 0 (Hexosephosphates)RN - 0 (Pentosephosphates)RN - 124-38-9 (Carbon Dioxide)RN - 56-65-5 (Adenosine Triphosphate)RN - 62-46-4 (Thioctic Acid)PT - Journal ArticleDA - 19661009IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:5329024
SO  - Biochim Biophys Acta 1966 Feb 7 ;112(2):189-203

16
UI  - 1052
AU  - Reid RA
AU  - Moyle J
AU  - Mitchell P
TI  - Synthesis of adenosine triphosphate by a protonmotive force in rat liver mitochondria
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - Animal
MH  - biosynthesis
MH  - cytology
MH  - Hydrogen-Ion Concentration
MH  - Liver
MH  - Male
MH  - metabolism
MH  - Mitochondria
MH  - Oligomycins
MH  - pharmacology
MH  - Phenylhydrazines
MH  - Rats
RP  - NOT IN FILE
SO  - Nature 1966 Oct 15 ;212(59):257-258

17
UI  - 19849
AU  - Shavit N
AU  - Boyer PD
TI  - Source of oxygen in adenosine triphosphate formed by illumination or by acid-base transition of chloroplasts
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - chloroplast
MH  - Chloroplasts
MH  - Oxygen
MH  - Oxygen Isotopes
MH  - Water
RP  - NOT IN FILE
NT  - UI - 67053248LA - engRN - 0 (Oxygen Isotopes)RN - 56-65-5 (Adenosine Triphosphate)RN - 7732-18-5 (Water)RN - 7782-44-7 (Oxygen)PT - Journal ArticleDA - 19670225IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:5928211
SO  - J Biol Chem 1966 Dec 10 ;241(23):5738-5740

18
UI  - 19848
AU  - Hill RD
AU  - Boyer PD
TI  - Inorganic orthophosphate activation and adenosine diphosphate as the primary phosphoryl acceptor in oxidative phosphorylation
MH  - ACTIVATION
MH  - Adenine Nucleotides
MH  - ADENINE-NUCLEOTIDES
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - Nucleotides
MH  - Oxidative Phosphorylation
MH  - Oxygen
MH  - Oxygen Isotopes
MH  - Phosphates
MH  - Phosphorus
MH  - Phosphorylation
RP  - NOT IN FILE
NT  - UI - 68086645LA - engRN - 0 (Adenine Nucleotides)RN - 0 (Oxygen Isotopes)RN - 0 (Phosphates)RN - 0 (Phosphorus Isotopes)RN - 56-65-5 (Adenosine Triphosphate)PT - Journal ArticleDA - 19680210IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:6070842
SO  - J Biol Chem 1967 Oct 10 ;242(19):4320-4323

19
UI  - 658
AU  - Hinkle PC
AU  - Penefsky HS
AU  - Racker E
TI  - Partial resolution of the enzymes catalyzine oxidative phosphorylation. XII. The H-2-18-O-inorganic phosphate and H-2-18-O-adenosine triphosphate exchange reactions in submitochondrial particles from beef heart
RP  - NOT IN FILE
NT  - UI - 67139788LA - engRN - 0 (Oligomycins)RN - 0 (Oxygen Isotopes)RN - 0 (Phosphates)RN - 0 (Phosphorus Isotopes)RN - 56-65-5 (Adenosine Triphosphate)PT - Journal ArticleDA - 19670706IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:6024769
SO  - J Biol Chem 1967 Apr 25 ;242(8):1788-1792

20
UI  - 1051
AU  - Mitchell P
TI  - Translocations through natural membranes
MH  - Adenine Nucleotides
MH  - Adenosinetriphosphatase
MH  - Amino Acids
MH  - Animal
MH  - Biological Transport
MH  - Diffusion
MH  - Electron Transport
MH  - Glucose
MH  - Hamsters
MH  - Hemoglobins
MH  - Intestinal Absorption
MH  - Kidney
MH  - Lipoproteins
MH  - Mathematics
MH  - Membranes
MH  - metabolism
MH  - Mitochondria
MH  - Models,Biological
MH  - Myoglobin
MH  - Oxygen
MH  - physiology
MH  - Potassium
MH  - Rats
MH  - Sodium
RP  - NOT IN FILE
SO  - Adv Enzymol Relat Areas Mol Biol 1967  ;29:33-87.():33-87

21
UI  - 1048
AU  - Mitchell P
TI  - Proton current flow in mitochondrial systems
MH  - Animal
MH  - Electron Transport
MH  - Hydrogen-Ion Concentration
MH  - In Vitro
MH  - Membrane Potentials
MH  - Mitochondria,Liver
MH  - Oxidative Phosphorylation
MH  - Protons
MH  - Rats
RP  - NOT IN FILE
SO  - Nature 1967 Jun 24 ;214(95):1327-1328

22
UI  - 1049
AU  - Mitchell P
TI  - Intramitochondrial pH: a statistical question of hydrogen ion concentration in a small element of space-time
MH  - Hydrogen
MH  - Hydrogen-Ion Concentration
MH  - Mitochondria
RP  - NOT IN FILE
SO  - Nature 1967 Apr 22 ;214(86):400

23
UI  - 1047
AU  - Mitchell P
AU  - Moyle J
TI  - Acid-base titration across the membrane system of rat-liver mitochondria. Catalysis by uncouplers
MH  - Animal
MH  - Anoxia
MH  - Catalysis
MH  - Dinitrophenols
MH  - Hydrogen-Ion Concentration
MH  - In Vitro
MH  - Kinetics
MH  - Liver
MH  - Male
MH  - Membrane Potentials
MH  - metabolism
MH  - Mitochondria
MH  - Models,Biological
MH  - Oxidative Phosphorylation
MH  - Permeability
MH  - pharmacology
MH  - Rats
RP  - NOT IN FILE
SO  - Biochem J 1967 Aug ;104(2):588-600

24
UI  - 1046
AU  - Mitchell P
TI  - Proton-translocation phosphorylation in mitochondria, chloroplasts and bacteria: natural fuel cells and solar cells
MH  - Adenosinetriphosphatase
MH  - Bacteria
MH  - Biological Transport
MH  - Cells
MH  - Chloroplasts
MH  - Electron Transport
MH  - Mitochondria
MH  - Models,Biological
MH  - Oxidation-Reduction
MH  - Oxidative Phosphorylation
MH  - Phosphorylation
MH  - physiology
MH  - Protons
RP  - NOT IN FILE
SO  - Fed Proc 1967 Sep ;26(5):1370-1379

25
UI  - 1050
AU  - Mitchell P
AU  - Moyle J
TI  - Chemiosmotic hypothesis of oxidative phosphorylation
MH  - Adenosine Triphosphate
MH  - Animal
MH  - biosynthesis
MH  - Chloroplasts
MH  - In Vitro
MH  - Liver
MH  - metabolism
MH  - Mitochondria
MH  - Nad
MH  - Oxidative Phosphorylation
MH  - pharmacology
MH  - Phosphorylation
MH  - Rats
MH  - Rotenone
MH  - Spectrophotometry
RP  - NOT IN FILE
SO  - Nature 1967 Jan 14 ;213(72):137-139

26
UI  - 668
AU  - Penefsky HS
TI  - Partial resolution of the enzymes catalyzing oxidative phosphorylation. XVI. Chemical modification of mitochondrial adenosine triphosphatase
RP  - NOT IN FILE
NT  - UI - 71013327LA - engRN - 0 (Carbon Isotopes)RN - 0 (Cyclohexanes)RN - 0 (Imides)RN - 0 (Iodine Isotopes)RN - 0 (Phosphorus Isotopes)RN - 0 (Sulfhydryl Compounds)RN - 53-84-9 (NAD)RN - 7439-95-4 (Magnesium)RN - 7553-56-2 (Iodine)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19701210IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:4319672
SO  - J Biol Chem 1967 Dec 25 ;242(24):5789-5795

27
UI  - 670
AU  - Schatz G
AU  - Penefsky HS
AU  - Racker E
TI  - Partial resolution of the enzymes catalyzing oxidative phosphorylation. XIV
RP  - NOT IN FILE
NT  - UI - 67165365LA - engRN - 0 (Immune Sera)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19670803IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:4226007
SO  - J Biol Chem 1967 May 25 ;242(10):2552-2560

28
UI  - 19847
AU  - Shavit N
AU  - Skye GE
AU  - Boyer PD
TI  - Occurrence and possible mechanism of 32P and 18O exchange reactions of photophosphorylation
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - mechanism
MH  - Oxygen
MH  - Oxygen Isotopes
MH  - phenazines
MH  - Phosphates
MH  - Phosphorus
MH  - Photophosphorylation
MH  - Water
RP  - NOT IN FILE
NT  - UI - 68048420LA - engRN - 0 (Ferrocyanides)RN - 0 (Oxygen Isotopes)RN - 0 (Phenazines)RN - 0 (Phosphates)RN - 0 (Phosphorus Isotopes)RN - 50-81-7 (Ascorbic Acid)RN - 56-65-5 (Adenosine Triphosphate)RN - 7732-18-5 (Water)RN - 7782-44-7 (Oxygen)PT - Journal ArticleDA - 19680119IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:6058949
SO  - J Biol Chem 1967 Nov 10 ;242(21):5125-5130

29
UI  - 261
AU  - Hachimori A
AU  - Nosoh Y
AU  - Sone N
TI  - Effects of photosynthetic inhibitors on the respiration of yeast mitochondria
MH  - CA69(15):56960e
MH  - inhibitor
MH  - Mitochondria
MH  - Respiration
RP  - NOT IN FILE
SO  - J Biochem (Tokyo) 1968  ;64():119-121

30
UI  - 604
AU  - Hatefi Y
AU  - Fakouh T
TI  - Control of beta-hydroxybutyrate and acetoacetate oxidation by inorganic phosphate and adenosine-5'-diphosphate in heart mitochondria
RP  - NOT IN FILE
NT  - UI - 68238498LA - engRN - 0 (Acetoacetates)RN - 0 (Adenine Nucleotides)RN - 0 (Dinitrophenols)RN - 0 (Guanine Nucleotides)RN - 0 (Hydroxybutyrates)RN - 0 (Ketoglutaric Acids)RN - 0 (Malates)RN - 0 (Phosphates)RN - 0 (Succinates)RN - 56-65-5 (Adenosine Triphosphate)RN - 7440-38-2 (Arsenic)RN - 85-61-0 (Coenzyme A)PT - Journal ArticleDA - 19680708IS - 0003-9861SB - IMCY - UNITED STATESJC - 6SK
UR  - PM:5649508
SO  - Arch Biochem Biophys 1968 Apr ;125(1):114-125

31
UI  - 868
AU  - Jackson JB
AU  - Crofts AR
AU  - von Stedingk LV
TI  - Ion transport induced by light and antibiotics IN CHROMATOPHORES FROM Rhodospirillum rubrum
RP  - NOT IN FILE
NT  - UI - 69060797LA - engRN - 0 (Antibiotics)RN - 0 (Phosphates)RN - 0 (Phosphorus Isotopes)RN - 1333-74-0 (Hydrogen)RN - 1404-88-2 (Tyrothricin)RN - 7440-09-7 (Potassium)RN - 7447-40-7 (Potassium Chloride)PT - Journal ArticleDA - 19690204IS - 0014-2956SB - IMCY - GERMANY, WESTJC - EMZ
UR  - PM:5725812
SO  - Eur J Biochem 1968 Oct 17 ;6(1):41-54

32
UI  - 9973
AU  - Jackson JB
AU  - Crofts AR
TI  - Energy-linked reduction of nicotinamide adenine dinucleotides in cells of Rhodospirillum rubrum
MH  - Cells
MH  - Cyanides
MH  - Cytochromes
MH  - Nad
MH  - Rhodospirillum
MH  - Rhodospirillum rubrum
RP  - NOT IN FILE
NT  - UI - 69038148LA - engRN - 0 (Cyanides)RN - 0 (Cytochromes)RN - 0 (Oxides)RN - 0 (Quinolines)RN - 53-84-9 (NAD)RN - 7782-41-4 (Fluorine)PT - Journal ArticleDA - 19690110IS - 0006-291XSB - IMCY - UNITED STATESJC - 9Y8
UR  - PM:4301663
SO  - Biochem Biophys Res Commun 1968 Sep 30 ;32(6):908-915

33
UI  - 18962
AU  - Junge W
AU  - Witt HT
TI  - On the ion transport system of photosynthesis--investigations on a molecular level
MH  - A
MH  - Adenosine Triphosphate
MH  - Biological Transport,Active
MH  - biosynthesis
MH  - Chlorophyll
MH  - Chloroplasts
MH  - Electron Transport
MH  - Hydrogen-Ion Concentration
MH  - ion
MH  - ion exchange
MH  - Ion Transport
MH  - Kinetics
MH  - Membranes
MH  - metabolism
MH  - Permeability
MH  - pharmacology
MH  - Photosynthesis
MH  - Potassium
MH  - spectrum analysis
MH  - transport
MH  - tyrothricin
RP  - NOT IN FILE
SO  - Z Naturforsch B 1968  ;23(2):244-254

34
UI  - 1044
AU  - Mitchell P
AU  - Moyle J
TI  - Proton translocation coupled to ATP hydrolysis in rat liver mitochondria
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - Animal
MH  - antagonists & inhibitors
MH  - Antibiotics
MH  - Calcium
MH  - drug effects
MH  - Edetic Acid
MH  - Hydrogen-Ion Concentration
MH  - Hydrolysis
MH  - Liver
MH  - Magnesium
MH  - Male
MH  - Membrane Potentials
MH  - metabolism
MH  - Mitochondria
MH  - Mitochondria,Liver
MH  - Osmosis
MH  - pharmacology
MH  - Phosphates
MH  - Phosphotransferases
MH  - Protons
MH  - Rats
RP  - NOT IN FILE
SO  - Eur J Biochem 1968 May ;4(4):530-539

35
UI  - 1045
AU  - Mitchell P
AU  - Moyle J
AU  - Smith L
TI  - Bromthymol blue as a pH indicator in mitochondrial suspensions
MH  - Animal
MH  - Antibiotics
MH  - Chemistry
MH  - Hydrogen-Ion Concentration
MH  - Indicators and Reagents
MH  - Membrane Potentials
MH  - Membranes
MH  - Methods
MH  - Mitochondria,Liver
MH  - Oxidation-Reduction
MH  - Rats
MH  - Spectrophotometry
MH  - Time Factors
RP  - NOT IN FILE
SO  - Eur J Biochem 1968 Mar ;4(1):9-19

36
UI  - 806
AU  - Rumberg B
AU  - Reinwald E
AU  - Schroder H
AU  - Siggel U
TI  - Correlation between electron flow, proton translocation and phosphorylation in chloroplasts
RP  - NOT IN FILE
NT  - UI - 69083196LA - engRN - 0 (Butyrophenones)RN - 0 (Cyanides)RN - 0 (Hydrazines)RN - 0 (Protons)RN - 1333-74-0 (Hydrogen)RN - 56-65-5 (Adenosine Triphosphate)RN - 642-15-9 (Antimycin A)PT - Journal ArticleDA - 19690225IS - 0028-1042SB - IMCY - GERMANY, WESTJC - NSW
UR  - PM:5703108
SO  - Naturwissenschaften 1968 Feb ;55(2):77-79

37
UI  - 807
AU  - Rumberg B
AU  - Siggel U
TI  - [Quantitative relationship between chlorophyll-b reaction, electron transport and phosphorylation during photosynthesis]
RP  - NOT IN FILE
NT  - UI - 68364428LA - gerRN - 0 (Amines)RN - 0 (Aniline Compounds)RN - 0 (Cyanides)RN - 0 (Cyclohexanes)RN - 1406-65-1 (Chlorophyll)RN - 56-65-5 (Adenosine Triphosphate)RN - 83-89-6 (Quinacrine)PT - Journal ArticleDA - 19681004IS - 0044-3174SB - IMCY - GERMANY, WESTJC - BZ2
UR  - PM:4385981
SO  - Z Naturforsch B 1968 Feb ;23(2):239-244

38
UI  - 18961
AU  - Schliephake W
AU  - Junge W
AU  - Witt HT
TI  - Correlation between field formation, proton translocation, and the light reactions in photosynthesis
MH  - Adenosine Triphosphate
MH  - ammonium compounds
MH  - biosynthesis
MH  - Chloroplasts
MH  - cresols
MH  - cytology
MH  - dyes
MH  - Electron Transport
MH  - FIELD
MH  - Hydrogen-Ion Concentration
MH  - indoles
MH  - Light
MH  - Membranes
MH  - metabolism
MH  - phenazines
MH  - Photosynthesis
MH  - plants,edible
MH  - proton
MH  - Protons
MH  - Quinones
MH  - sulfonic acids
MH  - translocation
RP  - NOT IN FILE
SO  - Z Naturforsch B 1968  ;23(12):1571-1578

39
UI  - 8262
AU  - Witt HT
AU  - Rumberg B
AU  - Junge W
TI  - Electron transfer, field changes, proton translocation, and phosphorylation in photosynthesis. Coupling in the thylakoid membrane
MH  - electron
MH  - FIELD
MH  - membrane
MH  - Phosphorylation
MH  - Photosynthesis
MH  - proton
MH  - thylakoid
MH  - TRANSFER
MH  - translocation
MH  - transport
T2  - Biochem. Sauerst., Colloq. Ges. Biol. Chem., 19th, 262-317. Edited by: Hess, B.. Springer-Verlag: Berlin, Ger
Y2  - -32676  
RP  - NOT IN FILE
SO  -  1968  ;():

40
UI  - 19846
AU  - Boyer PD
TI  - The inhibition of pyruvate kinase by ATP: a Mg++ buffer system for use in enzyme studies
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - Buffers
MH  - Magnesium
MH  - Pyruvate Kinase
RP  - NOT IN FILE
NT  - UI - 69157091LA - engRN - 0 (Buffers)RN - 0 (Glycerophosphates)RN - 56-65-5 (Adenosine Triphosphate)RN - 7439-95-4 (Magnesium)RN - EC 2.7.1.40 (Pyruvate Kinase)PT - Journal ArticleDA - 19690522IS - 0006-291XSB - IMCY - UNITED STATESJC - 9Y8
UR  - PM:5777786
SO  - Biochem Biophys Res Commun 1969 Mar 10 ;34(5):702-706

41
UI  - 18959
AU  - Emrich HM
AU  - Junge W
AU  - Witt HT
TI  - Further evidcence for an optical response of chloroplast bulk pigments to a light induced electrical field in photosynthesis
MH  - A
MH  - analysis
MH  - ANS
MH  - Carotenoids
MH  - Chlorophyll
MH  - chloroplast
MH  - Chloroplasts
MH  - Electric Conductivity
MH  - FIELD
MH  - Light
MH  - Membranes
MH  - metabolism
MH  - pharmacology
MH  - Photosynthesis
MH  - pigments
MH  - radiation effects
MH  - spectrum analysis
MH  - tyrothricin
RP  - NOT IN FILE
SO  - Z Naturforsch B 1969  ;24(9):1144-1146

42
UI  - 18958
AU  - Emrich HM
AU  - Junge W
AU  - Witt HT
TI  - An artificial indicator for electric phenomena in biological membranes and interfaces
MH  - ANS
MH  - Carotenoids
MH  - Chloroplasts
MH  - Electrophysiology
MH  - Fluorescent Dyes
MH  - indicator
MH  - interfaces
MH  - membrane
MH  - Membranes
MH  - physiology
MH  - spectrum analysis
RP  - NOT IN FILE
SO  - Naturwissenschaften 1969  ;56(10):514-515

43
UI  - 867
AU  - Jackson JB
AU  - Crofts AR
TI  - Bromothymol blue and bromocresol purple as indicators of pH changes in chromatophores of Rhodospirillum rubrum
RP  - NOT IN FILE
NT  - UI - 70001806LA - engRN - 0 (Antibiotics)RN - 0 (Buffers)RN - 0 (Ethers, Cyclic)RN - 0 (Indicators and Reagents)PT - Journal ArticleDA - 19691127IS - 0014-2956SB - IMCY - GERMANY, WESTJC - EMZ
UR  - PM:5823098
SO  - Eur J Biochem 1969 Sep ;10(2):226-237

44
UI  - 19845
AU  - Jones DH
AU  - Boyer PD
TI  - The apparent absolute requirement of adenosine diphosphate for the inorganic phosphate--water exchange of oxidative phosphorylation
MH  - Adenine Nucleotides
MH  - ADENINE-NUCLEOTIDES
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - Dinitrophenols
MH  - Nucleotides
MH  - Oligomycins
MH  - Oxidative Phosphorylation
MH  - Oxygen
MH  - Oxygen Isotopes
MH  - Phosphates
MH  - Phosphorus
MH  - Phosphorylation
MH  - Water
RP  - NOT IN FILE
NT  - UI - 70032787LA - engRN - 0 (Adenine Nucleotides)RN - 0 (Carbon Isotopes)RN - 0 (Dinitrophenols)RN - 0 (Oligomycins)RN - 0 (Oxygen Isotopes)RN - 0 (Phosphates)RN - 0 (Phosphonic Acids)RN - 0 (Phosphorus Isotopes)RN - 56-65-5 (Adenosine Triphosphate)RN - 7732-18-5 (Water)PT - Journal ArticleDA - 19700102IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:5350933
SO  - J Biol Chem 1969 Nov 10 ;244(21):5767-5772

45
UI  - 18960
AU  - Junge W
AU  - Witt HT
TI  - Analysis of electrical phenomena in membranes and interfaces by absorption changes
MH  - absorption
MH  - analysis
MH  - interfaces
MH  - membrane
MH  - Membrane Potentials
MH  - Membranes
MH  - membranes,artificial
RP  - NOT IN FILE
SO  - Nature 1969  ;222(198):1062-1062

46
UI  - 18963
AU  - Junge W
AU  - Schliephake WD
AU  - Witt HT
TI  - Experimental evidence for the chemiosmotic hypothesis
MH  - bioenergetics
MH  - proton
MH  - thylakoid
T2  - Progr. Photosyn. Res., Proc. Int. Congr., Meeting Date 1968, Volume 3, 1383-91. Edited by: Metzner, Helmut. Verlag C. Lichtenstern: Munich, Ger
Y2  - -32676  
RP  - NOT IN FILE
SO  -  1969  ;():

47
UI  - 1043
AU  - Mitchell P
AU  - Moyle J
TI  - Estimation of membrane potential and pH difference across the cristae membrane of rat liver mitochondria
MH  - Animal
MH  - Antibiotics
MH  - Biological Transport
MH  - Buffers
MH  - Calcium Chloride
MH  - Choline
MH  - Cyanides
MH  - Depression,Chemical
MH  - drug effects
MH  - Electrodes
MH  - Hydrogen-Ion Concentration
MH  - Liver
MH  - Mathematics
MH  - Membrane Potentials
MH  - Membranes
MH  - metabolism
MH  - Mitochondria
MH  - Mitochondria,Liver
MH  - Oxygen Consumption
MH  - Permeability
MH  - pharmacology
MH  - physiology
MH  - Potassium
MH  - Potentiometry
MH  - Protons
MH  - Rats
MH  - Surface-Active Agents
RP  - NOT IN FILE
SO  - Eur J Biochem 1969 Feb ;7(4):471-484

48
UI  - 563
AU  - Pedersen PL
AU  - Schnaitman CA
TI  - The oligomycin-sensitive adenosine diphosphate-adenosine triphosphate exchange in an inner membrane matrix fraction of rat liver mitochondria
RP  - NOT IN FILE
NT  - UI - 70004772LA - engRN - 0 (Adenine Nucleotides)RN - 0 (Antimetabolites)RN - 0 (Carbon Isotopes)RN - 0 (Cytosine Nucleotides)RN - 0 (Dinitrophenols)RN - 0 (Glycosides)RN - 0 (Oligomycins)RN - 0 (Phosphates)RN - 0 (Phosphorus Isotopes)RN - 0 (Uracil Nucleotides)RN - 56-65-5 (Adenosine Triphosphate)RN - 7439-95-4 (Magnesium)RN - EC 2.7 (Phosphotransferases)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 4.6.1.1 (Adenylate Cyclase)PT - Journal ArticleDA - 19691201IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:4241925
SO  - J Biol Chem 1969 Sep 25 ;244(18):5065-5074

49
UI  - 805
AU  - Rumberg B
AU  - Siggel U
TI  - pH changes in the inner phase of the thylakoids during photosynthesis
RP  - NOT IN FILE
NT  - UI - 70059399LA - engRN - 0 (Adenine Nucleotides)RN - 1404-88-2 (Tyrothricin)RN - 1406-65-1 (Chlorophyll)RN - 56-65-5 (Adenosine Triphosphate)PT - Journal ArticleDA - 19700130IS - 0028-1042SB - IMCY - GERMANY, WESTJC - NSW
UR  - PM:5358721
SO  - Naturwissenschaften 1969 Mar ;56(3):130-132

50
UI  - 839
AU  - Strotmann H
AU  - Berger S
TI  - Adenine nucleotide translocation across the membrane of isolated Acetabularia chloroplasts
RP  - NOT IN FILE
NT  - UI - 69162688LA - engRN - 0 (Adenine Nucleotides)RN - 0 (Carbon Isotopes)RN - 124-38-9 (Carbon Dioxide)RN - 1406-65-1 (Chlorophyll)RN - 56-65-5 (Adenosine Triphosphate)PT - Journal ArticleDA - 19690602IS - 0006-291XSB - IMCY - UNITED STATESJC - 9Y8
UR  - PM:5779146
SO  - Biochem Biophys Res Commun 1969 Apr 10 ;35(1):20-26

51
UI  - 8308
AU  - Witt HT
AU  - Rumberg B
AU  - Junge W
AU  - Doering G
AU  - Stiehl HH
AU  - Weikard J
AU  - Wolff C
TI  - Evidence for the coupling of electron transfer, field changes, proton translocation, and phosphorylation in photosynthesis
MH  - electron
MH  - FIELD
MH  - Phosphorylation
MH  - Photosynthesis
MH  - proton
MH  - TRANSFER
MH  - translocation
MH  - transport
T2  - Progr. Photosyn. Res., Proc. Int. Congr., Meeting Date 1968, Volume 3, 1361-73. Edited by: Metzner, Helmut. Verlag C. Lichtenstern: Munich, Ger
Y2  - -32676  
RP  - NOT IN FILE
SO  -  1969  ;():

52
UI  - 898
AU  - Beechey RB
TI  - Mitochondrial coupling factors
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - biosynthesis
MH  - Chemistry,Physical
MH  - Energy Transfer
MH  - enzymology
MH  - metabolism
MH  - Mitochondria
RP  - NOT IN FILE
SO  - Biochem J 1970 Feb ;116(4):6P-8P

53
UI  - 9957
AU  - Chance B
AU  - Crofts AR
AU  - Nishimura M
AU  - Price B
TI  - Fast membrane H+ binding in the light-activated state of Chromatium chromatophores
MH  - Antibiotics
MH  - Antimycin A
MH  - BINDING
MH  - Chlorophyll
MH  - H+
MH  - Hydrogen
MH  - Uncoupling Agents
RP  - NOT IN FILE
NT  - UI - 70163886LA - engRN - 0 (Antibiotics)RN - 0 (Culture Media)RN - 0 (Ethers, Cyclic)RN - 0 (Phenolphthaleins)RN - 0 (Uncoupling Agents)RN - 1333-74-0 (Hydrogen)RN - 1406-65-1 (Chlorophyll)RN - 642-15-9 (Antimycin A)RN - 7440-63-3 (Xenon)PT - Journal ArticleDA - 19700604IS - 0014-2956SB - IMCY - GERMANY, WESTJC - EMZ
UR  - PM:5439938
SO  - Eur J Biochem 1970 Apr ;13(2):364-374

54
UI  - 19884
AU  - Cross RL
AU  - Cross BA
AU  - Wang JH
TI  - Detection of a phosphorylated intermediate in mitochondrial oxidative phosphorylation
MH  - A
MH  - Antibiotics
MH  - Antimetabolites
MH  - Dinitrophenols
MH  - Oligomycins
MH  - Oxidative Phosphorylation
MH  - Phosphates
MH  - Phosphorus
MH  - Phosphorylation
MH  - Rotenone
MH  - succinate
MH  - Succinates
MH  - Uncoupling Agents
RP  - NOT IN FILE
NT  - UI - 71135298LA - engRN - 0 (Antibiotics)RN - 0 (Antimetabolites)RN - 0 (Dinitrophenols)RN - 0 (Oligomycins)RN - 0 (Phosphates)RN - 0 (Phosphorus Isotopes)RN - 0 (Succinates)RN - 0 (Uncoupling Agents)RN - 83-79-4 (Rotenone)PT - Journal ArticleDA - 19710426IS - 0006-291XSB - IMCY - UNITED STATESJC - 9Y8
UR  - PM:5503790
SO  - Biochem Biophys Res Commun 1970 Sep 10 ;40(5):1155-1161

55
UI  - 657
AU  - Datta A
AU  - Penefsky HS
TI  - Interaction of fluorescent probes with submitochondrial particles during oxidative phosphorylation
RP  - NOT IN FILE
NT  - UI - 70158475LA - engRN - 0 (Cyanides)RN - 0 (Fluorescent Dyes)RN - 0 (Naphthalenes)RN - 0 (Nitriles)RN - 0 (Phenylhydrazines)RN - 0 (Receptors, Drug)RN - 0 (Succinates)RN - 0 (Sulfonic Acids)RN - 0 (Uncoupling Agents)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19700526IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:4245220
SO  - J Biol Chem 1970 Apr 10 ;245(7):1537-1544

56
UI  - 804
AU  - Junge W
AU  - Rumberg B
AU  - Schroder H
TI  - The necessity of an electric potential difference and its use for photophosphorylation in short flash groups
RP  - NOT IN FILE
NT  - UI - 71032725LA - engRN - 56-65-5 (Adenosine Triphosphate)PT - Journal ArticleDA - 19710107IS - 0014-2956SB - IMCY - GERMANY, WESTJC - EMZ
UR  - PM:5479384
SO  - Eur J Biochem 1970 Jul ;14(3):575-581

57
UI  - 8309
AU  - Junge W
AU  - Emrich HM
AU  - Witt HT
TI  - Indication of a light induced electrical field by pigments incorporated in chloroplast membranes
MH  - A
MH  - chloroplast
MH  - electrochromism
MH  - FIELD
MH  - Light
MH  - membrane
MH  - Membranes
MH  - pigments
T2  - Phys. Principles Biol. Membranes, Proc. Coral Gables Conf., Meeting Date 1968, 383-96. Edited by: Snell, F. Gordon and Breach Sci. Publ.: New York, N. Y
Y2  - -32676  
RP  - NOT IN FILE
SO  -  1970  ;():

58
UI  - 18964
AU  - Junge W
TI  - Critical electric potential difference for photophosphorylation. Its relation to the chemiosomotic hypothesis and to the triggering requirements of the ATPase system
MH  - atp
MH  - ATPase
MH  - bioenergetics
MH  - Membrane Potential
MH  - Photophosphorylation
MH  - theory
RP  - NOT IN FILE
SO  - Eur J Biochem 1970  ;14():582-592

59
UI  - 628
AU  - Melandri BA
AU  - Baccarini-Melandri A
AU  - San Pietro A
AU  - Gest H
TI  - Role of phosphorylation coupling factor in light-dependent proton translocation by Rhodopseudomonas capsulata membrane preparations
RP  - NOT IN FILE
NT  - UI - 72070063LA - engRN - 0 (Oligomycins)RN - 0 (Phosphorus Isotopes)RN - 0 (Protons)RN - 0 (Uncoupling Agents)RN - 58-64-0 (Adenosine Diphosphate)RN - 60-00-4 (Edetic Acid)RN - 7439-95-4 (Magnesium)RN - 7440-38-2 (Arsenic)PT - Journal ArticleDA - 19720229IS - 0027-8424SB - IMCY - UNITED STATESJC - PV3
UR  - PM:5002093
SO  - Proc Natl Acad Sci U S A 1970 Oct ;67(2):477-484

60
UI  - 1041
AU  - Mitchell P
TI  - Aspects of the chemiosmotic hypothesis
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - Animal
MH  - Biological Transport
MH  - Cell Membrane Permeability
MH  - enzymology
MH  - metabolism
MH  - Mitochondria
MH  - Mitochondria,Liver
MH  - Models,Chemical
MH  - Osmosis
MH  - Oxidation-Reduction
MH  - Oxidative Phosphorylation
MH  - Rats
RP  - NOT IN FILE
SO  - Biochem J 1970 Feb ;116(4):5P-6P

61
UI  - 1042
AU  - Mitchell P
AU  - Moyle J
TI  - Inhibition of mitochondrial adenosine triphosphatase by adenosine diphosphate: release of adenosine diphosphate inhibition by aurovertin
MH  - Adenine Nucleotides
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosinetriphosphatase
MH  - Animal
MH  - antagonists & inhibitors
MH  - Antibiotics
MH  - Antimetabolites
MH  - Enzyme Activation
MH  - enzymology
MH  - Kinetics
MH  - Mitochondria,Liver
MH  - pharmacology
MH  - Rats
RP  - NOT IN FILE
SO  - Biochem J 1970 Feb ;116(4):10P-11P

62
UI  - 9956
AU  - Montal M
AU  - Nishimura M
AU  - Chance B
TI  - Uncoupling and charge transfer in bacterial chromatophores
MH  - Antibiotics
MH  - Bacterial Chromatophores
MH  - Potassium
MH  - Sodium
MH  - Sulfates
MH  - TRANSFER
MH  - Uncoupling Agents
RP  - NOT IN FILE
NT  - UI - 71050151LA - engRN - 0 (Antibiotics)RN - 0 (Boron Compounds)RN - 0 (Chlorides)RN - 0 (Nitrates)RN - 0 (Sulfates)RN - 0 (Uncoupling Agents)RN - 12125-02-9 (Ammonium Chloride)RN - 7440-09-7 (Potassium)RN - 7440-23-5 (Sodium)PT - Journal ArticleDA - 19710121IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:5484051
SO  - Biochim Biophys Acta 1970 Nov 3 ;223(1):183-188

63
UI  - 399
AU  - Akimenko VK
AU  - Minkov IB
AU  - Vinogradov AD
TI  - [Binding of soluble mitochondrial ATPase by particles deprived of ATPase activity]
RP  - NOT IN FILE
NT  - UI - 72076485LA - rusRN - 0 (Sulfhydryl Compounds)RN - 56-65-5 (Adenosine Triphosphate)RN - 67-64-1 (Acetone)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19720309IS - 0320-9725SB - IMCY - USSRJC - A28
UR  - PM:4257177
SO  - Biokhimiia 1971 May ;36(3):502-506

64
UI  - 562
AU  - Catterall WA
AU  - Pedersen PL
TI  - Adenosine triphosphatase from rat liver mitochondria. I. Purification, homogeneity, and physical properties
RP  - NOT IN FILE
NT  - UI - 71288578LA - engRN - 0 (Amino Acids)RN - 0 (Detergents)RN - 0 (Glycols)RN - 0 (Macromolecular Systems)RN - 0 (Sulfuric Acids)RN - 56-65-5 (Adenosine Triphosphate)RN - 60-00-4 (Edetic Acid)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19711112IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:4255092
SO  - J Biol Chem 1971 Aug ;246(16):4987-4994

65
UI  - 866
AU  - Jackson JB
AU  - Crofts AR
TI  - The kinetics of light induced carotenoid changes in Rhodopseudomonas spheroides and their relation to electrical field generation across the chromatophore membrane
RP  - NOT IN FILE
NT  - UI - 71090266LA - engRN - 0 (Carotenoids)RN - 56-65-5 (Adenosine Triphosphate)RN - 642-15-9 (Antimycin A)PT - Journal ArticleDA - 19710312IS - 0014-2956SB - IMCY - GERMANY, WESTJC - EMZ
UR  - PM:5540508
SO  - Eur J Biochem 1971 Jan 1 ;18(1):120-130

66
UI  - 2183
AU  - Junge W
AU  - Schmid R
TI  - The mechanism of action of valinomycin on the thylakoid membrane. Characterization of the electric current density
MH  - electrochromism
MH  - mechanism
MH  - membrane
MH  - thylakoid
MH  - thylakoid membrane
MH  - Valinomycin
RP  - IN FILE
NT  - "plants"; laut W. Junge ist die Arbeit falsch !
SO  - J Membr Biol 1971  ;4():179-192

67
UI  - 627
AU  - Melandri BA
AU  - Baccarini-Melandri A
AU  - San Pietro A
AU  - Gest H
TI  - Interchangeability of phosphorylation coupling factors in photosynthetic and respiratory energy conversion
RP  - NOT IN FILE
NT  - UI - 72015561LA - engRN - 0 (Phosphorus Isotopes)RN - 0 (Succinates)RN - 53-59-8 (NADP)PT - Journal ArticleDA - 19711216IS - 0036-8075SB - IMCY - UNITED STATESJC - UJ7
UR  - PM:4398683
SO  - Science 1971 Oct 29 ;174(8):514-516

68
UI  - 1040
AU  - Mitchell P
AU  - Moyle J
TI  - Activation and inhibition of mitochondrial adenosine triphosphatase by various anions and other agents
MH  - Adenosine
MH  - Adenosinetriphosphatase
MH  - Animal
MH  - Anions
MH  - antagonists & inhibitors
MH  - Antibiotics
MH  - Antimetabolites
MH  - Arsenic
MH  - Azides
MH  - Chromates
MH  - Diphosphates
MH  - Enzyme Activation
MH  - enzymology
MH  - Guanosine Triphosphate
MH  - Hydrogen-Ion Concentration
MH  - Hydrolysis
MH  - In Vitro
MH  - Kinetics
MH  - metabolism
MH  - Mitochondria,Liver
MH  - Oligomycins
MH  - pharmacology
MH  - Phosphates
MH  - Potassium Chloride
MH  - Rats
MH  - Sulfates
RP  - NOT IN FILE
SO  - J Bioenerg 1971 Feb ;2(1):1-11

69
UI  - 561
AU  - Pedersen PL
AU  - Catterall WA
TI  - Contribution of ATP synthesis from endogenous substrates to the oligomycin-sensitive ADP-ATP exchange activity of rat liver mitoplasts
RP  - NOT IN FILE
NT  - UI - 72065374LA - engRN - 0 (Adenine Nucleotides)RN - 0 (Carbon Isotopes)RN - 0 (Cyanides)RN - 0 (Oligomycins)RN - 0 (Phosphorus Isotopes)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 61-19-8 (Adenosine Monophosphate)RN - 7440-09-7 (Potassium)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19720222IS - 0006-291XSB - IMCY - UNITED STATESJC - 9Y8
UR  - PM:4256848
SO  - Biochem Biophys Res Commun 1971 Nov 5 ;45(3):809-815

70
UI  - 398
AU  - Akimenko VK
AU  - Minkov IB
AU  - Bakeeva LE
AU  - Vinogradov AD
TI  - [Isolation and properties of soluble ATPase from bovine heart mitochondria]
RP  - NOT IN FILE
NT  - UI - 72180198LA - rusRN - 56-65-5 (Adenosine Triphosphate)RN - 7439-95-4 (Magnesium)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19720714IS - 0320-9725SB - IMCY - USSRJC - A28
UR  - PM:4260175
SO  - Biokhimiia 1972 Mar ;37(2):348-359

71
UI  - 489
AU  - Brooks JC
AU  - Senior AE
TI  - Methods for purification of each subunit of the mitochondrial oligomycin-insensitive adenosine triphosphatase
RP  - NOT IN FILE
NT  - UI - 73132230LA - engRN - 0 (Amino Acids)RN - 0 (Guanidines)RN - 0 (Macromolecular Systems)RN - 0 (Oligomycins)RN - 151-21-3 (Sodium Dodecyl Sulfate)RN - 57-13-6 (Urea)RN - 7783-20-2 (Ammonium Sulfate)RN - EC 3.4.21.4 (Trypsin)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19730502IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:4266070
SO  - Biochemistry 1972 Dec 5 ;11(25):4675-4678

72
UI  - 656
AU  - Buckley NM
AU  - Penefsky ZJ
AU  - Litwak RS
TI  - Comparative force-frequency relationships in human and other mammalian ventricular myocardium
RP  - NOT IN FILE
NT  - UI - 72201630LA - engRN - 0 (Actins)RN - 0 (Myosin)RN - 56-65-5 (Adenosine Triphosphate)RN - 7440-09-7 (Potassium)RN - 7440-23-5 (Sodium)RN - 7440-70-2 (Calcium)PT - Journal ArticleDA - 19720807IS - 0031-6768SB - IMCY - GERMANY, WESTJC - OZX
UR  - PM:5064070
SO  - Pflugers Arch 1972  ;332(4):259-270

73
UI  - 969
AU  - Chance B
AU  - Lees H
AU  - Postgate JR
TI  - The meaning of &quot;reversed electron flow&quot; and &quot;high energy electron&quot; in biochemistry
MH  - Adenosine Triphosphate
MH  - Biochemistry
MH  - Electron Transport
MH  - Hydrolysis
MH  - metabolism
MH  - Nitrobacter
MH  - Nitrogen Fixation
MH  - Oxidation-Reduction
MH  - Succinates
RP  - NOT IN FILE
SO  - Nature 1972 Aug 11 ;238(5363):330-331

74
UI  - 9972
AU  - Deamer DW
AU  - Prince RC
AU  - Crofts AR
TI  - The response of fluorescent amines to pH gradients across liposome membranes
MH  - Antibiotics
MH  - Chlorophyll
MH  - Membranes
MH  - Phosphatidylcholines
MH  - Potassium
MH  - Sulfites
RP  - NOT IN FILE
NT  - UI - 72250128LA - engRN - 0 (Acridines)RN - 0 (Amines)RN - 0 (Antibiotics)RN - 0 (Fatty Alcohols)RN - 0 (Ferricyanides)RN - 0 (Phenazines)RN - 0 (Phosphatidylcholines)RN - 0 (Phosphoric Acids)RN - 0 (Sulfites)RN - 1406-65-1 (Chlorophyll)RN - 50-81-7 (Ascorbic Acid)RN - 7440-09-7 (Potassium)RN - 83-89-6 (Quinacrine)PT - Journal ArticleDA - 19720929IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:5049001
SO  - Biochim Biophys Acta 1972 Aug 9 ;274(2):323-335

75
UI  - 865
AU  - Dutton PL
AU  - Jackson JB
TI  - Thermodynamic and kinetic characterization of electron transfer components in situ in Rhodopseudomonas spheroides and Rhodospirillum rubrum
RP  - NOT IN FILE
NT  - UI - 73056891LA - engRN - 0 (Cytochromes)RN - 0 (Ferricyanides)RN - 0 (Sulfites)RN - 1406-65-1 (Chlorophyll)RN - 53-84-9 (NAD)RN - 642-15-9 (Antimycin A)RN - 9007-43-6 (Cytochrome c)PT - Journal ArticleDA - 19730208IS - 0014-2956SB - IMCY - GERMANY, WESTJC - EMZ
UR  - PM:4344828
SO  - Eur J Biochem 1972 Nov 7 ;30(3):495-510

76
UI  - 8306
AU  - Junge W
TI  - Brownian rotation of the cytochrome oxidase in the mitochondrial inner membrane
MH  - cytochrome
MH  - cytox
MH  - membrane
MH  - rotation
RP  - NOT IN FILE
SO  - FEBS Lett 1972  ;25():109-112

77
UI  - 667
AU  - Knowles AF
AU  - Penefsky HS
TI  - The subunit structure of beef heart mitochondrial adenosine triphosphatase. Isolation procedures
RP  - NOT IN FILE
NT  - UI - 73015881LA - engRN - 0 (Guanidines)RN - 0 (Macromolecular Systems)RN - 0 (Oxidative Phosphorylation Coupling Factors)RN - 0 (Peptides)RN - 151-21-3 (Sodium Dodecyl Sulfate)RN - 57-13-6 (Urea)RN - 7783-20-2 (Ammonium Sulfate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19721204IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:4263201
SO  - J Biol Chem 1972 Oct 25 ;247(20):6617-6623

78
UI  - 666
AU  - Knowles AF
AU  - Penefsky HS
TI  - The subunit structure of beef heart mitochondrial adenosine triphosphatase. Physical and chemical properties of isolated subunits
RP  - NOT IN FILE
NT  - UI - 73015882LA - engRN - 0 (Amino Acids)RN - 0 (Macromolecular Systems)RN - 0 (Oxidative Phosphorylation Coupling Factors)RN - 0 (Peptides)RN - 151-21-3 (Sodium Dodecyl Sulfate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19721204IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:4263202
SO  - J Biol Chem 1972 Oct 25 ;247(20):6624-6630

79
UI  - 622
AU  - Melandri BA
AU  - Baccarini-Melandri A
AU  - Fabbri E
TI  - Energy transduction in photosynthetic bacteria. IV. Light-dependent ATPase in photosynthetic membranes from Rhodopseudomonas capsulata
RP  - NOT IN FILE
NT  - UI - 73003007LA - engRN - 0 (Phenols)RN - 0 (Phosphorus Isotopes)RN - 0 (Uncoupling Agents)RN - 500-85-6 (Indophenol)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19721115IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:4262690
SO  - Biochim Biophys Acta 1972 Sep 20 ;275(3):383-394

80
UI  - 1037
AU  - Mitchell P
TI  - Self-electrophoretic locomotion in microorganisms: bacterial flagella as giant ionophores
MH  - Bacteria
MH  - Biological Transport
MH  - Cell Movement
MH  - Flagella
MH  - Ionophores
MH  - Membrane Potentials
MH  - metabolism
MH  - Models,Biological
MH  - Protons
MH  - Sodium
RP  - NOT IN FILE
SO  - FEBS Lett 1972 Nov 15 ;28(1):1-4

81
UI  - 1039
AU  - Mitchell P
TI  - Chemiosmotic coupling in energy transduction: a logical development of biochemical knowledge
MH  - Adenosinetriphosphatase
MH  - Biological Transport
MH  - development
MH  - Electron Transport
MH  - Energy Transfer
MH  - Membrane Potentials
MH  - Models,Biological
MH  - Osmosis
MH  - Oxidative Phosphorylation
MH  - Photophosphorylation
RP  - NOT IN FILE
SO  - J Bioenerg 1972 May ;3(1):5-24

82
UI  - 1004
AU  - Mitchell P
TI  - Chemiosmotic coupling in energy transduction: a logical development of biochemical knowledge
MH  - Adenosinetriphosphatase
MH  - development
RP  - NOT IN FILE
NT  - UI - 73042466LA - engRN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticlePT - ReviewDA - 19730118IS - 0449-5705SB - IMCY - ENGLANDJC - HIN
UR  - PM:4263930
SO  - J Bioenerg 1972 May ;3(1):5-24

83
UI  - 1038
AU  - Moyle J
AU  - Mitchell R
AU  - Mitchell P
TI  - Proton-translocating pyrophosphatase of Rhodospirillum rubrum
MH  - Anaerobiosis
MH  - Bacterial Chromatophores
MH  - cytology
MH  - Diphosphates
MH  - drug effects
MH  - enzymology
MH  - Hydrogen-Ion Concentration
MH  - Hydrolysis
MH  - metabolism
MH  - Oligomycins
MH  - pharmacology
MH  - Protons
MH  - Pyrophosphatases
MH  - Rhodospirillum
MH  - Rhodospirillum rubrum
RP  - NOT IN FILE
SO  - FEBS Lett 1972 Jun 15 ;23(2):233-236

84
UI  - 803
AU  - Siggel U
AU  - Renger G
AU  - Stiehl HH
AU  - Rumberg B
TI  - Evidence for electronic and ionic interaction between electron transport chains in chloroplasts
RP  - NOT IN FILE
NT  - UI - 72154271LA - engRN - 0 (Methylamines)RN - 0 (Quinones)RN - 1406-65-1 (Chlorophyll)RN - 330-54-1 (Diuron)RN - 56-65-5 (Adenosine Triphosphate)RN - 7782-44-7 (Oxygen)RN - 9005-25-8 (Starch)PT - Journal ArticleDA - 19720615IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:5016542
SO  - Biochim Biophys Acta 1972 Feb 28 ;256(2):328-335

85
UI  - 894
AU  - Skulachev VP
TI  - Solution of the problem of energy coupling in terms of chemiosmotic theory
MH  - Adenosine Triphosphate
MH  - Biological Transport
MH  - Electron Transport
MH  - Membrane Potentials
MH  - Osmosis
MH  - Oxidation-Reduction
MH  - Oxidative Phosphorylation
RP  - NOT IN FILE
SO  - J Bioenerg 1972 May ;3(1):25-38

86
UI  - 838
AU  - Strotmann H
AU  - von Gosseln C
TI  - [Photosystem I dependent phosphorylation of isolated chloroplasts with ascorbate-2,6-dichlorophenolindophenol as electron donor and methylviologen as electron acceptor]
RP  - NOT IN FILE
NT  - UI - 72259456LA - gerRN - 0 (Phenols)RN - 0 (Pyridinium Compounds)RN - 1333-74-0 (Hydrogen)RN - 4685-14-7 (Paraquat)RN - 50-81-7 (Ascorbic Acid)RN - 500-85-6 (Indophenol)RN - 56-65-5 (Adenosine Triphosphate)RN - 7782-44-7 (Oxygen)RN - EC 3.4.21.4 (Trypsin)PT - Journal ArticleDA - 19721005IS - 0044-3174SB - IMCY - GERMANY, WESTJC - XYU
UR  - PM:4403315
SO  - Z Naturforsch [B] 1972 Apr ;27(4):445-455

87
UI  - 626
AU  - Baccarini MA
AU  - Zannoni D
AU  - Melandri BA
TI  - Energy transduction in photosynthetic bacteria. VI. Respiratory sites of energy conservation in membranes from dark-grown cells of Rhodopseudomonas capsulata
RP  - NOT IN FILE
NT  - UI - 74027351LA - engRN - 0 (Cyanides)RN - 0 (Ferricyanides)RN - 0 (Phosphorus Radioisotopes)RN - 50-81-7 (Ascorbic Acid)RN - 642-15-9 (Antimycin A)RN - 83-79-4 (Rotenone)RN - 83-89-6 (Quinacrine)RN - 9007-43-6 (Cytochrome c)RN - EC 1.3.99.1 (Succinate Dehydrogenase)RN - EC 1.6. (NADH, NADPH Oxidoreductases)PT - Journal ArticleDA - 19740114IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:4148029
SO  - Biochim Biophys Acta 1973 Sep 26 ;314(3):298-311

88
UI  - 19843
AU  - Boyer PD
AU  - Cross RL
AU  - Momsen W
TI  - A new concept for energy coupling in oxidative phosphorylation based on a molecular explanation of the oxygen exchange reactions
MH  - A
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - Oligomycins
MH  - Oxidative Phosphorylation
MH  - Oxygen
MH  - Phosphorus
MH  - Phosphorus Radioisotopes
MH  - Phosphorylation
MH  - Uncoupling Agents
MH  - Water
RP  - NOT IN FILE
NT  - UI - 74009531LA - engRN - 0 (Oligomycins)RN - 0 (Phosphorus Radioisotopes)RN - 0 (Uncoupling Agents)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 7732-18-5 (Water)PT - Journal ArticleDA - 19731214IS - 0027-8424SB - IMCY - UNITED STATESJC - PV3
UR  - PM:4517936
SO  - Proc Natl Acad Sci U S A 1973 Oct ;70(10):2837-2839

89
UI  - 560
AU  - Bustamante E
AU  - Pedersen PL
TI  - Tetradifon: an oligomycin-like inhibitor of energy-linked activities of rat liver mitochondria
RP  - NOT IN FILE
NT  - UI - 73145380LA - engRN - 0 (Dinitrophenols)RN - 0 (Insecticides, Organochlorine)RN - 0 (Oligomycins)RN - 0 (Phosphorus Isotopes)RN - 0 (Serum Albumin, Bovine)RN - 0 (Sulfones)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 64-17-5 (Ethanol)RN - 7439-95-4 (Magnesium)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19730518IS - 0006-291XSB - IMCY - UNITED STATESJC - 9Y8
UR  - PM:4266412
SO  - Biochem Biophys Res Commun 1973 Mar 17 ;51(2):292-298

90
UI  - 275
AU  - Capaldi RA
TI  - On the subunit structure of oligomycin sensitive ATPase
RP  - NOT IN FILE
NT  - UI - 74022233LA - engRN - 0 (Bromides)RN - 0 (Guanidines)RN - 0 (Oligomycins)RN - 0 (Peptides)RN - 7439-95-4 (Magnesium)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19740105IS - 0006-291XSB - IMCY - UNITED STATESJC - 9Y8
UR  - PM:4270659
SO  - Biochem Biophys Res Commun 1973 Aug 21 ;53(4):1331-1337

91
UI  - 665
AU  - Chang T
AU  - Penefsky HS
TI  - Aurovertin, a fluorescent probe of conformational change in beef heart mitochondrial adenosine triphosphatase
RP  - NOT IN FILE
NT  - UI - 73156406LA - engRN - 0 (Antibiotics)RN - 0 (Succinates)RN - 0 (Uncoupling Agents)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19730608IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:4266808
SO  - J Biol Chem 1973 Apr 25 ;248(8):2746-2754

92
UI  - 864
AU  - Cogdell RJ
AU  - Jackson JB
AU  - Crofts AR
TI  - The effect of redox potential on the coupling between rapid hydrogen- ion binding and electron transport in chromatophores from Rhodopseudomonas spheroides
RP  - NOT IN FILE
NT  - UI - 73215560LA - engRN - 0 (Indicators and Reagents)RN - 0 (Phenanthrolines)RN - 0 (Phenols)RN - 1333-74-0 (Hydrogen)RN - 2001-95-8 (Valinomycin)RN - 642-15-9 (Antimycin A)PT - Journal ArticleDA - 19730912IS - 0449-5705SB - IMCY - ENGLANDJC - HIN
UR  - PM:4541536
SO  - J Bioenerg 1973 Jan ;4(1):211-227

93
UI  - 9970
AU  - Cogdell RJ
AU  - Prince RC
AU  - Crofts AR
TI  - Light induced H+ uptake catalysed by photochemical reaction centres from Rhodopseudomonas spheroides R26
MH  - Cytochrome c
MH  - H+
MH  - Hydrogen
MH  - Light
MH  - proton
MH  - Protons
MH  - Ubiquinone
RP  - NOT IN FILE
NT  - UI - 74008921LA - engRN - 0 (Naphthoquinones)RN - 0 (Phenanthrolines)RN - 0 (Protons)RN - 1333-74-0 (Hydrogen)RN - 1339-63-5 (Ubiquinone)RN - 9007-43-6 (Cytochrome c)PT - Journal ArticleDA - 19731214IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:4355316
SO  - FEBS Lett 1973 Sep 15 ;35(2):204-208

94
UI  - 19844
AU  - Cross RL
AU  - Boyer PD
TI  - Evidence for detection of AT 32 P bound at the coupling sites of mitochondrial oxidative phosphorylation
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - Dinitrophenols
MH  - Hexokinase
MH  - Oxidative Phosphorylation
MH  - P
MH  - Phosphates
MH  - Phosphorus
MH  - Phosphorylation
RP  - NOT IN FILE
NT  - UI - 73164212LA - engRN - 0 (Dinitrophenols)RN - 0 (Glucosephosphates)RN - 0 (Phosphates)RN - 0 (Phosphorus Isotopes)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 2.7.1.1 (Hexokinase)PT - Journal ArticleDA - 19730621IS - 0006-291XSB - IMCY - UNITED STATESJC - 9Y8
UR  - PM:4699564
SO  - Biochem Biophys Res Commun 1973 Mar 5 ;51(1):59-66

95
UI  - 19842
AU  - Dahms AS
AU  - Kanazawa T
AU  - Boyer PD
TI  - Source of the oxygen in the C-O-P linkage of the acyl phosphate in transport adenosine triphosphatases
MH  - Adenosine
MH  - ADENOSINE-TRIPHOSPHATASE
MH  - Adenosinetriphosphatase
MH  - Calcium
MH  - Oxygen
MH  - Oxygen Isotopes
MH  - Phosphates
MH  - Phosphorus
MH  - Phosphorus Radioisotopes
MH  - Potassium
MH  - Sodium
MH  - transport
MH  - Water
RP  - NOT IN FILE
NT  - UI - 74011236LA - engRN - 0 (Oxygen Isotopes)RN - 0 (Phosphates)RN - 0 (Phosphorus Radioisotopes)RN - 630-60-4 (Ouabain)RN - 7440-09-7 (Potassium)RN - 7440-23-5 (Sodium)RN - 7440-70-2 (Calcium)RN - 7732-18-5 (Water)RN - 7782-44-7 (Oxygen)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19731216IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:4270450
SO  - J Biol Chem 1973 Oct 10 ;248(19):6592-6595

96
UI  - 9971
AU  - Evans EH
AU  - Crofts AR
TI  - The relationship between delayed fluorescence and the H + gradient in chloroplasts
MH  - acetate
MH  - Acetates
MH  - chloroplast
MH  - Chloroplasts
MH  - fluorescence
MH  - Hydrogen
MH  - Indicators and Reagents
MH  - Sulfates
MH  - Valinomycin
RP  - NOT IN FILE
NT  - UI - 73180118LA - engRN - 0 (Acetates)RN - 0 (Indicators and Reagents)RN - 0 (Phenols)RN - 0 (Sulfates)RN - 1333-74-0 (Hydrogen)RN - 2001-95-8 (Valinomycin)PT - Journal ArticleDA - 19730717IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:4705125
SO  - Biochim Biophys Acta 1973 Jan 18 ;292(1):130-139

97
UI  - 602
AU  - Hatefi Y
AU  - Hanstein WG
TI  - Interactions of reduced and oxidized triphosphopyridine nucleotides with the electron-transport system of bovine heart mitochondria
RP  - NOT IN FILE
NT  - UI - 73253928LA - engRN - 0 (Antibiotics)RN - 0 (Succinates)RN - 0 (Uncoupling Agents)RN - 53-59-8 (NADP)RN - 53-84-9 (NAD)RN - 56-65-5 (Adenosine Triphosphate)RN - 85-61-0 (Coenzyme A)RN - 87-86-5 (Pentachlorophenol)RN - EC 1.6. (NADH, NADPH Oxidoreductases)PT - Journal ArticleDA - 19731113IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:4147216
SO  - Biochemistry 1973 Aug 28 ;12(18):3515-3522

98
UI  - 20980
AU  - Hirata H
AU  - Altendorf K
AU  - Harold FM
TI  - Role of an electrical potential in the coupling of metabolic energy to active transport by membrane vesicles of Escherichia coli
MH  - ACTIVE
MH  - coupling
MH  - Cyanides
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - Hydrazones
MH  - isotope
MH  - Lactates
MH  - membrane
MH  - membrane vesicles
MH  - transport
MH  - Valinomycin
RP  - NOT IN FILE
NT  - UI - 73223683LA - engRN - 0 (Carbon Isotopes)RN - 0 (Cyanides)RN - 0 (Hydrazones)RN - 0 (Lactates)RN - 0 (Quinolines)RN - 10028-17-8 (Tritium)RN - 12125-02-9 (Ammonium Chloride)RN - 147-85-3 (Proline)RN - 2001-95-8 (Valinomycin)PT - Journal ArticleDA - 19730926IS - 0027-8424SB - IMCY - UNITED STATES
UR  - PM:4578444
SO  - Proc Natl Acad Sci U S A 1973 Jun ;70(6):1804-1808

99
UI  - 863
AU  - Jackson JB
AU  - Cogdell RJ
AU  - Crofts AR
TI  - Some effects of o-phenanthroline on electron transport in chromatophores from photosynthetic bacteria
RP  - NOT IN FILE
NT  - UI - 73180130LA - engRN - 0 (Phenanthrolines)RN - 0 (Quinolines)RN - 7440-34-8 (Actinium)RN - 7440-63-3 (Xenon)PT - Journal ArticleDA - 19730717IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:4705131
SO  - Biochim Biophys Acta 1973 Jan 18 ;292(1):218-225

100
UI  - 862
AU  - Jackson JB
AU  - Dutton PL
TI  - The kinetic and redox potentiometric resolution of the carotenoid shifts in Rhodopseudomonas spheroides chromatophores: their relationship to electric field alterations in electron transport and energy coupling
RP  - NOT IN FILE
NT  - UI - 74075599LA - engRN - 0 (Carotenoids)RN - 0 (Cytochromes)RN - 0 (Hydrazones)RN - 0 (Nitriles)RN - 0 (Uncoupling Agents)RN - 2001-95-8 (Valinomycin)RN - 642-15-9 (Antimycin A)RN - 9007-43-6 (Cytochrome c)PT - Journal ArticleDA - 19740308IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:4358810
SO  - Biochim Biophys Acta 1973 Oct 19 ;325(1):102-113

101
UI  - 9955
AU  - Kobayashi Y
AU  - Nishimura M
TI  - Studies on ion transport in cells of photosynthetic bacteria. 3. The influence of uncouplers on hydrogen ion change
MH  - Anions
MH  - Bacteria
MH  - Cells
MH  - Hydrazones
MH  - Hydrogen
MH  - Ion Transport
MH  - Nitriles
MH  - transport
MH  - Uncoupling Agents
RP  - NOT IN FILE
NT  - UI - 74109617LA - engRN - 0 (Anions)RN - 0 (Chlorobenzenes)RN - 0 (Hydrazones)RN - 0 (Nitrates)RN - 0 (Nitriles)RN - 0 (Uncoupling Agents)RN - 1333-74-0 (Hydrogen)RN - 7782-50-5 (Chlorine)RN - 83-89-6 (Quinacrine)PT - Journal ArticleDA - 19740429IS - 0021-924XSB - IMCY - JAPANJC - HIF
UR  - PM:4205460
SO  - J Biochem (Tokyo ) 1973 Dec ;74(6):1233-1238

102
UI  - 960
AU  - Layton D
AU  - Azzi A
AU  - Graziotti P
TI  - The use of the fluorescent probe aurovertin, to monitor energy linked conformational changes in mitochondrial ATPases
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - Animal
MH  - diagnostic use
MH  - Energy Metabolism
MH  - Energy Transfer
MH  - enzymology
MH  - Fluorescent Dyes
MH  - In Vitro
MH  - metabolism
MH  - Mitochondria,Liver
MH  - Molecular Conformation
MH  - Oligomycins
MH  - pharmacology
MH  - Rats
MH  - Spectrophotometry,Ultraviolet
MH  - Succinates
MH  - Viscosity
RP  - NOT IN FILE
SO  - FEBS Lett 1973 Oct 1 ;36(1):87-92

103
UI  - 1036
AU  - Mitchell P
TI  - Performance and conservation of osmotic work by proton-coupled solute porter systems
MH  - Adenosinetriphosphatase
MH  - Biological Transport
MH  - Biological Transport,Active
MH  - Cell Membrane
MH  - Diffusion
MH  - Electron Transport
MH  - Escherichia coli
MH  - Glucose
MH  - Mathematics
MH  - metabolism
MH  - Models,Biological
MH  - Osmosis
MH  - Water
RP  - NOT IN FILE
SO  - J Bioenerg 1973 Jan ;4(1):63-91

104
UI  - 1033
AU  - Mitchell P
TI  - Hypothesis: cation-translocating adenosine triphosphatase models: how direct is the participation of adenosine triphosphate and its hydrolysis products in cation translocation?
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - Biological Transport
MH  - Calcium
MH  - Cell Membrane
MH  - enzymology
MH  - Hydrolysis
MH  - metabolism
MH  - Models,Biological
MH  - Phosphates
MH  - Potassium
MH  - Sodium
RP  - NOT IN FILE
SO  - FEBS Lett 1973 Jul 15 ;33(3):267-274

105
UI  - 1035
AU  - Moyle J
AU  - Mitchell P
TI  - The proton-translocating nicotinamide-adenine dinucleotide (phosphate) transhydrogenase of rat liver mitochondria
MH  - Acetoacetates
MH  - Animal
MH  - Biological Transport
MH  - Electron Transport
MH  - enzymology
MH  - Hydrogen-Ion Concentration
MH  - Hydroxybutyrates
MH  - Isocitrate Dehydrogenase
MH  - Isocitrates
MH  - Ketoglutaric Acids
MH  - Kinetics
MH  - Liver
MH  - Malate Dehydrogenase
MH  - Male
MH  - Membranes
MH  - metabolism
MH  - Mitochondria
MH  - Mitochondria,Liver
MH  - Nad
MH  - NADH,NADPH Oxidoreductases
MH  - Nadp
MH  - Oxidation-Reduction
MH  - Oxidoreductases
MH  - Oxygen Consumption
MH  - Potassium
MH  - Rats
MH  - Solubility
MH  - Spectrophotometry
MH  - Spectrophotometry,Ultraviolet
MH  - Time Factors
RP  - NOT IN FILE
SO  - Biochem J 1973 Mar ;132(3):571-585

106
UI  - 1034
AU  - Moyle J
AU  - Mitchell P
TI  - Proton translocation quotient for the adenosine triphosphatase of rat liver mitochondria
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - analysis
MH  - Animal
MH  - Centrifugation,Density Gradient
MH  - enzymology
MH  - Hydrogen
MH  - Hydrogen-Ion Concentration
MH  - Hydrolysis
MH  - Liver
MH  - metabolism
MH  - Mitochondria
MH  - Mitochondria,Liver
MH  - Oligomycins
MH  - Proteins
MH  - Protons
MH  - Rats
MH  - Sucrose
MH  - Time Factors
MH  - Valinomycin
MH  - Vibration
RP  - NOT IN FILE
SO  - FEBS Lett 1973 Mar 15 ;30(3):317-320

107
UI  - 559
AU  - Pedersen PL
TI  - Coupling of adenosine triphosphate formation in mitochondria to the formation of nucleoside triphosphates. Involvement of nucleoside diphosphokinase
RP  - NOT IN FILE
NT  - UI - 73187385LA - engRN - 0 (Cytosine Nucleotides)RN - 0 (Guanine Nucleotides)RN - 0 (Inosine Nucleotides)RN - 0 (Nucleotides)RN - 0 (Thymine Nucleotides)RN - 0 (Uracil Nucleotides)RN - 56-65-5 (Adenosine Triphosphate)RN - 86-01-1 (Guanosine Triphosphate)RN - EC 2.7 (Phosphotransferases)PT - Journal ArticleDA - 19730802IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:4708095
SO  - J Biol Chem 1973 Jun 10 ;248(11):3956-3962

108
UI  - 837
AU  - Strotmann H
AU  - Hesse H
AU  - Edelmann K
TI  - Quantitative determination of coupling factor CF1 of chloroplasts
RP  - NOT IN FILE
NT  - UI - 74014920LA - engRN - 0 (Glyceric Acids)RN - 0 (Organophosphorus Compounds)RN - 0 (Pentosephosphates)RN - 0 (Phosphorus Radioisotopes)RN - 0 (Plant Proteins)RN - 1406-65-1 (Chlorophyll)RN - 7439-95-4 (Magnesium)RN - EC 3.4.21.4 (Trypsin)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 4.1.1. (Carboxy-Lyases)PT - Journal ArticleDA - 19731219IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:4270536
SO  - Biochim Biophys Acta 1973 Aug 31 ;314(2):202-210

109
UI  - 9920
AU  - Thayer WS
AU  - Hinkle PC
TI  - Stoichiometry of adenosine triphosphate-driven proton translocation in bovine heart submitochondrial particles.
MH  - Adenosine
MH  - Adenosine Monophosphate
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - Animal
MH  - Cattle
MH  - Chromatography,Ion Exchange
MH  - Comparative Study
MH  - drug effects
MH  - Electron Transport
MH  - enzymology
MH  - Hydrogen-Ion Concentration
MH  - Kinetics
MH  - Mathematics
MH  - metabolism
MH  - Mitochondria,Muscle
MH  - Myocardium
MH  - Oligomycins
MH  - Oxidative Phosphorylation
MH  - Oxygen Consumption
MH  - pharmacology
MH  - Phosphorus Radioisotopes
MH  - Phosphotransferases
MH  - proton
MH  - Seasons
MH  - Spectrophotometry,Ultraviolet
MH  - Submitochondrial Particles
MH  - Time Factors
RP  - NOT IN FILE
SO  - J Biol Chem 1973 Aug 10 ;248(15):5395-5402

110
UI  - 603
AU  - You K
AU  - Hatefi Y
TI  - Inhibition by avidin of the ATP-Pi enchange activities associated with preparations of energy transfer factors A and A-D
RP  - NOT IN FILE
NT  - UI - 73196459LA - engRN - 0 (Antibiotics, Antifungal)RN - 0 (Oxidative Phosphorylation Coupling Factors)RN - 0 (Phosphates)RN - 0 (Propionates)RN - 0 (Succinates)RN - 0 (Uncoupling Agents)RN - 124-38-9 (Carbon Dioxide)RN - 1405-69-2 (Avidin)RN - 53-84-9 (NAD)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-85-5 (Biotin)RN - 85-61-0 (Coenzyme A)RN - 9006-59-1 (Ovalbumin)RN - EC 6. (Ligases)PT - Journal ArticleDA - 19730810IS - 0006-291XSB - IMCY - UNITED STATESJC - 9Y8
UR  - PM:4351134
SO  - Biochem Biophys Res Commun 1973 May 15 ;52(2):343-349

111
UI  - 20978
AU  - Altendorf K
AU  - Harold FM
AU  - Simoni RD
TI  - Impairment and restoration of the energized state in membrane vesicles of a mutant of Escherichia coli lacking adenosine triphosphatase
MH  - A
MH  - ACID
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - ADENOSINE-TRIPHOSPHATASE
MH  - Adenosinetriphosphatase
MH  - Amino Acids
MH  - ammonium compounds
MH  - Calcium
MH  - Carbon Radioisotopes
MH  - Dicyclohexylcarbodiimide
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - Magnesium
MH  - membrane
MH  - membrane vesicles
MH  - mutant
MH  - proton
MH  - Protons
MH  - Valinomycin
RP  - NOT IN FILE
NT  - UI - 74277416LA - engRN - 0 (Amino Acids)RN - 0 (Ammonium Compounds)RN - 0 (Carbon Radioisotopes)RN - 0 (Protons)RN - 0 (Radioisotopes)RN - 10028-17-8 (Tritium)RN - 147-85-3 (Proline)RN - 2001-95-8 (Valinomycin)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - 7439-95-4 (Magnesium)RN - 7440-17-7 (Rubidium)RN - 7440-70-2 (Calcium)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19741009IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:4276462
SO  - J Biol Chem 1974 Jul 25 ;249(14):4587-4593

112
UI  - 20979
AU  - Altendorf KH
AU  - Staehelin LA
TI  - Orientation of membrane vesicles from Escherichia coli as detected by freeze-cleave electron microscopy
MH  - Bacterial Proteins
MH  - buffer
MH  - Buffers
MH  - electron
MH  - electron microscopy
MH  - ELECTRON-MICROSCOPY
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - membrane
MH  - membrane vesicles
MH  - Microscopy
MH  - Phosphates
MH  - Potassium
MH  - protein
MH  - Proteins
MH  - Sodium
RP  - NOT IN FILE
NT  - UI - 74090302LA - engRN - 0 (Bacterial Proteins)RN - 0 (Buffers)RN - 0 (Phosphates)RN - 7440-09-7 (Potassium)RN - 7440-23-5 (Sodium)PT - Journal ArticleDA - 19740331IS - 0021-9193SB - IMCY - UNITED STATES
UR  - PM:4590489
SO  - J Bacteriol 1974 Feb ;117(2):888-899

113
UI  - 8303
AU  - Auslnder W
AU  - Junge W
TI  - Electric generator in the photosynthesis of green plants. II. Kinetic correlation between protolytic reactions and redox reactions
MH  - Kinetics
MH  - Photosynthesis
MH  - plant
MH  - Plants
MH  - redox
RP  - NOT IN FILE
SO  - Biochim Biophys Acta 1974  ;357():285-298

114
UI  - 621
AU  - Casadio R
AU  - Baccarini MA
AU  - Zannoni D
AU  - Melandri BA
TI  - Electrochemical proton gradient and phosphate potential in bacterial chromatophores
RP  - NOT IN FILE
NT  - UI - 75075533LA - engRN - 0 (Acridines)RN - 0 (Antibiotics)RN - 0 (Carboxylic Acids)RN - 0 (Phosphates)RN - 2001-95-8 (Valinomycin)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)PT - Journal ArticleDA - 19750408IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:4442600
SO  - FEBS Lett 1974 Dec 15 ;49(2):203-207

115
UI  - 655
AU  - Chang TM
AU  - Penefsky HS
TI  - Energy-dependent enhancement of aurovertin fluorescence. An indicator of conformational changes in beef heart mitochondrial adenosine triphosphatase
RP  - NOT IN FILE
NT  - UI - 74107410LA - engRN - 0 (Antibiotics)RN - 0 (Antibiotics, Antifungal)RN - 0 (Succinates)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 7439-95-4 (Magnesium)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19740425IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:4273518
SO  - J Biol Chem 1974 Feb 25 ;249(4):1090-1098

116
UI  - 19841
AU  - Chude O
AU  - Boyer PD
TI  - Protein-bound sulfhydryl groups and thiolesters in mitochondria and submitochondrial particles and their relationships to oxidative phosphorylation
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - Carbon Radioisotopes
MH  - Dinitrophenols
MH  - Edetic Acid
MH  - Mitochondria
MH  - Oligomycins
MH  - Oxidative Phosphorylation
MH  - Phosphorylation
MH  - Proteins
MH  - Submitochondrial Particles
MH  - succinate
MH  - Succinates
RP  - NOT IN FILE
NT  - UI - 74174283LA - engRN - 0 (Carbon Radioisotopes)RN - 0 (Dinitrophenols)RN - 0 (Guanidines)RN - 0 (Mercuribenzoates)RN - 0 (Oligomycins)RN - 0 (Proteins)RN - 0 (Receptors, Drug)RN - 0 (Succinates)RN - 0 (Sulfhydryl Compounds)RN - 10028-17-8 (Tritium)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 60-00-4 (Edetic Acid)RN - 70-18-8 (Glutathione)RN - EC 1.2.1.- (Glyceraldehyde-3-Phosphate Dehydrogenases)RN - EC 4.1.2.13 (Fructose-Bisphosphate Aldolase)PT - Journal ArticleDA - 19740726IS - 0003-9861SB - IMCY - UNITED STATESJC - 6SK
UR  - PM:4208772
SO  - Arch Biochem Biophys 1974 Feb ;160(2):366-371

117
UI  - 9968
AU  - Cogdell RJ
AU  - Crofts AR
TI  - H+ uptake by chromatophores from Rhodopseudomonas spheroides. The relation between rapid H+ uptake and the H+ pump
MH  - Antibiotics
MH  - Antimycin A
MH  - Carotenoids
MH  - H+
MH  - Hydrazones
MH  - Nitriles
MH  - Uncoupling Agents
MH  - Valinomycin
RP  - NOT IN FILE
NT  - UI - 74269772LA - engRN - 0 (Antibiotics)RN - 0 (Carboxylic Acids)RN - 0 (Carotenoids)RN - 0 (Hydrazones)RN - 0 (Nitriles)RN - 0 (Uncoupling Agents)RN - 2001-95-8 (Valinomycin)RN - 642-15-9 (Antimycin A)RN - 7440-63-3 (Xenon)PT - Journal ArticleDA - 19740911IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:4546206
SO  - Biochim Biophys Acta 1974 May 22 ;347(2):264-272

118
UI  - 897
AU  - Cori CF
TI  - Some highlights of the early period of bioenergetics
MH  - Adenosine Triphosphate
MH  - Animal
MH  - Anura
MH  - Biochemistry
MH  - Coenzymes
MH  - Energy Metabolism
MH  - Epinephrine
MH  - Fermentation
MH  - Glycolysis
MH  - Hexosephosphates
MH  - history
MH  - History of Medicine,20th Cent.
MH  - Iodoacetates
MH  - isolation &amp
MH  - Lactates
MH  - metabolism
MH  - Muscles
MH  - pharmacology
MH  - Phosphates
MH  - Phosphocreatine
MH  - purification
MH  - Rats
MH  - Trioses
RP  - NOT IN FILE
SO  - Mol Cell Biochem 1974 Nov 15 ;5(1-2):47-53

119
UI  - 9969
AU  - Crofts AR
AU  - Evans EH
AU  - Cogdell RJ
TI  - The relation between H+-uptake and electron flow in chromatophores from photosynthetic bacteria
MH  - Antimycin A
MH  - Bacteria
MH  - Carotenoids
MH  - Cytochromes
MH  - electron
MH  - Hydrogen
MH  - Indicators and Reagents
MH  - proton
MH  - Protons
MH  - review
RP  - NOT IN FILE
NT  - UI - 74162132LA - engRN - 0 (Carotenoids)RN - 0 (Cytochromes)RN - 0 (Indicators and Reagents)RN - 0 (Phenanthrolines)RN - 0 (Protons)RN - 1333-74-0 (Hydrogen)RN - 642-15-9 (Antimycin A)PT - Journal ArticlePT - ReviewDA - 19740715IS - 0077-8923SB - IMCY - UNITED STATESJC - 5NM
UR  - PM:4597309
SO  - Ann N Y Acad Sci 1974 Feb 18 ;227():227-243

120
UI  - 19883
AU  - Cross RL
AU  - de Sousa JT
AU  - Packer L
TI  - Thiophosphate labelling of mitochondria-lack of evidence for an acyl- phosphate intermediate in oxidative phosphorylation
MH  - Antibiotics
MH  - England
MH  - Oligomycins
MH  - Oxidative Phosphorylation
MH  - Phosphorylation
MH  - succinate
MH  - Succinates
RP  - NOT IN FILE
NT  - UI - 75060383LA - engRN - 0 (Antibiotics)RN - 0 (Ferricyanides)RN - 0 (Oligomycins)RN - 0 (Organophosphorus Compounds)RN - 0 (Succinates)RN - 0 (Sulfur Radioisotopes)RN - 0 (Thiophosphoric Acid Esters)PT - Journal ArticleDA - 19750310IS - 0449-5705SB - IMCY - ENGLANDJC - HIN
UR  - PM:4436295
SO  - J Bioenerg 1974  ;6(1):21-25

121
UI  - 904
AU  - de Vries WR
TI  - [Energy crisis in the muscles]
MH  - Adenosine Triphosphate
MH  - Energy Metabolism
MH  - Human
MH  - metabolism
MH  - Muscles
MH  - Proteins
RP  - NOT IN FILE
SO  - Tijdschr Ziekenverpl 1974 Sep 24 ;27(33):882-890

122
UI  - 21222
AU  - Drachev LA
AU  - Kaulen AD
AU  - Kondrashin AA
AU  - Liberman EA
AU  - Nemechek IB
TI  - [Electric current generation by cytochrome oxidase, H+-ATPase and bacteriorhodopsin]
MH  - Adenosinetriphosphatase
MH  - cytochrome
MH  - Cytochrome c
MH  - Cytochrome-c Oxidase
MH  - H+-ATPase
MH  - liposome
MH  - Liposomes
MH  - Phospholipids
MH  - pigments
MH  - retinal
RP  - NOT IN FILE
NT  - UI - 75057394LA - rusRN - 0 (Liposomes)RN - 0 (Phospholipids)RN - 0 (Retinal Pigments)RN - 9009-81-8 (Rhodopsin)RN - EC 1.9.3.1 (Cytochrome-c Oxidase)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19750319IS - 0002-3264SB - IMCY - USSR
UR  - PM:4373219
SO  - Dokl Akad Nauk SSSR 1974 Sep 11 ;218(2):481-484

123
UI  - 21223
AU  - Drachev LA
AU  - Jasaitis AA
AU  - Kaulen AD
AU  - Kondrashin AA
AU  - Liberman EA
AU  - Nemecek IB
AU  - Ostroumov SA
AU  - Semenov AY
AU  - Skulachev VP
TI  - Direct measurement of electric current generation by cytochrome oxidase, H+-ATPase and bacteriorhodopsin
MH  - Adenosinetriphosphatase
MH  - Bacterial Proteins
MH  - Bacteriorhodopsin
MH  - cytochrome
MH  - Cytochrome c
MH  - Cytochrome-c Oxidase
MH  - England
MH  - H+-ATPase
MH  - Hydrogen
MH  - liposome
MH  - Liposomes
MH  - Phospholipids
MH  - protein
MH  - Proteins
RP  - NOT IN FILE
NT  - UI - 74275166LA - engRN - 0 (Bacterial Proteins)RN - 0 (Liposomes)RN - 0 (Phospholipids)RN - 1333-74-0 (Hydrogen)RN - 9007-43-6 (Cytochrome c)RN - EC 1.9.3.1 (Cytochrome-c Oxidase)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19740919IS - 0028-0836SB - IMCY - ENGLAND
UR  - PM:4366965
SO  - Nature 1974 May 24 ;249(455):321-324

124
UI  - 21224
AU  - Drachev LA
AU  - Kaulen AD
AU  - Ostroumov SA
AU  - Skulachev VP
TI  - Electrogenesis by bacteriorhodopsin incorporated in a planar phospholipid membrane
MH  - A
MH  - Bacterial Proteins
MH  - Bacteriorhodopsin
MH  - Cyanides
MH  - Hydrazones
MH  - membrane
MH  - Phospholipids
MH  - pigments
MH  - protein
MH  - Proteins
MH  - retinal
RP  - NOT IN FILE
NT  - UI - 74301195LA - engRN - 0 (Bacterial Proteins)RN - 0 (Cyanides)RN - 0 (Hydrazones)RN - 0 (Phospholipids)RN - 0 (Retinal Pigments)PT - Journal ArticleDA - 19741024IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:4851814
SO  - FEBS Lett 1974 Feb 1 ;39(1):43-45

125
UI  - 21135
AU  - Ferguson SJ
AU  - John P
AU  - Lloyd WJ
AU  - Radda GK
AU  - Whatley FR
TI  - Selective and reversible inhibition of the ATPase of Micrococcus denitrificans by 7-chloro-4-nitrobenzo-2-oxa-1,3 diazole
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - ATPase
MH  - Dithiothreitol
RP  - NOT IN FILE
NT  - UI - 75014818LA - engRN - 0 (Nitro Compounds)RN - 0 (Oxazoles)RN - 3483-12-3 (Dithiothreitol)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19741216IS - 0006-3002SB - IMCY - NETHERLANDS
UR  - PM:4278263
SO  - Biochim Biophys Acta 1974 Sep 20 ;357(3):457-461

126
UI  - 228
AU  - Futai M
TI  - Orientation of membrane vesicles from Escherichia coli prepared by different procedures
MH  - ACID
MH  - Adenosinetriphosphatase
MH  - Cholic Acid
MH  - Cholic Acids
MH  - Dicyclohexylcarbodiimide
MH  - Edetic Acid
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - membrane
MH  - membrane vesicles
MH  - Oxidoreductases
MH  - Phosphorus
MH  - Phosphorus Radioisotopes
MH  - Surface-Active Agents
RP  - NOT IN FILE
NT  - UI - 74265678LA - engRN - 0 (Cholic Acids)RN - 0 (Ferricyanides)RN - 0 (Phosphorus Radioisotopes)RN - 0 (Surface-Active Agents)RN - 108-88-3 (Toluene)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 60-00-4 (Edetic Acid)RN - EC 1.6. (NADH, NADPH Oxidoreductases)RN - EC 3.2.1.17 (Muramidase)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19740906IS - 0022-2631SB - IMCY - UNITED STATES
UR  - PM:4152065
SO  - J Membr Biol 1974  ;15(1):15-28

127
UI  - 226
AU  - Futai M
AU  - Sternweis PC
AU  - Heppel LA
TI  - Purification and properties of reconstitutively active and inactive adenosinetriphosphatase from Escherichia coli
MH  - ACTIVE
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - Calcium
MH  - coupling
MH  - COUPLING FACTOR
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - Magnesium
MH  - Oxidative Phosphorylation
MH  - Oxidative Phosphorylation Coupling Factors
MH  - Oxidoreductases
MH  - Phosphorylation
MH  - purification
MH  - Sodium
RP  - NOT IN FILE
NT  - UI - 74306426LA - engRN - 0 (Oxidative Phosphorylation Coupling Factors)RN - 151-21-3 (Sodium Dodecyl Sulfate)RN - 56-65-5 (Adenosine Triphosphate)RN - 7439-95-4 (Magnesium)RN - 7440-70-2 (Calcium)RN - EC 1.6. (NADH, NADPH Oxidoreductases)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19741118IS - 0027-8424SB - IMCY - UNITED STATES
UR  - PM:4153028
SO  - Proc Natl Acad Sci U S A 1974 Jul ;71(7):2725-2729

128
UI  - 1209
AU  - Graber P
AU  - Witt HT
TI  - Extent of the electrical potential across the thylakoid membrane induced by continuous light in Chlorella cells
MH  - Cells
MH  - Light
MH  - membrane
MH  - thylakoid
MH  - thylakoid membrane
RP  - ON REQUEST (03/18/92)
SO  - Biochim Biophys Acta 1974  ;333():389-392

129
UI  - 18423
AU  - Halsey YD
AU  - Parson WW
TI  - Identification of ubiquinone as the secondary electron acceptor in the photosynthetic apparatus of Chromatium vinosum.
MH  - acceptor
MH  - Antibiotics
MH  - Antimetabolites
MH  - Chromatium
MH  - Chromatography,Thin Layer
MH  - chromatophores
MH  - Comparative Study
MH  - Cytochrome c
MH  - cytology
MH  - drug effects
MH  - electron
MH  - electron acceptor
MH  - Electron Transport
MH  - Energy Transfer
MH  - Kinetics
MH  - metabolism
MH  - Oxidation-Reduction
MH  - Petroleum
MH  - pharmacology
MH  - Photosynthesis
MH  - secondary
MH  - Spectrophotometry
MH  - Spectrophotometry,Ultraviolet
MH  - Structure-Activity Relationship
MH  - Temperature
MH  - Time Factors
MH  - Ubiquinone
RP  - NOT IN FILE
SO  - Biochim Biophys Acta 1974 Jun 28 ;347(3):404-416

130
UI  - 601
AU  - Hatefi Y
AU  - Stiggall DL
AU  - Galante Y
AU  - Hanstein WG
TI  - Mitochondrial ATP-Pi exchange complex
RP  - NOT IN FILE
NT  - UI - 75072566LA - engRN - 0 (Adenine Nucleotides)RN - 0 (Antimetabolites)RN - 0 (Arsenates)RN - 0 (Azides)RN - 0 (Cytochromes)RN - 0 (Phosphates)RN - 0 (Phosphorus Radioisotopes)RN - 0 (Ribonucleotides)RN - 0 (Uncoupling Agents)RN - 2001-95-8 (Valinomycin)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - 7440-09-7 (Potassium)RN - EC 1.3.99.1 (Succinate Dehydrogenase)RN - EC 1.6. (NADH, NADPH Oxidoreductases)RN - EC 1.6.2. (Cytochrome Reductases)RN - EC 1.9.3.1 (Cytochrome-c Oxidase)PT - Journal ArticleDA - 19750314IS - 0006-291XSB - IMCY - UNITED STATESJC - 9Y8
UR  - PM:4155298
SO  - Biochem Biophys Res Commun 1974 Nov 6 ;61(1):313-321

131
UI  - 1031
AU  - Mitchell P
TI  - A chemiosmotic molecular mechanism for proton-translocating adenosine triphosphatases
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - Bacteria
MH  - Binding Sites
MH  - Chloroplasts
MH  - Comparative Study
MH  - enzymology
MH  - Kinetics
MH  - Magnesium
MH  - Mathematics
MH  - metabolism
MH  - Mitochondria
MH  - Models,Biological
MH  - Osmosis
MH  - Oxygen Isotopes
MH  - Protein Binding
MH  - Thermodynamics
RP  - NOT IN FILE
SO  - FEBS Lett 1974 Jul 15 ;43(2):189-194

132
UI  - 654
AU  - Penefsky HS
TI  - Differential effects of adenylyl imidodiphosphate on adenosine triphosphate synthesis and the partial reactions of oxidative phosphorylation
RP  - NOT IN FILE
NT  - UI - 74172625LA - engRN - 0 (Anilino Naphthalenesulfonates)RN - 0 (Imides)RN - 0 (Phosphates)RN - 0 (Phosphoric Acids)RN - 0 (Phosphorus Radioisotopes)RN - 0 (Succinates)RN - 53-84-9 (NAD)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19740726IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:4364660
SO  - J Biol Chem 1974 Jun 10 ;249(11):3579-3585

133
UI  - 870
AU  - Repke KR
AU  - Dittrich F
AU  - Schon R
TI  - Chemical reaction mechanism for ATP synthesis and hydrolysis by ATP synthetase
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - biosynthesis
MH  - Energy Metabolism
MH  - Hydrolysis
MH  - Ligands
MH  - metabolism
MH  - Molecular Conformation
MH  - Multienzyme Complexes
MH  - Osmosis
MH  - Phosphates
MH  - Protons
MH  - Thermodynamics
RP  - NOT IN FILE
SO  - Acta Biol Med Ger 1974  ;33(1):K39-K47

134
UI  - 893
AU  - Repke KR
AU  - Schon R
TI  - Flip-flop model of energy interconversion by ATP synthetase
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - Biological Transport,Active
MH  - biosynthesis
MH  - Cell Membrane
MH  - Electrochemistry
MH  - Energy Metabolism
MH  - Energy Transfer
MH  - Hydrolysis
MH  - Ions
MH  - Magnesium
MH  - Membranes
MH  - metabolism
MH  - Models,Biological
MH  - Models,Chemical
MH  - Multienzyme Complexes
MH  - Osmosis
MH  - Phosphates
MH  - Potassium
MH  - Protons
MH  - Sodium
MH  - Thermodynamics
RP  - NOT IN FILE
SO  - Acta Biol Med Ger 1974  ;33(1):K27-K38

135
UI  - 885
AU  - Slater EC
TI  - Biological energy conversion
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - Bioelectric Energy Sources
MH  - Energy Metabolism
MH  - Energy Transfer
MH  - Human
MH  - metabolism
MH  - Mitochondria
MH  - physiology
RP  - NOT IN FILE
SO  - Bull Acad R Med Belg 1974  ;129(8-9):491-506

136
UI  - 9107
AU  - Strotmann H
AU  - Tischer W
AU  - Edelmann K
TI  - Specific binding of inhibitors by electron carriers of the photosynthetic electron transport chain
MH  - BINDING
MH  - carrier
MH  - electron
MH  - Electron Transport
MH  - inhibitor
MH  - transport
RP  - IN FILE
SO  - Ber Deut Bot Ges 1974  ;87():457-463

137
UI  - 1032
AU  - West IC
AU  - Mitchell P
TI  - The proton-translocating ATPase of Escherichia coli
MH  - Adenosinetriphosphatase
MH  - Anaerobiosis
MH  - Biological Transport,Active
MH  - Carbonate Dehydratase
MH  - Cell Membrane
MH  - Comparative Study
MH  - Cyanides
MH  - cytology
MH  - Dicyclohexylcarbodiimide
MH  - drug effects
MH  - Enzyme Activation
MH  - enzymology
MH  - Escherichia coli
MH  - Hydrazones
MH  - Hydrogen-Ion Concentration
MH  - Kinetics
MH  - Magnesium
MH  - metabolism
MH  - Organoids
MH  - pharmacology
MH  - Time Factors
MH  - Ultrasonics
MH  - Valinomycin
RP  - NOT IN FILE
SO  - FEBS Lett 1974 Mar 15 ;40(1):1-4

138
UI  - 1030
AU  - West IC
AU  - Mitchell P
TI  - Proton/sodium ion antiport in Escherichia coli
MH  - Biological Transport,Active
MH  - Cell Membrane
MH  - drug effects
MH  - Escherichia coli
MH  - Hydrogen-Ion Concentration
MH  - metabolism
MH  - Oxygen Consumption
MH  - pharmacology
MH  - Protons
MH  - Sodium
MH  - Uncoupling Agents
RP  - NOT IN FILE
SO  - Biochem J 1974 Oct ;144(1):87-90

139
UI  - 20977
AU  - Altendorf K
AU  - Zitzmann W
TI  - Identification of the DCCD-reactive protein of the energy transducing adenosinetriphosphatase complex from Escherichia coli
MH  - Adenosinetriphosphatase
MH  - Bacterial Proteins
MH  - COMPLEX
MH  - Dicyclohexylcarbodiimide
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - protein
MH  - Proteins
MH  - proton
MH  - Protons
RP  - NOT IN FILE
NT  - UI - 76210749LA - engRN - 0 (Bacterial Proteins)RN - 0 (Carbodiimides)RN - 0 (Protons)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19760823IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:132364
SO  - FEBS Lett 1975 Nov 15 ;59(2):268-272

140
UI  - 1265
AU  - Auslnder W
AU  - Junge W
TI  - Neutral red, a rapid indicator for pH changes in the inner phase of thylakoids
MH  - A
MH  - indicator
MH  - method
MH  - NEUTRALRED
MH  - pH
MH  - thylakoid
MH  - wox
RP  - IN FILE
SO  - FEBS Lett 1975  ;59(2)():310-315

141
UI  - 19839
AU  - Boyer PD
TI  - Energy transduction and proton translocation by adenosine triphosphatases
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - ADENOSINE-TRIPHOSPHATASE
MH  - Adenosinetriphosphatase
MH  - Phosphates
MH  - proton
MH  - translocation
RP  - NOT IN FILE
NT  - UI - 75093685LA - engRN - 0 (Phosphates)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19750513IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:122942
SO  - FEBS Lett 1975 Feb 1 ;50(2):91-94

142
UI  - 19838
AU  - Boyer PD
AU  - Stokes BO
AU  - Wolcott RG
AU  - Degani C
TI  - Coupling of "high-energy" phosphate bonds to energy transductions
AB  - Recent results suggest consideration of a new concept for oxidative phosphorylation in which a prime function of energy is to bring about release of ATP formed at the catalytic site by reversal of hydrolysis. Data with submitochondrial particles include properties of an uncoupler insensitive Pi=HOH exchange, a rapid reversible formation of bound ATP in presence of uncouplers, and predictable patterns of 32-Pi incorporation into ATP in rapid mixing experiments. ADP is confirmed as the primary Pi acceptor in mitochondrial ATP synthesis, but with chloroplasts ADP is also rapidly labeled. Other findings with pyrophosphatase and with transport ATPase harmonize with the new concept. Measurements of the reversal of ATP cleavage and binding by myosin suggest that oxygen exchanges result from reversible cleavage of ATP to ADP and Pi at the catalytic site and that the principal free energy change in ATP cleavage occurs in ATP binding. Reversal of conformational changes accompanying ATP binding and cleavage is proposed to drive the actin filament in contraction. Thus energy transductions linked to ATP in both mitochondria and muscle may occur primarily through protein conformational change
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - ADP
MH  - atp
MH  - ATP synthesis
MH  - ATPase
MH  - BINDING
MH  - chloroplast
MH  - Chloroplasts
MH  - conformational change
MH  - Hydrolysis
MH  - Mitochondria
MH  - Oligomycins
MH  - Oxidative Phosphorylation
MH  - Oxygen
MH  - Phosphates
MH  - Phosphorylation
MH  - Pyrophosphatases
MH  - review
MH  - Submitochondrial Particles
MH  - synthesis
MH  - transport
MH  - Uncoupling Agents
MH  - Water
RP  - NOT IN FILE
NT  - UI - 75168308LA - engRN - 0 (Myosin)RN - 0 (Oligomycins)RN - 0 (Phosphates)RN - 0 (Uncoupling Agents)RN - 56-65-5 (Adenosine Triphosphate)RN - 7732-18-5 (Water)RN - EC 3.6.1.- (Pyrophosphatases)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticlePT - ReviewDA - 19750829IS - 0014-9446SB - IMCY - UNITED STATESJC - EUV
UR  - PM:124270
SO  - Fed Proc 1975 Jul ;34(8):1711-1717

143
UI  - 19837
AU  - Boyer PD
TI  - A model for conformational coupling of membrane potential and proton translocation to ATP synthesis and to active transport
AB  - Acceptance of a membrane potential and/or a proton gradient as a possible means of transmitting energy from oxidations to ATP synthesis rests in part on a satisfactory hypothesis for how the potential or proton gradient could drive ATP synthesis. Recognition that energy input may drive ATP synthesis by change in binding of reactants at the catalytic site has led to the suggestions presented in this paper. These are that in oxidative phosphorylation and photophosphorylation, the requisite conformational changes may be coupled to exposure of charged groups to different sides of the membrane. The cycle of charged group exposure or movement may be driven by the membrane potential or, through protonation and deprotonation, may be coupled to proton translocation across the membrane. Effects of proton gradient and membrane potential may be additive. Similar conformational coupling suggestions may explain proton translocation coupled to ATP cleavage and active transport of metabolites coupled to membrane potential, proton gradients of ATP cleavage
MH  - A
MH  - ACTIVE
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthesis
MH  - BINDING
MH  - conformational change
MH  - membrane
MH  - Membrane Potential
MH  - model
MH  - Movement
MH  - Oxidative Phosphorylation
MH  - Phosphorylation
MH  - Photophosphorylation
MH  - proton
MH  - Protons
MH  - synthesis
MH  - translocation
MH  - transport
RP  - NOT IN FILE
NT  - UI - 76187898LA - engRN - 0 (Protons)RN - 56-65-5 (Adenosine Triphosphate)PT - Journal ArticleDA - 19760802IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:1225567
SO  - FEBS Lett 1975 Oct 15 ;58(1):1-6

144
UI  - 19840
AU  - Cross RL
AU  - Boyer PD
TI  - The rapid labeling of adenosine triphosphate by 32P-labeled inorganic phosphate and the exchange of phosphate oxygens as related to conformational coupling in oxidative phosphorylation
AB  - Evidence is presented that extends and amplifies the concept that in oxidative phosphorylation energy input serves to bring about release of ATP formed at a catalytic site by reversal of hydrolysis. The evidence with beef heart submitochondrial particles includes additional demonstration of uncoupler insensitive Pi leads to HOH exhchange, demonstration that this exchange is sensitive to the specific phosphorylation inhibitor, oligomycin, and demonstration that the small burst of uncoupler-insensitive ATP, rapidly labeled after addition of a tracer of 32Pi, behaves in a manner consistent with its participation as a membrane-bound intermediate in the Pi leads to HOH exchange. In addition, data are presented showing that addition of hexokinase plus glucose to submitochondrial particles in presence of ADP and Pi considerably lowers the Pi leads to HOH exchange but that further addition of cyanide or 2,4-dinitrophenol or both has little additional effect. Such data are compatible with no energy requirement for formation of bound ATP. However, with a large excess of hexokinase, the rate of the Pi leads to HOH exchange is further depressed. This could reflect some use of energy to promote formation of ATP at the catalytic site or to maintain the integrity of the phosphorylation system. Relationships of these findings to related information in the field are discussed
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - ADP
MH  - atp
MH  - Dinitrophenols
MH  - FIELD
MH  - Glucose
MH  - Hexokinase
MH  - Hydrolysis
MH  - INORGANIC-PHOSPHATE
MH  - Oligomycins
MH  - Oxidative Phosphorylation
MH  - Oxygen
MH  - Oxygen Isotopes
MH  - Phosphates
MH  - Phosphorus
MH  - Phosphorus Radioisotopes
MH  - Phosphorylation
MH  - Submitochondrial Particles
MH  - Uncoupling Agents
RP  - NOT IN FILE
NT  - UI - 75127922LA - engRN - 0 (Dinitrophenols)RN - 0 (Oligomycins)RN - 0 (Oxygen Isotopes)RN - 0 (Phosphates)RN - 0 (Phosphorus Radioisotopes)RN - 0 (Uncoupling Agents)RN - 56-65-5 (Adenosine Triphosphate)PT - Journal ArticleDA - 19750625IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:1168064
SO  - Biochemistry 1975 Jan 28 ;14(2):392-398

145
UI  - 21133
AU  - Ferguson SJ
AU  - Lloyd WJ
AU  - Radda GK
TI  - The mitochondrial ATPase. Selective modification of a nitrogen residue in the beta subunit
AB  - 1. When mitochondrial ATPase, which has been modified on a single tyrosine residue by 4-chloro-7-nitrobenzofurazan, is incubated at pH 9.0, the 7-nitrobenzofurazan group undergoes an intramolecular transfer to a nitrogen residue. The rate of this transfer is sensitive to the binding of adenine nucleotides to the enzyme. The resulting N- nitrobenzofurazan ATPase has little or no activity. 2. The fluorescence of the N-nitrobenzofurazan group in the modified ATPase is quenched on binding of ADP. 3. Electrophoresis of the modified enzyme in sodium dodecyl sulphate on a 10% polyacrylamide gel shows that the fluorescence of the N-nitrobenzofurazan chromophore is exclusively in the beta subunit. 4. The rate of transfer of the nitrobenzofurazan group from tyrosyl oxygen to nitrogen on the enzyme is compared with the rate of transfer between model compounds. 5. The interaction of the N-nitrobenzofurazan ATPase with aurovertin is reported
MH  - A
MH  - Adenine Nucleotides
MH  - ADENINE-NUCLEOTIDES
MH  - Adenosinetriphosphatase
MH  - ADP
MH  - ATPase
MH  - Aurovertins
MH  - BETA
MH  - BETA-SUBUNIT
MH  - BINDING
MH  - Electrophoresis
MH  - fluorescence
MH  - Macromolecular Systems
MH  - model
MH  - Nucleotides
MH  - Oxygen
MH  - pH
MH  - RESIDUE
MH  - Sodium
MH  - SUBUNIT
MH  - SYSTEM
MH  - SYSTEMS
MH  - TRANSFER
RP  - NOT IN FILE
NT  - UI - 75208996LA - engRN - 0 (Adenine Nucleotides)RN - 0 (Aurovertins)RN - 0 (Macromolecular Systems)RN - 151-21-3 (Sodium Dodecyl Sulfate)RN - 55520-40-6 (Tyrosine)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19751108IS - 0014-2956SB - IMCY - GERMANY, WEST
UR  - PM:238840
SO  - Eur J Biochem 1975 May ;54(1):127-133

146
UI  - 21134
AU  - Ferguson SJ
AU  - Lloyd WJ
AU  - Lyons MH
AU  - Radda GK
TI  - The mitochondrial ATPase. Evidence for a single essential tyrosine residue
AB  - 1. Evidence is presented which indicates that inactivation of the mitochondrial ATPase from bovine heart by the reagent 4-chloro-7- nitrobenzofurazan results from modification of one tyrosine residue per enzyme molecule. Activity can be restored by a variety of sulphydryl reagents. 2. In sodium dodecyl sulphate, the nitrogenzofurazan group on tyrosine is transfered to newly exposed sulphydryl groups on the enzyme. 3. The rate of transfer of the nitrobenzofurazan moiety from theenzyme to sulphydryl compounds is compared with that for transfer from the model compound N-acetyl-tyrosine-0(7-nitrobenzo-furazan) ethyl ester, the synthesis and properties of which are also described. 4. The ligands ATP and ADP exert a protective effect on the rate of reaction between the mitochondrial ATPase and 4-chloro-7-nitrobenzofurazan. The variation in rate of this reaction with change in pH has also been examined and a pKa of 9.5 estimated for the tyrosine residue. 5. The modification does not prevent substrate binding as judged by changes in the fluorescence of aurovertin, an antibiotic with specific affinity for mitochondiral ATPases. 6. When the ATPase activity of submitochondrial particles is inhibited by 4-chloro-7-nitrobenzo- furazan, there is a parallel decrease in the extent of the energy- linked fluorescence enhancement of 1-anilino-naphthalene-8-sulphonate induced by ATP hydrolysis. Both ATPase activity and the fluorescence enhancement are restored by sluphydryl reagents
MH  - A
MH  - Adenine Nucleotides
MH  - ADENINE-NUCLEOTIDES
MH  - Adenosinetriphosphatase
MH  - ADP
MH  - atp
MH  - ATPase
MH  - Aurovertins
MH  - BINDING
MH  - fluorescence
MH  - Hydrolysis
MH  - Ligands
MH  - model
MH  - Nucleotides
MH  - pH
MH  - RESIDUE
MH  - Sodium
MH  - Submitochondrial Particles
MH  - synthesis
MH  - TRANSFER
RP  - NOT IN FILE
NT  - UI - 75208995LA - engRN - 0 (Adenine Nucleotides)RN - 0 (Aurovertins)RN - 0 (Oxadiazoles)RN - 0 (Sulfhydryl Compounds)RN - 151-21-3 (Sodium Dodecyl Sulfate)RN - 509-14-8 (Tetranitromethane)RN - 55520-40-6 (Tyrosine)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19751108IS - 0014-2956SB - IMCY - GERMANY, WEST
UR  - PM:238839
SO  - Eur J Biochem 1975 May ;54(1):117-126

147
UI  - 126
AU  - Fillingame RH
TI  - Identification of the dicyclohexylcarbodiimide-reactive protein component of the adenosine 5'-triphosphate energy-transducing system of Escherichia coli
AB  - Membranes of Escherichia coli contain an adenosine 5'-triphosphate (ATP) energy-transducing system that is inhibited by treatment with dicyclohexylcarbodiimide (DCCD). The carbodiimide-reactive protein component of this system has been identified after treatment with [14C]DCCD. This protein has an apparent molecular weight of 9,000 as judged from acrylamide gel electrophoresis in the presence of sodium dodecyl sulfate and is extracted from the membrane with chloroform- methanol (2:1). These properties are similar to the analogous protein previously identified in mitochondria (Cattell et al., 1971). A mutant strain, RF-7, has been isolated which derives energy from oxidative phosphorylation in the presence of 5 mM DCCD. The ATP hydrolase activity of the membraned system in the mutant was considerably less sensitive to inhibition by DCCD than that in the wild type. The carbodiimide-reactive protein, which was easily labeled by [14C]DCCD in the wild type, was labeled much less rapidly in the carbodiimide- resistant mutant. It is thus concluded that the reaction of DCCD with this specific protein leads to inhibition of the ATP energy-transducing reactions. The mutation causing carbodiimide resistance in strain RF-7 was mapped. It is cotransduced with the uncA gene at a frequency exceeding 90%. The mutationally altered protein causing the carbodiimide resistance was not conclusively identified. However, reconstitution experiments indicate that the altered protein is not one of the subunits of the soluble ATP hydrolase activity, which can be removed from the membrane by washing with 1 mM tris(hydroxymethyl)aminomethane buffer lacking Mg2+. The carbodiimide- reactive protein remains with the membrane residue after removal of the soluble ATP hydrolase and is thus distinct from these subunits as well
RP  - NOT IN FILE
NT  - UI - 76046239LA - engRN - 0 (Bacterial Proteins)RN - 0 (Carbodiimides)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19760110IS - 0021-9193SB - IMCY - UNITED STATESJC - HH3
UR  - PM:126994
SO  - J Bacteriol 1975 Nov ;124(2):870-883

148
UI  - 653
AU  - Garrett NE
AU  - Penefsky HS
TI  - Physical and enzymatic properties of nucleotide-depleted beef heart mitochondrial adenosine triphosphatase
AB  - Tightly bound adenine nucleotides are removed from multiple binding sites on beef heart mitochondrial ATPase (F1) by chromatography on columns of Sephadex equilibrated with 50% glycerol. Release of nucleotides from the enzyme is associated with large decreases in sedimentation velocity (from 11.9 S to 8.4 S) which may be observed in concentrated solutions of polyols. Polyol-induced conformational changes are reversed when the enzyme is returned to dilute buffers. The nucleotide-depleted enzyme restores oxidative phosphorylation in F1- deficient submitochondrial particles. Reconstitution of nucleotide- depleted F1 with the ATP analog (adenylyl-imidodiphosphate (AMP-PNP), almost 5 moles of AMP-PNP per mole of enzyme, results in preparations with substantially inhibited ATPase activity which nevertheless restores oxidative phosphorylation and the 32Pi-ATP exchange reaction in F1-deficient submitochondrial particles. Incubation of the analog- labeled enzyme with ATP and Mg++ results in partial displacement of the analog and a time-dependent recovery of ATPase activity
RP  - NOT IN FILE
NT  - UI - 76098954LA - engRN - 0 (Adenine Nucleotides)RN - 25612-73-1 (Adenylyl Imidodiphosphate)RN - 56-65-5 (Adenosine Triphosphate)RN - 57-50-1 (Sucrose)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19760320IS - 0091-7419SB - IMCY - UNITED STATESJC - K75
UR  - PM:128661
SO  - J Supramol Struct 1975  ;3(5-6):469-478

149
UI  - 652
AU  - Garrett NE
AU  - Penefsky HS
TI  - Interaction of adenine nucleotides with multiple binding sites on beef heart mitochondrial adenosine triphosphatase
AB  - Beef heart mitochondrial ATPase (F1) contained 2 mol of ADP and 1 mol of ATP/mol of enzyme, which resisted removal by Sephadex chromatography with dilute buffers or repeated precipitation with ammonium sulfate. The native enzyme also contained two apparently equivalent binding sites, which participated in readily reversible binding of adenyl-5'- ylimidodiphosphate (AMP-P(NH)P), with a Kd of 1.3 mum. The failure of AMP-P(NH)P to compete effectively with ADP for binding sites on F1 may be related to the failure of the analog to inhibit oxidative phosphorylation. Virtually complete removal of all adenine nucleotides from F1 occurred when the enzyme was chromatographed on columns of Sephadex equilibrated with 50% glycerol. No loss in ATPase activity was observed following removal of nucleotides from the enzyme, which was then capable of binding more than 4 mol of ADP and almost 5 mol of AMP- P(NH)P/mol of protein. Subsequent chromatography on columns of Sephadex equilibrated with dilute buffers containing Mg2+ removed only 1.5 mol of ADP and no AMP-P(NH)P from the enzyme. Reconstitution of F1 with ADP or with almost 5 mol of AMP-P(NH)P resulted in preparations that exhibited an undiminished capacity to restore oxidative phosphorylation in F1-deficient submitochondrial particles
RP  - NOT IN FILE
NT  - UI - 76005545LA - engRN - 0 (Adenine Nucleotides)RN - 25612-73-1 (Adenylyl Imidodiphosphate)RN - 56-65-5 (Adenosine Triphosphate)RN - 56-81-5 (Glycerol)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19751204IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:125756
SO  - J Biol Chem 1975 Sep 10 ;250(17):6640-6647

150
UI  - 8690
AU  - Graber P
AU  - Witt HT
TI  - Direct measurement of the protons pumped into the inner phase of the functional membrane of photosynthesis per electron transfer
MH  - atp
MH  - electron
MH  - electron transfer
MH  - Electron-transfer
MH  - membrane
MH  - Photosynthesis
MH  - proton
MH  - Protons
MH  - TRANSFER
MH  - transport
RP  - ON REQUEST (05/29/92)
SO  - FEBS Lett 1975  ;59():184-189

151
UI  - 1207
AU  - Graber P
AU  - Witt HT
TI  - Electrical potential difference, pH gradient, and phosphorylation. Relation between the transmembrane electrical potential difference, pH gradient, and ATP formation in photosynthesis
MH  - atp
MH  - ATP FORMATION
MH  - pH
MH  - Phosphorylation
MH  - Photosynthesis
T2  - Proc. Int. Congr. Photosynth., 3rd, Meeting Date 1974, Volume 1, 427-36. Edited by: Avron, Mordhay. Am Elsevier: New York, N. Y
Y2  - -32676  
RP  - ON REQUEST (03/18/92)
SO  -  1975  ;():

152
UI  - 1208
AU  - Graber P
AU  - Witt HT
TI  - Effect of temperature on flash-induced transmembrane currents in chloroplasts of spinach
MH  - chloroplast
MH  - Chloroplasts
MH  - Spinach
MH  - Temperature
T2  - Proc. Int. Congr. Photosynth., 3rd, Meeting Date 1974, Volume 2, 951-6. Edited by: Avron, Mordhay. Am Elsevier: New York, N. Y
Y2  - -32676  
RP  - ON REQUEST (03/18/92)
SO  -  1975  ;():

153
UI  - 599
AU  - Hatefi Y
AU  - Hanstein WG
AU  - Galante Y
AU  - Stiggall DL
TI  - Mitochondrial ATP-Pi exchange complex and the site of uncoupling of oxidative phosphorylation
AB  - Five enzyme complexes, which are concerned with electron transport and oxidative phosphorylation, have been isolated from beef heart mitochondria. Enzyme complexes I, II, III and IV are the electron transfer complexes discovered in 1961. Complex V is an energy- conserving complex. It catalyzes ATP-Pi exchange and ATP hydrolysis. The exchange reaction is sensitive to uncouplers, rutamycin, valinomycin plus K-+, dicyclorexylcarboditmide, arsenate, azide, and adenylyl imidodiphosphate. It is also specific for ATP; ITP, GTP and UTP are essentially ineffective. Studies with the photoaffinity labeling uncoupler, 2-azido-4-nitrophenol (NPA), have shown that the mitochondrial uncoupler-binding sites are located exclusively in complex V. Complexes I, III and IV, which carry the three coupling sites of the respiratory chain, had negligible capacity for the binding of NPA, whereas the uncoupler-binding capacity of complex V appeared to be increased two- to threefold as compared to mitochondria. Complexes I, II, III, IV and V are obtained from the same batch of mitochondria by a simple fractionation procedure, which employs cholate, deoxycholate, ammonium acetate and ammonium sulfate. Studies with NPA have shown that mitochondria contain per milligram protein about 0.6 nmole of uniformly reacting uncoupler binding site. All of the uncouplers tested appeared to interact competitively with this site. Photoaffinity labeling with tritiated NPA has shown that a major portion of NPA binds to a polypeptide of molecular weight between 26,000 and 30,000. Other studies on the mechanism of uncoupling have shown that picrate is a membrane-impermeable uncoupler. It cannot uncouple mitochondria. However, it is an effective uncoupler of ATP synthesis and ATP-induced transhydrogenation or reverse electron transfer when used in conjunction with sonicated submitochondrial particles, which have an inside-out orientation of the inner membrane with respect to the medium. In these particles, picrate binds to the same uncoupler-binding site as NPA and other uncouplers. However, unlike the membrane-permeable uncouplers, picrate is a poor protonophore. It has a very small effect on the proton permeability of phosphorylating submitochondrial vesicles, even at two to three times the concentration needed for complete uncoupling. The increase in the proton permeability of submitochondrial vesicles caused by such high concentrations of picrate (500 mum) can be achieved with approximately 5 mum 2,4-dinitrophenol. At this concentration, dinitrophenol results in only about 20% uncoupling
RP  - NOT IN FILE
NT  - UI - 75168306LA - engRN - 0 (Phosphates)RN - 0 (Picrates)RN - 0 (Uncoupling Agents)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 1. (Oxidoreductases)PT - Journal ArticlePT - ReviewDA - 19750829IS - 0014-9446SB - IMCY - UNITED STATESJC - EUV
UR  - PM:1093889
SO  - Fed Proc 1975 Jul ;34(8):1699-1706

154
UI  - 600
AU  - Hatefi Y
TI  - Energy conservation and uncoupling in mitochondria
AB  - Energy conservation and uncoupling in mitochondria are examined in the light of three important new findings: (a) Studies with the photoaffinity-labeling uncoupler 2-azido-4-nitrophenol have shown that mitochondria contain a specific uncoupler binding site (apparently a polypeptide of Mr = 30,000 +/- 10%). (b) This site fractionates into an enzyme complex (complex V), which is capable of oligomycin- and uncoupler-sensitive ATP-Pi exchange. It is absent from electron transfer complexes I, III, and IV, which represent segments of the respiratory chain containing coupling sites 1, 2, and 3, respectively. (c) Trinitrophenol is a membrane-impermeable uncoupler (uncouples submitochondrial particles, but not mitochondria) and a poor protonophore. There is an excellent correlation between the uncoupling potencies and the affinities of uncouplers for the mitochondrial uncoupler-binding site. There is no correlation between uncoupling potency and protonophoric activity of uncouplers when a membrane- permeable uncoupler is compared with a membrane-impermeable one
RP  - NOT IN FILE
NT  - UI - 76049981LA - engRN - 0 (Cyanides)RN - 0 (Dinitrophenols)RN - 0 (Nitrophenols)RN - 0 (Uncoupling Agents)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - 7440-09-7 (Potassium)RN - 7440-31-5 (Tin)PT - Journal ArticlePT - ReviewDA - 19760116IS - 0091-7419SB - IMCY - UNITED STATESJC - K75
UR  - PM:1102805
SO  - J Supramol Struct 1975  ;3(3):201-213

155
UI  - 860
AU  - Jackson JB
AU  - Saphon S
AU  - Witt HT
TI  - The extent of the stimulated electrical potential decay under phosphorylating conditions and the H+/ATP ratio in Rhodopseudomonas sphaeroides chromatophores following short flash excitation
AB  - 1. In chromatophores from Rps. sphaeroides, the stimulation by ADP and Pi of the electric potential decay indicated by the carotenoid shift is greater than the stimulation of the decay of pH change indicated by the colour change of added cresol red under similar conditions. This difference is attributed to H+ consumption during the synthesis of ATP. The ratio of H+ translocated across the membrane to ATP synthesized was estimated to be approximately 1.7 H+/ATP. 2. The stimulation of the electrical potential decay by ADP and Pi was found to be a constant fraction (10%) of the total decay when the flash intensity was varied. No 'critical' or 'threshold' potential was observed. 3. The stimulated electrical potential decay after a second flash, given within a few seconds of the first, was related to the amplitude of the electrical potential produced by the second flash (10%) but neither to the dark time between the flashes, nor to the total extent of the electrical potential above the dark level. These results are consistent with two hypotheses (a) the chromatophores are a mixed population of vesicles, only a small fraction (10%) of which possess an active ATP synthesizing system (b) the activity of the ATP synthesizing system, though driven by a proton motive force, is controlled by electron transport processess. If alternative (a) is correct then the overall single turnover flash yield of 1 ATP per 1470 bacteriochlorophyll measured in (1) would mean that the yield of the active vesicles is approximately 10 ATP per 1470 bacteriochlorophyll or 30 ATP per vesicle. 4. The stimulation of the electrical potential decay by ADP and Pi is approximately 40% less in antimycin-treated chromatophores. It is shown that this is probably a consequence of antimycin-inhibited H+-release on the inside of the chromatophore vesicles following a flash
RP  - NOT IN FILE
NT  - UI - 76019224LA - engRN - 0 (Carotenoids)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 642-15-9 (Antimycin A)PT - Journal ArticleDA - 19760102IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:240445
SO  - Biochim Biophys Acta 1975 Oct 10 ;408(1):83-92

156
UI  - 18965
AU  - Junge W
TI  - Physical aspects of the electron transport and photophosphorylation in green plants
MH  - atp
MH  - electron
MH  - Electron Transport
MH  - Photophosphorylation
MH  - Plants
MH  - review
MH  - transport
RP  - NOT IN FILE
SO  - Ber Dtsch Bot Ges 1975  ;88():283-301

157
UI  - 797
AU  - Kagawa Y
AU  - Sone N
AU  - Hirata H
AU  - Yoshida M
TI  - [Oxidative phosphorylation--structure and function (author's transl)]
RP  - NOT IN FILE
NT  - UI - 76012782LA - jpnRN - 0 (Cytochromes)RN - 0 (Hemeproteins)RN - 0 (Phospholipids)RN - 0 (Uncoupling Agents)RN - 56-65-5 (Adenosine Triphosphate)RN - 9007-43-6 (Cytochrome c)RN - EC 1.6. (NADH, NADPH Oxidoreductases)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticlePT - ReviewDA - 19751204IS - 0039-9450SB - IMCY - JAPANJC - Q7D
UR  - PM:169549
SO  - Tanpakushitsu Kakusan Koso 1975 Mar ;20(4):318-351

158
UI  - 9907
AU  - Klingenberg M
TI  - Energetic aspects of transport of ADP and ATP through the mitochondrial membrane.
AB  - Evidence from various sources demonstrates that the release of ATP in exchange for the entry of ADP across the mitochondrial membrane is an active process requiring energy. The necessary energy may be derived from the same source of energy as that used for oxidative phosphorylation. The following results will be discussed:--(1) The exchange is asymmetric with respect to the specificity of ADP and ATP in 'energized' mitochondria. From the outside ADP is much preferred to ATP, but from the inside both exchange with equal specificity. This asymmetry is abolished by de-energization of the membrane. (2) The ADP-ATP exchange is about 50% electrogenic: about half the ATP released against ADP is protonated. The excess of negatively charged ATP might prevent ADP from entering mitochondria against a membrane potential. (3) The ratio of ATP to ADP across the inner mitochondrial membrane is higher outside than inside only in the energized state. Variation of the ATP/ADP ratio maintains this difference. (4) The ADP/ATP ratio apparently varies with changes in the membrane potential as measured by Rb+-distribution. The correlation factor between deltaE changes and this ratio is 0.5, in agreement with predictions from proton-stoichiometry measurements. The deltapH does not significantly change the distribution ratio. (5) By following the P/O ratio, one can show that energy derived from expelling ATP against the ADP/ATP gradient lowers the amount of ATP synthesis. The P/O ratio is lowered as the imbalance of ATP to ADP increases. (6) The energy difference of the phosphorylation potential of ATP is calculated by various methods to be about 8-12 kJ, depending on the conditions. This free energy is the result of the 'active' transport which corresponds to the release of ATP outside the mitochondria. (7) In vivo studies on the distribution of ADP and ATP inside and outside the mitochondria in liver show a corresponding ratio difference of about 15 as predicted from the in vitro studies.
MH  - Adenosine Diphosphate
MH  - Adenosine Monophosphate
MH  - Adenosine Triphosphate
MH  - ADP
MH  - Animal
MH  - atp
MH  - ATP synthesis
MH  - Atractyloside
MH  - Biological Transport
MH  - Biological Transport,Active
MH  - Bongkrekic Acid
MH  - Carrier Proteins
MH  - Cattle
MH  - drug effects
MH  - Energy Metabolism
MH  - Hydrogen-Ion Concentration
MH  - In Vitro
MH  - Kinetics
MH  - Liver
MH  - Membrane Potentials
MH  - Membranes
MH  - metabolism
MH  - Methods
MH  - Mitochondria
MH  - Mitochondria,Liver
MH  - Models,Biological
MH  - Myocardium
MH  - Oxidative Phosphorylation
MH  - Oxygen Consumption
MH  - pharmacology
MH  - Phosphates
MH  - Phosphorylation
MH  - Rats
MH  - Temperature
MH  - Thermodynamics
MH  - transport
RP  - NOT IN FILE
SO  - Ciba Found Symp 1975  ;(31):105-124

159
UI  - 19640
AU  - Lardy H
AU  - Reed P
AU  - Lin CH
TI  - Antibiotic inhibitors of mitochondrial ATP synthesis
AB  - Fourteen antibiotics have been found to inhibit oxidative phosphorylation and uncoupler-stimulated adenosinetriphosphatase in mitochondria. Four different types of binding sites for these inhibitors have been found. The first (1) binds aurovertin to purified MF1 ATPase in the stoichiometric ratio of two aurovertin molecules per molecule of ATPase. Site II is the locus for efrapeptin (A23871) and may be a catalytic site on purified ATPase. The remaining two sites have been demonstrated only in mitochondria or submitochondrial particles when the APTase is bound to other membrane components. Oligomycin, venturiciden, venturicidin X and ossamycin probably all bind at site III. Leucinostatin (A20668) binds at site IV. At low concentrations, this antibiotic acts like oligomycin; at higher concentrations it uncouples oxidative phosphorylation. Venturicidin appears to prevent leucinostation from binding at site IV for it allows uncoupling to occur at very low concentrations of the latter antibiotic. Venturicidin aglycone, which is a more effective inhibitor than its parent compound, does not exert this effect. It is concluded that sites III and IV are in juxtaposition and that when venturicidin binds at site III its sugar moiety projects into the area of site IV to prevent leucinostation from binding at its inhibitory site
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - ANTIBIOTIC
MH  - Antibiotics
MH  - atp
MH  - ATP synthesis
MH  - ATPase
MH  - aurovertin
MH  - Aurovertins
MH  - BINDING
MH  - BINDING SITE
MH  - Binding Sites
MH  - catalytic
MH  - inhibitor
MH  - inhibitors
MH  - lactones
MH  - membrane
MH  - Mitochondria
MH  - oligomycin
MH  - Oxidative Phosphorylation
MH  - peptide
MH  - peptides
MH  - Phosphorylation
MH  - review
MH  - Site
MH  - Submitochondrial Particles
MH  - synthesis
MH  - united states
MH  - venturicidin
RP  - NOT IN FILE
NT  - UI - 75168307LA - engRN - 0 (Antibiotics)RN - 0 (Antibiotics, Antifungal)RN - 0 (Aurovertins)RN - 0 (Lactones)RN - 0 (Peptides)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticlePT - ReviewDA - 19750829IS - 0014-9446SB - IMCY - UNITED STATESJC - EUV
UR  - PM:124269
SO  - Fed Proc 1975 Jul ;34(8):1707-1710

160
UI  - 619
AU  - Melandri AB
AU  - Fabbri E
AU  - Melandri BA
TI  - Energy transduction in photosynthetic bacteria. VIII. Activation of the energy-transducing ATPase by inorganic phosphate
AB  - ATPase activity and ATP-induced energization of photosynthetic membranes from Rhodopseudomonas capsulata are stimulated by phosphate; the maximum stimulatory effect occurs at a concentration between 1 and 2 mM. The sensitivity of the ATPase to oligomycin increases in the presence of phosphate since all the Pi-stimulated activity is inhibited by this antibiotic. Aurovertin, which has no effect on ATPase in the absence of phosphate, inhibits completely the activity elicited by this anion. The addition of Pi induces a substantial increase in the V of ATPase activity without changing the affinity of the enzyme for ATP or ADP. Arsenate, at the same concentrations, produces effects very similar to those of phosphate. The stimulation by arsenate of the transfer of energy from ATP to the membrane suggests a non-hydrolytic role of this anion as a modifier of the ATPase activity
RP  - NOT IN FILE
NT  - UI - 75146589LA - engRN - 0 (Antimetabolites)RN - 0 (Arsenates)RN - 0 (Hydrazones)RN - 0 (Nitriles)RN - 0 (Oligomycins)RN - 0 (Phosphates)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19750714IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:123766
SO  - Biochim Biophys Acta 1975 Jan 31 ;376(1):82-88

161
UI  - 620
AU  - Melandri AB
AU  - Fabbri E
AU  - Firstater E
AU  - Melandri BA
TI  - Energy transduction in photosynthetic bacteria. VII. Inhibition of the coupling ATPase by N-ethylmaleimide related to the energized state of the membrane
AB  - N-Ethylmaleimide, at millimolar concentrations, irreversibily inhibits photophosphorylation and ATPase activity of photosynthetic membranes from Rhodopseudomonas capsulata. The inhibitory effect of N- ethylmaleimide is evident only the membranes are preincubated with the inhibitor in the light and in the absence of phosphorylation substrates. ADP and orthophosphate (or arsenate) exert a protective effect against the inhibition if they are present during the preillumination stage. The energization of the membrane by ATP hydrolysis, measured as ATP-induced quenching of 9-aminoacridine fluorescence, also is inhibited irreversibly by N-ethylmaleimide. Uncouplers protect the ATPase from inhibition by N-ethylmaleimide at concentrations at which they inhibit photophosphorylation. The ATPase, as measured either in the dark or in the light, is also inhibited by carbonylcyanide p-trifluoromethoxypenylhydrazone in parallel with photophosphorylation. These results are interpreted as evidence that the high-energy state of the membrane induces a conformational change of the ATPase, making it sensitive to attack by N-ethylmaleimide; this conformational change might be related to the active state of the ATPase
RP  - NOT IN FILE
NT  - UI - 75146588LA - engRN - 0 (Hydrazones)RN - 0 (Nitriles)RN - 0 (Phosphates)RN - 128-53-0 (Ethylmaleimide)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19750714IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:123765
SO  - Biochim Biophys Acta 1975 Jan 31 ;376(1):72-81

162
UI  - 1027
AU  - Mitchell P
TI  - Protonmotive redox mechanism of the cytochrome b-c1 complex in the respiratory chain: protonmotive ubiquinone cycle
MH  - Binding Sites
MH  - Biological Transport
MH  - Cytochrome c
MH  - Cytochrome c1
MH  - Cytochromes
MH  - Diffusion
MH  - enzymology
MH  - Hydrogen-Ion Concentration
MH  - Kinetics
MH  - Mathematics
MH  - Membranes
MH  - metabolism
MH  - Mitochondria
MH  - Multienzyme Complexes
MH  - Osmosis
MH  - Oxidation-Reduction
MH  - Oxygen Consumption
MH  - Potentiometry
MH  - Protein Binding
MH  - Thermodynamics
MH  - Ubiquinone
RP  - NOT IN FILE
SO  - FEBS Lett 1975 Aug 1 ;56(1):1-6

163
UI  - 1029
AU  - Mitchell P
TI  - Proton translocation mechanisms and energy transduction by adenosine triphosphatases: an answer to criticisms
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - Biological Transport,Active
MH  - Energy Transfer
MH  - Hydrogen-Ion Concentration
MH  - Kinetics
MH  - metabolism
MH  - Mitochondria
MH  - Models,Biological
MH  - Oxidative Phosphorylation
MH  - Protein Conformation
RP  - NOT IN FILE
SO  - FEBS Lett 1975 Feb 1 ;50(2):95-97

164
UI  - 1026
AU  - Mitchell P
TI  - The protonmotive Q cycle: a general formulation
MH  - Biological Transport
MH  - Membranes
MH  - Models,Chemical
MH  - Oxidation-Reduction
MH  - Protons
MH  - Quinones
MH  - Thermodynamics
RP  - NOT IN FILE
SO  - FEBS Lett 1975 Nov 15 ;59(2):137-139

165
UI  - 1028
AU  - Moyle J
AU  - Mitchell P
TI  - Active/inactive state transitions of mitochondrial ATPase molecules influenced by Mg2+, anions and aurovertin
MH  - Adenosinetriphosphatase
MH  - Animal
MH  - Anions
MH  - Antibiotics
MH  - Aurovertins
MH  - Azides
MH  - Dinitrophenols
MH  - drug effects
MH  - Enzyme Activation
MH  - enzymology
MH  - Magnesium
MH  - metabolism
MH  - Mitochondria,Liver
MH  - pharmacology
MH  - Phosphates
MH  - Picrates
MH  - Rats
MH  - Sulfates
MH  - Sulfites
MH  - Time Factors
RP  - NOT IN FILE
SO  - FEBS Lett 1975 Aug 1 ;56(1):55-61

166
UI  - 557
AU  - Pedersen PL
TI  - Adenosine triphosphatase from rat liver mitochondria: separate sites involved in ATP hydrolysis and in the reversible, high affinity binding of ADP
RP  - NOT IN FILE
NT  - UI - 75204933LA - engRN - 0 (Azides)RN - 0 (Imides)RN - 56-65-5 (Adenosine Triphosphate)RN - 57-50-1 (Sucrose)RN - 58-64-0 (Adenosine Diphosphate)RN - 61-19-8 (Adenosine Monophosphate)RN - 7439-95-4 (Magnesium)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19750913IS - 0006-291XSB - IMCY - UNITED STATESJC - 9Y8
UR  - PM:125085
SO  - Biochem Biophys Res Commun 1975 May 19 ;64(2):610-616

167
UI  - 558
AU  - Pedersen PL
TI  - Interaction of homogeneous mitochondrial ATPase from rat liver with adenine nucleotides and inorganic phosphate
AB  - Mitochondrial ATPase from rat liver mitochondria contains multiple nucleotide binding sites. At low concentrations ADP binds with high affinity (1 mole/mole ATPase, KD = 1-2 muM). At high concentrations, ADP inhibits ATP hydrolysis presumably by competing with ATP for the active site (KI = 240-300 muM). As isolated, mitochondrial ATPase contains between 0.6 and 2.5 moles ATP/mole ATPase. This "tightly bound" ATP can be removed by repeated precipitations with ammonium sulfate without altering hydrolytic activity of the enzyme. However, the ATP-depleted enzyme must be redissolved in high concentrations of phosphate to retain activity. AMP-PNP (adenylyl imidodiphosphate) replaces tightly bound ATP removed from the enzyme and inhibits ATP hydrolysis. AMP-PNP has little effect on high affinity binding of ADP. Kinetics studies of ATP hydrolysis reveal hyperbolic velocity vs. ATP plots, provided assays are done in bicarbonate buffer or buffers containing high concentrations of phosphate. Taken together, these studies indicate that sites on the enzyme not directly associated with ATP hydrolysis bind ATP or ADP, and that in the absence of bound nucleotide, Pi can maintain the active form of the enzyme
RP  - NOT IN FILE
NT  - UI - 76049983LA - engRN - 0 (Aurovertins)RN - 0 (Phosphates)RN - 25612-73-1 (Adenylyl Imidodiphosphate)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 7439-95-4 (Magnesium)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19760116IS - 0091-7419SB - IMCY - UNITED STATESJC - K75
UR  - PM:127085
SO  - J Supramol Struct 1975  ;3(3):222-230

168
UI  - 861
AU  - Saphon S
AU  - Jackson JB
AU  - Witt HT
TI  - Electrical potential changes, H+ translocation and phosphorylation induced by short flash excitation in Rhodopseudomonas sphaeroides chromatophores
AB  - 1. The basal decay of the carotenoid shift of chromatophores from photosynthetic bacteria following short flash excitation is approximately biphasic. The decay indicates the dissipation of the transmembrane electrical potential. 2. The H+ efflux following rapid H+ binding after a flash, measured from the colour change of added cresol red, shows very similar kinetics to the carotenoid shift decay suggesting that the dissipation of the electric potential decay is a consequence of the H+ efflux. 3. The electric potential decay is stimulated when the chromatophore suspension is supplemented with ADP and Pi (in either the presence or absence of antimycin A). 4. The stimulated electric potential decay by ADP and Pi has a similar pH dependence to that of phosphorylation in continuous light. 5. The stimulation of the electric potential decay by ADP and Pi is reversed, by aurovertin, an antibiotic which inhibits phosphorylation. 6. The stimulation of the electric potential decay by ADP+Pi is also reversed by the inhibitors oligomycin and venturicidin. These inhibitors, but not aurovertin, also inhibit the fast phase of the decay under non- phosphorylating conditions. 7. Valinomycin accelerates the overall rate of decay of the electric potential, inhibits the ADP and Pi stimulated electric potential decay, and inhibits the flash-induced phosphorylation. The decay rate of the H+ efflux however, is slower in the presence of this ionophore. 8. Nigericin-type ionophores accelerate the overall decay rate of the H+ efflux and inhibit the ADP and Pi stimulated electric potential decay. The basal rate of the electric potential decay is unaffected by treatment with these ionophores. 9. When a coupling factor associated with the chromatophore ATPase is removed from the membrane, both the stimulation of the electric potential decay by ADP and Pi and ADP phosphorylation are inhibtied. Both reactions are completely restored after reconstitution with the crude coupling factor extract. The basal electric potential decay rate is not affected by the removal of coupling factor
RP  - NOT IN FILE
NT  - UI - 76019223LA - engRN - 0 (Aurovertins)RN - 0 (Carotenoids)RN - 0 (Oligomycins)RN - 2001-95-8 (Valinomycin)RN - 28380-24-7 (Nigericin)RN - 58-64-0 (Adenosine Diphosphate)RN - 642-15-9 (Antimycin A)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19760102IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:240444
SO  - Biochim Biophys Acta 1975 Oct 10 ;408(1):67-82

169
UI  - 859
AU  - Saphon S
AU  - Jackson JB
AU  - Lerbs V
AU  - Witt HT
TI  - The functional unit of electrical events and phosphorylation in chromatophores from Rhodopseudomonas sphaeroides
AB  - 1. From electron micrographs of chromatophores from Rhodopseudomonas spaeroides and from the estimated bacteriochlorophyll per chromatophore was estimated. The mean diameter of the chromatophore vesicles was 600 A. 2. The decay of the flash-induced electric potential across the chromatophore membrane measured by the carotenoid band shift was 20% accelerated by about one valinomycin molecule per 4700 bacteriochlorophyll, i.e. by one ionophore molecule per chromatophore. 3. The inhibition of the flash-induced ATP formation by valinomycin followed a similar pattern to the accelerated decay of the electric potential. 4. The single turnover flash yield of ATP synthesis gave a mean value of one ATP per 1470 bacteriochlorophyll or about 3 ATP per vesicle. 5. With regard to the partitioning of the ionophore between the membrane (85%) and aqueous phase (15%) we conclude that one molecule of valinomycin per chromatophore is sufficient to begin to collapse the electrical potential and inhibit ATP synthesis. It is therefore suggested that the membrane potential is an essential component of the energized state which is used for phosphorylation. The results correspond to those obtained for the 100-fold larger vesicles in chloroplasts (thylakoids) where one molecule of ioophore is also sufficient to quench both events
RP  - NOT IN FILE
NT  - UI - 76019222LA - engRN - 0 (Bacteriochlorophylls)RN - 2001-95-8 (Valinomycin)RN - 56-65-5 (Adenosine Triphosphate)PT - Journal ArticleDA - 19760102IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:1080674
SO  - Biochim Biophys Acta 1975 Oct 10 ;408(1):58-66

170
UI  - 18966
AU  - Schmid R
AU  - Junge W
TI  - Influence of extraction and recondensation of CF1 on the electric properties of the thylakoid membrane
MH  - atp
MH  - conductance
MH  - membrane
MH  - reconstitution
MH  - thylakoid
MH  - thylakoid membrane
T2  - Proc. Int. Congr. Photosynth., 3rd, Meeting Date 1974, Volume 2, 821-30. Edited by: Avron, Mordhay. Am Elsevier: New York, N. Y
Y2  - -32676  
RP  - NOT IN FILE
SO  -  1975  ;():

171
UI  - 2463
AU  - Schmid R
AU  - Junge W
TI  - Current-voltage studies on the thylakoid membrane in the presence of ionophores
MH  - Ionophores
MH  - membrane
MH  - thylakoid
MH  - thylakoid membrane
RP  - IN FILE
NT  - "plants";
SO  - Biochim Biophys Acta 1975  ;394():76-92

172
UI  - 496
AU  - Senior AE
TI  - Mitochondrial adenosine triphosphatase. Location of sulfhydryl groups and disulfide bonds in soluble enzyme from beef heart
AB  - The soluble beef heart mitochondrial ATPase (F1) contains eight sulfhydryl groups and two disulfide bonds. N-Ethylmaleimide has been used to radioactively label the sulfhydryl groups before and after cleavage of the disulfide bonds by dithiothreitol. After subjecting the labeled protein to polyacrylamide gel electrophoresis in sodium dodecyl sulfate and measuring radioactivity in each of the separated subunits the location of all the sulfhydryl groups and the disulfide bonds may be specified. The conclusions are supported by direct examination of depolymerized, unreduced, enzyme by polyacrylamide gel electrophoresis. The results also indicate that current ideas regarding the overall subunit structure of this enzyme may be incorrect, and this is discussed in light of new data presented here
RP  - NOT IN FILE
NT  - UI - 75109227LA - engRN - 0 (Disulfides)RN - 0 (Sulfhydryl Compounds)RN - 128-53-0 (Ethylmaleimide)RN - 151-21-3 (Sodium Dodecyl Sulfate)RN - 3483-12-3 (Dithiothreitol)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19750609IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:123157
SO  - Biochemistry 1975 Feb 25 ;14(4):660-664

173
UI  - 19760
AU  - Solioz M
AU  - Yen HC
AU  - Marris B
TI  - Release and uptake of gene transfer agent by Rhodopseudomonas capsulata
AB  - Many strains of Rhodopseudomonas capsulata are capable of exchanging genetic information via a recently discovered gene transfer process involving the release and subsequent uptake from the medium of particles containing genetic information (gene transfer agents, GTAs). No viral activities are observed to be associated with this system. An assay has been developed to quantitate gene transfer in R. capsulata. Conditions are described for which the number of cells acquiring a new genetic trait is direcly proportional to the number of GTAs and independent of the number of receipient cells. These conditions were used for the assay of the uptake and release of GTAs by cells. The maximum fraction of recipients that acquire a given genetic marker is approximately 4 X 10(-4). Free GTA appears in a growing culture in one or two abrupt waves near the time of transition from exponential to stationary phase. During these waves, the titer of GTA for a given marker may reach 2 X 103/ml. A comparison of the frequency of single- and double-marker transfers suggests that most of the cells in early- stationary-phase cultures are active recipients. The ultraviolet inactivation spectrum of GTA resembles that of the small ribonucleic acid phages. The inactivation cross section section beta, for GTA was calculated to be 1.7 X 10(-16) cm2/photon at 265 nm
MH  - A
MH  - ACTIVE
MH  - Cells
MH  - Cyanides
MH  - rhodopseudomonas
MH  - Time
MH  - TRANSFER
RP  - NOT IN FILE
NT  - UI - 75211173LA - engRN - 0 (Cyanides)PT - Journal ArticleDA - 19751108IS - 0021-9193SB - IMCY - UNITED STATESJC - HH3
UR  - PM:1150627
SO  - J Bacteriol 1975 Aug ;123(2):651-657

174
UI  - 192
AU  - Sone N
AU  - Yoshida M
AU  - Hirata H
AU  - Kagawa Y
TI  - Purification and properties of a dicyclohexylcarbodiimide-sensitive adenosine triphosphatase from a thermophilic bacterium
AB  - 1. A stable ATPase complex with sensitivity to dicyclohexylcarbodiimide (TFo-F1) was purified from the membranes of the thermophilic aerobic bacterium PS3, by ion exchange chromatography in the presence of Triton X-100. 2. The ATPase of TFo-F1 was maximal at 70 degrees at pH 8.6 and was stable after monomerization in 4 M urea and 0.5% Triton X-100 at 25 degrees. The activity was dependent on Mg2+, Co2+, or Mn2+, and it became insensitive to dicyclohexylcarbodiimide when Ca2+ or Cd2+ was added instead. 3. TFo-F1 required P-lipids of this bacterium contained branched fatty acyl groups but no unsaturated groups and were stable against oxidation and heat. 4. Studies by electron microscopy, gel electrophoresis, and use of anti-ATPase antibody and [3H]acetyl-ATPase indicated that the TFo-F1 complex was composed of an ATPase moiety (TF1, five different subunits) and a hydrophobic moiety (TFo, three different subunits. TFo conferred TF1 with sensitivity to dicyclohexylcarbodiimide. 5. Vesicles catalyzing 32Pi-ATP exchange and ATP-driven enhancement of fluorescence of anilinonaphthalene sulfonate were reconstituted by dialyzing pure TFo-F1 and P-lipids together, and were active even at 50-75 degrees. The vesicles reconstituted from TFo- F1 and bacterial P-lipids were more stable than those reconstituted from TFo-F1 and soybean P-lipids
RP  - NOT IN FILE
NT  - UI - 76025018LA - engRN - 0 (Anilino Naphthalenesulfonates)RN - 0 (Carbodiimides)RN - 0 (Cations, Divalent)RN - 0 (Macromolecular Systems)RN - 0 (Phospholipids)RN - 0 (Polyethylene Glycols)RN - 0 (Uncoupling Agents)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19751223IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:240843
SO  - J Biol Chem 1975 Oct 10 ;250(19):7917-7923

175
UI  - 193
AU  - Yoshida M
AU  - Sone N
AU  - Hirata H
AU  - Kagawa Y
TI  - A highly stable adenosine triphosphatase from a thermophillie bacterium. Purification, properties, and reconstitution
AB  - 1. A highly stable ATPase (TF1) was purified to a monodispersed state from the membranes of a thermophilic bacterium PS3. Its molecular weight was 380,000, and it was composed of five subunits alpha, beta, gamma, sigma', and sigma with molecular weights of 56,000, 53,000, 32,000, 15,500, and 11,000, respectively. 2. TF1 was stable against dissociating agents such as 5.5 M urea and 4.0 M LiCl, organic solvents, such as 60% acetone, heavy metals, and detergents. Low concentrations of all these agents stimulated its activity at 60 degrees. 3. TF1 was not cold-labile and showed a maximal activity at 70 degrees. Its CD spectrum revealed that its conformation changed between 81 and 96 degrees, and that its contents of alpha helices and beta structures were 27.3 and 12.8%, respectively, at 75 degrees. 4. TF1 was completely dissociated by treatment with dodecyl sulfate at 60 degrees and then with 7.1 M urea. The dissociated TF1 was reconstituted by treatment with Dowex 1-X2, and then dialysis. 5. [3H]Acetyl-TF1 bound to TF1-depleted membranes. TF1 only catalyzed 32Pi-ATP exchange and showed sensitivity to inhibitors of energy transfer when bound to the membranes. 6. A hydrophobic membrance component (TFo) was isolated which rendered TF1 sensitive to inhibitors of energy transfer. It was composed of three subunits (with molecular weights of 19,000, 13,500, and 5,400) and P-lipids
RP  - NOT IN FILE
NT  - UI - 76025017LA - engRN - 0 (Amino Acids)RN - 0 (Cations, Divalent)RN - 0 (Macromolecular Systems)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19751223IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:240842
SO  - J Biol Chem 1975 Oct 10 ;250(19):7910-7916

176
UI  - 191
AU  - Yoshida M
AU  - Sone N
AU  - Hirata H
AU  - Kagawa Y
TI  - ATP synthesis catalyzed by purified DCCD-sensitive ATPase incorporated into reconstituted purple membrane vesicles
RP  - NOT IN FILE
NT  - UI - 76088021LA - engRN - 0 (Carbodiimides)RN - 370-86-5 (Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone)RN - 53026-44-1 (Bacteriorhodopsin)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19760219IS - 0006-291XSB - IMCY - UNITED STATESJC - 9Y8
UR  - PM:1031
SO  - Biochem Biophys Res Commun 1975 Dec 15 ;67(4):1295-1300

177
UI  - 21221
AU  - Drachev LA
AU  - Frolov VN
AU  - Kaulen AD
AU  - Kondrashin AA
AU  - Samuilov VD
AU  - Semenov AY
AU  - Skulachev VP
TI  - Generation of electric current by chromatophores of Rhodospirillum rubrum and reconstitution of electrogenic function in subchromatophore pigment-protein complexes
AB  - Lipoprotein complexes, containing (1) bacteriochlorophyll reaction centers, (2) bacteriochlorophyll light-harvesting antenna or (3) both reaction centers and antenna, have been isolated from chromatophores of non-sulphur purple bacteria Rhodospirillum rubrum by detergent treatments. The method of reconstituting the proteoliposomes containing these complexes is described. Being associtated with planas azolectin membrane, ptoteoliposomes as well as intact chromatophores were found to generate a light-dependent transmembrane electric potential difference measured by Ag/AgC1 electrodes and voltmeter. The direction of the electric field inproteoliposomes can be regulated by the addition of antenna complexes to the reconstitution mixture. The reaction center complex proteoliposomes generate an electric field of a direction opposite to that in chromatophores, whereas proteoliposomes containing reaction center complexes and a sufficient amount of antenna complexes produce a potential difference as in chromatophores. ATP and inorganic pyrophosphate, besides light, were shown to be usable as energy sources for electric generation in chromatophores associated with planar membrane
MH  - A
MH  - atp
MH  - Bacteria
MH  - Bacterial Proteins
MH  - Carbonyl Cyanide m-Chlorophenyl Hydrazone
MH  - chromatophore
MH  - chromatophores
MH  - COMPLEX
MH  - Electrodes
MH  - electrogenic
MH  - FIELD
MH  - function
MH  - Light
MH  - Lipoproteins
MH  - liposome
MH  - Liposomes
MH  - membrane
MH  - method
MH  - pigments
MH  - protein
MH  - Proteins
MH  - proteoliposome
MH  - reaction center
MH  - reconstitution
MH  - Rhodospirillum
MH  - Rhodospirillum rubrum
RP  - NOT IN FILE
NT  - UI - 77000322LA - engRN - 0 (Bacterial Proteins)RN - 0 (Lipoproteins)RN - 0 (Liposomes)RN - 0 (Pigments)RN - 1405-97-6 (Gramicidin)RN - 555-60-2 (Carbonyl Cyanide m-Chlorophenyl Hydrazone)PT - Journal ArticleDA - 19761121IS - 0006-3002SB - IMCY - NETHERLANDS
UR  - PM:61042
SO  - Biochim Biophys Acta 1976 Sep 13 ;440(3):637-660

178
UI  - 21220
AU  - Drachev LA
AU  - Frolov VN
AU  - Kaulen AD
AU  - Liberman EA
AU  - Ostroumov SA
AU  - Plakunova VG
AU  - Semenov AY
AU  - Skulachev VP
TI  - Reconstitution of Biological Molecular generators of electric current. Bacteriorhodopsin
AB  - 1. Photoinduced generation of electric current by bacteriorhodopsin, incorporated into the planar phospholipid membrane, has been directly measured with conventional electrometer techniques. 2. Two methods for bacteriorhodopsin incorporation have been developed: (a) formation of planar membrane from a mixture of decane solution of phospholipids and of the fraction of violet fragments of the Halobacterium halobium membrane (bacteriorhodopsin sheets), and (b) adhesion of bacteriorhodopsin-containing reconstituted spherical membranes (proteoliposomes) to the planar membrane in the presence of Ca2+ or some other cations. In both cases, illumination was found to induce electric current generation directed across the planar membrane, an effect which was measured by macroelectrodes immersed into electrolyte solutions on both sides of the membrane. 3. The maximal values of the transmembrane electric potential were of about 150 mV at a current of about 10(-11) A. The electromotive force measured by means of counterbalancing the photoeffect by an external battery, was found to reach the value of 300 mV. 4. The action spectrum of the photoeffect coincides with the bacteriorhodopsin absorption spectrum (maximum about 570 nm). 5. Both components of the electrochemical potential of H+ ions (electric potential and delta pH) across the planar membrane affect the bacteriorhodopsin photoelectric response in a fashion which could be expected if bacteriorhodopsin were a light-dependent electrogenic proton pump. 6. La3+ ions were shown to inhibit operation of those bacteriorhodopsin which pump out H+ ions from the La3+-containing compartment. 7. The photoeffect, mediated by proteoliposomes associated with thick planar membrane, is decreased by gramicidin A at concentrations which do not influence the planar membrane resistance in the light. On the contrary, a protonophorous uncoupler, trichlorocarbonylcyanidephenylhydrazone, decreases the photoeffect only if it is added at a concentration lowering the light resistance. The dark resistance is shown to be higher than the light one, and decreases to the light level by gramicidin. 8. A simple equivalent electric scheme consistent with the above results has been proposed
MH  - A
MH  - absorption
MH  - ACID
MH  - action spectra
MH  - Bacteriorhodopsin
MH  - Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone
MH  - carotenoid
MH  - Carotenoids
MH  - Cations
MH  - delta
MH  - DELTA-PH
MH  - Edetic Acid
MH  - electrogenic
MH  - H+
MH  - Halobacterium
MH  - ion
MH  - Ions
MH  - Light
MH  - liposome
MH  - Liposomes
MH  - membrane
MH  - Membranes
MH  - method
MH  - Methods
MH  - pH
MH  - Phospholipids
MH  - Proteolipids
MH  - proteoliposome
MH  - proton
MH  - Proton Pump
MH  - reconstitution
MH  - Solutions
MH  - spectra
RP  - NOT IN FILE
NT  - UI - 77051306LA - engRN - 0 (Lectins)RN - 0 (Liposomes)RN - 0 (Proteolipids)RN - 1405-97-6 (Gramicidin)RN - 36-88-4 (Carotenoids)RN - 370-86-5 (Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone)RN - 53026-44-1 (Bacteriorhodopsins)RN - 60-00-4 (Edetic Acid)RN - 7439-91-0 (Lanthanum)PT - Journal ArticleDA - 19770129IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:62754
SO  - J Biol Chem 1976 Nov 25 ;251(22):7059-7065

179
UI  - 21219
AU  - Drachev LA
AU  - Jasaitis AA
AU  - Kaulen AD
AU  - Kondrashin AA
AU  - Chu LV
AU  - Semenov AY
AU  - Severina II
AU  - Skulachev VP
TI  - Reconstitution of biological molecular generators of electric current. Cytochrome oxidase
AB  - 1. Direct measurement of the electric current generation by cytochrome oxidase has been carried out. To this end, two procedures were used. The simpler one consists in formation of planar artificial membrane from the mixture of decane solution of soya bean phospholipids and beef heart cytochrome oxidase. Addition of cytochrome c and ascorbate to one of the two compartments separated by the cytochrome oxidase-containing planar membrane was found to result in a transmembrane electric potential difference being formed (plus on cytochrome c side of the membrane). Maximal values of potential differences obtained by this method were about 40 mV. Much higher potentials were observed when another ("photeoliposome-planar membrane") method was applied. In this case cytochrome oxidase was reconstituted with phospholipid to form proteoliposomes which adhered to planar phospholipid membrane in the presence of Ca2+ ions. Addition of cytochrome c and ascorbate to the proteoliposome-containing compartment gives rise to generation of an electric potential difference across the planar membrane, which reached 100 mV at a current of about 1 X 10(-11) A (minus in the proteoliposome- free compartment). The electromotive force of this generator was estimated as being about 0.2 V. If ascorbate and proteoliposomes were added into different compartments, a penetrating hydrogen atom carrier (phenazine methosulfate, (PMS) or tetramethyl-p-phenylenediamine (TMPD)) was required for a membrane potential to be formed. Generation of an electric potential difference of the opposite direction (plus in the proteoliposome-free compartment) was revealed in experiments with cytochrome oxidase proteoliposome containing cytochrome c in their interior. In this case, addition of PMS or TMPD was necessary. 2. In the suspension of cytochrome oxidase proteoliposome the uptake of a cationic penetrant (tetraphenyl phosphonium cation) was found to be coupled with electron transfer via external cytochrome c. Electron transfer via intraproteoliposomal cytochrome c induced the uptake of anionic penetrants (tetraphenyl borate and phenyldicarbaundecaborane anions). 3. All the above effects were sensitive to cyanide and protonophorous uncouplers. 4. In proteoliposomes containing both cytochrome oxidase and bacteriorhodopsin, the light- and oxidation- dependent generations of membrane potential have been revealed. 5. The data obtained are in agreement with Mitchell's idea of transmembrane electron flow in the cytochrome oxidase segment of the respiratory chain
MH  - A
MH  - Anions
MH  - Bacteriorhodopsin
MH  - Cyanides
MH  - cytochrome
MH  - Cytochrome c
MH  - Cytochrome-c Oxidase
MH  - electron
MH  - electron transfer
MH  - Electron-transfer
MH  - Hydrogen
MH  - ion
MH  - Ions
MH  - Light
MH  - liposome
MH  - Liposomes
MH  - membrane
MH  - Membrane Potential
MH  - method
MH  - Phospholipids
MH  - PMS
MH  - Proteolipids
MH  - proteoliposome
MH  - reconstitution
MH  - TRANSFER
RP  - NOT IN FILE
NT  - UI - 77051308LA - engRN - 0 (Cyanides)RN - 0 (Lectins)RN - 0 (Liposomes)RN - 0 (Phospholipids)RN - 0 (Proteolipids)RN - 53026-44-1 (Bacteriorhodopsins)RN - 9007-43-6 (Cytochrome c)RN - EC 1.9.3.1 (Cytochrome-c Oxidase)PT - Journal ArticleDA - 19770129IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:186452
SO  - J Biol Chem 1976 Nov 25 ;251(22):7072-7076

180
UI  - 858
AU  - Edwards PA
AU  - Jackson JB
TI  - The control of the adenosine triphosphatase of Rhodospirillum rubrum chromatophores by divalent cations and the membrane high energy state
AB  - 1. The rate of ATP hydrolysis, catalysed by Rhodospirilum rubrum chromatophores is accelerated by low concentrations and inhibited by high concentrations of uncoupling agent. 2. The inhibition at high concentrations of uncoupling agent is potentiated by the presence of free magnesium ions. At low uncoupler concentrations magnesium has no effect on the rate of ATP hydrolysis. 3. Inhibition of ATP hydrolysis by high concentrations of uncoupling agent and free magnesium ions is reversed by illumination. Illumination has less effect at low magnesium concentrations. 4. Free calcium ions inhibit ATP hydrolysis independently of the coupled state of the membrane. 5. Under coupled conditions, magnesium ions can overcome the inhibition induced by calcium. The two ions complete for the same site on the enzyme. 6. Inhibition by free magnesium in highly uncoupled chromatophores and inhibition by free calcium are both non-competitive with respect to the divalent cation-ATP substrate. 7. These data are consistent with a model in which divalent cations can bind to a site on the enzyme which is distinct from the substrate site. The regulation of the enzyme activity by the high energy state of the membrane is dependent on the occupant of this site
RP  - NOT IN FILE
NT  - UI - 76117851LA - engRN - 370-86-5 (Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone)RN - 56-65-5 (Adenosine Triphosphate)RN - 7439-95-4 (Magnesium)RN - 7440-70-2 (Calcium)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19760430IS - 0014-2956SB - IMCY - GERMANY, WESTJC - EMZ
UR  - PM:129328
SO  - Eur J Biochem 1976 Feb 2 ;62(1):7-14

181
UI  - 21130
AU  - Ferguson SJ
AU  - Lloyd WJ
AU  - Radda GK
AU  - Slater EC
TI  - On the role of the essential tyrosine residue in the mitochondrial ATPase
MH  - Adenosinetriphosphatase
MH  - ATPase
MH  - Aurovertins
MH  - Dithiothreitol
MH  - RESIDUE
RP  - NOT IN FILE
NT  - UI - 76161417LA - engRN - 0 (Aurovertins)RN - 3483-12-3 (Dithiothreitol)RN - 55520-40-6 (Tyrosine)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19760706IS - 0006-3002SB - IMCY - NETHERLANDS
UR  - PM:130937
SO  - Biochim Biophys Acta 1976 Apr 9 ;430(1):189-193

182
UI  - 21131
AU  - Ferguson SJ
AU  - John P
AU  - Lloyd WJ
AU  - Radda GK
AU  - Whatley FR
TI  - The ATPase as an irreversible component in electron transport linked ATP synthesis
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - atp
MH  - ATP synthesis
MH  - ATPase
MH  - electron
MH  - Electron Transport
MH  - Nad
MH  - synthesis
MH  - transport
RP  - NOT IN FILE
NT  - UI - 76210856LA - engRN - 53-84-9 (NAD)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19760823IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:179878
SO  - FEBS Lett 1976 Mar 1 ;62(3):272-275

183
UI  - 21132
AU  - Ferguson SJ
AU  - Lloyd WJ
AU  - Radda GK
TI  - On the nature of the energised state of submitochondrial particles; investigations with N-aryl naphthalene sulphonate probes
AB  - 1. A further investigation has been made of the way in which the fluorescent probes 1-anilino-naphthalene-8-sulphonate and 2-(N-methyl- anilino) naphthalene-6-sulphonate report on the energised state of bovine heart submitochondrial particles. 2. A comparison of the probe responses to energisation with ATP or to a potassium diffusion potential has been made. The fluorescence enhancements seen in these two cases have different characteristics, and in view of this it is questioned whether a substrate generated energised state of a submitochondrial particle can be equated with a trans-membrane potassium diffusion potential. 3. Substitution of ITP for ATP reduces the rate at which either of the probes respond to energisation. In contrast reducing the ATPase activity of the particles by treatment with the covalent ATPase inhibitors 4-chloro-7-nitrobenzofurazan or N,N'-dicyclohexyl-carbodiimide has no effect on this rate. This finding that the rate of the fluorescence changes is directly sensitive to events at the level of the ATPase, but not to the total ATPase activity, suggests that this rate may not be controlled by a delocalised energised state. Reduction of ATPase activity decreases the extent of the fluorescence enhancement and a relationship between the change in probe fluorescence and ATPase activity is given. 4. The results in this paper are discussed in the context of the mechanisms which have been proposed to account for the fluorescence enhancements of N-aryl naphthalene sulphonate probes upon energisation of submitochondrial particles
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - atp
MH  - ATPase
MH  - Diffusion
MH  - diffusion potential
MH  - fluorescence
MH  - inhibitor
MH  - mechanism
MH  - MECHANISMS
MH  - Potassium
MH  - Submitochondrial Particles
RP  - NOT IN FILE
NT  - UI - 76115006LA - engRN - 0 (Anilino Naphthalenesulfonates)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19760423IS - 0006-3002SB - IMCY - NETHERLANDS
UR  - PM:129165
SO  - Biochim Biophys Acta 1976 Feb 16 ;423(2):174-188

184
UI  - 21129
AU  - Ferguson SJ
AU  - Lloyd WJ
AU  - Radda GK
TI  - A method for determining the adenosine triphosphatase content of energy- transducing membranes. reaction of 4-chloro-7-nitrobenzofurazan with the adenosine triphosphatase of bovine heart submitochondrial particles
AB  - 1. Modification of a single amino acid residue by introduction of the nitrobenzofurazan group inactivates mitochondrial ATPase (adenosine triphosphatase) when membrane-bound in submitochondrial particles. The similarity between the reactions of both membrane-bound and isolated ATPase with 4-chloro-7-nitrobenzofurazan indicates that the single essential tryosine residue identified in the isolated enzyme [Ferguson, Loyd, Lyons & Radda (1975) Eur. J. Biochem. 54, 117-126] Is also a feature of the membrane-bound ATPase. 2. A procedure is presented for estimating the ATPase content of the inner mitochondrial membrane. It is based on the specificity of the incorporation of the nitrobenzofurazan group, and the ready removal of this group by compounds that contain a thiol group. This method indicates that 8.5% of the membrane protein is ATPase. The procedure should be applicable to the titration of the energy-transducing ATPases of bacterial plasma membranes and of the thylakoid membranes of chloroplasts. 3. Combination of the data obtained on the ATPase content of the bovine heart inner mitochondrial membrane with a titration of the cytochrome bc1 complex with antimycin indicates that these two components of the membrane are present in approximately equal amounts
MH  - A
MH  - ACID
MH  - Adenosine
MH  - ADENOSINE-TRIPHOSPHATASE
MH  - Adenosinetriphosphatase
MH  - Antimycin A
MH  - ATPase
MH  - chloroplast
MH  - Chloroplasts
MH  - COMPLEX
MH  - cytochrome
MH  - Cytochromes
MH  - Dithiothreitol
MH  - England
MH  - membrane
MH  - Membranes
MH  - method
MH  - protein
MH  - RESIDUE
MH  - Submitochondrial Particles
MH  - thylakoid
MH  - thylakoid membrane
RP  - NOT IN FILE
NT  - UI - 77065163LA - engRN - 0 (Cytochromes)RN - 3483-12-3 (Dithiothreitol)RN - 642-15-9 (Antimycin A)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19770129IS - 0264-6021SB - IMCY - ENGLAND
UR  - PM:136962
SO  - Biochem J 1976 Nov ;159(2):347-353

185
UI  - 125
AU  - Fillingame RH
TI  - Purification of the carbodiimide-reactive protein component of the ATP energy-transducing system of Escherichia coli
AB  - The ATP-energy transducing system in membranes of Escherichia coli is inhibited by dicyclohexylcarbodiimide. The protein component of this complex with which carbodiimides covalently react to inhibit function was previously identified by labeling wild type and dicyclohexylcarbodiimide-resistant mutants with dicyclohexyl[14C]carbodiimide (Fillingame, R. H. (1975) J. Bacteriol. 124, 870-883). This specific carbodiimide-reactive protein has now been purified. The protein was extracted from the membrane with chloroform:methanol and chromatographed on DEAE-cellulose and hydroxypropyl Spehadex G-50 in this sulvent mixture. The resultant 700- fold purification yielded a protein that was homogeneous on dodecyl sulfate-acrylamide gel electrophoresis and virtually free of phospholipid. It remained soluble in neutral chloroform:methanol throughout the purification procedure. The amino acid composition of the purified protein was extraordinary in that only 16% of the amino acids present could be considered polar. Histidine, serine, cysteine, and tryptophan were not found. Abnormally high contents of methionine, glycine, alanine, and leucine were present. One mole of lysine and threonine were found/mole of dicyclohexyl[14C]carbodiimide bound. The minimum molecular weight based on the amino acid composition was 8400. The specific carbodiimide-reactive protein has also been purified without prior modification by dicyclohexylcarbodiimide. The unmodified protein eluted from DEAE-cellulose at a higher salt concentration than the dicyclohexylcarbodiimide-modified form, which suggested that the reaction with the carbodiimide neutralized the negative charge. Only one-third of the total carbodiimide-reactive protein in the membrane was modified by dicyclohexylcarbodiimide under conditions which maximally inhibited adenosine triphosphatase activity. These results rais the possibility that the carbodiimide-reactive protein may be present as an oligomer in the energy-transducing complex. The purification of the unmodified carbodiimide-reactive protein should permit assessment of tis biological function, particularly its role in the protein-translocation process that is catalyzed by this energy- transducing complex
RP  - NOT IN FILE
NT  - UI - 77028924LA - engRN - 0 (Amino Acids)RN - 0 (Carbodiimides)RN - 0 (Membrane Proteins)RN - 0 (Proteolipids)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)PT - Journal ArticleDA - 19770103IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:789371
SO  - J Biol Chem 1976 Nov 10 ;251(21):6630-6637

186
UI  - 973
AU  - Gadkari D
AU  - Stolp H
TI  - Energy metabolism of Bdellovibrio bacteriovorus. II. P/O ratio and ATP pool turnover rate
AB  - The P/O ratio of Bdellovibrio bacteriovorus, strain Bd 109 Sa, was evaluated by two different methods based on the determination of energy-rich phosphate bonds and either NADH oxidation or oxygen-uptake. P/O values calculated on the basis of NADH oxidation were up to 6, which has to be regarded as being overestimated. P/O values calculated from energy-rich phosphate bonds and oxygen uptake were around 2. The P/O values determined for Escherichia coli B were similar. The loss of phosphorylation efficiency at one site is discussed. The ATP pool turnover rate of Bdellovibrio was 8/min during endogenous respiration and 24/min during substrate respiration. The corresponding values in Escherichia coli B were 3/min and 38/min
MH  - Adenosine Triphosphate
MH  - Bacteria
MH  - Energy Metabolism
MH  - Escherichia coli
MH  - metabolism
MH  - Methods
MH  - Nad
MH  - Oxidative Phosphorylation
MH  - Oxygen
MH  - Oxygen Consumption
MH  - Phosphorylation
MH  - Respiration
MH  - Species Specificity
RP  - NOT IN FILE
SO  - Arch Microbiol 1976 May 3 ;108(1):125-132

187
UI  - 9902
AU  - Gould JM
TI  - Inhibition by triphenyltin chloride of a tightly-bound membrane component involved in photophosphorylation.
AB  - At very low concentrations (less than 1 muM) triphenyltin chloride inhibits ATP formation and coupled electron transport in isolated spinach chloroplasts. Basal (-Pi) and uncoupled electron transport are not affected by triphenyltin. The membrane-bount ATP in equilibrium Pi exchange and Mg2+-dependent ATPase activities of chloroplasts are also completely sensitive to triphenyltin, although the Ca2+-dependent and Mg2+-dependent ATPase activities of the isolated coupling factor protein are insensitive to triphenyltin. The light-driven proton pump in chloroplasts is stimulated (up to 60%) by low levels of triphenyltin. Indeed, the amount of triphenyltin necessary to inhibit ATP formation or stimulate proton uptake is dependent upon the amount of chloroplasts present in the reaction mixture, with an apparent stoichiometry of 2-2.5 triphenyltin molecules/100 chlorophyll molecules at 50% inhibition of ATP formation and half-maximal stimulation of proton uptake. Chloroplasts partially stripped of coupling factor by an EDTA was are no longer able to accumulate protons in the light. However, low levels of triphenyltin can effectively restore this ability. The amount of triphenyltin required for the restoration of net proton uptake is also dependent upon the amount of chloroplasts, with a stoichiometry of 4-5 triphenyltin molecules/100 chlorophyll molecules at 50% reconstitution. On the basis of this and other evidence it is concluded that triphenyltin chloride inhibits phosphorylation.Atp in equilibrium Pi exchange and membrane-bound ATPase activities in chloroplasts by specifically blocking the transport of protons through a membrane-bound carrier or channel located in a hydrophobic region of the membrane at or near the functional binding site for the coupling factor.
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - atp
MH  - ATPase
MH  - BINDING
MH  - Binding Sites
MH  - Cell Membrane
MH  - Chlorophyll
MH  - chloroplast
MH  - Chloroplasts
MH  - COUPLING FACTOR
MH  - Dithiothreitol
MH  - drug effects
MH  - Edetic Acid
MH  - electron
MH  - Electron Transport
MH  - Enzyme Activation
MH  - Kinetics
MH  - Light
MH  - Magnesium
MH  - metabolism
MH  - Organotin Compounds
MH  - pharmacology
MH  - Phosphates
MH  - Photophosphorylation
MH  - Plants
MH  - proton
MH  - Proton Pump
MH  - Protons
MH  - Spinach
MH  - SPINACH-CHLOROPLASTS
MH  - Support,U.S.Gov't,Non-P.H.S.
MH  - Support,U.S.Gov't,P.H.S.
MH  - transport
RP  - NOT IN FILE
SO  - Eur J Biochem 1976 Mar 1 ;62(3):567-575

188
UI  - 21018
AU  - Graber P
AU  - Witt HT
TI  - Relations between the electrical potential, pH gradient, proton flux and phosphorylation in the photosynthetic membrane
AB  - The transmembrane electrical potential (deltaphi), the proton flux (H+), the rate of electron transport (e), the pH gradient (deltapH) and the rate of phosphorylation (ATP) were measured in chloroplasts of spinach. Photosynthesis was excited periodically with flashes of variable frequencies and intensities. A new method is described for determining the rate of electron transport and proton flux. Under conditions where the rate of electron transport and proton flux are not pH controlled the following correlations were found in the range 50 mV less than or equal to deltaphi less than or equal to 125 mV and 1.8 less than or equal to deltapH less than or equal to 2.7: (1) The pH gradient, deltapH, increases with H+ independently of Phout between 7- 9. (2) The rate of phosphorylation, ATP, depends exponentially on deltapH (at constant deltaphi) and is independent of pHout between 7-9. (3) The rate of phosphorylation, ATP, depends also on deltaphi (at constant deltapH and at constant proton flux H+). (4) The proton flux via the ATPase pathway, Hp+, depends non-linearly on the ratio of the proton concentrations: Hp+ approximately (Hin+/Hout+)b, (b=2.3--2.6). The proton flux via the basal pathway, Hb+, depends linearly on the ratio of the proton concentrations: Hb+ approximately (Hin/Hout). (5) The ratio deltaH+/ATP (e/ATP, i.e. the ratio of the total proton flux, Hp+ + Hb+, and the rate of ATP formation, ATP, depends strongly on deltaphi and on deltapH. The ratio is deltaH+/ATP approximately 3 (e/ATP approximately 1.5) at deltapH 2.7 and deltaphi = 125 mV. (6) It is supposed that the reason for the dependence of deltaH+/ATP on deltaphi anddeltapH is the different functional dependence of the basal proton flux Hb+ and the phosphorylating proton flux Hp+ on deltapH and deltaphi. The calculation of deltaH+/ATP on the basis of this assumption is in fair agreement with the experimental values. Also the "threshold" effects can be explained in this way. (7) The ratio of deltaHp+/ATP, i.e. the ratio of the phosphorylating proton flux Hp+ and ATP, is deltaHp+/ATP APPROXIMATELY 2.4
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP FORMATION
MH  - ATPase
MH  - chloroplast
MH  - Chloroplasts
MH  - CONSTANT
MH  - DEPENDENCE
MH  - electron
MH  - Electron Transport
MH  - H+
MH  - membrane
MH  - method
MH  - pH
MH  - Phosphorylation
MH  - Photosynthesis
MH  - proton
MH  - Spinach
MH  - transport
RP  - NOT IN FILE
NT  - UI - 76115004LA - engRN - 56-65-5 (Adenosine Triphosphate)PT - Journal ArticleDA - 19760423IS - 0006-3002SB - IMCY - NETHERLANDS
UR  - PM:2316
SO  - Biochim Biophys Acta 1976 Feb 16 ;423(2):141-163

189
UI  - 18967
AU  - Junge W
TI  - Flash kinetic spectrophotometry in the study of plant pigments
MH  - Kinetics
MH  - Photosynthesis
MH  - pigments
MH  - plant
MH  - review
MH  - Spectrophotometry
MH  - spectroscopy
T2  - Chem. Biochem. Plant Pigm., 2nd Ed., Volume 2, 233-333. Edited by: Goodwin, Trevor W. Academic: London, Engl
Y2  - -32676  
RP  - NOT IN FILE
SO  -  1976  ;():

190
UI  - 19834
AU  - Kayalar C
AU  - Rosing J
AU  - Boyer PD
TI  - 2,4-Dinitrophenol causes a marked increase in the apparent Km of Pi and of ADP for oxidative phosphorylation
MH  - A
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - ADP
MH  - Dinitrophenols
MH  - Oxidative Phosphorylation
MH  - Phosphates
MH  - Phosphorylation
RP  - NOT IN FILE
NT  - UI - 77044870LA - engRN - 0 (Dinitrophenols)RN - 0 (Phosphates)RN - 58-64-0 (Adenosine Diphosphate)PT - Journal ArticleDA - 19770103IS - 0006-291XSB - IMCY - UNITED STATESJC - 9Y8
UR  - PM:985515
SO  - Biochem Biophys Res Commun 1976 Oct 4 ;72(3):1153-1159

191
UI  - 487
AU  - Leimgruber RM
AU  - Senior AE
TI  - Removal of "tightly bound" nucleotides from phosphorylating submitochondrial particles
AB  - Phosphorylating submitochondrial particles from beef heart (ETPH) prepared here contained about 2.4 nmol of ATP and 1.9 nmol of ADP/mg of protein after repeated washing of the particles. Essentially all of the "tightly bound " ATP and ADP was removed by trypsin treatment. The trypsin-treated ETPH had increased ATPase activity, undiminished NADH oxidase and succinate oxidase activity, but energy-coupling activity (ATP-driven reversed electron transfer) was abolished. Removal of half the ATP and ADP occurred at low levels of trypsin and was associated with loss of half of the coupling activity. Gel filtration of ETPH in high ionic strength buffer also removed ADP and ATP from the particles, resulting in loss of energy-coupling activity, while ATPase activity was increased. The results support the contention that the tightly bound ADP is essential in energy coupling in mitochondria. Tightly bound ATP may also play an essential role
RP  - NOT IN FILE
NT  - UI - 77051314LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.4.21.4 (Trypsin)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19770129IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:136446
SO  - J Biol Chem 1976 Nov 25 ;251(22):7110-7113

192
UI  - 488
AU  - Leimgruber RM
AU  - Senior AE
TI  - Removal of "tightly bound" nucleotides from soluble mitochondrial adenosine triphosphatase (F1)
AB  - Soluble mitochondrial ATPase (F1) from beef heart prepared in this laboratory contained approximately 1.8 mol of ADP and 0 mol of ATP/mol of F1 which were not removed by repeated precipitation of the enzyme with ammonium sulfate solution or by gel filtration in low ionic strength buffer containing EDTA. This enzyme had full coupling activity. Treatment of the enzyme with trypsin (5 mug/mg of F1 for 3 min) reduced the "tightly bound" ADP to zero, abolished coupling activity, but had no effect on the ATPase activity, stability, or membrane-binding capability of the F1. When the trypsin concentration was varied between 0 and 5 mug/mg of F1, tightly bound ADP was removed to varying degrees, and a correlation was seen between amount of residual tightly bound ADP and residual coupling activity. Gel filtration of the native F1 in high ionic strength buffer containing EDTA also caused complete loss of tightly bound ADP and coupling ability, whereas ATPase activity, stability, and membrane-binding capability were retained. The ADP-depleted F1 preparations were unable to rebind normal amounts of ADP or any ATP in simple reloading experiments. The results strongly suggest that tightly bound ADP is required for ATP synthesis and for energy-coupled ATP hydrolysis on F1. The results also suggest that ATP synthesis and energy-linked ATP hydrolysis rather than involving one nucleotide binding site on F1, involve a series or "cluster" of sites. The ATP hydrolysis site may represent one component of this cluster. The results show that nonenergy-coupled ATP hydrolysis on F1 can occur in the absence of tightly bound ADP or ATP
RP  - NOT IN FILE
NT  - UI - 77051313LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.4.21.4 (Trypsin)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19770129IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:136445
SO  - J Biol Chem 1976 Nov 25 ;251(22):7103-7109

193
UI  - 19765
AU  - Lundin A
AU  - Richardsson A
AU  - Thore A
TI  - Continous monitoring of ATP-converting reactions by purified firefly luciferase
MH  - Adenosine
MH  - Adenosine Triphosphate
RP  - NOT IN FILE
NT  - UI - 77042643LA - engRN - 0 (Luciferins)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 1.13.12.- (Luciferase)RN - EC 3.6.1.5 (Apyrase)PT - Journal ArticleDA - 19761230IS - 0003-2697SB - IMCY - UNITED STATESJC - 4NK
UR  - PM:10755
SO  - Anal Biochem 1976 Oct ;75(2):611-620

194
UI  - 618
AU  - Melandri BA
AU  - Melandri AB
TI  - Coupling factors ATPases from photosynthetic bacteria
RP  - NOT IN FILE
NT  - UI - 76260184LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - 7439-95-4 (Magnesium)RN - 7440-70-2 (Calcium)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19761020IS - 0449-5705SB - IMCY - ENGLANDJC - HIN
UR  - PM:134033
SO  - J Bioenerg 1976 Apr ;8(2):109-119

195
UI  - 21141
AU  - Michel H
AU  - Oesterhelt D
TI  - Light-induced changes of the pH gradient and the membrane potential in H. halobium
MH  - Carbonyl Cyanide m-Chlorophenyl Hydrazone
MH  - membrane
MH  - Membrane Potential
MH  - pH
RP  - NOT IN FILE
NT  - UI - 76210953LA - engRN - 555-60-2 (Carbonyl Cyanide m-Chlorophenyl Hydrazone)RN - 956-48-9 (2,6-Dichloroindophenol)PT - Journal ArticleDA - 19760823IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:6333
SO  - FEBS Lett 1976 Jun 1 ;65(2):175-178

196
UI  - 1024
AU  - Mitchell P
TI  - Possible molecular mechanisms of the protonmotive function of cytochrome systems
MH  - Bacteria
MH  - Chloroplasts
MH  - Cytochrome-c Oxidase
MH  - Cytochromes
MH  - Electron Transport
MH  - Kinetics
MH  - Membrane Potentials
MH  - metabolism
MH  - Mitochondria
MH  - Models,Biological
MH  - Molecular Conformation
MH  - Oxidation-Reduction
MH  - Protons
MH  - Ubiquinone
RP  - NOT IN FILE
SO  - J Theor Biol 1976 Oct 21 ;62(2):327-367

197
UI  - 1025
AU  - Mitchell P
TI  - Vectorial chemistry and the molecular mechanics of chemiosmotic coupling: power transmission by proticity
MH  - Adenosinetriphosphatase
MH  - Biological Transport
MH  - Cells
MH  - Chemistry
MH  - enzymology
MH  - Kinetics
MH  - Magnesium
MH  - Membranes
MH  - metabolism
MH  - Mitochondria
MH  - Models,Biological
MH  - Osmosis
MH  - Oxidation-Reduction
MH  - Oxidative Phosphorylation
MH  - Oxygen Consumption
MH  - Photophosphorylation
MH  - Thermodynamics
RP  - NOT IN FILE
SO  - Biochem Soc Trans 1976  ;4(3):399-430

198
UI  - 1003
AU  - Mitchell P
TI  - Vectorial chemistry and the molecular mechanics of chemiosmotic coupling: power transmission by proticity
MH  - Adenosinetriphosphatase
MH  - Chemistry
MH  - Magnesium
RP  - NOT IN FILE
NT  - UI - 77069358LA - engRN - 7439-95-4 (Magnesium)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticlePT - ReviewDA - 19770224IS - 0300-5127SB - IMCY - ENGLANDJC - E48
UR  - PM:137147
SO  - Biochem Soc Trans 1976  ;4(3):399-430

199
UI  - 556
AU  - Pedersen PL
TI  - ATP-dependent reactions catalyzed by inner membrane vesicles of rat liver mitochondria. Kinetics, substrate specificity, and bicarbonate sensitivity
AB  - Three ATP-dependent reactions catalyzed by the inner membrane of rat liver mitochondria and the ATPase reaction catalyzed by purified mitochondrial ATPase (F1), were studied with respect to kinetic properties, substrates specificity, and sensitivity to bicarbonate. The ATP-dependent transhydrogenase reaction (reduction of NADP+ by NADH) catalyzed by inner membrane vesicles displays typical Michaelis-Menten kinetics in both Tris-Cl and Tris-bicarbonate buffers, with Km (ATP) values of 0.035 mM and 0.054 mM respectively. The Vmax of transhydrogenase activity (25 nmol min-1 mg-1) is the same in Tris- bicarbonate or Tris-Cl buffer. ITP and GTP readily substitute for ATP in the transhydrogenase reaction. The ATP-P1 exchange reaction catalyzed by inner membrane vesicles displays typical Michaelis-Menten kinetics in both Tris-Cl and Tris-bicarbonate buffers with Km (ATP) values of 1.0 mM and 1.4 mM respectively. The Vmax of exchange (200 nmol min-1 mg-1) is the same in either buffer. ITP and GTP do not effectively replace ATP in the exchange reaction
RP  - NOT IN FILE
NT  - UI - 76120594LA - engRN - 0 (Bicarbonates)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 1.6. (NADH, NADPH Oxidoreductases)RN - EC 3.1.3.41 (4-Nitrophenylphosphatase)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19760430IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:175067
SO  - J Biol Chem 1976 Feb 25 ;251(4):934-940

200
UI  - 3394
AU  - Petty KM
AU  - Dutton PL
TI  - Properties of the Flash-Induced Proton Binding Encountered in Membranes of Rhodopseudomonas sphaeroides: A Functional pK on the Ubisemiquinone?
MH  - A
MH  - Bacteria
MH  - BINDING
MH  - membrane
MH  - Membranes
MH  - proton
MH  - quinone
MH  - reaction center
MH  - rhodopseudomonas
MH  - wox
RP  - IN FILE
NT  - K
SO  - Arch Biochem Biophys 1976  ;172():335-345

201
UI  - 21145
AU  - Petty KM
AU  - Dutton PL
TI  - Ubiquinone-cytochrome b electron and proton transfer: a functional pK on cytochrome b50 in Rhodopseudomonas sphaeroides membranes
MH  - A
MH  - Antimycin A
MH  - cytochrome
MH  - Cytochromes
MH  - electron
MH  - membrane
MH  - Membranes
MH  - proton
MH  - Proton transfer
MH  - Proton-transfer
MH  - Protons
MH  - rhodopseudomonas
MH  - sphaeroides
MH  - TRANSFER
MH  - Ubiquinone
RP  - NOT IN FILE
NT  - UI - 76159515LA - engRN - 0 (Cytochromes)RN - 0 (Protons)RN - 1339-63-5 (Ubiquinone)RN - 642-15-9 (Antimycin A)PT - Journal ArticleDA - 19760520IS - 0003-9861SB - IMCY - UNITED STATES
UR  - PM:4015
SO  - Arch Biochem Biophys 1976 Feb ;172(2):346-353

202
UI  - 883
AU  - Raven JA
AU  - Smith FA
TI  - The evolution of chemiosmotic energy coupling
MH  - Adenosine Triphosphate
MH  - Biological Transport,Active
MH  - biosynthesis
MH  - Cell Membrane
MH  - Energy Metabolism
MH  - Evolution
MH  - Fermentation
MH  - Halobacterium
MH  - Light
MH  - metabolism
MH  - Oxidation-Reduction
MH  - Photophosphorylation
MH  - Photosynthesis
RP  - NOT IN FILE
SO  - J Theor Biol 1976 Apr ;57(2):301-312

203
UI  - 19835
AU  - Rosing J
AU  - Smith DJ
AU  - Kayalar C
AU  - Boyer PD
TI  - Medium ADP and not ADP already tightly bound to phylakoid membranes forms the initial ATP in chloroplast phosphorylation
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - ADP
MH  - atp
MH  - chloroplast
MH  - membrane
MH  - Membranes
MH  - Phosphorylation
RP  - NOT IN FILE
NT  - UI - 77044744LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)PT - Journal ArticleDA - 19761223IS - 0006-291XSB - IMCY - UNITED STATESJC - 9Y8
UR  - PM:10896
SO  - Biochem Biophys Res Commun 1976 Sep 7 ;72(1):1-8

204
UI  - 19836
AU  - Smith DJ
AU  - Stokes BO
AU  - Boyer PD
TI  - Probes of initial phosphorylation events in ATP synthesis by chloroplasts
AB  - Rapid mixing and quenching techniques have been used with chloroplasts activated by an acid-base transition or by light to assess the nature and characteristics of the substances initially labeled by inorganic [32P]phosphate during ATP synthesis. With light-activated chloroplast fragments, but not with acid-base-activated preparations, an initial rapid labeling of a small amount of ADP is observed. With the acid-base activated preparations a slower continued labeling of ADP occurs that is uncoupler-sensitive, that does not proceed via [gamma-32]ATP of the medium and for which medium ADP furnishes the AMP moiety. The results point to ADP as the initial acceptor of phosphate for ATP synthesis, with a slow side reaction in which bound ATP phosphorylates bound AMP to give a bound ADP. The phosphorylation of bound ADP by medium [32P]phosphate in the absence of added ADP is confirmed, but the reaction is too slow to serve as an intermediate in photophosphorylation. The appearance of label from [32P]phosphate in ATP in the acid-base transition at 25 degrees shows a lag of only about 3 to 7 ms, consistent with the absence of any phosphorylated intermediate. The lag is followed by a linear rate of [gamma-32]ATP formation that is about as fast as that observed in steady photophosphorylation, consistent with a proton gradient serving for transmission of energy from electron transfer reactions to the ATP- synthesizing complex
MH  - A
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - ADP
MH  - atp
MH  - ATP synthesis
MH  - chloroplast
MH  - Chloroplasts
MH  - COMPLEX
MH  - electron
MH  - Light
MH  - Phosphorylation
MH  - Photophosphorylation
MH  - proton
MH  - synthesis
MH  - TRANSFER
RP  - NOT IN FILE
NT  - UI - 76213293LA - engRN - 12125-02-9 (Ammonium Chloride)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)PT - Journal ArticleDA - 19760902IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:932027
SO  - J Biol Chem 1976 Jul 25 ;251(14):4165-4171

205
UI  - 796
AU  - Sone N
AU  - Yoshida M
AU  - Hirata H
AU  - Okamoto H
AU  - Kagawa Y
TI  - Electrochemical potential of protons in vesicles reconstituted from purified, proton-translocating adenosine triphosphatase
AB  - Measurements were made of the difference in the electrochemical potential of protons (delta-mu H+) across the membrane of vesicles restituted from the ATPase complex (TF0.F1) purified from a thermophilic bacterium and P-lipids. Two fluorescent dyes, anilinonaphthalene sulfonate (ANS) and 9-aminoacridine (9AA) were used as probes for measuring the membrane potential (delta psi) and pH difference across the membrane (delta pH), respectively. In the presence of Tris buffer the maximal delta psi ans no delta pH were produced, while in the presence of the permeant anion NO-3 the maximal delta pH and a low delta psi were produced by the addition of ATP. When thATP concentration was 0.24 mm, the delta psi was 140-150 mV (positive inside) in Tris buffer, and the delta pH was 2.9-3.5 units (acidic inside) in the presence of NO-3. Addition of a saturating amount of ATP produced somewhat larger delta psi and delta pH values, and the delta - muH+attained was about 310mV. By trapping pH indicators in the vesicles during their reconstitution it was found that the pH inside the vesicles was pH 4-5 during ATP hydrolysis. The effects of energy transfer inhibitors, uncouplers, ionophores, and permeant anions on these vesicles were studied
RP  - NOT IN FILE
NT  - UI - 77096875LA - engRN - 0 (Liposomes)RN - 1333-74-0 (Hydrogen)RN - 28380-24-7 (Nigericin)RN - 370-86-5 (Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - 7440-09-7 (Potassium)RN - 77-86-1 (Tromethamine)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19770315IS - 0022-2631SB - IMCY - UNITED STATESJC - J4E
UR  - PM:13221
SO  - J Membr Biol 1976 Dec 28 ;30(2):121-134

206
UI  - 555
AU  - Soper JW
AU  - Pedersen PL
TI  - Adenosine triphosphatase of rat liver mitochondria: detergent solubilization of an oligomycin- and dicyclohexylcarbodiimide-sensitive form of the enzyme
AB  - The hydrolytic activity of the ATPase bound to purified inner membrane vesicles of rat liver mitochondria can be increased threefold by washing extensively with a high ionic strength phosphate buffer. The specific ATPase activities of such phosphate-washed membranes are the highest reported to date for a mitochondrial membrane preparation (21- 24 mumol of ATP hydrolyzed min-1 mg-1 in bicarbonate buffer at 37 degrees C). Deoxycholate (0.1 mg/mg of protein) extracts from these membranes a soluble, cold-stable ATPase complex which exhibits a specific activity under optimal assay conditions of 12 mumol of ATP hydrolyzed min-1 mg-1. This complex is not sedimented by centrifugation at 201000 g for 90 min, and readily passes through a 250-A Millipore filter. The ATPase activity of the soluble complex is inhibited 95% by 2.4 muM oligomycin. In addition, inhibitions of 60% or better are obtained in the presence of 1-8 muM dicyclohexylcarbodiimide, p- chloromercuribenzoate, venturicidin, and aurovertin. While a similar complex may be extracted with Triton X-100 this preparation is always lower in both specific activity and in inhibitor sensitivities than the complex extracted with deoxycholate. Detergents of the Tween and Brij series and other detergents of the Triton series are also much less effective than deoxycholate in solubilizing the oligomycin-sensitive. ATPase complex of rat liver. It is concluded that deoxycholate is superior to other detergents as an extractant of the oligomycin- sensitive ATPase complex of rat liver mitochondria, and that the complex extracted with deoxycholate possesses a closer similarity to the membrane-associated ATPase than does the complex extracted with Triton X-100. These studies document the first report of a detergent- solubilized, oligomycin-sensitive ATPase preparation from rat liver mitochondria
RP  - NOT IN FILE
NT  - UI - 76232204LA - engRN - 0 (Antibiotics)RN - 0 (Carbodiimides)RN - 0 (Cholic Acids)RN - 0 (Oligomycins)RN - 0 (Polyethylene Glycols)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 83-44-3 (Deoxycholic Acid)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19760925IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:132962
SO  - Biochemistry 1976 Jun 15 ;15(12):2682-2690

207
UI  - 836
AU  - Strotmann H
AU  - Bickel S
AU  - Huchzermeyer B
TI  - Energy-dependent release of adenine nucleotides tightly bound to chloroplast coupling factor CF1
RP  - NOT IN FILE
NT  - UI - 76118244LA - engRN - 0 (Adenine Nucleotides)RN - 0 (Phosphates)RN - 0 (Plant Proteins)RN - 1406-65-1 (Chlorophyll)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 61-19-8 (Adenosine Monophosphate)PT - Journal ArticleDA - 19760419IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:2497
SO  - FEBS Lett 1976 Jan 15 ;61(2):194-198

208
UI  - 21017
AU  - Witt HT
AU  - Schlodder E
AU  - Graber P
TI  - Membrane-bound ATP synthesis generated by an external electrical field
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthesis
MH  - Chlorophyll
MH  - FIELD
MH  - synthesis
MH  - Uncoupling Agents
RP  - NOT IN FILE
NT  - UI - 77048843LA - engRN - 0 (Uncoupling Agents)RN - 1406-65-1 (Chlorophyll)RN - 330-54-1 (Diuron)RN - 56-65-5 (Adenosine Triphosphate)PT - Journal ArticleDA - 19770128IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:992038
SO  - FEBS Lett 1976 Oct 15 ;69(1):272-276

209
UI  - 890
AU  - Yaguzhinsky LS
AU  - Boguslavsky LI
AU  - Volkov AG
AU  - Rakhmaninova AB
TI  - Synthesis of ATP coupled with action of membrane protonic pumps at the octane-water interface
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - Alkanes
MH  - Bacteriorhodopsin
MH  - biosynthesis
MH  - Chemistry
MH  - Energy Metabolism
MH  - Membrane Potentials
MH  - metabolism
MH  - Mitochondria
MH  - Protons
MH  - Water
RP  - NOT IN FILE
SO  - Nature 1976 Feb 12 ;259(5543):494-496

210
UI  - 598
AU  - You KS
AU  - Hatefi Y
TI  - Purification and properties of a low molecular weight protein factor of mitochondrial energy-linked functions
AB  - 1. A soluble protein with a molecular weight of 11-12-10(3) has been isolated from bovine-heart mitochondria, which stimulates the following ATP-dependent reactions of submitochondrial particles treated with 0.6 mM EDTA and 1 M NH4OH: reverse electron transfer from succinate to NAD, transhydrogenation from NADH to NADP, and ATP-Pi exchange. The factor has no effect on the NADH oxidase, succinate oxidase and ATPase activities of the particles. 2. The stimulatory effect of the factor in the ATP-dependent reduction of NAD by succinate is 12 mumol-min-1-mg-1 of the factor protein. However, the NH4OH-EDTA treated particles are saturated for maximal activation of the above reaction by very small amounts of the factor (about 20-40 mug factor per mg particle). 3. Electrophoresis of the factor preparation on polyacrylamide gels showed a single protein band plus a nonprotein material which moved at the dye front and was weakly stained with Coomassie Blue. The protein was shown to be required for activation of the particles; whether the fast- moving, nonprotein material is also required is not known. 4. The factor is inhibited by mercurials and N-ethylmaleimide. The former, but not the latter, inhibition is completely reversed by 1,4- dithiothreitol. 5. The NH4OH-EDTA treated particles are also stimulated by rutamycin up to about 0.1 nmol of rutamycin per mg particle; higher rutamycin concentrations inhibit. Depending on the particle preparation, the factor stimulates up to about 3 nmol per mg particle, but does not inhibit at higher concentrations. In addition, under certain conditions in which appropriate concentrations of rutamycin fail to stimulate the particles, the factor still does
RP  - NOT IN FILE
NT  - UI - 76161202LA - engRN - 0 (Proteins)RN - 0 (Succinates)RN - 1404-59-7 (Rutamycin)RN - 3483-12-3 (Dithiothreitol)RN - 53-59-8 (NADP)RN - 53-84-9 (NAD)RN - 554-77-8 (4-Chloromercuribenzenesulfonate)RN - 56-65-5 (Adenosine Triphosphate)PT - Journal ArticleDA - 19760706IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:4097
SO  - Biochim Biophys Acta 1976 Mar 12 ;423(3):398-412

211
UI  - 20976
AU  - Altendorf K
AU  - Lukas M
AU  - Kohl B
AU  - Muller CR
AU  - Sandermann H
TI  - Isolation and purification of bacterial membrane proteins by the use of organic solvents: the lactose permease and the carbodiimide-reactive protein of the adenosinetriphosphatase complex of Escherichia coli
MH  - Adenosinetriphosphatase
MH  - Bacterial Proteins
MH  - COMPLEX
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - Lactose
MH  - membrane
MH  - Membrane Proteins
MH  - protein
MH  - Proteins
MH  - Proteolipids
MH  - purification
MH  - SOLVENT
MH  - transport
RP  - NOT IN FILE
NT  - UI - 78008907LA - engRN - 0 (Bacterial Proteins)RN - 0 (Carbodiimides)RN - 0 (Membrane Proteins)RN - 0 (Membrane Transport Proteins)RN - 0 (Proteolipids)RN - 0 (Solvents)RN - 63-42-3 (Lactose)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19771130IS - 0091-7419SB - IMCY - UNITED STATES
UR  - PM:143553
SO  - J Supramol Struct 1977  ;6(2):229-238

212
UI  - 20975
AU  - Altendorf K
TI  - Purification of the DCCD-reactive protein of the energy-transducing adenosine triphosphatase complex from Escherichia coli
MH  - Adenosine
MH  - ADENOSINE-TRIPHOSPHATASE
MH  - Adenosinetriphosphatase
MH  - Bacterial Proteins
MH  - COMPLEX
MH  - Dicyclohexylcarbodiimide
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - protein
MH  - Proteins
MH  - purification
RP  - NOT IN FILE
NT  - UI - 77116176LA - engRN - 0 (Bacterial Proteins)RN - 0 (Carbodiimides)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19770430IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:138598
SO  - FEBS Lett 1977 Feb 1 ;73(2):271-275

213
UI  - 7038
AU  - Bockris JO
AU  - Reddy AKN
TI  - Modern Electrochemistry
MH  - Electrochemistry
PB  - New York: Plenum Press
RP  - IN FILE
SO  -  1977  ;(2):

214
UI  - 8316
AU  - Boyer PD
AU  - Chance B
AU  - Ernster L
AU  - Mitchell P
AU  - Racker E
AU  - Slater EC
TI  - Oxidative Phosphorylation and Photophosphorylation
MH  - bioenergetics
MH  - Oxidative Phosphorylation
MH  - Phosphorylation
MH  - Photophosphorylation
MH  - review
RP  - IN FILE
NT  - Ju Rev
SO  - Annu Rev Biochem 1977  ;46():955-1026

215
UI  - 19833
AU  - Boyer PD
AU  - de Meis L
AU  - Gloria Costa CM
AU  - Hackney DD
TI  - Dynamic reversal of enzyme carboxyl group phosphorylation as the basis of the oxygen exchange catalyzed by sarcoplasmic reticulum adenosine triphosphatase
AB  - Millisecond mixing and quenching experiments demonstrate an apparent t1/2 for the labeling of phosphorylated sarcoplasmic reticulum ATPase by 32Pi at pH 6 and 30 degrees C of 30 to 40 ms. Under the same conditions, the rate of exchange of water oxygens with inorganic phosphate (Pi) is about 40 mol of H2O exchanged with Pi per 10(6) g of protein per s. Theoretical equations are developed for the expected 32P- labeling pattern given various comparative rates of flux between Pi and the Michaelis complex and between the Michaelis complex and phosphorylated enzyme. The results show that the rapid reversal of the formation of the phosphorylated enzyme is a major source of the oxygen exchange and are consistent with such reversal being the only source
MH  - A
MH  - Adenosine
MH  - ADENOSINE-TRIPHOSPHATASE
MH  - Adenosinetriphosphatase
MH  - ATPase
MH  - COMPLEX
MH  - INORGANIC-PHOSPHATE
MH  - Oxygen
MH  - pH
MH  - Phosphorylation
MH  - Sarcoplasmic Reticulum
MH  - Water
RP  - NOT IN FILE
NT  - UI - 77087741LA - engRN - 0 (Phosphoproteins)RN - 7782-44-7 (Oxygen)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19770331IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:137741
SO  - Biochemistry 1977 Jan 11 ;16(1):136-140

216
UI  - 9897
AU  - Brand MD
AU  - Lehninger AL
TI  - H+/ATP ratio during ATP hydrolysis by mitochondria: modification of the chemiosmotic theory.
AB  - The stoichiometry of H+ ejection by mitochondria during hydrolysis of a small pulse of ATP (the H+/ATP ratio) has been reexamined in the light of our recent observation that the stoichiometry of H+ ejection during mitochondrial electron transport (the H+/site ratio) was previously underestimated. We show that earlier estimates of the H+/ATP ratio in intact mitochondria were based upon an invalid correction for scaler H+ production and describe a modified method for determination of this ratio which utilizes mersalyl or N-ethylmaleimide to prevent complicating transmembrane movements of phosphate and H+. This method gives a value for the H+/ATP ratio of 2.0 without the need for questionable corrections, compared with a value of 3.0 for the H+/site ratio also obtained by pulse methods. A modified version of the chemiosmotic theory is presented, in which 3 H+ are ejected per pair of electrons traversing each energy-conserving site of the respiratory chain. Of these, 2 H+ return to the matrix through the ATPase to form ATP from ADP and phosphate, and 1 H+ returns through the combined action of the phosphate and adenine nucleotide exchange carriers of the inner membrane to allow the energy-requiring influx of Pi and ADP3- and efflux of ATP4-. Thus, up to one-third of the energy input into synthesis of extramitochondrial ATP may be required for transport work. Since other methods suggest that the H+/site significantly exceeds 3.0, an alternative possibility is that 4 h+ are ejected per site, followed by return of 3 H+ through the ATPase and 1 H+ through the operation of the proton-coupled membrane transport systems.
MH  - Adenine Nucleotides
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - ADP
MH  - Animal
MH  - atp
MH  - ATPase
MH  - Biological Transport,Active
MH  - Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone
MH  - Cations,Monovalent
MH  - chemiosmotic theory
MH  - drug effects
MH  - electron
MH  - Electron Transport
MH  - Electrons
MH  - Ethylmaleimide
MH  - H+
MH  - Hydrogen
MH  - Hydrogen-Ion Concentration
MH  - Hydrolysis
MH  - Light
MH  - metabolism
MH  - Methods
MH  - Mitochondria
MH  - Mitochondria,Liver
MH  - Models,Biological
MH  - Movement
MH  - Osmolar Concentration
MH  - Oxidative Phosphorylation
MH  - pharmacology
MH  - Phosphates
MH  - Rats
MH  - Support,U.S.Gov't,Non-P.H.S.
MH  - Support,U.S.Gov't,P.H.S.
MH  - transport
RP  - NOT IN FILE
SO  - Proc Natl Acad Sci U S A 1977 May ;74(5):1955-1959

217
UI  - 20826
AU  - Cain K
AU  - Griffiths DE
TI  - Studies of energy-linked reactions. Localization of the site of action of trialkyltin in yeast mitochondria
AB  - Ligand-binding studies with labelled triethyltin on yeast mitochondrial membranes showed the presence of high-affinity sites (KD = 0.6 micronM; 1.2 +/- 0.3 nmol/mg of protein) and low-affinity sites (KD less than 45 micronM; 70 +/- 20 nmol/mg of protein). The dissociation constant of the high-affinity site is in good agreement with the concentration of triethyltin required for inhibition of mitochondrial ATPase (adenosine triphosphatase) and oxidative phosphorylation. The high-affinity site is not competed for by oligomycin or venturicidin, indicating that triethyltin reacts at a different site from these inhibitors of oxidative phosphorylation. Fractionation of the mitochondrial membrane shows a specific association of the high-affinity sites with the ATP synthase complex. During purification of ATP synthase (oligomycin- sensitive ATPase) there is a 5-6-fold purification of oligomycin- and triethyltin-sensitive ATPase activity concomitant with a 7-9-fold increase in high-affinity triethyltin-binding sites. The purified yeast oligomycin-sensitive ATPase complex contains approximately six binding sites for triethyltin/mol of enzyme complex. It is concluded that specific triethyltin-binding sites are components of the ATP synthase complex, which accounts for the specific inhibition of ATPase and oxidative phosphorylation by triethyltin
MH  - A
MH  - Adenosine
MH  - ADENOSINE-TRIPHOSPHATASE
MH  - Adenosinetriphosphatase
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BINDING
MH  - Binding Sites
MH  - COMPLEX
MH  - CONSTANT
MH  - England
MH  - membrane
MH  - Membranes
MH  - Mitochondria
MH  - Oligomycins
MH  - Oxidative Phosphorylation
MH  - Phosphorylation
MH  - protein
MH  - purification
MH  - Site
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 77201446LA - engRN - 0 (Oligomycins)RN - 0 (Trialkyltin Compounds)RN - 0 (Venturicidins)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19770718IS - 0264-6021SB - IMCY - ENGLAND
UR  - PM:141273
SO  - Biochem J 1977 Mar 15 ;162(3):575-580

218
UI  - 20824
AU  - Cain K
AU  - Hyams RL
AU  - Griffiths DE
TI  - Studies on energy-linked reactions: inhibition of oxidative phosphorylation and energy-linked reactions by dibutyltin dichloride
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone
MH  - Organotin Compounds
MH  - Oxidative Phosphorylation
MH  - Oxidoreductases
MH  - Phosphorylation
MH  - succinate
MH  - Succinates
RP  - NOT IN FILE
NT  - UI - 78024230LA - engRN - 0 (Organotin Compounds)RN - 0 (Succinates)RN - 0 (Sulfhydryl Compounds)RN - 370-86-5 (Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 1.6. (NADH, NADPH Oxidoreductases)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19771229IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:144065
SO  - FEBS Lett 1977 Oct 1 ;82(1):23-28

219
UI  - 625
AU  - Casadio R
AU  - Melandri BA
TI  - The behavior of 9-aminoacridine as an indicator of transmembrane pH difference in liposomes of natural bacterial phospholipids
AB  - The behavior of 9-aminoacridine as an indicator of pH differences artificially set across a membrane has been reexamined in liposomes prepared from bacterial phospholipids extracted from chromatophores of Rhodopseudomonas capsulata grown photoheterotrophically. The dye behaves as an ideal indicator for pH differences lower than about three units; at higher pH's the expected linear dependence of Q/(100-Q) vs. pH is no longer strictly observed. Similarly a linear dependence upon the volume of the liposomes added has been verified. The amine ceases to respond to pH changes when the pH of the external medium exceeds the value of 10, corresponsing to the pKa of 9-aminoacridine. The apparent volume of the inner phase of liposomes, as calculated from fluorescence quenching, but not the slope of dependence of fluorescence on pH, appears to be affected by several factors, including the ionic composition, the osmolarity of the external medium, and the microscopic structure of the liposomes. Millimolar concentrations of earth-alkaline cations diminish the apparent internal volume of liposomes, in agreement with the complexing effect of these ions on phospholipid bilayers. The osmotic response of the apparent inner volume has also been verified; this parameter decreases linearly with the reciprocal of the external osmolarity, as expected from the van't Hoff relation; an osmolarity exceeding 0.3 M is, however, necessary in order to observe this effect
RP  - NOT IN FILE
NT  - UI - 77227761LA - engRN - 0 (Acridines)RN - 0 (Indicators and Reagents)RN - 0 (Liposomes)RN - 0 (Phospholipids)PT - Journal ArticleDA - 19770917IS - 0145-479XSB - IMCY - ENGLANDJC - HIO
UR  - PM:18457
SO  - J Bioenerg Biomembr 1977 Feb ;9(1):17-29

220
UI  - 9964
AU  - Cogdell RJ
AU  - Celis S
AU  - Celis H
AU  - Crofts AR
TI  - Reaction centre carotenoid band shifts
MH  - Carotenoids
RP  - NOT IN FILE
NT  - UI - 77246769LA - engRN - 0 (Carotenoids)RN - 0 (Ferricyanides)PT - Journal ArticleDA - 19771028IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:302224
SO  - FEBS Lett 1977 Aug 1 ;80(1):190-194

221
UI  - 9967
AU  - Crofts AR
AU  - Crowther D
AU  - Celis H
AU  - De Celis SA
AU  - Tierney G
TI  - Proton pumps in bacterial photosynthesis
MH  - Cytochrome c
MH  - England
MH  - Lipoproteins
MH  - Liposomes
MH  - Photosynthesis
MH  - proton
MH  - Proton Pump
RP  - NOT IN FILE
NT  - UI - 78003969LA - engRN - 0 (Lipoproteins)RN - 0 (Liposomes)RN - 9007-43-6 (Cytochrome c)PT - Journal ArticleDA - 19771125IS - 0300-5127SB - IMCY - ENGLANDJC - E48
UR  - PM:20371
SO  - Biochem Soc Trans 1977  ;5(2):491-495

222
UI  - 21218
AU  - Drachev LA
AU  - Kaulen AD
AU  - Skulachev VP
TI  - [Temporal characteristics of bacteriorhodopsin as a molecular biological generator of current]
AB  - Generation of electric potential difference by bacteriorhodopsin proteoliposomes incorporated into the phospholipid-impregnated collodion film has been studied. It is shown that illumination of this film by continuous light gives rise to the generation of an electric potential difference across the film (plus on the bacteriorhodopsin- free side), which can be as high as 300 mV. Short unsaturating flash inducing single turn-over of bacteriorhodopsin generates the potential difference which is a function of the flash intensity (70 mV at 3 mjoule light). The flash-induced photoelectric response consists of four phases. (1) Very fast (tau less than 1 microsec) generation of a potential difference (minus in the bacteriorhodopsin-free compartment). The amplitude of this phase is rather small (1--5 mV). (2) Fast phase of positive charging of the bacteriorhodopsin-free compartment (tau = 25--50 microsec). (3) Slow phase of positive charging of this compartment (tau = 6--12 msec) Amplitude of the second phase is to that of the third as 1 : 2. (4) A very slow phase of discharge of the flash- induced potential difference (tau = 1 sec at 10(8) ohm X cm2 film resistance). The third phase was specifically inhibited by La3+. Both the second and the third phases are decelerated by substitution of D2O in 4.5--5 and 2 times, respectively, while the amplitude of the first phase increases. Prolonged storage of the system in the dark (tua = 20-- 25 min) causes the decrease in the amplitudes of the second and the third phases as if the amount of active bacteriorhodopsin molecules were increased by factor 2. Such an inhibition was reversed by 30--60 sec illumination of the system. The dark adaptation is accompanied by some increase in the first phase amplitude. Comparison of these data with results of other studies on bacteriorhodopsin suggests that (1) the first phase is due to the photoinduced change in the retinal dipole; (2) the second phase corresponds to H+ transfer from Schiff base to the water solution in the proteoliposome interior; 3) the third phase represents H+ transfer from the incubation mixture to Schiff base; (4) the dark adaptation is a result of transition of photoelectrochemically active all-trans-retinal to the inactive 13-cis- retinal
MH  - A
MH  - ACTIVE
MH  - Bacteriorhodopsin
MH  - BASE
MH  - carotenoid
MH  - Carotenoids
MH  - flash
MH  - function
MH  - H+
MH  - Light
MH  - liposome
MH  - Liposomes
MH  - proteoliposome
MH  - retinal
MH  - Schiff base
MH  - Schiff-base
MH  - SYSTEM
MH  - Time
MH  - TRANSFER
MH  - Water
RP  - NOT IN FILE
NT  - UI - 79199347LA - rusRN - 0 (Liposomes)RN - 36-88-4 (Carotenoids)RN - 53026-44-1 (Bacteriorhodopsins)PT - Journal ArticleDA - 19790901IS - 0026-8984SB - IMCY - USSR
UR  - PM:618349
SO  - Mol Biol (Mosk ) 1977 Nov ;11(6):1377-1387

223
UI  - 19830
AU  - Ernster L
AU  - Carlsson C
AU  - Boyer PD
TI  - Reconstituted mitochondrial oligomycin-sensitive ATPase (F0F1) with intermediate Pi in equilibrium HOH exchange but no Pi in equilibrium ATP exchange activity
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - atp
MH  - ATPase
MH  - Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone
MH  - F0F1
MH  - Oligomycins
MH  - Phosphates
MH  - Water
RP  - NOT IN FILE
NT  - UI - 78084784LA - engRN - 0 (Oligomycins)RN - 0 (Phosphates)RN - 28380-24-7 (Nigericin)RN - 370-86-5 (Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone)RN - 56-65-5 (Adenosine Triphosphate)RN - 7732-18-5 (Water)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19780310IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:145953
SO  - FEBS Lett 1977 Dec 15 ;84(2):283-286

224
UI  - 21125
AU  - Ferguson SJ
AU  - Sorgato MC
TI  - The phosphorylation potential generated by respiring bovine heart submitochondrial particles
AB  - A phosphorylation potential deltaGp, where deltaGp = deltaGo' + RT2.303 log ([ATP]/([ADP][Pi])), of approx. 44.3 kJ.mol-1 (10.6 kcal.mol-1) was generated by submitochondrial particles that were oxidizing either NADH or succinate. Addition of adenylyl imidodiphosphate, which should suppress adenosine triphosphatase activity of any uncoupled particles, did not raise the phosphorylation potential. Raising the Pi concentration slightly increased the magnitude of the value for [ATP]/[ADP], but this did not fully compensate for the increased Pi concentration, so that the phosphorylation potential decreased slightly as the Pi concentration was raised. The phosphorylation potential developed by submitochondrial particles is lower than that generated by phosphorylating membrane vesicles from some bacteria, and is also less than that developed externally by mitochondria, but is strikingly close to the phosphorylation potential that is generated internally by mitochondria
MH  - A
MH  - Adenosine
MH  - ADENOSINE-TRIPHOSPHATASE
MH  - Bacteria
MH  - England
MH  - membrane
MH  - membrane vesicles
MH  - Mitochondria
MH  - Nad
MH  - Phosphorylation
MH  - Submitochondrial Particles
MH  - succinate
MH  - Succinates
MH  - vesicles
RP  - NOT IN FILE
NT  - UI - 78080229LA - engRN - 0 (Succinates)RN - 53-84-9 (NAD)PT - Journal ArticleDA - 19780223IS - 0264-6021SB - IMCY - ENGLAND
UR  - PM:202265
SO  - Biochem J 1977 Nov 15 ;168(2):299-303

225
UI  - 21128
AU  - Ferguson SJ
TI  - Some aspects of adenosine triphosphatase mechanisms [proceedings]
MH  - Adenosine
MH  - ADENOSINE-TRIPHOSPHATASE
MH  - Adenosinetriphosphatase
MH  - Calcium
MH  - England
MH  - mechanism
MH  - MECHANISMS
MH  - Potassium
MH  - Sodium
RP  - NOT IN FILE
NT  - UI - 78043916LA - engRN - 7440-09-7 (Potassium)RN - 7440-23-5 (Sodium)RN - 7440-70-2 (Calcium)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19780127IS - 0300-5127SB - IMCY - ENGLAND
UR  - PM:144622
SO  - Biochem Soc Trans 1977  ;5(5):1281-1283

226
UI  - 21126
AU  - Ferguson SJ
AU  - Harris DA
AU  - Radda GK
TI  - The adenosine triphosphatase-inhibitor content of bovine heart submitochondrial particles. Influence of the inhibitor on adenosine triphosphate-dependent reactions
AB  - 1. The activity of the ATPase (adenosine triphosphatase) of phosphorylating particles prepared by sonication of bovine heart mitochondria in the presence of MgCl2 and ATP is influenced by the isolation method for the mitochondria used in the preparation of particles. Type-I particles, made from mitochondria isolated in a medium lacking succinate, have a lower ATPase activity than to Type-II particles, which are prepared from mitochondria isolated in a medium containing succinate. 2. Centrifugation under appropriate energized conditions increases the ATPase activity of Type-I particles almost to that of the Type-II particles. The ATPase activity of Type-II particles was only slightly stimulated by this procedure. These data are interpreted as indicating a higher content of the ATPase-inhibitor protein in the Type-I particles. 3. A comparison was made of the ATP- driven enhancement of 8-anilinonaphthalene-1-sulphonate fluorescence and the exchange of the endogenous tightly bound nucleotides of the ATPase in Type-I and Type-II particles. The effect of exogenous inhibitor protein on both these reactions was also studied. 4. The time- scale on which the inhibitor protein can exchange between ATPase molecules is discussed
MH  - A
MH  - Adenine Nucleotides
MH  - ADENINE-NUCLEOTIDES
MH  - Adenosine
MH  - ADENOSINE-TRIPHOSPHATASE
MH  - Adenosinetriphosphatase
MH  - atp
MH  - ATPase
MH  - BOUND NUCLEOTIDES
MH  - England
MH  - Enzyme Inhibitors
MH  - fluorescence
MH  - inhibitor
MH  - INHIBITOR PROTEIN
MH  - method
MH  - Mitochondria
MH  - Nucleotides
MH  - protein
MH  - Submitochondrial Particles
MH  - succinate
MH  - Succinates
MH  - Time
RP  - NOT IN FILE
NT  - UI - 77157185LA - engRN - 0 (Adenine Nucleotides)RN - 0 (Anilino Naphthalenesulfonates)RN - 0 (Enzyme Inhibitors)RN - 0 (Succinates)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19770525IS - 0264-6021SB - IMCY - ENGLAND
UR  - PM:139891
SO  - Biochem J 1977 Feb 15 ;162(2):351-357

227
UI  - 597
AU  - Frigeri L
AU  - Galante YM
AU  - Hanstein WG
AU  - Hatefi Y
TI  - Effects of arginine binding reagents on ATPase and ATP-Pi exchange activities of mitochondrial ATP synthetase complex (complex V)
RP  - NOT IN FILE
NT  - UI - 77187779LA - engRN - 0 (Aldehydes)RN - 0 (Butanones)RN - 0 (Uncoupling Agents)RN - 107-22-2 (Glyoxal)RN - 431-03-8 (Diacetyl)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 7004-12-8 (Arginine)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19770723IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:140868
SO  - J Biol Chem 1977 May 25 ;252(10):3147-3152

228
UI  - 215
AU  - Futai M
TI  - Reconstitution of ATPase activity from the isolated alpha, beta, and gamma subunits of the coupling factor, F1, of Escherichia coli
MH  - Adenine Nucleotides
MH  - ADENINE-NUCLEOTIDES
MH  - Adenosinetriphosphatase
MH  - ATPase
MH  - Bacterial Proteins
MH  - coupling
MH  - COUPLING FACTOR
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1
MH  - Magnesium
MH  - membrane
MH  - Membrane Proteins
MH  - Nucleotides
MH  - protein
MH  - Proteins
MH  - reconstitution
MH  - SUBUNIT
RP  - NOT IN FILE
NT  - UI - 78103217LA - engRN - 0 (Adenine Nucleotides)RN - 0 (Bacterial Proteins)RN - 0 (Membrane Proteins)RN - 0 (Peptides)RN - 7439-95-4 (Magnesium)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19780310IS - 0006-291XSB - IMCY - UNITED STATES
UR  - PM:146491
SO  - Biochem Biophys Res Commun 1977 Dec 21 ;79(4):1231-1237

229
UI  - 8646
AU  - Graber P
AU  - Schlodder E
AU  - Witt HT
TI  - Conformational change of the chloroplast ATPase induced by a transmembrane electric field and its correlation to phosphorylation
MH  - A
MH  - atp
MH  - ATPase
MH  - chloroplast
MH  - conformational change
MH  - FIELD
MH  - mechanism
MH  - Phosphorylation
MH  - regulation
MH  - voltage
RP  - IN FILE
NT  - J ATP 4
SO  - Biochim Biophys Acta 1977  ;461():426-440

230
UI  - 20825
AU  - Griffiths DE
AU  - Cain K
AU  - Hyams RL
TI  - Studies of energy-linked reactions. Inhibition of oxidative phosphorylation by DL-8-methyldihydrolipoate
AB  - 1. DL-8-Methyldihydrolipoate was shown to be a potent inhibitor of mitochondrial oxidative phosphorylation and ATP-driven energy-linked reactions. 2. ADP-stimulated respiration utilizing pyruvate + malate and succinate in both ox heart and rat liver mitochondria is inhibited; oxidative phosphorylation using pyruvate + malate, succinate and ascorbate + NNN'N'-tetramethyl-p-phenylenediamine as substrates is also inhibited; uncoupler-stimulated respiration is unaffected regardless of the substrate used. 3. Mitochondrial oligomycin-sensitive adenosine triphosphatase is inhibited in both the membrane-bound form and the purified detergent-dispersed preparation. 4. ATP-driven transhydrogenase and the ATP-driven energy-linked reduction of NAD+ by succinate in ox heart submitochondrial particles are inhibited, whereas the respiratory-chain-driven transhydrogenase is unaffected. 5. DL-8- Methyl-lipoate has no immediate effect on the above reactions, demonstrating the requirement for the reduced form for inhibition. 6. The inhibitory properties of DL-8-methyldihydrolipoate are analogous to those of oligomycin and provide further evidence of a role for lipoic acid in oxidative phosphorylation
MH  - A
MH  - ACID
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - ADENOSINE-TRIPHOSPHATASE
MH  - Adenosinetriphosphatase
MH  - England
MH  - Liver
MH  - Mitochondria
MH  - Oxidative Phosphorylation
MH  - Phosphorylation
MH  - Respiration
MH  - Submitochondrial Particles
MH  - succinate
RP  - NOT IN FILE
NT  - UI - 77241788LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - 62-46-4 (Thioctic Acid)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19770917IS - 0264-6021SB - IMCY - ENGLAND
UR  - PM:142482
SO  - Biochem J 1977 Jun 15 ;164(3):699-704

231
UI  - 20827
AU  - Griffiths DE
AU  - Cain K
AU  - Hyams RL
TI  - Oxidative phosphorylation: a new biological function for lipoic acid
MH  - A
MH  - ACID
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - England
MH  - function
MH  - Organotin Compounds
MH  - Oxidative Phosphorylation
MH  - Phosphorylation
RP  - NOT IN FILE
NT  - UI - 77247122LA - engRN - 0 (Organotin Compounds)RN - 56-65-5 (Adenosine Triphosphate)RN - 62-46-4 (Thioctic Acid)PT - Journal ArticleDA - 19771020IS - 0300-5127SB - IMCY - ENGLAND
UR  - PM:330275
SO  - Biochem Soc Trans 1977  ;5(1):205-207

232
UI  - 9965
AU  - Holmes NG
AU  - Crofts AR
TI  - The carotenoid shift in Rhodopseudomonas sphaeroides. Change induced under continuous illumination
AB  - The spectrum of the carotenoid shift generated under continuous illumination in the GIC mutant of Rhodopseudomonas sphaeroides, which has a single carotenoid, has been examined under a variety of conditions expected to alter the size of the membrane potential. If the difference spectrum observed was due to a species with the spectrum of the bulk pigment, it would correspond to a change of a variable proportion of the pigment to a form absorbing at a higher wavelength. The maximal change induced by light could be described as a shift of about 10% of the pigment by 7 nm to the red, assuming that the shifted species was spectrally identical to the bulk carotenoid. It is concluded that the changes seen are not easily compatible with a progressive red shift in the whole spectrum with increasing applied potential as would be expected from a simple linear electrochromic mechanism; alternative hypotheses are discussed
MH  - Carotenoids
MH  - Light
MH  - mechanism
RP  - NOT IN FILE
NT  - UI - 77221904LA - engRN - 0 (Carotenoids)PT - Journal ArticleDA - 19770922IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:301749
SO  - Biochim Biophys Acta 1977 Jul 7 ;461(1):141-150

233
UI  - 9966
AU  - Holmes NG
AU  - Crofts AR
TI  - The carotenoid shift in Rhodopseudomonas sphaeroides. The flash induced change
AB  - A mutant, Rhodopseudomonas sphaeroides GIC, having only one major carotenoid, neurosporene, is described. The spectrum of the carotenoid shift in this mutant is analysed and it is concluded that only 7-11% of the pigment is involved under conditions of steady-state illumination and that this pigment undergoes a shift of 7 nm. The spectrum of the carotenoid shift under conditions of multi-flash illumination is examined for changes in shape concordant with a progressive red shift of the pigment with increasing membrane potential; the spectra of the fast change after each of three flashes does not agree well with predictions from a model involving a progressive shift of the pigment, the slow change shows qualitative agreement with such a model but the small size of the signal and the presence of more than one phase makes analysis of this phase more difficult. No separate pool of carotenoid, that might correspond to that postulated to participate in the carotenoid shift, could be identified by fourth derivative analysis of, or curve fitting to, the spectrum of the neurosporene
MH  - analysis
MH  - Antimycin A
MH  - Carotenoids
MH  - model
RP  - NOT IN FILE
NT  - UI - 77134795LA - engRN - 0 (Carotenoids)RN - 642-15-9 (Antimycin A)PT - Journal ArticleDA - 19770527IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:300248
SO  - Biochim Biophys Acta 1977 Mar 11 ;459(3):492-505

234
UI  - 833
AU  - Huchzermeyer B
AU  - Strotmann H
TI  - Acid/base-induced exchange of adenine nucleotides on chloroplast coupling factor (CF1)
RP  - NOT IN FILE
NT  - UI - 78056860LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 7439-95-4 (Magnesium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19780127IS - 0341-0382SB - IMCY - GERMANY, WESTJC - XYX
UR  - PM:22169
SO  - Z Naturforsch [C] 1977 Sep ;32(9-10):803-809

235
UI  - 13890
AU  - Izawa S
TI  - 16. Inhibitors of electron transport
MH  - electron
MH  - Electron Transport
MH  - inhibitor
MH  - inhibitors
MH  - Photosynthesis
MH  - physiology
MH  - plant
MH  - transport
T2  - Photosynthesis I; Encyclopedia of Plant Physiology; New Ser.;
A2  - Trebst A
A2  - Avron M
Y2  - -32676  
PB  - Berlin: Springer
RP  - IN FILE
SO  -  1977  ;():266-282

236
UI  - 8294
AU  - Junge W
TI  - Membrane potentials in photosynthesis
MH  - bioenergetics
MH  - membrane
MH  - Membrane Potential
MH  - Membrane Potentials
MH  - Photosynthesis
MH  - review
RP  - NOT IN FILE
SO  - Annu Rev Plant Physiol 1977  ;28():503-536

237
UI  - 793
AU  - Kagawa Y
AU  - Ohno K
AU  - Yoshida M
AU  - Takeuchi Y
AU  - Sone N
TI  - Proton translocation by ATPase and bacteriorhodopsin
AB  - Stable membrane proteins and lipids are convenient to study biomembranes. Two stable proton translocating proteins were purified and reconstituted into vesicles capable of proton translocation. One was a thermostable ATPase (TF0-F1) of thermophilic bacterium PS3 and the other was rhodopsin of Halobacterium halobium. TF0-F1 was composed of a proton pump moiety (TF1) and a proton channel moiety (TF0). TF1 was the first membrane ATPase which was crystallized and reconstituted from its five polypeptides. Like TF0 and TF1, the rhodopsin in purple membrane was highly stable against dissociating agents, acids and alkali. Phospholipids of these biomembranes were also stable and contained no unsaturated fatty acyl groups. The molecular species of the phospholipids of PS3 were determined by mass chromatography. Measurements were made of the difference in electrochemical potential of protons (deltamicronH+) across the membrane of the reconstituted vesicles. The deltamicronH+ attained was 312 mV in TF0-F1 vesciles and was 230 mV in the rhodopsin vesicles. To conclude that electron transport components are not necessary for ATP synthesis in energy yielding biomembranes, two experiments were performed: The ATP synthesis was observed i) on acid-base treatment of TF0-F1 vesicles, and ii) on illumination of the rhodopsin-TF0-F1 vesicles
RP  - NOT IN FILE
NT  - UI - 77162671LA - engRN - 0 (Carotenoids)RN - 0 (Liposomes)RN - 0 (Phospholipids)RN - 0 (Proteolipids)RN - 370-86-5 (Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone)RN - 53026-44-1 (Bacteriorhodopsin)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19770622IS - 0014-9446SB - IMCY - UNITED STATESJC - EUV
UR  - PM:15875
SO  - Fed Proc 1977 May ;36(6):1815-1818

238
UI  - 19831
AU  - Kayalar C
AU  - Rosing J
AU  - Boyer PD
TI  - An alternating site sequence for oxidative phosphorylation suggested by measurement of substrate binding patterns and exchange reaction inhibitions
AB  - Catalysis by beef heart submitochondrial particles of the medium Pi in equilibrium HOH, Pi in equilibrium ATP, and the ATP in equilibrium HOH exchanges is strongly inhibited while the ATPase and intermediate Pi in equilibrium HOH exchange are accelerated when medium ADP is removed by pyruvate kinase action. Arsenate readily blocks completely the Pi in equilibrium ATP and medium Pi in equilibrium HOH exchange reactions, but not the ATP in equilibrium HOH exchange reaction. The residual ATP in equilibrium HOH exchange in presence of arsenate is inhibited by 2,4- dinitrophenol. These results and other data are explained by an alternating site model for oxidative phosphorylation. In this model during net oxidative phosphorylation ATP is formed at one site but is transitorily tightly bound and not released until ADP and Pi bind at a second site and the membrane ATPase complex is energized. Under conditions of net ATP hydrolysis, ATP binding at one site is accompanied by hydrolysis of the transitorily tightly bound ATP as a second site. Attractive features are only one site of input for conformational energization of the membrane ATPase, a single conformational transition that accounts for both the promotion of ADP and Pi binding in a competent mode and the release of tightly bound ATP, and a symmetry of catalytic sites. The Pi in equilibrium ATP exchange is not inhibited by increase in MgADP and MgATP at constant ratios, and the energy-linked ADP in equilibrium ATP exchange is not inhibited by increased concentrations of MgATP and Pi at a constant ratio. Such exchange patterns indicate a random binding and release of ADP and Pi
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - ADP
MH  - atp
MH  - ATPase
MH  - BINDING
MH  - Carrier Proteins
MH  - Catalysis
MH  - COMPLEX
MH  - Dinitrophenols
MH  - Hydrolysis
MH  - Magnesium
MH  - membrane
MH  - model
MH  - Oxidative Phosphorylation
MH  - Phosphates
MH  - Phosphorylation
MH  - Proteins
MH  - Pyruvate Kinase
MH  - Submitochondrial Particles
RP  - NOT IN FILE
NT  - UI - 77165165LA - engRN - 0 (Arsenates)RN - 0 (Carrier Proteins)RN - 0 (Dinitrophenols)RN - 0 (Phosphates)RN - 56-65-5 (Adenosine Triphosphate)RN - 7439-95-4 (Magnesium)PT - Journal ArticleDA - 19770611IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:856791
SO  - J Biol Chem 1977 Apr 25 ;252(8):2486-2491

239
UI  - 18358
AU  - Lundin A
AU  - Thore A
AU  - Baltscheffsky M
TI  - Sensitive measurement of flash induced photophosphorilation in bacterial chromatophores by firefly luciferase
MH  - ATPase
MH  - Bacterial Chromatophores
MH  - chromatophore
MH  - chromatophores
MH  - flash
MH  - luciferase
RP  - NOT IN FILE
SO  - FEBS Lett 1977  ;79(1):73-76

240
UI  - 9954
AU  - Matsuura K
AU  - Nishimura M
TI  - Light- and diffusion-potential-induced shift of carotenoid spectrum in reconstituted vesicles of Rhodopseudomonas sphaeroides
AB  - Proteoliposomes were reconstituted from detergent-solubilized pigment.protein complexes of chromatophores of Rhodopseudomonas sphaeroides and soybean phospholipids. The reconstituted vesicles showed a photooxidation of reaction center bacteriochlorophyll and a light-induced spectral shift of carotenoid to longer wave-lengths. The red shift similar to that in intact cells or chromatophores, indicates the generation of local fields in the membrane of proteoliposomes. When inside-positive membrane potential was induced by adding valinomycin and potassium salt, a shift of carotenoid spectrum to shorter wavelengths was observed. Therefore, the reconstituted vesicles, at least in the major part of population, produced the light-induced local field in the same direction as in intact cells, which is inside negative. Sidedness of the membrane structure and the direction of electric field formation in reconstituted vesicles were opposite to those in chromatophores (inside-out vesicles
MH  - Carotenoids
MH  - Cells
MH  - COMPLEX
MH  - FIELD
MH  - Light
MH  - Liposomes
MH  - Phospholipids
MH  - Potassium
MH  - proteoliposome
MH  - Valinomycin
RP  - NOT IN FILE
NT  - UI - 78080814LA - engRN - 0 (Carotenoids)RN - 0 (Liposomes)RN - 2001-95-8 (Valinomycin)RN - 7440-09-7 (Potassium)PT - Journal ArticleDA - 19780310IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:304357
SO  - Biochim Biophys Acta 1977 Dec 23 ;462(3):700-705

241
UI  - 1069
AU  - Matsuura K
AU  - Nishimura M
TI  - Sidedness of membrane structures in Rhodopseudomonas sphaeroides. Electrochemical titration of the spectrum changes of carotenoid in spheroplasts, spheroplast membrane vesicles and chromatophores
MH  - membrane vesicles
MH  - rhodopseudomonas
RP  - NOT IN FILE
SO  - Biochim Biophys Acta 1977  ;459():483-491

242
UI  - 617
AU  - Melandri BA
AU  - Casadio R
AU  - Melandri AB
TI  - Bacterial photosynthetic phosphorylation under conditions of limited electron flow
RP  - NOT IN FILE
NT  - UI - 78003970LA - engRN - 370-86-5 (Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19771125IS - 0300-5127SB - IMCY - ENGLANDJC - E48
UR  - PM:20372
SO  - Biochem Soc Trans 1977  ;5(2):495-499

243
UI  - 1002
AU  - Mitchell P
TI  - A commentary on alternative hypotheses of protonic coupling in the membrane systems catalysing oxidative and photosynthetic phosphorylation
MH  - Adenosinetriphosphatase
MH  - Nucleotides
MH  - Phosphorylation
MH  - Phosphotransferases
RP  - NOT IN FILE
NT  - UI - 77204840LA - engRN - 0 (Adenine Nucleotides)RN - 0 (Anhydrides)RN - EC 2.7 (Phosphotransferases)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticlePT - ReviewDA - 19770825IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:17549
SO  - FEBS Lett 1977  ;78(1):1-20

244
UI  - 1022
AU  - Mitchell P
TI  - Vectorial chemiosmotic processes
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - Animal
MH  - Biological Transport
MH  - Cytochromes
MH  - Electron Transport
MH  - Hydrogen-Ion Concentration
MH  - Membrane Potentials
MH  - Membranes
MH  - metabolism
MH  - NADH,NADPH Oxidoreductases
MH  - Osmosis
MH  - Oxidative Phosphorylation
RP  - NOT IN FILE
SO  - Annu Rev Biochem 1977  ;46:996-1005.():996-1005

245
UI  - 1126
AU  - Nishimura M
AU  - Yamamoto Y
AU  - Takahama U
AU  - Shimizu M
AU  - Matsuura K
TI  - Biological conversion of light energy into electrochemical potential
MH  - Light
T2  - Biol. Sol. Energy Convers., [Pap. Conf.], Meeting Date 1976, 143- 8. Edited by: Mitsui, Akira; Miyachi, Shigetoh; San Pietro, Anthony. Academic: New York, N. Y
Y2  - -32676  
RP  - NOT IN FILE
SO  -  1977  ;():

246
UI  - 651
AU  - Penefsky HS
TI  - Reversible binding of Pi by beef heart mitochondrial adenosine triphosphatase
AB  - Beef heart mitochondrial ATPase (F1) exhibited a single binding site for Pi. The interaction with Pi was reversible, partially dependent on the presence of divalent metal ions, and characterized by a dissociation constant at pH 7.5 of 80 micronM. A variety of substances known to influence oxidative phosphorylation or the activity of the soluble ATPase (F1) also influenced Pi binding by the enzyme. Thus aurovertin, an inhibitor of oxidative phosphorylation, which was bound tightly by F1 and inhibited ATPase activity, enhanced Pi binding via a 4-fold increase in the affinity of the enzyme for Pi (KD = 20 micronM) but did not alter binding stoichiometry. Anions such as SO4(2-), SO3(2- ), chromate, and 2,4-dinitrophenolate, which stimulated ATPase activity of F1, also enhanced Pi binding. Inhibitors of ATPase activity such as nickel/bathophenanthroline and the protein ATPase inhibitor of Pullman and Monroy (Pullman, M. E., and Monroy, G. C. (1963) J. Biol. Chem. 238, 3762-3769) inhibited Pi binding. The adenine nucleotides ADP, ATP, and the ATP analog adenylyl imidodiphosphate as well as the Pi analog arsenate, also inhibited Pi binding. The observations suggest that the Pi binding site was located in or near an adenine nucleotide binding site on the molecule
RP  - NOT IN FILE
NT  - UI - 77165226LA - engRN - 0 (Adenine Nucleotides)RN - 0 (Anions)RN - 0 (Aurovertins)RN - 0 (Phenanthrolines)RN - 0 (Phosphates)RN - 7440-38-2 (Arsenic)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19770622IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:16006
SO  - J Biol Chem 1977 May 10 ;252(9):2891-2899

247
UI  - 855
AU  - Petty KM
AU  - Jackson JB
AU  - Dutton PL
TI  - Kinetics and stoichiometry of proton binding in Phodopseudomonas sphaeroides chromatophores
RP  - NOT IN FILE
NT  - UI - 78084788LA - engRN - 2001-95-8 (Valinomycin)RN - 642-15-9 (Antimycin A)PT - Journal ArticleDA - 19780310IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:23313
SO  - FEBS Lett 1977 Dec 15 ;84(2):299-303

248
UI  - 19832
AU  - Rosing J
AU  - Kayalar C
AU  - Boyer PD
TI  - Evidence for energy-dependent change in phosphate binding for mitochondrial oxidative phosphorylation based on measurements of medium and intermediate phosphate-water exchanges
AB  - Characteristics of the exchange reactions catalyzed by beef heart submitochondrial particles give new insight into energy transducing steps of oxidative phosphorylation. The uncoupler-insensitive portion of the total Pi in equilibrium HOH exchange in presence of ATP, ADP, and Pi is the intermediate Pi in equilibrium HOH exchange, that is the exchange occurring with Pi formed by hydrolysis of ATP prior to release of Pi from the catalytic site. The exchange of medium Pi with HOH is as sensitive to uncouplers as the Pi in equilibrium ATP exchange and net oxidative phosphorylation, demonstrating a requirement of an uncoupler- sensitive energized state, probably a transmembrane potential or proton gradient, for bringing medium Pi to the reactive state. The covalent bond forming and breaking step at the catalytic site (ADP + Pi in equilibrium ATP + HOH) appears relatively insensitive to uncouplers. Thus to the extent that uncouplers dissipate transmembrane proton- motive force, it is unlikely that such a force is used to drive ATP formation by direct protonations of Pi oxygens. When only Pi and ADP are added and formation of ATP from added ADP by adenylate kinase and subsequent ATP hydrolysis are adequately blocked, no Pi in equilibrium HOH exchange can be observed, demonstrating a requirement of energization by ATP binding and cleavage for such an exchange. This uncoupler-insensitive energization is suggested to represent a conformationally energized state that can be used reversibly to develop a transmembrane protonmotive force accompanying ADP and Pi release. Rates of various exchanges as estimated by improved procedures are compatible with all oxygen exchanges occurring by dynamic reversal of ATP hydrolysis at the catalytic site
MH  - A
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - ADP
MH  - atp
MH  - ATP FORMATION
MH  - BINDING
MH  - Dinitrophenols
MH  - Glucose
MH  - Hexokinase
MH  - Hydrolysis
MH  - Oxidative Phosphorylation
MH  - Oxygen
MH  - Phosphates
MH  - Phosphorylation
MH  - proton
MH  - Submitochondrial Particles
RP  - NOT IN FILE
NT  - UI - 77165164LA - engRN - 0 (Dinitrophenols)RN - 0 (Phosphates)RN - 50-99-7 (Glucose)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 2.7.1.1 (Hexokinase)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19770611IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:140165
SO  - J Biol Chem 1977 Apr 25 ;252(8):2478-2485

249
UI  - 9963
AU  - Saphon S
AU  - Crofts AR
TI  - The H+/e ration in chloroplasts in 2. Possible errors in its determination
MH  - chloroplast
MH  - Chloroplasts
RP  - NOT IN FILE
NT  - UI - 78056861LA - engPT - Journal ArticleDA - 19780127IS - 0341-0382SB - IMCY - GERMANY, WESTJC - XYX
UR  - PM:22170
SO  - Z Naturforsch [C] 1977 Sep ;32(9-10):810-816

250
UI  - 21127
AU  - Shaughnessy JP
AU  - Ferguson SJ
AU  - Harris DA
AU  - Radda GK
TI  - Interaction of mitochondrial adenosine triphosphatase with nucleotides and with glycerol [proceedings]
MH  - Adenosine
MH  - ADENOSINE-TRIPHOSPHATASE
MH  - Adenosinetriphosphatase
MH  - England
MH  - Nucleotides
RP  - NOT IN FILE
NT  - UI - 78044026LA - engRN - 0 (Ribonucleotides)RN - 56-81-5 (Glycerol)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19780127IS - 0300-5127SB - IMCY - ENGLAND
UR  - PM:144631
SO  - Biochem Soc Trans 1977  ;5(5):1539-1540

251
UI  - 790
AU  - Sone N
AU  - Takeuchi Y
AU  - Yoshida M
AU  - Ohno K
TI  - Formations of electrochemical proton gradient and adenosine triphosphate in proteoliposomes containing purified adenosine triphosphatase and bacteriorhodopsin
AB  - Proteoliposome vesicles containing both bacteriorhodopsin of Halobacterium halobium and H+-translocating ATPase [EC 3.6,1.3] of a thermophilic bacterium, PS3, (TF0-F1) were reconstituted by either the dialysis method or the sonication method. Generation of the electrochemical proton gradient (deltamuH+) in these vesicles was measured using 9-aminoacridine for estimation of the chemical (deltapH) component and 8-anilinonaphthalene sulfonate for the electrical (deltaphi) component). In illuminated bacteriorhodopsin-vesicles the deltamuH+ reached 180-190 mV when reconstituted by the dialysis method and 210-220 mV when reconstituted by the sonication method. Vesicles reconstituted from both TF0-F1 and bacteriorhodopsin by the dialysis method generated a deltapH+ of about 200 mV on addition of ATP, while vesicles prepared by the sonication method generated very little deltamuH+, if any. These vesicles generated similar deltamuH+ on illumination to that found in bacteriorhodopsin-vesicles. Using vesicles reconstituted from both TF0-F1 and bacteriorhodopsin by the dialysis method, light dependent ATP synthesis was measured in relation to deltamuH+ formation. It was necessary to generate a deltamuH+ of above 170 mV for demonstration of appreciable formation of ATP and the greater the deltamuH+, the faster the rate of ATP synthesis
RP  - NOT IN FILE
NT  - UI - 78087507LA - engRN - 0 (Anilino Naphthalenesulfonates)RN - 0 (Carotenoids)RN - 0 (Liposomes)RN - 0 (Proteolipids)RN - 53026-44-1 (Bacteriorhodopsin)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19780310IS - 0021-924XSB - IMCY - JAPANJC - HIF
UR  - PM:23379
SO  - J Biochem (Tokyo ) 1977 Dec ;82(6):1751-1758

252
UI  - 795
AU  - Sone N
AU  - Yoshida M
AU  - Hirata H
AU  - Kagawa Y
TI  - Reconstitution of vesicles capable of energy transformation from phospholipids and adenosine triphosphatase of a thermophilic bacterium
AB  - 1. A stable ATPase [EC 3.6.1.3] complex (TF0-F1) from the thermophilic bacterium PS3 was reconstituted into vesicles capable of energy transformation,measured as ATP-dependent enhancement of fluorescence of 8-anilinonoaphthalene-1-sulfonate. 2. The factors necessary for obtaining highly active vesicles were investigated. Cholate and deoxycholate were both required for solubilization of TF0-F1 and P- lipids, and removal of the detergents by dialysis resulted in vesicle formation. Medium of around pH 8 and low ionic strength containing 2.5 mM MgSO4 was found suitable for dialysis. The optimal temperature for reconstitution was 30 degrees with soybean P-lipids and 45 degree with PS3 P-lipids. The optimal ratio of protein to lipid was about 1/50. 3. The vesicles obtained under these conditions were mainly 100-200 nm in diameter, covered with 9.5 nm spheres, and had a bouyant density of 1.06 in sucrose andan internal volume of about 0.5 mul per mg of P- lipids
RP  - NOT IN FILE
NT  - UI - 77140893LA - engRN - 0 (Anilino Naphthalenesulfonates)RN - 0 (Cholic Acids)RN - 0 (Phospholipids)RN - 7439-95-4 (Magnesium)RN - 83-44-3 (Deoxycholic Acid)RN - 9004-54-0 (Dextrans)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19770527IS - 0021-924XSB - IMCY - JAPANJC - HIF
UR  - PM:14954
SO  - J Biochem (Tokyo ) 1977 Feb ;81(2):519-528

253
UI  - 792
AU  - Sone N
AU  - Yoshida M
AU  - Hirata H
AU  - Kagawa Y
TI  - Adenosine triphosphate synthesis by electrochemical proton gradient in vesicles reconstituted from purified adenosine triphosphatase and phospholipids of thermophilic bacterium
AB  - Vesicles were reconstituted from a purified dicyclohexyl-carbodiimide- sensitive ATPase complex (TF0-F1) and phospholipids of a thermophilic bacterium PS3. These vesicles synthesized ATP from ADP and Pi with energy from an electrochemical proton gradient (delta-micronH+) formed by a pH gradient and an electrical potential across their membranes. Maximal ATP synthesis was achieved by incubating the vesicles in malonate at pH 5.5 with valinomycin, and then rapidly transferring them to a solution of pH 8.4 and 150 mM K+. Under these conditons ATP synthesis continued at a decreasing rate for 30 s at 40 degrees. Appreciable formation of ATP (40 to 150 nmol/mg of TF0-F1) occurred at an initial delta-micronH+ above 205 mV and moderate formation at an initial value above 180 mV. ATP hydrolysis by the vesicles produced a delta-micronH+, and the additions of 32Pi and hexokinase to them resulted in 32Pi esterification. Analysis of the time courses of 32Pi esterification and decays of the pH difference and membrane potential, followed using 9-aminoacridine and 8-anilinonaphthalene-1-sulfonate, respectively, as probes, showed a relationship between delta-micronH+ and the rate of ATP synthesis. These results demonstrate that purified TF0-F1 is itself a reversible H+-translocating ATPase of oxidative phosphorylation
RP  - NOT IN FILE
NT  - UI - 77165235LA - engRN - 0 (Liposomes)RN - 0 (Phosphates)RN - 0 (Phospholipids)RN - 0 (Protons)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 2.7.1.1 (Hexokinase)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19770622IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:16011
SO  - J Biol Chem 1977 May 10 ;252(9):2956-2960

254
UI  - 834
AU  - Strotmann H
AU  - Bickel-Sandkotter S
TI  - Energy-dependent exchange of adenine nucleotides on chloroplast coupling factor (CF1)
AB  - 1. [14C]ADP is incorporated into washed broken chloroplasts in the light. The bound labelled nucleotides which cannot be removed by washing are almost exclusively related to coupling factor CF1. [14C]ADP binding exhibits a monophasic concentration curve with a Km of 2 micronM. 2. By illumination of the chloroplasts, previously incorporated labelled nucleotides are released. A fast release is obtained in the presence of unlabelled ADP and ATP, indicating an energy-dependent exchange. A slow and incomplete release is induced by light in the absence of unlabelled adenine nucleotides. Obviously, under those conditions, an adenine nucleotide depleted CF1 conformation is established. 3. Re-binding of [14C]ADP by depleted membranes is an energy-independent process. Even after solubilization of adenylate- depleted CF1, [14C]ADP is incorporated into the protein. By re-binding of ADP in the dark, CF1 is converted to a non-exchangeable form. 4. Energy-dependent adenine nucleotide exchange on CF1 is suggested to include three different conformational states of the enzyme: (1) a stable, non-exchangeable form which contains firmly bound nucleotides, is converted to (2), an unstable form containing loosely bound adenine nucleotides. This conformation allows adenylate exchange; it is in equilibrium with (3) a metastable, adenylate-depleted form. The transition from state (1) to state (2) is the energy-requiring step
RP  - NOT IN FILE
NT  - UI - 77157861LA - engRN - 0 (Oxidative Phosphorylation Coupling Factors)RN - 0 (Plant Proteins)RN - 370-86-5 (Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 61-19-8 (Adenosine Monophosphate)PT - Journal ArticleDA - 19770611IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:856262
SO  - Biochim Biophys Acta 1977 Apr 11 ;460(1):126-135

255
UI  - 21269
AU  - Symons M
AU  - Swysen C
AU  - Sybesma C
TI  - The light-induced carotenoid absorbance changes in Rhodopseudomonas sphaeroides: an analysis and interpretation of the band shifts
AB  - An analysis has been made of the spectrum of the carotenoid absorption band shift generated by continuous illumination of chromatophores of the GlC-mutant of Rhodopseudomonas sphaeroides at room temperature by means of three computer programs. There appears to be at least two pools of the same carotenoid, only one of which, comprising about 20% of the total carotenoid content, is responsible for the light-induced absorbance changes. The 'remaining' pool absorbs at wavelengths which were about 5 nm lower than those at which the 'changing' pool absorbs. This difference in absorption wavelength could indicate that the two pools are influenced differently by permanent local electric fields. The electrochromic origin of the absorbance changes has been demonstrated directly; the isosbestic points of the absorption difference spectrum move to shorter wavelengths upon lowering of the light-induced electric field. Band shifts up to 1.7 nm were observed. A comparison of the light-induced absorbance changes with a KCl- valinomycin-induced diffusion potential has been used to calibrate the electrochromic shifts. The calibration value appeared to be 137 +/- 6 mV per nm shift
MH  - A
MH  - absorption
MH  - analysis
MH  - carotenoid
MH  - Carotenoids
MH  - chromatophore
MH  - chromatophores
MH  - Diffusion
MH  - diffusion potential
MH  - FIELD
MH  - rhodopseudomonas
MH  - spectra
MH  - sphaeroides
MH  - Temperature
RP  - NOT IN FILE
NT  - UI - 78080815LA - engRN - 36-88-4 (Carotenoids)PT - Journal ArticleDA - 19780310IS - 0006-3002SB - IMCY - NETHERLANDS
UR  - PM:304358
SO  - Biochim Biophys Acta 1977 Dec 23 ;462(3):706-717

256
UI  - 881
AU  - Thauer RK
AU  - Jungermann K
AU  - Decker K
TI  - Energy conservation in chemotrophic anaerobic bacteria
MH  - Adenosine Triphosphate
MH  - Anaerobiosis
MH  - Bacteria
MH  - biosynthesis
MH  - Cell Membrane
MH  - development
MH  - Energy Metabolism
MH  - enzymology
MH  - Fermentation
MH  - Glucose
MH  - growth &amp
MH  - Hydrogenation
MH  - metabolism
RP  - NOT IN FILE
SO  - Bacteriol Rev 1977 Mar ;41(1):100-180

257
UI  - 835
AU  - Tischer W
AU  - Strotmann H
TI  - Relationship between inhibitor binding by chloroplasts and inhibition of photosynthetic electron transport
AB  - The binding of radioactively labelled atrazin, metribuzin and phenmedipham by broken chloroplasts was studied. From the double- reciprocal plots (bound vs. free inhibitors) a high affinity binding reaction is graphically isolated which is related to the inhibition of photosynthetic electron transport. It is concluded that the specific binding sites correspond to the electron carrier molecules which are attacked by the inhibitors. The relative concentration of specific binding sites is 1 per 300-500 chlorophyll molecules. The binding of the labelled substances is competitively inhibited by each of the indicated unlabelled substances, by DCMU and by several pyridazinone derivatives. These results suggest that triazines, triazinones, pyridazinones, biscarbamates and phenylureas interfere with the same electron carrier of the photosynthetic electron transport chain, according to the same molecular mechanism
RP  - NOT IN FILE
NT  - UI - 77157860LA - engRN - 0 (Herbicides, Carbamate)RN - 0 (Triazines)RN - 1912-24-9 (Atrazine)RN - 330-54-1 (Diuron)PT - Journal ArticleDA - 19770611IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:856261
SO  - Biochim Biophys Acta 1977 Apr 11 ;460(1):113-125

258
UI  - 21228
AU  - Van Dam K
AU  - Wiechmann AH
AU  - Westerhoff HV
AU  - Hellingwerf KJ
TI  - Proton gradients across energy-transducing membranes
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - England
MH  - membrane
MH  - Membranes
MH  - proton
RP  - NOT IN FILE
NT  - UI - 77247160LA - engRN - 56-65-5 (Adenosine Triphosphate)PT - Journal ArticleDA - 19771020IS - 0300-5127SB - IMCY - ENGLAND
UR  - PM:19321
SO  - Biochem Soc Trans 1977  ;5(1):28-29

259
UI  - 18968
AU  - Wagner R
AU  - Junge W
TI  - Gated proton conduction via the coupling factor of photophosphorylation modified by N,N-orthophenyldimaleimide
MH  - atp
MH  - conductance
MH  - COUPLING FACTOR
MH  - gating
MH  - Photophosphorylation
MH  - proton
RP  - NOT IN FILE
SO  - Biochim Biophys Acta 1977  ;462():259-272

260
UI  - 857
AU  - Webster GD
AU  - Edwards PA
AU  - Jackson JB
TI  - Inhibition of the proton-translocating adenosine triphosphatase from chromatophores of photosynthetic bacteria by free bivalent cations and adenosine triphosphate [proceedings]
RP  - NOT IN FILE
NT  - UI - 78044021LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - 7439-95-4 (Magnesium)RN - 7440-70-2 (Calcium)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19780127IS - 0300-5127SB - IMCY - ENGLANDJC - E48
UR  - PM:144630
SO  - Biochem Soc Trans 1977  ;5(5):1527-1529

261
UI  - 856
AU  - Webster GD
AU  - Edwards PA
AU  - Jackson JB
TI  - Interconversion of two kinetically distinct states of the membrane- bound and solubilised H+-translocating ATPase from Rhodospirillum rubrum
RP  - NOT IN FILE
NT  - UI - 77162626LA - engRN - 0 (Sulfites)RN - 370-86-5 (Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone)RN - 7439-95-4 (Magnesium)RN - 7440-70-2 (Calcium)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19770611IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:15868
SO  - FEBS Lett 1977 Apr 1 ;76(1):29-35

262
UI  - 1202
AU  - Witt HT
AU  - Schlodder E
AU  - Graber P
TI  - Conformational change, ATP generation and turnover rate of the chloroplast ATPase analyzed by energization with an external electric field
MH  - atp
MH  - ATPase
MH  - chloroplast
MH  - conformational change
MH  - FIELD
RP  - ON REQUEST (03/18/92)
SO  - Dev Bioenerg Biomembr 1977  ;1():447-457

263
UI  - 794
AU  - Yoshida M
AU  - Okamoto H
AU  - Sone N
AU  - Hirata H
AU  - Kagawa Y
TI  - Reconstitution of thermostable ATPase capable of energy coupling from its purified subunits
AB  - Purified dicyclohexylcarbodiimide-sensitive ATPase (TF0-F1) from thermophilic bacterium PS3 is composed of a water soluble part with ATP hydrolytic activity (TF1) and a water insoluble moiety (TF0). All of the five subunits (alpha, beta, gamma, delta, and epsilon) of TF1 were isolated. TF1 was reconstituted from the five subunits, which catalyzed an ATP-32Pi exchange and an ATP-driven enhancement of fluorescence of 1- anilinonaphthalene-8-sulfonate, when adsorbed on proteoliposome inlaid with TF0 (TF3-vesicles). Subunit epsilon and/or delta became firmly bound to TF0-vesicles and there was no preferential sequence in the binding. Both subunits were required for binding of the remaining subunits of TF1 to TF0-vesicles, but they did not modify the high H+ - permeability of TF0-vesicles. The addition of gamma but they did not modify the high H+-permeability of TFO-vesicles. The addition of gamma subunit together with epsilon and delta subunits caused a marked decrease of H+ -permeability of TF0-vesicles, similar to that induced by TF1. We conclude tentatively that the epsilon and delta subunits connect TF0 and the other subunits forming a part of a proton pathway, gamma is a gate of proton flow coupled to ATP hydrolysis (or synthesis), and alpha and beta subunits contain the active site for energy transformation. A possible model of subunit structure of TF1 is proposed
RP  - NOT IN FILE
NT  - UI - 77149025LA - engRN - 0 (Protons)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19770512IS - 0027-8424SB - IMCY - UNITED STATESJC - PV3
UR  - PM:139610
SO  - Proc Natl Acad Sci U S A 1977 Mar ;74(3):936-940

264
UI  - 791
AU  - Yoshida M
AU  - Sone N
AU  - Hirata H
AU  - Kagawa Y
TI  - Reconstitution of adenosine triphosphatase of thermophilic bacterium from purified individual subunits
AB  - 1. Five subunits (alpha, beta, gamma, delta, and epsilon) of an ATPase from a thermophilic bacterium PS3 were purified in the presence of 8 M urea by ion exchange chromatography. Then the ATPase activity was reconstituted by mixing the subunit solutions and incubating them at 20- 45 degrees, at pH 6.3 to 7.0. 2. Mixtures containing beta + gamma or alpha + beta + delta regained ATP-hydrolyzing activity, but mixtures of alpha + beta and beta + delta did not. Combinations not including beta were all inactive. 3. The ATPase activity reconstituted from alpha + beta + delta was thermolabile and insensitive to NaN3, whereas the activities obtained from mixtures containing beta and gamma were thermostable and sensitive to NaN3, like the native ATPase. 4. The assemblies containing both beta and gamma subunits had the same mobility as the native ATPase molecule on gel electrophoresis, those without the gamma subunit moved more rapidly toward the anode. 5. Subunits epsilon and delta did not inhibit the ATPase activity of either the assembly (alpha + beta + gamma) or the native ATPase
RP  - NOT IN FILE
NT  - UI - 77187831LA - engRN - 0 (Azides)RN - 0 (Peptides)RN - 0 (Sulfites)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19770723IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:140872
SO  - J Biol Chem 1977 May 25 ;252(10):3480-3485

265
UI  - 21216
AU  - Abdulaev NG
AU  - Feigina MY
AU  - Kiselev AV
AU  - Ovchinnikov YA
AU  - Drachev LA
AU  - Kaulen AD
AU  - Khitrina LV
AU  - Skulachev VP
TI  - Products of limited proteolysis of bacteriorhodopsin generate a membrane potential
MH  - A
MH  - Bacteriorhodopsin
MH  - carotenoid
MH  - Carotenoids
MH  - liposome
MH  - Liposomes
MH  - membrane
MH  - Membrane Potential
MH  - Peptide Fragments
RP  - NOT IN FILE
NT  - UI - 78214996LA - engRN - 0 (Liposomes)RN - 0 (Peptide Fragments)RN - 36-88-4 (Carotenoids)RN - 53026-44-1 (Bacteriorhodopsins)RN - EC 3.4.22.2 (Papain)RN - EC 3.4.24.27 (Thermolysin)PT - Journal ArticleDA - 19780925IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:668882
SO  - FEBS Lett 1978 Jun 15 ;90(2):190-194

266
UI  - 554
AU  - Amzel LM
AU  - Pedersen PL
TI  - Adenosine triphosphatase from rat liver mitochondria. Crystallization and x-ray diffraction studies of the F1-component of the enzyme
AB  - The homogeneous rat liver F1-ATPase preparation of Catterall and Pedersen (Catterall, W.A., and Pedersen, P.L. (1971) J. Biol. Chem. 246, 4987-4994) has been crystallized from a solution containing phosphate and ATP by precipitation with ammonium sulfate. Most of the resultant crystals are cubes of approximately 0.3 to 0.6 mm per side. X- ray precession photographs show that the crystals are rhombohedral, space group R32 (D37 NO155) with hexagonal cell dimensions a = 148 A, c = 368 A. The molecular weight of the asymmetric unit of the crystals is 190,000 or about half the molecular weight (384,000) of the rat liver enzyme indicating that the crystallographic 2-fold axes of symmetry coincide with a molecular symmetry axis. The crystals diffract to at least 3.5 A and therefore this is the first report of an ATPase preparation in which crystals suitable for x-ray analysis have been obtained
RP  - NOT IN FILE
NT  - UI - 78130065LA - engRN - 0 (Isoenzymes)RN - 0 (Macromolecular Systems)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19780524IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:147272
SO  - J Biol Chem 1978 Apr 10 ;253(7):2067-2069

267
UI  - 8292
AU  - Auslnder W
AU  - Junge W
TI  - The proton pump in photosynthesis of green plant: quantitative measurement of the rapid pH-changes in the internal volume of thylakoids via neutral red
MH  - NEUTRALRED
MH  - Photosynthesis
MH  - plant
MH  - proton
MH  - Proton Pump
MH  - thylakoid
RP  - NOT IN FILE
SO  - Rev Bioenerg Biomembr 1978  ;2():31-44

268
UI  - 19882
AU  - Cross RL
AU  - Kohlbrenner WE
TI  - The mode of inhibition of oxidative phosphorylation by efrapeptin (A23871). Evidence for an alternating site mechanism for ATP synthesis
AB  - Results are presented that confirm and extend earlier findings that efrapeptin is a potent inhibitor of oxidative phosphorylation. Binding of efrapeptin is shown to be reversible, and a dissociation constant for the enzyme-inhibitor complex is estimated to be 10(-8) M under conditions for either ATP synthesis or hydrolysis. Fifty per cent inhibition of the ATP hydrolysis activity of submitochondrial particles is obtained at a ratio of 0.56 mol of inhibitor/mol of enzyme. Studies of efrapeptin binding under pseudo-first order conditions show that the onset of inhibition is first order with respect to efrapeptin. Combined with the inhibition titer, these results indicate that there is one inhibitor binding site per molecule of enzyme. Steady state velocity studies using a substrate regenerating system show that efrapeptin is competitive with both ADP and phosphate during ATP synthesis. However, during ATP hydrolysis, a distinctly different mode of inhibition is indicated with respect to ATP. Data are presented which suggest that ATP promotes the binding of efrapeptin to the enzyme. Indications that efrapeptin is a catalytic site inhibitor make these results difficult to reconcile with a simple mechanistic scheme involving a single independnet catalytic site for ATP synthesis and hydrolysis. Our results are discussed in terms of support for catalytic cooperativity between adjacent subunits as recently proposed by Kayalar et al. (Kayalar, C., Rosing, J., and Boyer, P. D. (1977) J. Biol. Chem. 252, 2486-2491)
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - ADP
MH  - Antibiotics
MH  - atp
MH  - ATP synthesis
MH  - BINDING
MH  - COMPLEX
MH  - Hydrolysis
MH  - mechanism
MH  - Oxidative Phosphorylation
MH  - P
MH  - Phosphorylation
MH  - Submitochondrial Particles
MH  - SUBUNIT
MH  - synthesis
RP  - NOT IN FILE
NT  - UI - 78218130LA - engRN - 0 (Antibiotics)RN - 0 (Peptides)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19780915IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:149791
SO  - J Biol Chem 1978 Jul 25 ;253(14):4865-4873

269
UI  - 21217
AU  - Drachev LA
AU  - Kaulen AD
AU  - Skulachev VP
TI  - Time resolution of the intermediate steps in the bacteriorhodopsin- linked electrogenesis
MH  - Bacteriorhodopsin
MH  - BASE
MH  - carotenoid
MH  - Carotenoids
MH  - Deuterium
MH  - intermediate
MH  - liposome
MH  - Liposomes
MH  - resolution
MH  - Retinaldehyde
MH  - Schiff base
MH  - Schiff Bases
MH  - Schiff-base
MH  - Time
RP  - NOT IN FILE
NT  - UI - 78127257LA - engRN - 0 (Liposomes)RN - 0 (Schiff Bases)RN - 116-31-4 (Retinaldehyde)RN - 36-88-4 (Carotenoids)RN - 53026-44-1 (Bacteriorhodopsins)RN - 7439-91-0 (Lanthanum)RN - 7782-39-0 (Deuterium)PT - Journal ArticleDA - 19780508IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:24553
SO  - FEBS Lett 1978 Mar 1 ;87(1):161-167

270
UI  - 124
AU  - Fillingame RH
AU  - Wopat AE
TI  - Carbodiimide-resistant mutant of Escherichia coli: suppression of resistance to dicyclohexylcarbodiimide by growth on glucose or glycerol
AB  - We have previously reported on the isolation of a mutant strain of Escherichia coli, RF-7, that has a dicyclohexylcarbodiimide (DCCD)- resistant, membrane-associated adenosine triphosphatase (ATPase) activity (R. H. Fillingame, J. Bacteriol. 124:870--883, 1975). We report here that the DCCD resistance of the ATPase of this mutant varies significantly, depending upon the carbon source used for growth. When strain RF-7 was grown aerobically on either glycerol or glucose or anaerobically on glucose rather than on a combination of succinate, acetate, and malate, ATPase activity was more sensitive to inhibition by DCCD because the carbodiimide-reactive proteolipid reacted more readily with DCCD
RP  - NOT IN FILE
NT  - UI - 78193953LA - engRN - 0 (Acetates)RN - 0 (Carbodiimides)RN - 0 (Malates)RN - 0 (Succinates)RN - 50-99-7 (Glucose)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-81-5 (Glycerol)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19780814IS - 0021-9193SB - IMCY - UNITED STATESJC - HH3
UR  - PM:149107
SO  - J Bacteriol 1978 May ;134(2):687-689

271
UI  - 64
AU  - Fillingame RH
AU  - Knoebel K
AU  - Wopat AE
TI  - Method for isolation of Escherichia coli mutants with defects in the proton-translocating sector of the membrane adenosine triphosphatase complex
AB  - A technique for selecting mutants of Escherichia coli in which the proton-translocating sector of the adenosine triphosphatase (ATPase) complex has been inactivated is reported. The procedure uses a strain of E. coli (NR-70) lacking the extrinsic (F1) sector of the ATPase complex and which in consequently permeable to protons (B. P. Rosen, J. Bacteriol. 116:1124--1129, 1973). After growing strain NR-70 under noninducing conditions for the lac operon, cells were mutagenized and plated on minimal medium containing low concentrations of lactose. Several mutants of strain NR-70 were isolated as large colonies on these plates, apparently because they could concentrate lactose more efficiently. A description of one of the mutants, strain KW-1, is reported here. The most distinguishing difference in growth properties of the two strains was that, when transferred to medium containing low concentrations of lactose, strain KW-1 induced the lac operon with a shorter lag time than strain NR-70. The mutation in strain KW-1 leading to more rapid growth on lactose was cotransducible with the asn and unc loci, at 83 min on the E. coli genetic map. Intact cells of strain KW-1 actively transported L-proline as well as did wild-type cells, whereas cells of strain NR-70 were markedly deficient in L-proline transport. The improvement in the transport capacity of strain KW-1 correlated with a marked decrease in proton permeability relative to that of strain NR-70. Based on an acid-base pulse technique that measured the proton conductance of the membranes of intact cells, strain NR-70 was at least 10 times more permeable to protons than was the wild type, whereas strain KW-1 was only 2 times more permeable. The transport properties and proton conductance were also compared with membrane vesicles prepared by osmotic shock. With either D-lactate or ascorbate- N-methylphenazonium methosulfate as respiratory substrates, vesicles of strain KW-1 transported L-proline much more rapidly than did vesicles of strain NR-70, but still at rates less rapid than those of the wild type. The passive proton conductance of the membrane vesicles was quantitated by measuring the rate of H+ influx into vesicles in response to a valinomycin-generated K+ diffusion potential. The proton permeability of vesicles of strain KW-1 was reduced 1.5-fold relative to vesicles of strain NR-70, but these vesicles were still four times more permeable to protons than was the wild type. Vesicles of strain KW- 1 corresponded to wild-type vesicles treated with 0.5 micrometer carbonylcyanide m-chlorophenylhydrazone (CCCP) and vesicles of strain NR-70 corresponded to wild-type vesicles treated with 1.4 micrometer CCCP. Treatment of wild-type vesicles with these concentrations of CCCP caused decreases in transport comparable to those observed in the mutants. Strain KW-1 lacked ATPase activity. Cross-reacting material to F1-ATPase was not found in strain KW-1 by double immunodiffusion analysis
RP  - NOT IN FILE
NT  - UI - 79048278LA - engRN - 0 (Multienzyme Complexes)RN - 1333-74-0 (Hydrogen)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19790124IS - 0021-9193SB - IMCY - UNITED STATESJC - HH3
UR  - PM:152309
SO  - J Bacteriol 1978 Nov ;136(2):570-581

272
UI  - 1201
AU  - Graber P
AU  - Saphon S
TI  - Conformational changes of the membrane-bound ATPase of bacterial chromatophores revealed by fluorescence changes of fluorescamine- labelled coupling factors
MH  - ATPase
MH  - Bacterial Chromatophores
MH  - chromatophore
MH  - chromatophores
MH  - conformational change
MH  - coupling
MH  - COUPLING FACTOR
MH  - fluorescence
RP  - ON REQUEST (03/18/92)
SO  - Z Naturforsch ,C: Biosci 1978  ;33C():421-427

273
UI  - 1200
AU  - Graber P
AU  - Schlodder E
AU  - Witt HT
TI  - Control of the rate of ATP synthesis by conformational changes in the chloroplast ATPase induced by the transmembrane electric field
MH  - atp
MH  - ATP synthesis
MH  - ATPase
MH  - chloroplast
MH  - conformational change
MH  - FIELD
MH  - synthesis
T2  - Proc. Int. Congr. Photosynth., 4th, Meeting Date 1977, 197-210. Edited by: Hall, D. O.; Coombs, J.; Goodwin, T. W. Biochem. Soc.: Colchester, Engl
Y2  - -32676  
RP  - ON REQUEST (03/18/92)
SO  -  1978  ;():

274
UI  - 1199
AU  - Graber P
AU  - Zickler A
AU  - Akerlund HE
TI  - Electric evidence for the isolation of inside-out vesicles from spinach chloroplasts
MH  - chloroplast
MH  - Chloroplasts
MH  - Spinach
MH  - SPINACH-CHLOROPLASTS
RP  - ON REQUEST (03/18/92)
SO  - FEBS Lett 1978  ;96():233-237

275
UI  - 2079
AU  - Grber P
AU  - Zickler A
AU  - Akerlund H
TI  - Electric evidence for the isolation of inside-out vesicles from spinach chloroplasts
MH  - chloroplast
MH  - Chloroplasts
MH  - inside-out
MH  - light-gradient
MH  - photoelectric measurements
MH  - Spinach
MH  - SPINACH-CHLOROPLASTS
MH  - vesicles
RP  - IN FILE
NT  - "light-gradient";
SO  - FEBS Lett 1978  ;96():233-237

276
UI  - 878
AU  - Green DE
AU  - Blondin GA
TI  - Molecular mechanism of mitochondrial energy coupling
MH  - Adenosine Triphosphate
MH  - Anions
MH  - Binding Sites
MH  - biosynthesis
MH  - Cations
MH  - Electron Transport
MH  - Energy Metabolism
MH  - Ionophores
MH  - metabolism
MH  - Mitochondria
MH  - Models,Biological
MH  - Oxidative Phosphorylation Coupling Factors
MH  - Proteins
MH  - Support,U.S.Gov't,P.H.S.
MH  - Uncoupling Agents
RP  - NOT IN FILE
SO  - Bioscience 1978 Jan ;28(1):18-24

277
UI  - 19828
AU  - Hackney DD
AU  - Boyer PD
TI  - Subunit interaction during catalysis. Implications of concentration dependency of oxygen exchanges accompanying oxidative phosphorylation for alternating site cooperativity
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - Catalysis
MH  - Hexokinase
MH  - Oxidative Phosphorylation
MH  - Oxygen
MH  - Oxygen Isotopes
MH  - Phosphates
MH  - Phosphorus
MH  - Phosphorus Radioisotopes
MH  - Phosphorylation
MH  - SUBUNIT
RP  - NOT IN FILE
NT  - UI - 78150941LA - engRN - 0 (Oxygen Isotopes)RN - 0 (Phosphates)RN - 0 (Phosphorus Radioisotopes)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 2.7.1.1 (Hexokinase)PT - Journal ArticleDA - 19780628IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:641061
SO  - J Biol Chem 1978 May 10 ;253(9):3164-3170

278
UI  - 21227
AU  - Hellingwerf KJ
AU  - Schuurmans JJ
AU  - Westerhoff HV
TI  - Demonstration of coupling between the protonmotive force across bacteriorhodopsin and the flow through its photochemical cycle
MH  - Bacteriorhodopsin
MH  - carotenoid
MH  - Carotenoids
MH  - coupling
RP  - NOT IN FILE
NT  - UI - 79024948LA - engRN - 0 (Cardiolipins)RN - 36-88-4 (Carotenoids)RN - 53026-44-1 (Bacteriorhodopsins)PT - Journal ArticleDA - 19781220IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:29778
SO  - FEBS Lett 1978 Aug 15 ;92(2):181-186

279
UI  - 871
AU  - Hinkle PC
AU  - McCarty RE
TI  - How cells make ATP
MH  - Adenosine Triphosphate
MH  - Animal
MH  - Bacteria
MH  - biosynthesis
MH  - Cell Physiology
MH  - Cells
MH  - Chloroplasts
MH  - Energy Metabolism
MH  - Membrane Potentials
MH  - metabolism
MH  - Mitochondria
MH  - Oxidative Phosphorylation
MH  - Photosynthesis
MH  - physiology
MH  - Plants
MH  - Protons
MH  - Thermodynamics
RP  - NOT IN FILE
SO  - Sci Am 1978 Mar ;238(3):104-3

280
UI  - 8288
AU  - Junge W
AU  - McGeer A
AU  - Auslnder W
TI  - Calibration of flash induced pH changes inside thylakoids and kinetic resolution of proton ejection and consumption
MH  - Kinetics
MH  - NEUTRALRED
MH  - pH
MH  - proton
MH  - resolution
MH  - thylakoid
T2  - Front. Biol. Energ., [Pap. Int. Symp.], Volume 1, 275-83. Edited by: Dutton, P. Leslie; Leigh, Jack S.; Scarpa, Antonio. Academic: New York, N. Y
Y2  - -32676  
RP  - NOT IN FILE
SO  -  1978  ;():

281
UI  - 212
AU  - Kanazawa H
AU  - Saito S
AU  - Futai M
TI  - Coupling factor ATPase from Escherichia coli. An uncA mutant (uncA401) with defective alpha subunit
AB  - Inactive coupling factor ATPase (F1) was prepared from an uncoupled mutant (uncA401) of Escherichia coli. Reconstitution of ATPase activity was observed when alpha subunit from wild-type F1 was added to the dissociated inactive F1 and the mixture was dialyzed against buffer containing ATP and Mg2+. ATPase was also reconstituted when the mixture of alpha subunit (wild type) and crude extract from the mutant was dialyzed against the same buffer. These results indicate that the mutant is defective in alpha subunit, suggesting that the uncA401 locus carries the structural gene for alpha subunit, and that this polypeptide plays an essential role in ATPase activity in F1 molecule
MH  - Adenosinetriphosphatase
MH  - ALPHA-SUBUNIT
MH  - atp
MH  - ATPase
MH  - buffer
MH  - coupling
MH  - COUPLING FACTOR
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1
MH  - Macromolecular Systems
MH  - mutant
MH  - reconstitution
MH  - SUBUNIT
MH  - SYSTEM
MH  - SYSTEMS
RP  - NOT IN FILE
NT  - UI - 79109487LA - engRN - 0 (Macromolecular Systems)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19790428IS - 0021-924XSB - IMCY - JAPAN
UR  - PM:153904
SO  - J Biochem (Tokyo ) 1978 Dec ;84(6):1513-1517

282
UI  - 664
AU  - Kasahara M
AU  - Penefsky HS
TI  - High affinity binding of monovalent Pi by beef heart mitochondrial adenosine triphosphatase
RP  - NOT IN FILE
NT  - UI - 78194124LA - engRN - 0 (Phosphates)RN - 7439-95-4 (Magnesium)RN - 7439-96-5 (Manganese)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19780828IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:149125
SO  - J Biol Chem 1978 Jun 25 ;253(12):4180-4187

283
UI  - 21124
AU  - Kell DB
AU  - John P
AU  - Ferguson SJ
TI  - On the current-voltage relationships of energy-transducing membranes: phosphorylating membrane vesicles from Paracoccus denitrificans [proceedings]
MH  - England
MH  - membrane
MH  - membrane vesicles
MH  - Membranes
MH  - Nad
MH  - Rotenone
MH  - vesicles
RP  - NOT IN FILE
NT  - UI - 79128482LA - engRN - 53-84-9 (NAD)RN - 83-79-4 (Rotenone)PT - Journal ArticleDA - 19790516IS - 0300-5127SB - IMCY - ENGLAND
UR  - PM:217778
SO  - Biochem Soc Trans 1978  ;6(6):1292-1295

284
UI  - 21121
AU  - Kell DB
AU  - Ferguson SJ
AU  - John P
TI  - Measurement by a flow dialysis technique of the steady-state proton- motive force in chromatophores from Rhodospirillum rubrum. Comparison with phosphorylation potential
AB  - 1. In the light a transmembrane electrical potential of 100 mV has been estimated to occur in chromatophores from Rhodospirillum rubrum. The potential was determined by measuring the steady-state distribution of the permeant SCN- across the chromatophore membrane using a flow dialysis technique. The potential was not observed in the dark, nor in the presence of antimycin. It was dissipated on the addition of carbonyl cyanide p-trifluoromethoxyphenylhydrazone. The potential was reduced by between 15 and 20 mV when ADP and Pi were added. Hydrolysis of ATP by the chromatophores generated a membrane potential of about 80 mV. 2. Using a flow dialysis technique light-dependent uptake of methylamine was observed only in the presence of concentrations of SCN- that were 500-fold higher than were used to measure the membrane potential. It is concluded that the pH gradient across the illuminated chromatophore membrane is insignificant except in the presence of relatively high concentrations of a permeant anion like thiocyanate. Further evidence that a negligible pH gradient was generated by the chromatophores is that addition of K+ and nigericin to illuminated chromatophores did not stimulate uptake of SCN-. 3. In the light of chromatophores established and maintained a phosphorylation potential of up to 14 kcal/mol. If a phosphorylation potential of this magnitude is to be poised against a proton-motive force that comprises solely a membrane potential of approx. 100 mV, then at least five protons must be translocated for each ATP synthesised via a chemiosmotic mechanism
MH  - A
MH  - Adenine Nucleotides
MH  - ADENINE-NUCLEOTIDES
MH  - ADP
MH  - atp
MH  - Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone
MH  - chromatophore
MH  - chromatophores
MH  - Dialysis
MH  - Hydrolysis
MH  - Light
MH  - mechanism
MH  - membrane
MH  - Membrane Potential
MH  - Nucleotides
MH  - pH
MH  - Phosphates
MH  - Phosphorylation
MH  - proton
MH  - Proton-Motive Force
MH  - Protons
MH  - Rhodospirillum
MH  - Rhodospirillum rubrum
RP  - NOT IN FILE
NT  - UI - 78144772LA - engRN - 0 (Adenine Nucleotides)RN - 0 (Bacteriochlorophylls)RN - 0 (Phosphates)RN - 0 (Protons)PT - Journal ArticleDA - 19780628IS - 0006-3002SB - IMCY - NETHERLANDS
UR  - PM:416847
SO  - Biochim Biophys Acta 1978 Apr 11 ;502(1):111-126

285
UI  - 21122
AU  - Kell DB
AU  - John P
AU  - Sorgato MC
AU  - Ferguson SJ
TI  - Continuous monitoring of the electrical potential across energy- transducing membranes using ion-selective electrodes. Application to submitochondrial particles and chromatophores
MH  - chromatophore
MH  - chromatophores
MH  - Electrodes
MH  - membrane
MH  - Membranes
MH  - Submitochondrial Particles
MH  - Thiocyanates
RP  - NOT IN FILE
NT  - UI - 78107145LA - engRN - 0 (Nitrates)RN - 0 (Thiocyanates)PT - Journal ArticleDA - 19780417IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:414935
SO  - FEBS Lett 1978 Feb 15 ;86(2):294-298

286
UI  - 21118
AU  - Kell DB
AU  - John P
AU  - Ferguson SJ
TI  - The protonmotive force in phosphorylating membrane vesicles from Paracoccus denitrificans. Magnitude, sites of generation and comparison with the phosphorylation potential
MH  - ACID
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - England
MH  - membrane
MH  - membrane vesicles
MH  - Nad
MH  - Phosphorylation
MH  - proton
MH  - Protons
MH  - Site
MH  - succinate
MH  - Succinates
MH  - Thiocyanates
MH  - vesicles
RP  - NOT IN FILE
NT  - UI - 79020814LA - engRN - 0 (Protons)RN - 0 (Succinates)RN - 0 (Thiocyanates)RN - 50-81-7 (Ascorbic Acid)RN - 53-84-9 (NAD)RN - 56-65-5 (Adenosine Triphosphate)PT - Journal ArticleDA - 19781202IS - 0264-6021SB - IMCY - ENGLAND
UR  - PM:212022
SO  - Biochem J 1978 Jul 15 ;174(1):257-266

287
UI  - 19881
AU  - Kohlbrenner WE
AU  - Cross RL
TI  - Efrapeptin prevents modification by phenylglyoxal of an essential arginyl residue in mitochondrial adenosine triphosphatase
AB  - Studies of phenylglyoxal incorporation by beef-heart mitochondrial ATPase reveal one fast-reacting arginyl residue/enzyme molecule. Modification of this group proceeds at a rate which parallels the loss of enzymatic activity. Efrapeptin protects the arginyl residue from reaction with phenylglyoxal. The data suggest that efrapeptin binds at the catalytic site and blocks accessibility of an essential arginine at the adenine nucleotide binding site. The detection of a single, fast- reacting, essential arginine on an enzyme with multiple copies of the catalytic subunit, provides further evidence in support of the alternating site mechanism for ATP synthesis proposed by Kayalar et al. (Kayalar, C., Rosing, J., and Boyer, P.D. (1977) J.Biol. Chem. 252, 2486--2491)
MH  - A
MH  - Adenosine
MH  - ADENOSINE-TRIPHOSPHATASE
MH  - Adenosinetriphosphatase
MH  - Antibiotics
MH  - atp
MH  - ATP synthesis
MH  - ATPase
MH  - BINDING
MH  - mechanism
MH  - nucleotide binding
MH  - SUBUNIT
MH  - synthesis
RP  - NOT IN FILE
NT  - UI - 79027244LA - engRN - 0 (Aldehydes)RN - 0 (Antibiotics)RN - 0 (Peptides)RN - 107-22-2 (Glyoxal)RN - 7004-12-8 (Arginine)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19781220IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:151685
SO  - J Biol Chem 1978 Nov 10 ;253(21):7609-7611

288
UI  - 486
AU  - Leimgruber RM
AU  - Senior AE
TI  - Tightly-bound ATP and ADP in reconstituted submitochondrial particles
RP  - NOT IN FILE
NT  - UI - 79041656LA - engRN - 0 (Oligomycins)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19781220IS - 0006-291XSB - IMCY - UNITED STATESJC - 9Y8
UR  - PM:152109
SO  - Biochem Biophys Res Commun 1978 Aug 14 ;83(3):837-842

289
UI  - 1021
AU  - Moyle J
AU  - Mitchell P
TI  - Measurements of mitochondrial comes from H+/O quotients: effects of phosphate and N-ethylmaleimide
MH  - Animal
MH  - Antimycin A
MH  - Biological Transport
MH  - Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone
MH  - Cytochrome c
MH  - drug effects
MH  - Electron Transport
MH  - Ethylmaleimide
MH  - Hydrogen-Ion Concentration
MH  - metabolism
MH  - Mitochondria,Liver
MH  - Oxygen Consumption
MH  - pharmacology
MH  - Phosphates
MH  - Potassium
MH  - Rats
MH  - Valinomycin
RP  - NOT IN FILE
SO  - FEBS Lett 1978 Jun 15 ;90(2):361-365

290
UI  - 854
AU  - Packham NK
AU  - Berriman JA
AU  - Jackson JB
TI  - The charging capacitance of the chromatophore membrane
RP  - NOT IN FILE
NT  - UI - 78191240LA - engRN - 0 (Bacteriochlorophylls)RN - 0 (Cytochromes)RN - 27061-78-5 (Alamethicin)RN - 7439-95-4 (Magnesium)PT - Journal ArticleDA - 19780814IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:658407
SO  - FEBS Lett 1978 May 15 ;89(2):205-210

291
UI  - 553
AU  - Pedersen PL
AU  - Hullihen J
TI  - Adenosine triphosphatase of rat liver mitochondria. Capacity of the homogeneous F1 component of the enzyme to restore ATP synthesis in urea- treated membranes
RP  - NOT IN FILE
NT  - UI - 78130084LA - engRN - 0 (Macromolecular Systems)RN - 0 (Oligomycins)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - 57-13-6 (Urea)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19780524IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:147273
SO  - J Biol Chem 1978 Apr 10 ;253(7):2176-2183

292
UI  - 8688
AU  - Saphon S
AU  - Graber P
TI  - External proton uptake, internal proton release and internal pH changes in chromatophores from Rps. sphaeroides following single turnover flashes
MH  - atp
MH  - chromatophore
MH  - chromatophores
MH  - pH
MH  - proton
MH  - proton release
MH  - sphaeroides
RP  - ON REQUEST (05/29/92)
SO  - Z Naturforsch 1978  ;33C():715-722

293
UI  - 789
AU  - Sone N
AU  - Yoshida M
AU  - Hirata H
AU  - Kagawa Y
TI  - Resolution of the membrane moiety of the H+-ATPase complex into two kinds of subunits
AB  - The H+-translocating ATPase complex from the thermophilic bacterium PS3 (TF0-F1) is composed of a water-soluble part with ATP-hydrolyzing activity (TF1) and a membrane moiety with H+-conducting activity (TF0). TF0 was obtained by treating TF0-F1 with urea and removing contaminations on a carboxymethyl-cellulose column. This TF0 contained only two kinds of subunits, band 6 protein (13,500 daltons) and band 8 protein (5400 daltons), and it was active in H+ conduction and TF1 binding when reconstituted into proteoliposomes (TF0 vesicles). The binding of TF1 to TF0 present in vesicles restored energy-transducing activities, such as ATP-32Pi exchange, dicyclohexylcarbodiimide- sensitive ATPase, and ATP-dependent enhancement of 8-anilinonaphthalene- 1-sulfonate fluorescence. Treatments such as protease digestion and chemical modification with acetic anhydride, succinic anhydride, or diazobenzenesulfonic acid destroyed the TF1-binding activity, which was caused by band 6 protein. Band 8 protein was a proteolipid that reacted specifically with dicylcohexyl-carbodiimide and seemed to play a central role in H+ conduction through the membrane
RP  - NOT IN FILE
NT  - UI - 79033809LA - engRN - 0 (Bacterial Proteins)RN - 0 (Liposomes)RN - 0 (Macromolecular Systems)RN - 0 (Membrane Proteins)RN - 0 (Proteolipids)RN - 0 (Protons)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19781227IS - 0027-8424SB - IMCY - UNITED STATESJC - PV3
UR  - PM:151864
SO  - Proc Natl Acad Sci U S A 1978 Sep ;75(9):4219-4223

294
UI  - 21120
AU  - Sorgato MC
AU  - Ferguson SJ
TI  - Measurements of the components of the protonmotive force generated by cytochrome oxidase in submitochondrial particles
MH  - ACID
MH  - cytochrome
MH  - Cytochrome c
MH  - Cytochrome-c Oxidase
MH  - Ethylmaleimide
MH  - proton
MH  - Protons
MH  - Submitochondrial Particles
MH  - Valinomycin
RP  - NOT IN FILE
NT  - UI - 78191289LA - engRN - 0 (Protons)RN - 128-53-0 (Ethylmaleimide)RN - 2001-95-8 (Valinomycin)RN - 27215-51-6 (Tetramethylphenylenediamine)RN - 50-81-7 (Ascorbic Acid)RN - EC 1.9.3.1 (Cytochrome-c Oxidase)PT - Journal ArticleDA - 19780828IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:207580
SO  - FEBS Lett 1978 Jun 1 ;90(1):178-182

295
UI  - 21123
AU  - Sorgato MC
AU  - Ferguson SJ
AU  - Kell DB
TI  - On the current-voltage relationships of energy-transducing membranes: submitochondrial particles [proceedings]
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - England
MH  - membrane
MH  - Membranes
MH  - Nad
MH  - Rotenone
MH  - Submitochondrial Particles
MH  - succinate
MH  - Succinates
RP  - NOT IN FILE
NT  - UI - 79128485LA - engRN - 0 (Succinates)RN - 53-84-9 (NAD)RN - 56-65-5 (Adenosine Triphosphate)RN - 83-79-4 (Rotenone)PT - Journal ArticleDA - 19790516IS - 0300-5127SB - IMCY - ENGLAND
UR  - PM:217779
SO  - Biochem Soc Trans 1978  ;6(6):1301-1302

296
UI  - 21119
AU  - Sorgato MC
AU  - Ferguson SJ
AU  - Kell DB
AU  - John P
TI  - The protonmotive force in bovine heart submitochondrial particles. Magnitude, sites of generation and comparison with the phosphorylation potential
AB  - 1. The magnitude of the protonmotive force in respiring bovine heart submitochondrial particles was estimated. The membrane-potential component was determined from the uptake of S14CN-ions, and the pH- gradient component from the uptake of [14C]methylamine. In each case a flow-dialysis technique was used to monitor uptake. 2. With NADH as substrate the membrane potential was approx. 145mV and the pH gradient was between 0 and 0.5 unit when the particles were suspended in a Pi/Tris reaction medium. The addition of the permeant NO3-ion decreased the membrane potential with a corresponding increase in the pH gradient. In a medium containing 200mM-sucrose, 50mM-KCl and Hepes as buffer, the total protonmotive force was 185mV, comprising a membrane potential of 90mV and a pH gradient of 1.6 units. Thus the protonmotive force was slightly larger in the high-osmolarity medium. 3. The phosphorylation potential (= deltaG0' + RT ln[ATP]/[ADP][Pi]) was approx. 43.1 kJ/mol (10.3kcal/mol) in all the reaction media tested. Comparison of this value with the protonmotive force indicates that more than 2 and up to 3 protons must be moved across the membrane for each molecule of ATP synthesized by a chemiosmotic mechanism. 4. Succinate generated both a protonmotive force and a phosphorylation potential that were of similar magnitude to those observed with NADH as substrate. 5. Although oxidation of NADH supports a rate of ATP synthesis that is approximately twice that observed with succinate, respiration with either of these substrates generated a very similar protonmotive force. Thus there seemed to be no strict relation between the size of the protonmotive force and the phosphorylation rate. 6. In the presence of antimycin and/or 2-n-heptyl-4-hydroxyquinoline N-oxide, ascorbate oxidation with either NNN'N'-tetramethyl-p-phenylenediamine or 2,3,5,6-tetramethyl-p-phenylenediamine as electron mediator generated a membrane potential of approx. 90mV, but no pH gradient was detected, even in the presence of NO3-. These data are discussed with reference to the proposal that cytochrome oxidase contains a proton pump
MH  - A
MH  - ACID
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthesis
MH  - buffer
MH  - cytochrome
MH  - electron
MH  - England
MH  - mechanism
MH  - membrane
MH  - Membrane Potential
MH  - Nad
MH  - pH
MH  - Phosphorylation
MH  - proton
MH  - Proton Pump
MH  - Protons
MH  - Respiration
MH  - Site
MH  - Submitochondrial Particles
MH  - succinate
MH  - Succinates
MH  - synthesis
RP  - NOT IN FILE
NT  - UI - 79020813LA - engRN - 0 (Protons)RN - 0 (Succinates)RN - 50-81-7 (Ascorbic Acid)RN - 53-84-9 (NAD)RN - 56-65-5 (Adenosine Triphosphate)PT - Journal ArticleDA - 19781202IS - 0264-6021SB - IMCY - ENGLAND
UR  - PM:212021
SO  - Biochem J 1978 Jul 15 ;174(1):237-256

297
UI  - 596
AU  - Stiggall DL
AU  - Galante YM
AU  - Hatefi Y
TI  - Preparation and properties of an ATP-Pi exchange complex (complex V) from bovine heart mitochondria
RP  - NOT IN FILE
NT  - UI - 78087561LA - engRN - 0 (Oligomycins)RN - 0 (Phosphates)RN - 0 (Uncoupling Agents)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19780321IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:146039
SO  - J Biol Chem 1978 Feb 10 ;253(3):956-964

298
UI  - 9921
AU  - van dD
AU  - Janssen DB
AU  - van WP
TI  - Hydrolysis and synthesis of ATP by membrane-bound ATPase from a motile Streptococcus.
AB  - ATPase was detected in the membranes of a motile Streptococcus. Maximal enzymic activity was observed at pH 8 and ATP/Mg2+ ratio of 2. Mn2+ and Ca2+ could replace Mg2+ to some extent. Besides ATP, GTP and ITP were substrates. The enzyme was inhibited by N,N'-dicyclohexylcarbodiimide but not by sodium azide, uncouplers or bathophenanthroline. An electrochemical gradient of protons, which was artificially imposed across the membranes of Streptococcus cells by manipulation of either the K+ diffusion potential or the transmembrane pH gradient, led to ATP synthesis. ATP synthesis was abolished by proton conductors, an inhibitor of the ATPase or an increase in the extracellular K+ concentration. A comparison between the phosphate potential and the electrochemical proton gradient showed that the data found are in agreement with a stoichiometry of 2 protons translocated per molecule ATP synthesized.
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - atp
MH  - ATP synthesis
MH  - ATPase
MH  - biosynthesis
MH  - Cell Membrane
MH  - Cells
MH  - Diffusion
MH  - diffusion potential
MH  - Hydrogen-Ion Concentration
MH  - Hydrolysis
MH  - Membrane Potentials
MH  - Membranes
MH  - metabolism
MH  - Movement
MH  - pharmacology
MH  - physiology
MH  - proton
MH  - Protons
MH  - Sodium
MH  - Streptococcus
MH  - Uncoupling Agents
MH  - Valinomycin
RP  - NOT IN FILE
SO  - Arch Microbiol 1978 Oct 4 ;119(1):31-36

299
UI  - 19829
AU  - Vinkler C
AU  - Rosen G
AU  - Boyer PD
TI  - Light-driven ATP formation from 32Pi by chloroplast thylakoids without detectable labeling of ADP, as measured by rapid mixing and acid quench techniques
AB  - The labeling of ATP and ADP in the first few milliseconds of exposure of chloroplast thylakoids to light, 32Pi, and ADP has been measured. At least 4 mol of ATP/mol of coupling factor 1 (CF1)-ATPase can be formed without detectable labeling of membrane-bound or free ADP. Such results are consistent with ADP and not AMP as the primary acceptor of Pi in photophosphorylation. Evidence is presented demonstrating that quenching with perchloric acid, as used in these and earlier experiments, is satisfactory for the measurement of the amount and nature of membrane-bound nucleotides
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - ADP
MH  - atp
MH  - ATP FORMATION
MH  - chloroplast
MH  - COUPLING FACTOR
MH  - Light
MH  - Nucleotides
MH  - Phosphates
MH  - Photophosphorylation
MH  - thylakoid
RP  - NOT IN FILE
NT  - UI - 78130129LA - engRN - 0 (Phosphates)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)PT - Journal ArticleDA - 19780524IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:632282
SO  - J Biol Chem 1978 Apr 25 ;253(8):2507-2510

300
UI  - 853
AU  - Webster GD
AU  - Jackson JB
TI  - Affinity chromatography of H+-translocating adenosine triphosphatase isolated by chloroform extraction of Rhodospirillum rubrum chromatophores. Modification of binding affinity by divalent cations and activating anions
AB  - 1. ATPase isolated from Rhodospirillum rubrum by chloroform extraction and purified by gel filtration or affinity chromatography shows three bands (alpha, beta and gamma) upon electrophoresis in sodium dodecyl sulphate. 2. Ca2+-ATPase activity of the preparation is inhibited by aurovertin and efrapeptin but not by oligomycin. Activity may be inhibited by treatment with 4-chloro-7-nitrobenzofurazan and subsequently restored by dithiothreitol. 3. The enzyme fails to reconstitute photophosphorylation in chromatophores depleted of ATPase by sonic irradiation. 4. Most of the active protein from the crude chloroform extract binds to an affinity chromatography column bearing an immobilised ADP analogue but not to a column bearing immobilised pyrophosphate. 5. In the absence of divalent cations, a component with a very high specific activity for Ca2+-ATPase is eluted from the column by 1.6 mM ATP. This protein migrates asa single band on 5% polyacrylamide gel electrophoresis and only possesses three subunits. At 12 mM ATP an inactive protein is eluted which does not run on acid or alkali polyacrylamide gels and shows a complex subunit structure. 6. ATPase preparations prepared by acetone extraction or by sonic irradiation of chromatophores may also be purified 10-fold by affinity chromatography. 7. The inclusion of 5 mM MgCl2 or CaCl2 during affinity chromatography of chloroform ATPase increases the capacity of the column for the enzyme and demands a higher eluting concentration of ATP. 8. When the enzyme is more than 90% inhibited by efrapeptin or 4- chloro-7-nitrobenzofurazan, the binding characteristics of the enzyme are not affected. 9. 10 mM Na2SO3, which greatly stimulates the Ca2+- and Mg2+-dependent ATPase activity of the enzyme and increases Ki (ADP) for Ca2+-ATPase from 50 to 850 micron, prevents binding to the affinity column. Binding may be restored by the addition of divalent cations. 10. Na2SO3 increases the rate of ATP hydrolysis, ATP-driven H+ translocation and ATP-driven transhydrogenase in chromatophores. 11. It is proposed that anions such as sulphite convert the chromatophore ATPase into a form which is a more efficient energy transducer
RP  - NOT IN FILE
NT  - UI - 78211069LA - engRN - 0 (Chlorides)RN - 0 (Macromolecular Systems)RN - 7439-95-4 (Magnesium)RN - 7440-70-2 (Calcium)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19780930IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:27212
SO  - Biochim Biophys Acta 1978 Jul 6 ;503(1):135-154

301
UI  - 8164
AU  - Williams RJP
TI  - The Multifarious Couplings of Energy Transduction
MH  - bioenergetics
MH  - coupling
MH  - review
RP  - IN FILE
NT  - Ju Rev
SO  - Biochim Biophys Acta 1978  ;505():1-44

302
UI  - 888
AU  - Williams RJP
TI  - The history and the hypotheses concerning ATP-formation by energised protons
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - Biological Transport
MH  - chemiosmotic theory
MH  - Diffusion
MH  - Electron Transport
MH  - Energy Metabolism
MH  - Hydrogen-Ion Concentration
MH  - local coupling
MH  - Membranes
MH  - metabolism
MH  - Models,Biological
MH  - Nad
MH  - Oxidation-Reduction
MH  - Oxygen Consumption
MH  - Protons
RP  - NOT IN FILE
SO  - FEBS Lett 1978 Jan 1 ;85(1):9-19

303
UI  - 4113
AU  - Wraight CA
AU  - Cogdell RJ
AU  - Chance B
TI  - Ion transport and electrochemical gradients in photosynthetic bacteria
MH  - Bacteria
MH  - cyt b6f
MH  - ion
MH  - Ion Transport
MH  - pmf
MH  - transport
T2  - The Photosynthetic Bacteria
A2  - Clayton RK
A2  - Sistrom WR
Y2  - -32676  
PB  - New York: Academic Press
RP  - IN FILE
SO  -  1978  ;():471-511

304
UI  - 788
AU  - Yoshida M
AU  - Sone N
AU  - Hirata H
AU  - Kagawa Y
TI  - Evidence for three alpha subunits in one molecule of F1-ATPase from thermophilic bacterium PS3
RP  - NOT IN FILE
NT  - UI - 79082236LA - engRN - 0 (Macromolecular Systems)RN - 69-78-3 (Dithionitrobenzoic Acid)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19790226IS - 0006-291XSB - IMCY - UNITED STATESJC - 9Y8
UR  - PM:153137
SO  - Biochem Biophys Res Commun 1978 Sep 14 ;84(1):117-122

305
UI  - 18723
AU  - Baltscheffsky M
AU  - Lundin A
TI  - Flash-induced increase of ATPase activity in Rhodospirillum rubrum chromatophores.
MH  - ACTIVATION
MH  - ATP FORMATION
MH  - ATPase
MH  - chromatophore
MH  - chromatophores
MH  - flash
MH  - luciferase
MH  - Rhodospirillum
MH  - Rhodospirillum rubrum
A2  - Mukohata Y
A2  - Packer L
Y2  - -32676  
T3  - Cation Flux across Biomembranes
PB  - Academic Press
RP  - IN FILE
SO  -  1979  ;():209-218

306
UI  - 123
AU  - Beechey RB
AU  - Linnett PE
AU  - Fillingame RH
TI  - Isolation of carbodiimide-binding proteins from mitochondria and Escherichia coli
RP  - NOT IN FILE
NT  - UI - 79220700LA - engRN - 0 (Carbodiimides)RN - 0 (Carrier Proteins)RN - 0 (Proteolipids)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19790917IS - 0076-6879SB - IMCY - UNITED STATESJC - MVA
UR  - PM:156852
SO  - Methods Enzymol 1979  ;55():426-434

307
UI  - 21214
AU  - Bol'shakov VI
AU  - Drachev AL
AU  - Drachev LA
AU  - Kalamkarov GR
AU  - Kaulen AD
TI  - [Community of properties of bacterial and visual rhodopsins: light energy conversion to electric potential difference]
MH  - Bacteriorhodopsin
MH  - carotenoid
MH  - Carotenoids
MH  - Light
MH  - pigments
MH  - retinal
RP  - NOT IN FILE
NT  - UI - 80112373LA - rusRN - 0 (Retinal Pigments)RN - 36-88-4 (Carotenoids)RN - 53026-44-1 (Bacteriorhodopsins)RN - 9009-81-8 (Rhodopsin)PT - Journal ArticleDA - 19800417IS - 0002-3264SB - IMCY - USSR
UR  - PM:527472
SO  - Dokl Akad Nauk SSSR 1979  ;249(6):1462-1466

308
UI  - 19827
AU  - Boyer PD
AU  - Stempel K
TI  - Rapid nucleotide labeling and 18O exchange probes of intermediate states in electron-transport-coupled phosphorylation
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - Oxygen
MH  - Oxygen Isotopes
MH  - Phosphates
MH  - Phosphorylation
RP  - NOT IN FILE
NT  - UI - 79220668LA - engRN - 0 (Oxygen Isotopes)RN - 0 (Phosphates)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)PT - Journal ArticleDA - 19790917IS - 0076-6879SB - IMCY - UNITED STATESJC - MVA
UR  - PM:459844
SO  - Methods Enzymol 1979  ;55():245-261

309
UI  - 624
AU  - Casadio R
AU  - Melandri BA
AU  - Piretti MV
AU  - Serrazanetti GP
TI  - Phospholipid composition of photosynthetic membranes of Rhodopseudomonas capsulata
AB  - The phospholipids and the fatty acids present in membranes of cells of Rhodopseudomonas capsulata, grown photosynthetically in anaerobiosis, were analyzed by thin layer chromatography and gas chromatography-mass spectrometry. The three phospholipids detected, phosphatidylcholine, phosphatidylethanolamine and phosphatidylglycerol, contained about 80% of a single monounsaturated C18 fatty acid, cis-vaccenic acid. These membranes offer therefore a naturally occurring model system endowed with an extremely simplified phospholipid complement. The data indicate moreover that the biosynthetic pathway of unsaturated fatty acids present in these facultative aerobic bacteria proceeds only via the 3- hydroxydecanoyl acyl carrier protein dehydratase (E.C. 4.2.1.60)
RP  - NOT IN FILE
NT  - UI - 81093297LA - engRN - 0 (Fatty Acids)RN - 0 (Membrane Lipids)RN - 0 (Phospholipids)PT - Journal ArticleDA - 19810324IS - 0021-2938SB - IMCY - ITALYJC - GYW
UR  - PM:553902
SO  - Ital J Biochem 1979 May ;28(3):183-193

310
UI  - 19826
AU  - Choate GL
AU  - Hutton RL
AU  - Boyer PD
TI  - Occurrence and significance of oxygen exchange reactions catalyzed by mitochondrial adenosine triphosphatase preparations
AB  - The capacity of various ATPase preparations from beef heart mitochondria to catalyze exchange of phosphate oxygens with water has been evaluated. Oligomycin-sensitive ATPase preparations retain a capacity for considerable intermediate Pi equilibrium HOH exchange per Pi formed during ATP hydrolysis at relatively high ATP concentration (5 mM). Submitochondrial particles prepared by an ammonia-Sephadex procedure with 5 mM ATP showed more rapid ATPase, less oligomycin sensitivity, and less capacity for intermediate exchange. With these particles, intermediate Pi equilibrium HOH exchange per Pi formed was increased as ATP concentration was decreased. The purified, soluble ATPase from mitochondria catalyzed little or no intermediate Pi equilibrium HOH exchange at 5 mM ATP but showed pronounced increase in capacity for such exchange as ATP concentration was lowered. The ATPase also showed a weak catalysis of an ADP-stimulated medium Pi equilibrium HOH exchange. The results support the alternating catalytic site model for ATP synthesis or cleavage. They also demonstrate that a transmembrane protonmotive force is not necessary for oxygen exchange reactions. At lower ATP concentrations, ADP and Pi formed at a catalytic site appear to remain bound and continue to allow exchange of Pi oxygens until ATP binds at another site on the enzyme
MH  - A
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - ADENOSINE-TRIPHOSPHATASE
MH  - Adenosinetriphosphatase
MH  - ADP
MH  - atp
MH  - ATP synthesis
MH  - ATPase
MH  - Catalysis
MH  - Hydrolysis
MH  - Mitochondria
MH  - model
MH  - Oligomycins
MH  - Oxygen
MH  - Oxygen Isotopes
MH  - Submitochondrial Particles
MH  - synthesis
MH  - Water
RP  - NOT IN FILE
NT  - UI - 79109565LA - engRN - 0 (Oligomycins)RN - 0 (Oxygen Isotopes)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19790425IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:153910
SO  - J Biol Chem 1979 Jan 25 ;254(2):286-290

311
UI  - 551
AU  - Cintron NM
AU  - Pedersen PL
TI  - A protein inhibitor of the mitochondrial adenosine triphosphatase complex of rat liver. Purification and characterization
AB  - A heat-stable protein has been purified from rat liver mitochondria which inhibits the ATP hydrolytic activity of both the soluble and membrane-bound mitochondrial F1-ATPase. The overall purification is about 2400-fold with the major purification step consisting of Sephadex "affinity" chromatography. The purified rat liver inhibitor is homogeneous as assessed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis with an apparent molecular weight of 12,300. Amino acid analysis reveals a high content of glutamic acid, lysine, and arginine and the absence of cysteine, proline and methionine. Whether tested with the rat liver or bovine heart ATPase, the liver inhibitor is equally as potent and specific as the heart inhibitor preparation of Pullman and Monroy (Pullman, M.E., and Monroy, G.C. (1963) J. Biol. Chem. 238, 3762-3769). Although the results presented show that the rat liver ATPase inhibitor resembles closely the ATPase inhibitors from other tissues with respect to specific activity and reaction specificity, it is important to note that the rat liver inhibitor is almost 2000 daltons larger than the bovine heart inhibitor, about 5000 daltons larger than ATPase inhibitors of yeast, and contains significantly more lysine residues than both the bovine heart and yeast inhibitors
RP  - NOT IN FILE
NT  - UI - 79151103LA - engRN - 0 (Amino Acids)RN - 0 (Proteins)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19790629IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:155068
SO  - J Biol Chem 1979 May 10 ;254(9):3439-3443

312
UI  - 495
AU  - Downie JA
AU  - Senior AE
AU  - Cox GB
AU  - Gibson F
TI  - Solubilization of adenosine triphosphatase from membranes of Escherichia coli: effect of p-aminobenzamidine
AB  - The five subunits of the membrane-bound adenosine triphosphatase (F1) from Escherichia coli were identified on electrophoretograms of membranes which had been washed with a low-ionic-strength buffer containing the protease inhibitor p-aminobenzamidine. All of the subunits of the membrane-bound F1 appeared to have the same molecular weights and isoelectric points as those of the soluble F1, as judged by two-dimensional electrophoresis. p-Aminobenzamidine inhibited the solubilization of F1 rebound to F1-depleted membranes, and was found to inhibit the membrane-bound adenosine triphosphatase activity to a much greater extent than the solubilized activity. It is therefore unlikely that p-aminobenzamidine inhibits the solubilization of F1 by inhibiting a protease, as suggested previously by Cox et al. (G.B. Cox, J.A. Downie, D.R.H. Fayle, F. Gibson, and J. Radik, J. Bacteriol. 133:287-- 292, 1978)
RP  - NOT IN FILE
NT  - UI - 79172996LA - engRN - 0 (Amidines)RN - 0 (Benzamidines)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19790716IS - 0021-9193SB - IMCY - UNITED STATESJC - HH3
UR  - PM:155691
SO  - J Bacteriol 1979 Apr ;138(1):87-91

313
UI  - 21213
AU  - Drachev LA
AU  - Kaulen AD
AU  - Semenov AY
AU  - Severina II
AU  - Skulachev VP
TI  - Lipid-impregnated filters as a tool for studying the electric current- generating proteins
MH  - A
MH  - Adenosinetriphosphatase
MH  - Bacteriorhodopsin
MH  - membrane
MH  - Membrane Proteins
MH  - Phospholipids
MH  - protein
MH  - Proteins
MH  - Pyrophosphatases
RP  - NOT IN FILE
NT  - UI - 80040342LA - engRN - 0 (Membrane Proteins)RN - 0 (Phospholipids)RN - 0 (Proteins)RN - 53026-44-1 (Bacteriorhodopsins)RN - EC 3.6.1.- (Pyrophosphatases)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19791218IS - 0003-2697SB - IMCY - UNITED STATES
UR  - PM:158996
SO  - Anal Biochem 1979 Jul 1 ;96(1):250-262

314
UI  - 21212
AU  - Drachev LA
AU  - Kaulen AD
AU  - Samuilov VD
AU  - Severina II
AU  - Semenov AI
TI  - [Incorporation of proteoliposomes and chromatophores into membranes based on filters]
MH  - acetate
MH  - Acetates
MH  - Bacteriorhodopsin
MH  - chromatophore
MH  - chromatophores
MH  - liposome
MH  - Liposomes
MH  - membrane
MH  - Membranes
MH  - Phospholipids
MH  - Proteolipids
MH  - proteoliposome
RP  - NOT IN FILE
NT  - UI - 80065799LA - rusRN - 0 (Acetates)RN - 0 (Liposomes)RN - 0 (Nitrates)RN - 0 (Phospholipids)RN - 0 (Proteolipids)RN - 53026-44-1 (Bacteriorhodopsins)RN - 9002-84-0 (Polytetrafluoroethylene)RN - 9004-34-6 (Cellulose)PT - Journal ArticleDA - 19800228IS - 0006-3029SB - IMCY - USSR
UR  - PM:508817
SO  - Biofizika 1979 Nov ;24(6):1035-1042

315
UI  - 21211
AU  - Drachev LA
AU  - Kaulen AD
AU  - Samuilov VD
AU  - Severina II
AU  - Semenov AI
TI  - [Filters as selective electrodes for synthetic penetrating ions]
MH  - Bacteriorhodopsin
MH  - Electrodes
MH  - liposome
MH  - Liposomes
MH  - Phospholipids
RP  - NOT IN FILE
NT  - UI - 80065800LA - rusRN - 0 (Boron Compounds)RN - 0 (Liposomes)RN - 0 (Phospholipids)RN - 4358-26-3 (Tetraphenylborate)RN - 53026-44-1 (Bacteriorhodopsins)PT - Journal ArticleDA - 19800228IS - 0006-3029SB - IMCY - USSR
UR  - PM:508818
SO  - Biofizika 1979 Nov ;24(6):1043-1047

316
UI  - 21117
AU  - Ferguson SJ
AU  - Sorgato MC
AU  - Kell DB
AU  - John P
TI  - Comparative aspects of the energetics of oxidative phosphorylation in bacteria and mitochondria
MH  - Bacteria
MH  - England
MH  - Mitochondria
MH  - Oxidative Phosphorylation
MH  - Phosphorylation
RP  - NOT IN FILE
NT  - UI - 80069850LA - engPT - Journal ArticleDA - 19800215IS - 0300-5127SB - IMCY - ENGLAND
UR  - PM:41780
SO  - Biochem Soc Trans 1979 Oct ;7(5):870-874

317
UI  - 21138
AU  - Ferguson SJ
AU  - Jones OT
AU  - Kell DB
AU  - Sorgato MC
TI  - Comparison of permeant ion uptake and carotenoid band shift as methods for determining the membrane potential in chromatophores from Rhodopseudomonas sphaeroides Ga
AB  - 1. A comparison was made of two methods for estimating the membrane potential in chromatophores from Rhodopseudomonas sphaeroides Ga. Illuminated chromatophores generated a potential that is apparently much larger when estimated on the basis of the red-band shift of carotenoids rather than from the extent of uptake of the permeant SCN- ion. 2. In contrast, when the chromatophores were oxidizing NADH or succinate the uptake of SCN- indicated a larger membrane potential than was estimated from the carotenoid band shift. 3. The extent of SCN- uptake and the carotenoid-band shift respond differently to changes in the ionic composition of the reaction medium. 4. The effects of antimycin on the carotenoid band shift and SCN- uptake are reported. 5. It is concluded that the carotenoid band shift and the uptake of SCN- are responding to different aspects of the energized state
MH  - A
MH  - Antimycin A
MH  - carotenoid
MH  - Carotenoids
MH  - chromatophore
MH  - chromatophores
MH  - England
MH  - ion
MH  - membrane
MH  - Membrane Potential
MH  - method
MH  - Methods
MH  - Nad
MH  - rhodopseudomonas
MH  - sphaeroides
MH  - succinate
MH  - Succinates
MH  - Thiocyanates
RP  - NOT IN FILE
NT  - UI - 80020175LA - engRN - 0 (Succinates)RN - 0 (Thiocyanates)RN - 36-88-4 (Carotenoids)RN - 53-84-9 (NAD)RN - 642-15-9 (Antimycin A)PT - Journal ArticleDA - 19791121IS - 0264-6021SB - IMCY - ENGLAND
UR  - PM:226068
SO  - Biochem J 1979 Apr 15 ;180(1):75-85

318
UI  - 62
AU  - Fillingame RH
TI  - Isolation of carbodiimide-resistant ATPase mutants from Escherichia coli
RP  - NOT IN FILE
NT  - UI - 79220745LA - engRN - 0 (Carbodiimides)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19790917IS - 0076-6879SB - IMCY - UNITED STATESJC - MVA
UR  - PM:156863
SO  - Methods Enzymol 1979  ;56():163-172

319
UI  - 122
AU  - Foster DL
AU  - Fillingame RH
TI  - Energy-transducing H+-ATPase of Escherichia coli. Purification, reconstitution, and subunit composition
RP  - NOT IN FILE
NT  - UI - 79239428LA - engRN - 0 (Macromolecular Systems)RN - 0 (Oxidative Phosphorylation Coupling Factors)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19791026IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:38249
SO  - J Biol Chem 1979 Sep 10 ;254(17):8230-8236

320
UI  - 594
AU  - Galante YM
AU  - Wong SY
AU  - Hatefi Y
TI  - Composition of complex V of the mitochondrial oxidative phosphorylation system
RP  - NOT IN FILE
NT  - UI - 80049917LA - engRN - 0 (Carrier Proteins)RN - 0 (Membrane Proteins)RN - 0 (Multienzyme Complexes)RN - 0 (Oligomycins)RN - 0 (Oxidative Phosphorylation Coupling Factors)RN - 0 (Phosphates)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19800128IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:159305
SO  - J Biol Chem 1979 Dec 25 ;254(24):12372-12378

321
UI  - 19822
AU  - Gresser M
AU  - Cardon J
AU  - Rosen G
AU  - Boyer PD
TI  - Demonstration and quantitation of catalytic and noncatalytic bound ATP in submitochondrial particles during oxidative phosphorylation
AB  - Techniques are described for studying the labeling of ADP and ATP bound to the ATP synthase complex of beef heart submitochondrial particles catalyzing oxidative phosphorylation. These suffice for measurements of bound nucleotides during the time required for a single turnover, during steady state net ATP synthesis, or under quasiequilibrium conditions of ATP formation and hydrolysis. Results show that the "tightly bound" ATP associated with isolated submitochondrial particles does not become labeled by medium [32P]Pi rapidly enough to qualify as an intermediate in ATP synthesis. In contrast to chloroplast preparations, little or no bound [32P]Pi committed to ATP formation is present on particles during steady state synthesis. Also, highly active particles synthesizing ATP from [32P]Pi and filtered after EDTA addition have no detectable bound [32P]ATP even though several ATPs have been made per synthase complex. However, under quasiequilibrium conditions membrane-bound ADP and ATP are present whose labeling characteristics qualify them as intermediates in ATP synthesis. In addition, a hexokinase-accessibility approach shows the presence of a steady level of bound ATP. Lack of detection of bound intermediates under other conditions is regarded as reflecting the ready reversibility of oxidative phosphorylation, with consequent facile cleavage of bound ATP and release of bound Pi
MH  - A
MH  - ACTIVE
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - ADP
MH  - atp
MH  - ATP FORMATION
MH  - ATP synthase
MH  - ATP synthesis
MH  - BOUND NUCLEOTIDES
MH  - chloroplast
MH  - COMPLEX
MH  - Hexokinase
MH  - Hydrolysis
MH  - Nucleotides
MH  - Oxidative Phosphorylation
MH  - Phosphorylation
MH  - Submitochondrial Particles
MH  - SYNTHASE
MH  - synthesis
MH  - Time
RP  - NOT IN FILE
NT  - UI - 80049653LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 2.7.1.1 (Hexokinase)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19800119IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:159294
SO  - J Biol Chem 1979 Nov 10 ;254(21):10649-10653

322
UI  - 19824
AU  - Hackney DD
AU  - Rosen G
AU  - Boyer PD
TI  - Subunit interaction during catalysis: alternating site cooperativity in photophosphorylation shown by substrate modulation of [18O]ATP species formation
AB  - Pronounced substrate modulation of incorporation of water oxygen into ATP formed by photophosphorylation is observed, as measured by 31P NMR analysis of products formed from ADP and highly 18O-labeled Pi. A marked increase occurs in oxygen exchange per ATP formed as ADP or Pi concentration is decreased. This is explainable by the binding-change mechanism for ATP synthesis, in which the energy-linked release of ATP from one site requires the binding of ADP and Pi at an alternate site. Analysis of the distribution of 18O-labeled species arising from the ATP formed eliminates explanations for substrate modulation based on preexisting or induced enzyme heterogeneity. Furthermore, the results, together with other related findings, make participation of control sites unlikely. The occurrence of alternating site catalysis cooperativity in ATP synthesis by chloroplasts thus appears to be reasonably well established
MH  - A
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - ADP
MH  - analysis
MH  - atp
MH  - ATP synthesis
MH  - BINDING
MH  - Catalysis
MH  - chloroplast
MH  - Chloroplasts
MH  - Macromolecular Systems
MH  - mechanism
MH  - Oxygen
MH  - Phosphates
MH  - Photophosphorylation
MH  - SUBUNIT
MH  - synthesis
MH  - Water
RP  - NOT IN FILE
NT  - UI - 80034933LA - engRN - 0 (Macromolecular Systems)RN - 0 (Phosphates)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 7782-44-7 (Oxygen)PT - Journal ArticleDA - 19791229IS - 0027-8424SB - IMCY - UNITED STATESJC - PV3
UR  - PM:291029
SO  - Proc Natl Acad Sci U S A 1979 Aug ;76(8):3646-3650

323
UI  - 9904
AU  - Hill TL
TI  - Steady-state coupling of four membrane systems in mitochondrial oxidative phosphorylation.
AB  - According to Alexandre, Reynafarje, and Lehninger, four different membrane systems are involved, with definite stoichiometry, in the mitochondrial synthesis of ATP by electron transport, via proton transport. We adopt this model and pursue some of its thermodynamic consequences. At steady state, each of the four systems must have the same flux J through the membrane and the overall thermodynamic force X for oxidative phosphorylation is the sum of the four separate forces. From these properties, using an empirical linear flux-force relation for each system, it is easy to obtain J as a function of X. In turn, X depends on the inside [NAD+]/[NADH] and the outside [ATP]/[ADP][Pi] quotients (and on the pH inside). Thus, J is related to these quotients. The relationship we derive is similar to that described by Erecinska and Wilson, as deduced from a quite different model of oxidative phosphorylation. Proton transport is involved explicitly in three of the four systems of the present model. However, because of the steady-state stoichiometric coupling of the four systems, proton transport does not appear in the overall reaction. On the other hand, Erecinska and Wilson use, in their model, a direct connection between electron transport and ATP synthesis. The present paper demonstrates that J can be related to the quotients mentioned above without this direct connection.
MH  - atp
MH  - ATP synthesis
MH  - electron
MH  - Electron Transport
MH  - Intracellular Membranes
MH  - metabolism
MH  - Mitochondria
MH  - model
MH  - Models,Biological
MH  - Oxidative Phosphorylation
MH  - Phosphorylation
MH  - proton
MH  - transport
MH  - ultrastructure
RP  - NOT IN FILE
SO  - Proc Natl Acad Sci U S A 1979 May ;76(5):2236-2238

324
UI  - 19823
AU  - Hutton RL
AU  - Boyer PD
TI  - Subunit interaction during catalysis. Alternating site cooperativity of mitochondrial adenosine triphosphatase
AB  - ATP concentration modulates oxygen exchange catalyzed by purified, soluble mitochondrial ATPase during ATP hydrolysis so that water oxygen incorporation into each Pi formed increases markedly as ATP concentration is lowered. This behavior is readily explained by catalytic cooperativity between subunits of the ATPase. However, other reasonable explanations also need consideration. A new approach for assessing these various explanations is used, based on measurement of the [18O]Pi species formed by hydrolysis of ATP highly labeled with 18O in the gamma-phosphoryl group. The results and other supporting data give what appears to be the most compelling evidence yet attained for alternating site catalytic cooperativity in an enzymic catalysis
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - ADENOSINE-TRIPHOSPHATASE
MH  - Adenosinetriphosphatase
MH  - atp
MH  - ATPase
MH  - Catalysis
MH  - COUPLING FACTOR
MH  - Hydrolysis
MH  - Macromolecular Systems
MH  - Oxidative Phosphorylation
MH  - Oxidative Phosphorylation Coupling Factors
MH  - Oxygen
MH  - Oxygen Isotopes
MH  - Phosphorylation
MH  - SUBUNIT
MH  - Water
RP  - NOT IN FILE
NT  - UI - 80027413LA - engRN - 0 (Macromolecular Systems)RN - 0 (Oxidative Phosphorylation Coupling Factors)RN - 0 (Oxygen Isotopes)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19791218IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:158596
SO  - J Biol Chem 1979 Oct 25 ;254(20):9990-9993

325
UI  - 1065
AU  - Itoh S
AU  - Matsuura K
AU  - Masamoto K
AU  - Nishimura M
TI  - Estimation of the surface potential in photosynthetic membranes
MH  - Membranes
T2  - Cation Fluxes in Biomembranes., [Proc. Symp.], Meeting Date 1978
A2  - Mukohata Y
A2  - Packer L
Y2  - -32676  
PB  - New York: Academic Press
RP  - NOT IN FILE
SO  -  1979  ;():229-242

326
UI  - 19825
AU  - Janson CA
AU  - Degani C
AU  - Boyer PD
TI  - The formation of enzyme-bound and medium pyrophosphate and the molecular basis of the oxygen exchange reaction of yeast inorganic pyrophosphatase
AB  - Yeast inorganic pyrophosphatase, with 10 mM 32Pi and 10 mM Mg2+ present at pH 7.3 TO 7.6, rapidly forms enzyme-bound pyrophosphate equivalent to about 5% of the total catalytic sties on the two enzyme subunits. The enzyme thus appears to bind PPi so as to favor thermodynamically its formation from Pi. The enzyme catalyzes a measurable equilibrium formation of free PPi at a much slower rate. Under similar conditions, the enzyme catalyzes a rapid exchange of oxygen atoms between Pi and water with the relative activation by metals being Mg2+ greater than Zn2+ greater than Co2+ greater than Mn2+. Millisecond mixing and quenching experiments demonstrate that the rate of formation and cleavage of the enzyme-bound PPi is rapid enough to explain most or all of the oxygen exchange reaction
MH  - A
MH  - ACTIVATION
MH  - Cations
MH  - Diphosphates
MH  - Magnesium
MH  - Oxygen
MH  - pH
MH  - Pyrophosphatases
MH  - SUBUNIT
MH  - Water
RP  - NOT IN FILE
NT  - UI - 79173075LA - engRN - 0 (Cations, Divalent)RN - 0 (Diphosphates)RN - 7439-95-4 (Magnesium)RN - 7782-44-7 (Oxygen)RN - EC 3.6.1.- (Pyrophosphatases)PT - Journal ArticleDA - 19790725IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:220217
SO  - J Biol Chem 1979 May 25 ;254(10):3743-3749

327
UI  - 1267
AU  - Junge W
AU  - Auslnder W
AU  - McGeer AJ
AU  - Runge T
TI  - The buffering capacity of the internal phase of thylakoids and the magnitude of the pH changes inside under flashing light
MH  - atp
MH  - Buffers
MH  - Light
MH  - method
MH  - NEUTRALRED
MH  - P
MH  - pH
MH  - proton
MH  - thylakoid
MH  - wox
RP  - IN FILE
SO  - Biochim Biophys Acta 1979  ;546():121-141

328
UI  - 786
AU  - Kagawa Y
AU  - Sone N
AU  - Hirata H
AU  - Yoshida M
TI  - Structure and function of H+-ATPase
AB  - (1) Extensive studies on proton-translocating ATPase (H+-ATPase) revealed that H+-ATPase is an energy transforming device universally distributed in membranes of almost all kinds of cells. (2) Crystallization of the catalytic portion (F1) of H+-ATPase showed that F1 is a hexagonal molecule with a central hole. The diameter of F1 is about 90 A and its molecular weight is about 380,000. (3) Use of thermophilic F1 permits the complete reconstitution of F1 from its five subunits (alpha, beta, gamma, delta, epsilon) and demonstration of the gate function of the gamma delta epsilon-complex, the catalytic function of beta (supported by alpha and gamma), and the H+- translocating functions of all five subunits. (4) Studies using purified thermostable F0 showed that F0 is an H+-channel portion of H+- ATPase. The direct measurement of H+-flux through F0, sequencing of DCCD-binding protein, and isolation of F1-binding protein are described. (5) The subunit stoichiometry of F1 may be alpha 3 beta 3 gamma delta epsilon. (6) Reconstitution of stable H+-ATPase-liposomes revealed that ATP is directly synthesized by the flow of H+ driven by an electrochemical potential gradient and that H+ is translocated by ATP hydrolysis. This rules out functions for all the hypothetical components that do not belong to H+-ATPase in H+-driven ATP synthesis. The roles of conformation change and other phenomena in ATP synthesis are also discussed
RP  - NOT IN FILE
NT  - UI - 81142212LA - engRN - 0 (Ion Channels)RN - 0 (Liposomes)RN - 0 (Macromolecular Systems)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewDA - 19810526IS - 0145-479XSB - IMCY - UNITED STATESJC - HIO
UR  - PM:233471
SO  - J Bioenerg Biomembr 1979 Aug ;11(3-4):39-78

329
UI  - 210
AU  - Kanazawa H
AU  - Futai M
TI  - Physiological change in the ionophore-portion of proton-translocating ATPase in an uncoupled mutant of Escherichia coli
MH  - Adenosinetriphosphatase
MH  - ATPase
MH  - Dicyclohexylcarbodiimide
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - mutant
MH  - succinate
MH  - Succinates
RP  - NOT IN FILE
NT  - UI - 80024755LA - engRN - 0 (Succinates)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19791227IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:39783
SO  - FEBS Lett 1979 Sep 15 ;105(2):275-277

330
UI  - 211
AU  - Kanazawa H
AU  - Miki T
AU  - Tamura F
AU  - Yura T
AU  - Futai M
TI  - Specialized transducing phage lambda carrying the genes for coupling factor of oxidative phosphorylation of Escherichia coli: increased synthesis of coupling factor on induction of prophage lambda asn
AB  - Studies were made of the synthesis of the coupling factor complex (F1-- F0) of oxidative phosphorylation after prophage induction of a set of Escherichia coli strains lysogenic for defective transducing phage lambda asn, lambda uncA, or lambda bglC. The transducing phages had been isolated from a strain of E. coli carrying prophage lambda cI857 S7 within the bglB gene located near the unc gene cluster [Miki, T., Hiraga, S., Nagata, T. & Yura, T. (1978) Proc. Natl. Acad. Sci. USA 75, 5099--5103]. When lysogenic cells carrying lambda asn and lambda cI857 S7 were induced at high temperature, synthesis of the F1-ATPase portion of the complex increased to severalfold that of the noninduced cells. In contrast, no increase was observed upon thermoinduction of cells carrying lambda uncA or lambda bglC. The number of membrane sites that could bind purified F1-ATPase also increased significantly upon induction by lambda asn but not by lambda uncA or lambda bglC. In addition, F1-depleted membranes prepared from lambda asn-induced bacteria required more dicyclohexylcarbodiimide to seal the proton pathway than did those from noninduced bacteria. These results strongly suggest that lambda asn carries a set of bacterial genes coding for all the F1 polypeptides (the alpha, beta, gamma, delta, and probably the epsilon subunits) and at least some of the genes involved in formation of F0 polypeptides. Although lambda uncA carries the structural gene (uncA) for the alpha subunit of F1-ATPase, it apparently does not carry the whole set of F1--F0 genes
MH  - A
MH  - Adenosinetriphosphatase
MH  - ALPHA-SUBUNIT
MH  - Bacteria
MH  - Cells
MH  - COMPLEX
MH  - coupling
MH  - COUPLING FACTOR
MH  - Dicyclohexylcarbodiimide
MH  - EPSILON-SUBUNIT
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - membrane
MH  - Membranes
MH  - Oxidative Phosphorylation
MH  - Phosphorylation
MH  - proton
MH  - Site
MH  - SUBUNIT
MH  - synthesis
MH  - Temperature
RP  - NOT IN FILE
NT  - UI - 79180145LA - engRN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19790725IS - 0027-8424SB - IMCY - UNITED STATES
UR  - PM:155817
SO  - Proc Natl Acad Sci U S A 1979 Mar ;76(3):1126-1130

331
UI  - 209
AU  - Kanazawa H
AU  - Futai M
TI  - [Proton translocating ATPase of Escherichia coli (author's transl)]
MH  - Adenosinetriphosphatase
MH  - ATPase
MH  - Bacterial Proteins
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - membrane
MH  - Membrane Proteins
MH  - protein
MH  - Proteins
MH  - proton
MH  - Protons
MH  - review
RP  - NOT IN FILE
NT  - UI - 80116645LA - jpnRN - 0 (Bacterial Proteins)RN - 0 (DNA, Viral)RN - 0 (Membrane Proteins)RN - 0 (Protons)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticlePT - ReviewDA - 19800425IS - 0037-1017SB - IMCY - JAPAN
UR  - PM:160921
SO  - Seikagaku 1979 Oct 25 ;51(10):1139-1144

332
UI  - 21137
AU  - Kell DB
TI  - On the functional proton current pathway of electron transport phosphorylation. An electrodic view
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - electron
MH  - Electron Transport
MH  - Phosphorylation
MH  - proton
MH  - review
MH  - transport
MH  - Uncoupling Agents
RP  - NOT IN FILE
NT  - UI - 80000505LA - engRN - 0 (Uncoupling Agents)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)PT - Journal ArticlePT - ReviewDA - 19791121IS - 0006-3002SB - IMCY - NETHERLANDS
UR  - PM:38839
SO  - Biochim Biophys Acta 1979 Jul 3 ;549(1):55-99

333
UI  - 19879
AU  - Kohlbrenner WE
AU  - Cross RL
TI  - The mode of inhibition of oxidative phosphorylation by efrapeptin (A23871): measurement of substrate effects on rates of inactivation by a tight-binding inhibitor
MH  - A
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - Antibiotics
MH  - Magnesium
MH  - Oxidative Phosphorylation
MH  - Phosphates
MH  - Phosphorylation
RP  - NOT IN FILE
NT  - UI - 80086566LA - engRN - 0 (Antibiotics)RN - 0 (Peptides)RN - 0 (Phosphates)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 7439-95-4 (Magnesium)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19800228IS - 0003-9861SB - IMCY - UNITED STATESJC - 6SK
UR  - PM:160214
SO  - Arch Biochem Biophys 1979 Dec ;198(2):598-607

334
UI  - 21290
AU  - Linnett PE
AU  - Beechey RB
TI  - Inhibitors of the ATP synthetase system
MH  - atp
MH  - inhibitor
MH  - inhibitors
MH  - review
MH  - SYSTEM
RP  - IN FILE
NT  - ATP 7
SO  - Methods Enzymol 1979  ;55():472-518

335
UI  - 245
AU  - Ludwig B
AU  - Capaldi RA
TI  - Immunoprecipitation of ATP synthetase from beef heart mitochondria
RP  - NOT IN FILE
NT  - UI - 79231460LA - engRN - 0 (Immune Sera)RN - 0 (Membrane Proteins)RN - 0 (Multienzyme Complexes)RN - 0 (Oxidative Phosphorylation Coupling Factors)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 2.7 (Phosphotransferases)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19790917IS - 0006-291XSB - IMCY - UNITED STATESJC - 9Y8
UR  - PM:223550
SO  - Biochem Biophys Res Commun 1979 Apr 27 ;87(4):1159-1167

336
UI  - 1066
AU  - Matsuura K
AU  - Masamoto K
AU  - Itoh S
AU  - Nishimura M
TI  - Effect of surface potential on the intramembrane electrical field measured with carotenoid spectral shift in chromatophores from Rhodopseudomonas sphaeroides
MH  - FIELD
MH  - rhodopseudomonas
RP  - NOT IN FILE
SO  - Biochim Biophys Acta 1979  ;547():91-102

337
UI  - 1017
AU  - Mitchell P
TI  - Keilin's respiratory chain concept and its chemiosmotic consequences
MH  - Bacteria
MH  - Chloroplasts
MH  - Cytochromes
MH  - Electron Transport
MH  - Energy Metabolism
MH  - England
MH  - History of Medicine,20th Cent.
MH  - Membrane Potentials
MH  - metabolism
MH  - Mitochondria
MH  - Nobel Prize
MH  - Osmolar Concentration
MH  - Oxidation-Reduction
MH  - Oxidative Phosphorylation
MH  - Photophosphorylation
MH  - physiology
MH  - Protons
RP  - NOT IN FILE
SO  - Science 1979 Dec 7 ;206(4423):1148-1159

338
UI  - 1019
AU  - Mitchell P
TI  - The Ninth Sir Hans Krebs Lecture. Compartmentation and communication in living systems. Ligand conduction: a general catalytic principle in chemical, osmotic and chemiosmotic reaction systems
AB  - Chemical reactions, like osmotic reactions, are transport processes when looked at in detail. Chemical catalysis by enzymes or catalytic carriers, and osmotic catalysis by porters, may be conceived as occurring by specific ligand-conduction mechanisms. In chemiosmotic reaction systems, the pathways of specific ligand conduction are spatially orientated through anisotropic enzyme and catalytic carrier complexes in which the reactions of chemical group transfer occur as vectorial diffusion processes of group translocation down gradients of group potential that represent real spatially-directed fields of chemical force. Thus, it is easier to explain biochemistry in terms of transport than it is to explain transport in terms of biochemistry
MH  - Animal
MH  - Biochemistry
MH  - Biological Transport
MH  - Catalysis
MH  - Cell Physiology
MH  - Cells
MH  - Citric Acid Cycle
MH  - Diffusion
MH  - Electron Transport
MH  - Energy Transfer
MH  - Enzymes
MH  - Ligands
MH  - metabolism
MH  - Mitochondria
MH  - Osmolar Concentration
MH  - Oxidation-Reduction
MH  - Oxidative Phosphorylation
MH  - Oxygen Consumption
MH  - physiology
RP  - NOT IN FILE
SO  - Eur J Biochem 1979 Mar 15 ;95(1):1-20

339
UI  - 1020
AU  - Mitchell P
AU  - Moyle J
AU  - Mitchell R
TI  - Measurement of translocation of H+/O in mitochondria and submitochondrial vesicles
MH  - Electrochemistry
MH  - Electrodes
MH  - Hydrogen-Ion Concentration
MH  - instrumentation
MH  - metabolism
MH  - Methods
MH  - Mitochondria
MH  - Oxygen Consumption
MH  - Submitochondrial Particles
RP  - NOT IN FILE
SO  - Methods Enzymol 1979  ;55:627-40.():627-640

340
UI  - 1018
AU  - Mitchell P
AU  - Moyle J
TI  - Respiratory-chain protonmotive stoicheiometry
MH  - Animal
MH  - Electron Transport
MH  - Hydrogen-Ion Concentration
MH  - Kinetics
MH  - Mathematics
MH  - metabolism
MH  - Mitochondria
MH  - Oxygen Consumption
RP  - NOT IN FILE
SO  - Biochem Soc Trans 1979 Oct ;7(5):887-894

341
UI  - 19880
AU  - Nalin CM
AU  - Cross RL
AU  - Lucas JJ
AU  - Kohlbrenner WE
TI  - Lack of evidence for covalently-bound carbohydrates in energy- transducing ATPases from mitochondria, bacteria, and chloroplasts
MH  - A
MH  - Adenosinetriphosphatase
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Bacteria
MH  - Carbohydrates
MH  - chloroplast
MH  - Chloroplasts
MH  - H(+)-Transporting ATP Synthase
MH  - Mitochondria
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 80004140LA - engRN - 0 (Carbohydrates)RN - 11028-71-0 (Concanavalin A)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19791129IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:157886
SO  - FEBS Lett 1979 Aug 15 ;104(2):209-214

342
UI  - 552
AU  - Pedersen PL
AU  - Hullihen J
TI  - Resolution and reconstitution of ATP synthesis and ATP-dependent functions of liver mitochondria
RP  - NOT IN FILE
NT  - UI - 79220728LA - engRN - 0 (Oligomycins)RN - 0 (Oxidative Phosphorylation Coupling Factors)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - 57-13-6 (Urea)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19790917IS - 0076-6879SB - IMCY - UNITED STATESJC - MVA
UR  - PM:156857
SO  - Methods Enzymol 1979  ;55():736-741

343
UI  - 661
AU  - Penefsky HS
TI  - Preparation of nucleotide-depleted F1 and binding of adenine nucleotides and analogs to the depleted enzyme
RP  - NOT IN FILE
NT  - UI - 79220690LA - engRN - 0 (Adenine Nucleotides)RN - 0 (Oxidative Phosphorylation Coupling Factors)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19790917IS - 0076-6879SB - IMCY - UNITED STATESJC - MVA
UR  - PM:156846
SO  - Methods Enzymol 1979  ;55():377-380

344
UI  - 663
AU  - Penefsky HS
TI  - ATPases associated with electron transport
RP  - NOT IN FILE
NT  - UI - 79220674LA - engRN - 0 (Macromolecular Systems)RN - 0 (Oligomycins)RN - 0 (Oxidative Phosphorylation Coupling Factors)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19790917IS - 0076-6879SB - IMCY - UNITED STATESJC - MVA
UR  - PM:156833
SO  - Methods Enzymol 1979  ;55():297-303

345
UI  - 662
AU  - Penefsky HS
TI  - Preparation of beef heart mitochondrial ATPase
RP  - NOT IN FILE
NT  - UI - 79220676LA - engRN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19790917IS - 0076-6879SB - IMCY - UNITED STATESJC - MVA
UR  - PM:156834
SO  - Methods Enzymol 1979  ;55():304-308

346
UI  - 650
AU  - Penefsky HS
TI  - Mitochondrial ATPase
AB  - Considerable progress has been made in recent years in our understanding of the phosphorylating apparatus in mitochondria, chloroplasts, and bacteria. It has become clear that the structure and the function of the ATP synthesizing apparatus in these widely divergent organisms is similar if not virtually identical. The subunit composition of F1, its molecular architecture, the location and function of substrate binding sites, as well as putative control sites, understanding of the component parts of the oligomycin-sensitive ATPase complex, and the role of these components in the function of the complex all are under active investigation in many laboratories. The developing information and the new insights provided have begun to permit experimental approaches, at the molecular level, to the mode of action of the ATPase in electron-transport-coupled ATP synthesis
RP  - NOT IN FILE
NT  - UI - 87072850LA - engRN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewDA - 19870105IS - 0065-258XSB - IMCY - UNITED STATESJC - 2LM
UR  - PM:162556
SO  - Adv Enzymol Relat Areas Mol Biol 1979  ;49():223-280

347
UI  - 852
AU  - Petty K
AU  - Jackson JB
AU  - Dutton PL
TI  - Factors controlling the binding of two protons per electron transferred through the ubiquinone and cytochrome b/c2 segment of Rhodopseudomonas sphaeroides chromatophores
AB  - 1. On every turnover, 2.0 protons can be bound by the membrane for each single electron moving through the Q-b/c2 oxidoreductase. 2. One proton (H+II) binding reaction is, and one (H+I) is not, sensitive to antimycin. 3. The redox states of electron transfer components other than the proton binding agents can affect both the rate of proton uptake and the apparent pK values on the agents binding the protons. 4. The presence of valinomycin under certain well-defined conditions can strongly influence the value of the measured pK on the H+II binding agent
RP  - NOT IN FILE
NT  - UI - 79187879LA - engRN - 0 (Cytochromes)RN - 1339-63-5 (Ubiquinone)RN - 9007-43-6 (Cytochrome c)PT - Journal ArticleDA - 19790829IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:36140
SO  - Biochim Biophys Acta 1979 Apr 11 ;546(1):17-42

348
UI  - 850
AU  - Petty KM
AU  - Jackson JB
TI  - Kinetic factors limiting the synthesis of ATP by chromatophores exposed to short flash excitation
AB  - ATP synthesis was measured after chromatophores from Rhodopseudomonas capsulata had been subjected to illumination by single turnover flashes fired at variable frequencies. Three processes were examined, which under different conditions can limit the net yield of ATP. (1) A process with an apparent relaxation time of 10-20 ms. This reaction probably limits the rate of ATP synthesis in continuous illumination. It has similar time dependence to the stimulation of the carotenoid shift decay by ADP after a single flash. (2) An active state of the ATPase only persists when the chromatophores are excited more often than once in 10 s. This state decays with similar kinetics to the entire carotenoid shift decay. Full activation is achieved after two flashes. (1) and (2) are not significantly affected by concentrations of antimycin A sufficient to block electron flow through the cytochrome b/c2 oxidoreductase and abolish phase III in the generation of the carotenoid shift. (3) In the presence of antimycin A, after the third, fourth and subsequent flashes ATP synthesis is limited by the quantity of reducing equivalents transported through the reaction centre rather than by the level of the electrochemical proton gradient
RP  - NOT IN FILE
NT  - UI - 80020986LA - engRN - 0 (Carotenoids)RN - 56-65-5 (Adenosine Triphosphate)RN - 642-15-9 (Antimycin A)RN - 9007-43-6 (Cytochrome c)PT - Journal ArticleDA - 19791220IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:226128
SO  - Biochim Biophys Acta 1979 Sep 11 ;547(3):474-483

349
UI  - 851
AU  - Petty KM
AU  - Jackson JB
TI  - Correlation between ATP synthesis and the decay of the arotenoid band shift after single flash activation of chromatophores from Rhodopseudomonas capsulata
AB  - ATP synthesis and the acceleration of the decay of the carotenoid absorption band shift after single flash excitation of Rhodopseudomonas capsulata chromatophores were compared. The two processes behave similarly with respect to: (1) ADP and Pi concentration; (2) inhibition by efrapeptin and venturicidin, and (3) inhibition by valinomycin/K+ and by ionophores. Taken together with earlier evidence for the electrochromic nature of the carotenoid band shift the data support the contention that positive charge moves outwards across the chromatophore membrane during ATP synthesis and justify the method for determination of the H+/ATP ratio (Petty, K.M. and Jackson, J.B. (1979) FEBS Lett. 97, 367-372). The ability of nucleotide diphosphates in the presence of Pi and Mg2+ to give rise to the acceleration of the carotenoid shift decay closely correlates with the rate of phosphorylation of the nucleotides in steady-state light. Nucleotide triphosphates enhance the decay in parallel with their rate of hydrolysis. Adenylyl imidodiphosphate is itself without effect on the decay of the carotenoid shift and it does not prevent the ADP-induced acceleration. The analogue does prevent the ATP effect but only after repeated flashes
RP  - NOT IN FILE
NT  - UI - 80020985LA - engRN - 0 (Carotenoids)RN - 25612-73-1 (Adenylyl Imidodiphosphate)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 1.13.12.- (Luciferase)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19791220IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:158382
SO  - Biochim Biophys Acta 1979 Sep 11 ;547(3):463-473

350
UI  - 19821
AU  - Rosen G
AU  - Gresser M
AU  - Vinkler C
AU  - Boyer PD
TI  - Assessment of total catalytic sites and the nature of bound nucleotide participation in photophosphorylation
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - Hexokinase
MH  - Photophosphorylation
RP  - NOT IN FILE
NT  - UI - 80049654LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 2.7.1.1 (Hexokinase)PT - Journal ArticleDA - 19800119IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:500602
SO  - J Biol Chem 1979 Nov 10 ;254(21):10654-10661

351
UI  - 895
AU  - Rottenberg H
TI  - Non-equilibrium thermodynamics of energy conversion in bioenergetics
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - Animal
MH  - Biological Transport
MH  - Cattle
MH  - Chloroplasts
MH  - Energy Metabolism
MH  - Kinetics
MH  - Mathematics
MH  - Membranes
MH  - metabolism
MH  - Methods
MH  - Mitochondria
MH  - Mitochondria,Heart
MH  - Nad
MH  - Oxidation-Reduction
MH  - Oxidative Phosphorylation
MH  - Submitochondrial Particles
MH  - Support,U.S.Gov't,P.H.S.
MH  - Thermodynamics
RP  - NOT IN FILE
SO  - Biochim Biophys Acta 1979 Dec 13 ;549(3-4):225-253

352
UI  - 830
AU  - Schlimme E
AU  - de Groot EJ
AU  - Schott E
AU  - Strotmann H
AU  - Edelmann K
TI  - Photophosphorylation of base-modified nucleotide analogs by spinach chloroplasts
RP  - NOT IN FILE
NT  - UI - 80047213LA - engRN - 0 (Ribonucleotides)PT - Journal ArticleDA - 19800124IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:499499
SO  - FEBS Lett 1979 Oct 1 ;106(1):251-256

353
UI  - 492
AU  - Senior AE
TI  - Tightly bound magnesium in mitochondrial adenosine triphosphatase from beef heart
AB  - Tightly bound magnesium was found in soluble, purified ATPase (F1) from beef heart mitochondria in the amount of 1 mol/mol of F1. Iron, zinc, cobalt, manganese, calcium, sodium, copper, and potassium were not tightly bound at stoichiometric levels. Removal of magnesium by chelating agents caused loss of ATPase activity. Removal of tightly bound nucleotide by gel filtration in 50% glycerol- or 60 mM K2SO4- containing buffers did not remove magnesium. Cold dissociation did release magnesium when complete denaturation was accomplished. The results suggest that magnesium is an integral part of F1, that it is required for activity, and that magnesium and nucleotides are tightly bound at separate sites. The idea that the tightly bound nucleotides are not complexed with cations suggests certain structural requirements at their binding sites which might account for the unusual properties of the sites
RP  - NOT IN FILE
NT  - UI - 80049754LA - engRN - 0 (Oxidative Phosphorylation Coupling Factors)RN - 60-00-4 (Edetic Acid)RN - 7439-95-4 (Magnesium)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19800119IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:159299
SO  - J Biol Chem 1979 Nov 25 ;254(22):11319-11322

354
UI  - 493
AU  - Senior AE
AU  - Fayle DR
AU  - Downie JA
AU  - Gibson F
AU  - Cox GB
TI  - Properties of membranes from mutant strains of Escherichia coli in which the beta-subunit of the adenosine triphosphatase is abnormal
AB  - Five uncoupled mutant strains of Escherichia coli carrying mutations in the uncD gene have been studied. In each of these mutant strains the beta-subunit of the F1 portion of the membrane-bound adenosine triphosphatase is abnormal. In one of the mutant strains (carrying the uncD12 allele) in F1-ATPase aggregate was formed which was purified and found to have low ATPase activity. ATPase activity was absent in the other four strains and the abnormal beta-subunits were tightly bound to the membranes. However, membranes from these strains exhibited various proton permeabilities as indicated by NADH-dependent atebrin- fluorescence quenching and bound different amounts of normal F1-ATPase. The amounts of reconstitution of energy-linked reactions after the addition of normal F1-ATPase also varied depending on the mutant allele. It is apparent that considerable phenotypic variations can occur between strains carrying mutations in the same unc gene
RP  - NOT IN FILE
NT  - UI - 80020150LA - engRN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19791121IS - 0264-6021SB - IMCY - ENGLANDJC - 9YO
UR  - PM:158358
SO  - Biochem J 1979 Apr 15 ;180(1):111-118

355
UI  - 494
AU  - Senior AE
AU  - Downie JA
AU  - Cox GB
AU  - Gibson F
AU  - Langman L
AU  - Fayle DR
TI  - The uncA gene codes for the alpha-subunit of the adenosine triphosphatase of Escherichia coli. Electrophoretic analysis of uncA mutant strains
AB  - Four mutant strains of Escherichia coli which lack membrane-bound adenosine triphosphatase activity were shown by genetic-complementation tests to carry mutations in the uncA gene. A soluble inactive F1-ATPase aggregate was released from the membranes of three of the uncA mutant strains by low-ionic-strength washing, and purified by procedures developed for the purification of F1-ATPase from normal strains. Analysis of the subunit structure by two-dimensional gel electrophoresis indicated that the F1-ATPase in strains carrying the uncA401 or uncA453 alleles had a subunit structure indistinguishable from normal F1-ATPase. In contrast, the F1-ATPase from the strain carrying the uncA447 allele contained an alpha-subunit of normal molecular weight, but abnormal net charge. Membranes from strains carrying the uncA450 allele did not have F1-ATPase aggregates that could be solubilized by low-ionic-strength washing. However, a partial dipolid strain carrying both the uncA+ and uncA450 alleles formed an active F1-ATPase aggregate which could be solubilized by low-ionic- strength washing of the membranes and which contained two types of alpha-subunit, one of which was normal and the other had abnormal net charge. It is concluded that the uncA gene codes for the alpha-subunit of the adenosine triphosphatase
RP  - NOT IN FILE
NT  - UI - 80020148LA - engRN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19791121IS - 0264-6021SB - IMCY - ENGLANDJC - 9YO
UR  - PM:158357
SO  - Biochem J 1979 Apr 15 ;180(1):103-109

356
UI  - 787
AU  - Sone N
AU  - Yoshida M
AU  - Hirata H
AU  - Kagawa Y
TI  - Carbodiimide-binding protein of H+-translocating ATPase and inhibition of H+ conduction by dicyclohexylcarbodiimide
AB  - H+-Translocating ATPase, which catalyzes ATP synthesis in biomembranes, is composed of a head piece (F1) and a membrane moiety (F0). Using highly-purified F0 from a thermophilic bacterium PS3 (TF0), the following results were obtained. 1. Inhibition by N,N'- dicyclohexylcarbodiimide (DCCD) of H+ conduction through TF0 followed pseudo-first-order kinetics. The second-order rate constant for inhibitor-enzyme interaction was 5 times 10(3) M(-1)-min(-1). 2. H+ conductivity blocked by DCCD was proportional to the amount of DCCD incorporated in the band 8 protein of TF0. When only one-third of the band 8 protein was labeled with DCCD, TF0 hardly transported any H+. 3. By extracting TF0 with chloroform-methanol, the band 8 protein was obtained as a proteolipid. Polyacrylamide gel electrophoresis with dodecyl sulfate and urea showed that the molecular weight was about 6,000. 4. The amino acid composition of band 8 protein indicated that this protein contained an extremely high percentage of hydrophobic amino acids (0.29 in polarity) and was devoid of histidine, tryptophan, cysteine, and lysine. Its minimum molecular weight was 6,500. 5. The role of band 8 protein (DCCD-binding protein) in H+ conduction through TF0 is discussed on the basis of these results
RP  - NOT IN FILE
NT  - UI - 79130378LA - engRN - 0 (Amino Acids)RN - 0 (Carbodiimides)RN - 0 (Carrier Proteins)RN - 2001-95-8 (Valinomycin)RN - 370-86-5 (Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 7440-09-7 (Potassium)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19790526IS - 0021-924XSB - IMCY - JAPANJC - HIF
UR  - PM:33978
SO  - J Biochem (Tokyo ) 1979 Feb ;85(2):503-509

357
UI  - 595
AU  - Stiggall DL
AU  - Galante YM
AU  - Hatefi Y
TI  - Preparation and properties of complex V
RP  - NOT IN FILE
NT  - UI - 79220677LA - engRN - 0 (Cytochromes)RN - 0 (Oxidative Phosphorylation Coupling Factors)RN - 0 (Phosphates)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19790917IS - 0076-6879SB - IMCY - UNITED STATESJC - MVA
UR  - PM:156835
SO  - Methods Enzymol 1979  ;55():308-321

358
UI  - 832
AU  - Strotmann H
AU  - Bickel-Sandkotter S
AU  - Edelmann K
AU  - Eckstein F
AU  - Schlimme E
AU  - Boos KS
AU  - Lustorff J
TI  - Thiophosphate analogs of ADP and ATP as substrates in partial reactions of energy conversion in chloroplasts
RP  - NOT IN FILE
NT  - UI - 79082976LA - engRN - 0 (Thionucleotides)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)PT - Journal ArticleDA - 19790324IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:758935
SO  - Biochim Biophys Acta 1979 Jan 11 ;545(1):122-130

359
UI  - 831
AU  - Strotmann H
AU  - Bickel-Sandkotter S
AU  - Shoshan V
TI  - Kinetic analysis of light-dependent exchange of adenine nucleotides on chloroplast coupling factor CF1
RP  - NOT IN FILE
NT  - UI - 79191868LA - engRN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19790901IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:156126
SO  - FEBS Lett 1979 May 15 ;101(2):316-320

360
UI  - 1196
AU  - Tiemann R
AU  - Renger G
AU  - Graber P
AU  - Witt HT
TI  - The plastoquinone pool as possible hydrogen pump in photosynthesis
MH  - Hydrogen
MH  - Photosynthesis
RP  - ON REQUEST (03/18/92)
SO  - Biochim Biophys Acta 1979  ;546():498-519

361
UI  - 21215
AU  - Voitsitskii VM
AU  - Drachev LA
AU  - Kaulen AD
AU  - Skulachev VP
TI  - [Formation of the electrical potential difference by bacteriorhodopsin membrane preparations associated with the lipid-water interface surface]
MH  - Bacteriorhodopsin
MH  - carotenoid
MH  - Carotenoids
MH  - Lipids
MH  - membrane
MH  - Water
RP  - NOT IN FILE
NT  - UI - 79148060LA - rusRN - 0 (Membrane Lipids)RN - 36-88-4 (Carotenoids)RN - 53026-44-1 (Bacteriorhodopsins)RN - 7732-18-5 (Water)PT - Journal ArticleDA - 19790626IS - 0002-3264SB - IMCY - USSR
UR  - PM:428314
SO  - Dokl Akad Nauk SSSR 1979  ;245(3):726-729

362
UI  - 783
AU  - Arana JL
AU  - Yoshida M
AU  - Kagawa Y
AU  - Vallejos RH
TI  - Functional arginine residues and carboxyl groups in the adenosine triphosphatase of the thermophilic bacterium PS-3
AB  - Treatment of purified ATPase of the thermophilic bacterium PS-3 with the arginine reagent phenylglyoxal or with Woodward's reagent K, gave complete inactivation of the enzyme. The inactivation rates followed apparent first-order kinetics. The apparent order of reaction with respect to inhibitor concentrations gave values near to 1 with both reagents, suggesting that inactivation was a consequence of modifying one arginine or carboxyl group per active site. ADP and ATP strongly protected the thermophilic ATPase against both reagents. GDP and IDP protected less, whilst CTP did not protect. Experiments in which the incorporation of [14C]phenylglyoxal into the enzyme was measured show that extrapolation of incorporation to 100% inactivation of the enzyme gives 8-9 mol [14C]phenylglyoxal per mol ATPase, whilst ADP or ATP prevent modification of about one arginine per mol
RP  - NOT IN FILE
NT  - UI - 81040053LA - engRN - 0 (Indicators and Reagents)RN - 1074-12-0 (Phenylglyoxal)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 7004-12-8 (Arginine)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19810129IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:6448637
SO  - Biochim Biophys Acta 1980 Nov 5 ;593(1):11-16

363
UI  - 9951
AU  - Arata H
AU  - Nishimura M
TI  - Thermodynamics of electron transfer and its coupling to vectorial processes in biological membranes
AB  - A method is developed to express the flux of an electron transfer reaction as a function of the conjugate force, the redox potential difference, throughout the nonlinear region. The flux can be expressed by a product of the hyperbolic sine of the force, a factor ("redox- poising parameter") determined by the redox potentials of subsystem (in certain cases by local pH's and pK's of subsystems), and some constants. This is analogous to the expression of the flux of a diffusion process by the product of its force and the concentration of the diffusing species. The redox-poising parameter corresponds to the concentration term. The expression is applied to redox chains in which electron transfers are coupled to vectorial processes such as proton translocation or electric current
MH  - Diffusion
MH  - electron
MH  - Membranes
MH  - proton
MH  - Thermodynamics
MH  - TRANSFER
RP  - NOT IN FILE
NT  - UI - 81257030LA - engPT - Journal ArticleDA - 19811025IS - 0006-3495SB - IMCY - UNITED STATESJC - A5S
UR  - PM:7260302
SO  - Biophys J 1980 Nov ;32(2):791-806

364
UI  - 21210
AU  - Drachev LA
AU  - Kalamkarov GR
AU  - Kaulen AD
AU  - Ostrovsky MA
AU  - Skulachev VP
TI  - Animal rhodopsin as a photogenerator of an electric potential that increases photoreceptor membrane permeability
MH  - A
MH  - Animal
MH  - Bacteriorhodopsin
MH  - carotenoid
MH  - Carotenoids
MH  - ion
MH  - Ion Channels
MH  - membrane
MH  - Permeability
MH  - pigments
MH  - retinal
RP  - NOT IN FILE
NT  - UI - 81044384LA - engRN - 0 (Ion Channels)RN - 0 (Retinal Pigments)RN - 36-88-4 (Carotenoids)RN - 53026-44-1 (Bacteriorhodopsins)RN - 9009-81-8 (Rhodopsin)PT - Journal ArticleDA - 19810126IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:6253316
SO  - FEBS Lett 1980 Sep 22 ;119(1):125-131

365
UI  - 207
AU  - Dunn SD
AU  - Futai M
TI  - Reconstitution of a functional coupling factor from the isolated subunits of Escherichia coli F1 ATPase
MH  - A
MH  - Adenosinetriphosphatase
MH  - ATPase
MH  - Aurovertins
MH  - COLI F1 ATPASE
MH  - coupling
MH  - COUPLING FACTOR
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - Macromolecular Systems
MH  - Oxidative Phosphorylation
MH  - Oxidative Phosphorylation Coupling Factors
MH  - Phosphorylation
MH  - reconstitution
MH  - SUBUNIT
MH  - SYSTEM
MH  - SYSTEMS
RP  - NOT IN FILE
NT  - UI - 80072064LA - engRN - 0 (Aurovertins)RN - 0 (Macromolecular Systems)RN - 0 (Oxidative Phosphorylation Coupling Factors)RN - 0 (Ribonucleotides)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19800226IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:6444218
SO  - J Biol Chem 1980 Jan 10 ;255(1):113-118

366
UI  - 121
AU  - Fillingame RH
TI  - The proton-translocating pumps of oxidative phosphorylation
RP  - NOT IN FILE
NT  - UI - 80263997LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewDA - 19801024IS - 0066-4154SB - IMCY - UNITED STATESJC - 6DJ
UR  - PM:6157352
SO  - Annu Rev Biochem 1980  ;49():1079-1113

367
UI  - 58
AU  - Foster DL
AU  - Mosher ME
AU  - Futai M
AU  - Fillingame RH
TI  - Subunits of the H+-ATPase of Escherichia coli. Overproduction of an eight-subunit F1F0-ATPase following induction of a lambda-transducing phage carrying the unc operon
AB  - The proton-translocating ATPase complex (F1F0) of Escherichia coli was purified after inductin of a lambda-transducing phage (lambda asn5) carrying the ATPase genes of th unc operon. ATPase activity of membranes prepared from the induced lambda-unc lysogen was 6-fold greater than the activity of membranes prepared from strains lacking the unc-transducing phage, confirming the report of Kanazawa et al. (1979) Proc. Natl. Acad. Sci. U. S. A. 76, 1126-1130). The F1F0-ATPase complex was purified in comparable yield from either enriched membranes or control membranes using a modification of the procedure reported by Foster and Fillingame ((1979) J. Biol. Chem. 254, 8230-8236). EAch of the eight subunits that had been reported as components of the F1F0 complex from wild type E. coli was overproduced in the lambda-unc lysogen. All eight subunits co-purified in the same stoichiometric proportion as in the complex purified from wild type E. coli. We conclude that all eight subunits are likely coded by the small segment of chromosomal DNA carried by the lambda-transducing phage. These experiments provide the first evidence that all eight polypeptides are authentic subunits of the ATPase complex rather than contaminants that fortuitously co-purify
RP  - NOT IN FILE
NT  - UI - 81069901LA - engRN - 0 (Macromolecular Systems)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - 5 T32 GM07215/GM/NIGMSID - GM-7215/GM/NIGMSID - GM023105/GM/NIGMSDA - 19810219IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:6160157
SO  - J Biol Chem 1980 Dec 25 ;255(24):12037-12041

368
UI  - 204
AU  - Futai M
AU  - Kanazawa H
AU  - Takeda K
AU  - Kagawa Y
TI  - Reconstitution of ATPase from the isolated subunits of coupling factor F1's of Escherichia coli and thermophilic bacterium PS3
MH  - Adenosinetriphosphatase
MH  - ATPase
MH  - Bacteria
MH  - coupling
MH  - COUPLING FACTOR
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1
MH  - Macromolecular Systems
MH  - PS3
MH  - reconstitution
MH  - SUBUNIT
MH  - SYSTEM
MH  - SYSTEMS
RP  - NOT IN FILE
NT  - UI - 81062423LA - engRN - 0 (Macromolecular Systems)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19810116IS - 0006-291XSB - IMCY - UNITED STATES
UR  - PM:6449197
SO  - Biochem Biophys Res Commun 1980 Sep 16 ;96(1):227-234

369
UI  - 9962
AU  - Govindjee R
AU  - Ebrey TG
AU  - Crofts AR
TI  - The quantum efficiency of proton pumping by the purple membrane of Halobacterium halobium
AB  - The quantum yield of H+ release in purple membrane (PM) sheets, and H+ uptake in phospholipid (egg phosphatidylcholine, PC) vesicles containing PM, was measured in single turnover light flashes using a pH- sensitive dye, p-nitrophenol, with rhodopsin as an actinometer. We have also calculated the ratio of H+ released per M412 formed (an unprotonated Shiff-base intermediate formed during the photocycle). In PM sheets, the quantum yield of H+ release depends on the medium. The quantum yield of M412 is independent of salt concentration. The ratio H+/M412 is approximately 1.8 M KC; and approximately 0.64 in 10 mM KCl. Direct measurements of the quantum yield of H+ give approximately 0.7 when the PM is suspended in 0.5 M KC; and 0.25 in 10 mM KCl. Using a quantum yield for M412 formation of 0.3 (Becher and Ebrey, 1977 Biophys J. 17:185.), these measurements also give a H+/M412 approximately 2 at high salt. In PM/PC vesicles, the H+/M412 is approximately 2 at all salt concentrations. The M412 decay is biphasic and the dye absorption change is monophasic. The dissipation of the proton gradient is very slow, taking on the order of seconds. Addition of nigericin (H+/K+ antiporter) drastically reduces the pH changes observed in PM/PC vesicles. This and the observation that the proton relaxation time is much longer than the photochemical cycling time suggest that the protons are pumped across the membrane and there is no contribution as a result of reversible binding and release of protons on just one side of the membrane
MH  - Bacteriorhodopsin
MH  - BINDING
MH  - Carotenoids
MH  - H+
MH  - Halobacterium
MH  - Light
MH  - Liposomes
MH  - Phosphatidylcholines
MH  - proton
MH  - Protons
MH  - Time
RP  - NOT IN FILE
NT  - UI - 81256976LA - engRN - 0 (Carotenoids)RN - 0 (Liposomes)RN - 0 (Phosphatidylcholines)RN - 53026-44-1 (Bacteriorhodopsin)PT - Journal ArticleDA - 19811025IS - 0006-3495SB - IMCY - UNITED STATESJC - A5S
UR  - PM:7260274
SO  - Biophys J 1980 May ;30(2):231-242

370
UI  - 21016
AU  - Graber P
TI  - Phosphorylation of 1,N6-etheno-ADP in flash groups and the concomitant decay kinetics of the absorption change at 515 nm
MH  - absorption
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - flash
MH  - Kinetics
MH  - Phosphorylation
MH  - proton
MH  - Protons
RP  - NOT IN FILE
NT  - UI - 81062507LA - engRN - 0 (Ethenoadenosine Triphosphate)RN - 0 (Protons)RN - 38806-39-2 (1,N(6)-ethenoadenosine diphosphate)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)PT - Journal ArticleDA - 19810126IS - 0006-291XSB - IMCY - UNITED STATES
UR  - PM:6254525
SO  - Biochem Biophys Res Commun 1980 Oct 16 ;96(3):1232-1239

371
UI  - 1191
AU  - Graber P
TI  - Proton translocation at the photosynthetic membrane
MH  - membrane
MH  - proton
MH  - translocation
RP  - ON REQUEST (03/18/92)
SO  - Dev Bioenerg Biomembr 1980  ;4():19-31

372
UI  - 19820
AU  - Hackney DD
AU  - Stempel KE
AU  - Boyer PD
TI  - Oxygen-18 probes of enzymic reactions of phosphate compounds
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - Enzymes
MH  - Oxygen
MH  - Oxygen Isotopes
MH  - Phosphates
MH  - Water
RP  - NOT IN FILE
NT  - UI - 80187384LA - engRN - 0 (Enzymes)RN - 0 (Oxygen Isotopes)RN - 0 (Phosphates)RN - 56-65-5 (Adenosine Triphosphate)RN - 7732-18-5 (Water)PT - Journal ArticleDA - 19800728IS - 0076-6879SB - IMCY - UNITED STATESJC - MVA
UR  - PM:7374458
SO  - Methods Enzymol 1980  ;64():60-83

373
UI  - 205
AU  - Kanazawa H
AU  - Horiuchi Y
AU  - Takagi M
AU  - Ishino Y
AU  - Futai M
TI  - Coupling factor F1 ATPase with defective beta subunit from a mutant of Escherichia coli
AB  - The defective coupling factor F1 ATPase from a mutant strain (KF11) of Escherichia coli was purified to a practically homogeneous form. The final specific activity of Mg2+-ATPase was 6-9 units/mg protein, which is about 10-15 times lower than that of F1 ATPase from the wild-type strain. The mutant F1 had a ratio of Ca2+-ATPase to Mg2+-ATPase of about 3.5, whereas the wild-type F1 had ratio of about 0.8. The mutant F1 was more unstable than wild-type F1: on storage at -80 degrees C for 2 weeks, about 80% of its activity (dependent on Ca2+ or Mg2+) was lost, whereas none of the activity of the wild-type F1 was lost. The following results indicate that the mutation is in the beta subunit. (i) High Mg2+-ATPase activity (about 20 units/mg protein) was reconstituted when the beta subunit from wild type F1 was added to dissociated mutant F1 and the mixture was dialyzed against buffer containing ATP and Mg2+. (ii) Low ATPase activity having the same ratio of Ca2+-ATPase to Mg2+-ATPase as the mutant F1 was reconstituted when a mixture of the beta subunit from the mutant F1 and the alpha and gamma subunits from wild-type F1 was dialyzed against the same buffer. (iii) Tryptic peptide analysis of the beta subunit of the mutant showed a difference in a single peptide compared with the wild-type strain
MH  - A
MH  - Adenosinetriphosphatase
MH  - analysis
MH  - atp
MH  - ATPase
MH  - BETA-SUBUNIT
MH  - buffer
MH  - Ca(2+)-Transporting ATPase
MH  - coupling
MH  - COUPLING FACTOR
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - Magnesium
MH  - mutant
MH  - protein
MH  - SUBUNIT
MH  - Time
MH  - Trypsin
RP  - NOT IN FILE
NT  - UI - 81026297LA - engRN - 0 (Peptides)RN - 7439-95-4 (Magnesium)RN - EC 3.4.21.4 (Trypsin)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.38 (Ca(2+)-Transporting ATPase)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19801218IS - 0021-924XSB - IMCY - JAPAN
UR  - PM:6448252
SO  - J Biochem (Tokyo ) 1980 Sep ;88(3):695-703

374
UI  - 208
AU  - Kanazawa H
AU  - Futai M
TI  - Release of the alpha subunit of coupling factor F1 ATPase from membranes of an uncoupled mutant of Escherichia coli
MH  - Adenosinetriphosphatase
MH  - ALPHA-SUBUNIT
MH  - ATPase
MH  - Bacterial Proteins
MH  - coupling
MH  - COUPLING FACTOR
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - membrane
MH  - Membrane Proteins
MH  - Membranes
MH  - mutant
MH  - Oxidative Phosphorylation
MH  - Oxidative Phosphorylation Coupling Factors
MH  - Peptide Fragments
MH  - Phosphorylation
MH  - protein
MH  - Proteins
MH  - SUBUNIT
RP  - NOT IN FILE
NT  - UI - 80113232LA - engRN - 0 (Bacterial Proteins)RN - 0 (Membrane Proteins)RN - 0 (Oxidative Phosphorylation Coupling Factors)RN - 0 (Peptide Fragments)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19800417IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:6444383
SO  - FEBS Lett 1980 Jan 1 ;109(1):104-106

375
UI  - 20919
AU  - Kanazawa H
AU  - Tamura F
AU  - Mabuchi K
AU  - Miki T
AU  - Futai M
TI  - Organization of unc gene cluster of Escherichia coli coding for proton- translocating ATPase of oxidative phosphorylation
AB  - The proton-translocating ATPase (F1-F0) of oxidative phosphorylation (ATP phosphohydrolase, EC 3.6.1.3) is coded for by a set of structural genes comprising the unc operon in Escherichia coli. We have analyzed several new transducing phages and plasmids carrying various lengths of the DNA segments of the unc operon by complementation assay using 14 new unc- mutants and representatives of previously described strains which were made available to us. Transducing phages carrying parts of the unc gene cluster were isolated: lambda uncA-9 and lambda glmS phages converted only some of the unc- mutants to the Unc+, as determined by complementation assays. A new hybrid plasmid (pMCR533) carrying part of the unc operon was constructed by inserting the HindIII fragment of lambda asn-5 DNA (a phage carrying the entire unc operon) into the unique HindIII site of pBR322. This plasmid transformed eight unc- strains to Unc+, including uncB402 and uncA401, but did not complement uncD11 or four other strains. Two minichromosomes which carry the E. coli replication origin were also tested: plasmid pNH05 transformed the uncB402 but not the uncA401 strain to Unc+, whereas plasmid pMCF1 transformed none of the mutants tested. Analysis of the DNAs from these transducing phages and plasmids with restriction endonucleases suggested that all of the structural genes for the F1-F0 complex are localized within a DNA segment of approximately 4.5 megadaltons containing two EcoRI sites. The approximate locations of the unc- mutations were mapped on this DNA segment
MH  - A
MH  - Adenosinetriphosphatase
MH  - analysis
MH  - atp
MH  - ATPase
MH  - COMPLEX
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - mutant
MH  - Oxidative Phosphorylation
MH  - Phosphorylation
MH  - proton
MH  - Protons
MH  - Site
RP  - NOT IN FILE
NT  - UI - 81175078LA - engRN - 0 (Plasmids)RN - 0 (Protons)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19810613IS - 0027-8424SB - IMCY - UNITED STATES
UR  - PM:6261234
SO  - Proc Natl Acad Sci U S A 1980 Dec ;77(12):7005-7009

376
UI  - 956
AU  - Lappin SA
AU  - Malpress FH
TI  - Proton turnover at mitochondrial coupling sites
MH  - Adenosine Triphosphate
MH  - biosynthesis
MH  - Intracellular Membranes
MH  - metabolism
MH  - Mitochondria
MH  - Oxidative Phosphorylation
MH  - Protons
RP  - NOT IN FILE
SO  - J Theor Biol 1980 May 21 ;84(2):181-184

377
UI  - 19632
AU  - Lardy HA
TI  - Antibiotic inhibitors of mitochondrial energy transfer
MH  - A
MH  - ANTIBIOTIC
MH  - Antibiotics
MH  - aurovertin
MH  - Aurovertins
MH  - cyclic
MH  - energy
MH  - Energy Transfer
MH  - England
MH  - inhibitor
MH  - inhibitors
MH  - oligomycin
MH  - Oligomycins
MH  - peptide
MH  - peptides
MH  - review
MH  - tentoxin
MH  - TRANSFER
MH  - venturicidin
MH  - venturicidins
RP  - NOT IN FILE
NT  - UI - 81077484LA - engRN - 0 (Aminoglycosides)RN - 0 (Antibiotics)RN - 0 (Aurovertins)RN - 0 (Oligomycins)RN - 0 (Peptides)RN - 0 (Peptides, Cyclic)RN - 0 (Venturicidins)RN - 11015-84-2 (ossamycin)RN - 28540-82-1 (tentoxin)RN - 56645-91-1 (efrapeptin)RN - 76600-38-9 (leucinostatin A)PT - Journal ArticlePT - ReviewDA - 19810224IS - 0163-7258SB - IMCY - ENGLANDJC - P44
UR  - PM:7003608
SO  - Pharmacol Ther 1980  ;11(3):649-660

378
UI  - 484
AU  - Martin SS
AU  - Senior AE
TI  - Membrane adenosine triphosphatase activities in rat pancreas
AB  - The membrane ATPase activities present in rat pancreas were studied to investigate the possible role of ATPase enzymes in HCO3(-) secretion in the pancreas. It was found that all the HCO3(-)-sensitive (anion- sensitive) ATPase activity was accountable as pancreatic mitochondrial ATPase, thus supporting the view that a distinct plasma membrane 'bicarbonate-ATPase' is not involved in HCO3(-) secretion in pancreas. A remarkably high Mg+- and CA2+-requiring ATPase activity (30 mumol ATP hydrolysed/min per mg) was found in the plasma membrane fraction (rho = 1.10-1.13). This activity has been characterized in some detail. It is inhibited by p-fluorosulfonylbenzoyladenosine, an affinity label analogue of ATP and the analogue appears to label covalently a protein of Mr approximately 35 000. The (Ca2+ + Mg2+)-ATPase activity did not form a 'phosphorylated-intermediate' and was vanadate-insensitive. These and other tests have served to demonstrate that the (Ca2+ + Mg2+)- ATPase activity is different in properties from (Na+ + K+)-ATPase, Ca2+- ATPase, (H+ + K+)-ATPase or mitochondrial H+-ATPase. Apart from the (Ca2+ + Mg2+)-ATPase of plasma membrane and mitochondrial ATPase, the only other membrane ATPase activities noted were (Na+ + K+)-ATPase, which occurred in the same fractions as the (Ca2+ + Mg2+)-AtPase at rho = 1.10-1.13 and was of surprisingly low activity, and an ATPase activity in light membrane fractions (rho - 1.08-1.09) derived from zymogen granule membranes. At this time, therefore, there is no obvious candidate for an ATPase activity at the luminal surface of pancreatic cells which is directly involved in ion transport, but the results presented here direct attention to the high activity (Ca2+ + Mg2+)- ATPase in the plasma membrane fraction
RP  - NOT IN FILE
NT  - UI - 81040082LA - engRN - 0 (5'-(4-fluorosulfonylbenzoyl)adenosine)RN - 0 (Bicarbonates)RN - 58-61-7 (Adenosine)RN - 7439-95-4 (Magnesium)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.37 (Na(+)-K(+)-Exchanging ATPase)RN - EC 3.6.1.38 (Ca(2+)-Transporting ATPase)PT - Journal ArticleID - GM-25349/GM/NIGMSDA - 19810126IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:6252965
SO  - Biochim Biophys Acta 1980 Nov 4 ;602(2):401-418

379
UI  - 1062
AU  - Matsuura K
AU  - Nishimura M
TI  - Ionophore-sensitive spectral shift of carotenoid induced by ferricyanide in chromatophores from Rhodopseudomonas sphaeroides
MH  - rhodopseudomonas
RP  - NOT IN FILE
SO  - Plant Cell Physiol 1980  ;21():481-485

380
UI  - 9952
AU  - Matsuura K
AU  - Masamoto K
AU  - Itoh S
AU  - Nishimura M
TI  - Surface potential on the periplasmic side of the photosynthetic membrane of Rhodopseudomonas sphaeroides
MH  - Carotenoids
MH  - Magnesium
MH  - Octoxynol
MH  - Polyethylene Glycols
MH  - Sodium
RP  - NOT IN FILE
NT  - UI - 80242646LA - engRN - 0 (Carotenoids)RN - 0 (Polyethylene Glycols)RN - 7487-88-9 (Magnesium Sulfate)RN - 7647-14-5 (Sodium Chloride)RN - 9002-93-1 (Octoxynol)PT - Journal ArticleDA - 19801021IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:6967328
SO  - Biochim Biophys Acta 1980 Aug 5 ;592(1):121-129

381
UI  - 9953
AU  - Matsuura K
AU  - Ishikawa T
AU  - Nishimura M
TI  - Correlation of membrane-potential-sensing carotenoid to pigment-protein complex II in Rhodopseudomonas sphaeroides
AB  - The changes in carotenoid absorbance induced by illumination or by a diffusion potential were larger in chromatophores from cells cultured under low light intensity than those in chromatophores from high-light culture in a photosynthetic bacterium, Rhodopseudomonas sphaeroides. The carotenoid molecules which are associated with the pigment-protein complex (with the infrared bacteriochlorophyll peaks at 800 and 850 nm) (complex II) probably respond to the electrical field changes in the chromatophore membrane
MH  - Bacteria
MH  - Bacterial Proteins
MH  - Carotenoids
MH  - Cells
MH  - COMPLEX
MH  - Diffusion
MH  - diffusion potential
MH  - FIELD
MH  - Light
MH  - Potassium
MH  - Potassium Chloride
MH  - Proteins
RP  - NOT IN FILE
NT  - UI - 80198376LA - engRN - 0 (Bacterial Proteins)RN - 0 (Bacteriochlorophylls)RN - 0 (Carotenoids)RN - 7447-40-7 (Potassium Chloride)PT - Journal ArticleDA - 19800815IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:6966511
SO  - Biochim Biophys Acta 1980 May 9 ;590(3):339-344

382
UI  - 616
AU  - Melandri BA
AU  - Venturoli G
AU  - De Santis A
AU  - Baccarini-Melandri A
TI  - The induction kinetics of bacterial photophosphorylation. Threshold effects by the phosphate potential and correlation with the amplitude of the carotenoid absorption band shift
AB  - 1. ATP synthesis (monitored by luciferin-luciferase) can be elicited by a single turnover flash of saturating intensity in chromatophores from Rhodopseudomonas capsulata, Kb1. The ATP yield from the first to the fourth turnover is strongly influenced by the phosphate potential: at high phosphate potential (-11.5 kcal/mol) no ATP is formed in the first three turnovers while at lower phosphate potential (-8.2 kcal/mol) and the yield in the first flash is already one half of the maximum, which is reached after 2-3 turnovers. 2. The response to ionophores indicates that the driving force for ATP synthesis in the first 20 turnovers is mainly given by a membrane potential. The amplitude of the carotenoid band shift shows that during a train of flashes an increasing delta psi is built up, which reaches a stationary level after a few turnovers; at high phosphate potential, therefore, more turnovers of the same photosynthetic unit are required to overcome an energetic threshold. 3. After several (six to seven) flashes the ATP yield becomes constant, independently from the phosphate potential; the yield varies, however, as a function of dark time (td) between flashes, with an optimum for td = 160-320 ms. 4. The decay kinetics of the high energy state generated by a long (125 ms) flash have been studied directly measuring the ATP yield produced in post-illumination by one single turnover flash, under conditions of phosphate potential (-10 kcal/mol), which will not allow ATP formation by one single turnover. The high energy state decays within 20 s after the illumination. The decay rate is strongly accelerated by 10(-8) M valinomycin. 5. Under all the experimental conditions described, the amplitude of the carotenoid signal correlates univocally with the ATP yield per flash, demonstrating that this signal monitores accurately an energetic state of the membrane directly involved in ATP synthesis. 6. Although values of the carotenoid signal much larger than the minimal threshold are present, relax slowly, and contribute to the energy input for phosphorylation, no ATP is formed unless electron flow is induced by a single turnover flash. 7. The conclusions drawn are independent from the assumption that a delta psi between bulk phases is evaluable from the carotenoid signal
RP  - NOT IN FILE
NT  - UI - 80242654LA - engRN - 0 (Carotenoids)RN - 2001-95-8 (Valinomycin)RN - 28380-24-7 (Nigericin)RN - 56-65-5 (Adenosine Triphosphate)PT - Journal ArticleDA - 19801021IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:7397138
SO  - Biochim Biophys Acta 1980 Aug 5 ;592(1):38-52

383
UI  - 21140
AU  - Michel H
AU  - Oesterhelt D
TI  - Electrochemical proton gradient across the cell membrane of Halobacterium halobium: effect of N,N'-dicyclohexylcarbodiimide, relation to intracellular adenosine triphosphate, adenosine diphosphate, and phosphate concentration, and influence of the potassium gradient
AB  - The proton motive force across the cell membrane of halobacterial cells has been estimated and compared to intracellular values of ATP, ADP, and inorganic phosphate concentrations with respect to the chemiosmotic hypothesis. The accumulation of 14C-labeled indicator substances, triphenylmethylphosphonium for the membrane potential and 5,5- dimethyloxazolidine-2,4-dione for the pH difference between the cell interior and the medium, has been measured in the cells. Values up to 270 mV for the proton motive force have been found in cells pretreated with N,N'-dicyclohexylcarbodiimide (DCCD, 10(-4) M, 30 degrees C, 12 h). Upon illumination a high membrane potential is generated, which is then gradually replaced by a large pH difference. Cells treated with lower DCCD concentrations show only an enhancement of membrane potential upon illumination; the pH difference remains at a low level. Under anaerobic dark conditions, untreated cells maintain a proton motive force of 120-140 mV, which is equilibrated with the intracellular levels of ATP, ADP, and inorganic phosphate. The pH gradient is 1 unit at pH 6 but 0 at pH 8. The membrane potential is low (60-80 mV) at pH 6 and high (120-130 mV) at pH 8. We propose that the proton translocating ATPase compensates for the lowered pH difference at high external pH values by enhancing the membrane potential. The concentration difference of the potassium ions influences the proton motive force and the intracellular ATP levels, apparently via its action on the membrane potential. When the difference of the chemical potential of the potassium ion, expressed in millivolts, exceeds the preexisting membrane potential, the intracellular ATP level is enhanced. When the difference of the chemical potential of the potassium ion (millivolts) is smaller than the membrane potential, the ATP level is decreased
MH  - A
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - ADP
MH  - atp
MH  - ATPase
MH  - Cell Membrane
MH  - Cells
MH  - Dicyclohexylcarbodiimide
MH  - Halobacterium
MH  - indicator
MH  - INORGANIC-PHOSPHATE
MH  - ion
MH  - Ions
MH  - M
MH  - membrane
MH  - Membrane Potential
MH  - Onium Compounds
MH  - pH
MH  - Phosphates
MH  - Potassium
MH  - proton
MH  - Trityl Compounds
RP  - NOT IN FILE
NT  - UI - 81039997LA - engRN - 0 (Carbodiimides)RN - 0 (Onium Compounds)RN - 0 (Phosphates)RN - 0 (Trityl Compounds)RN - 15912-74-0 (triphenylmethylphosphonium)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 7440-09-7 (Potassium)PT - Journal ArticleDA - 19810116IS - 0006-2960SB - IMCY - UNITED STATES
UR  - PM:7426619
SO  - Biochemistry 1980 Sep 30 ;19(20):4607-4614

384
UI  - 21139
AU  - Michel H
AU  - Oesterhelt D
TI  - Electrochemical proton gradient across the cell membrane of Halobacterium halobium: comparison of the light-induced increase with the increase of intracellular adenosine triphosphate under steady-state illumination
AB  - The increase of the proton motive force mediated by bacteriorhodopsin is compared to the intracellular ATP concentration under steady-state illumination. The membrane potential was measured via the accumulation of the lipophilic ion [14C]triphenylmethylphosphonium and the pH gradient via the accumulation of the weak acid 5,5-dimethyloxazolidine- 2,4-dione. Light causes a parallel increase of ATP level and membrane potential at an external pH of 8. In contrast, at pH 6 an increase of the intracellular ATP concentration occurs without a corresponding increase of the proton motive force. If the extracellular NaCl concentration is reduced and not replaced by other ions, no membrane potential at all can be measured at pH 6-7; nonetheless, light-induced ATP synthesis occurs. A significant enhancement of the pH gradient occurs only at irradiances higher than those required for the attainment of the maximal intracellular ATP concentration. In the presence of the uncoupler carbonyl cyanide m-chlorophenylhydrazone it is possible to obtain light-induced ATP synthesis without a measurable proton motive force in basal salt at pH 8. It is concluded that if bacteriorhodopsin acts as a proton pump, then the pumped protons can be used for ATP synthesis before they equilibrate with the protons in the extracellular bulk phase
MH  - A
MH  - ACID
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthesis
MH  - Bacteriorhodopsin
MH  - Cell Membrane
MH  - Dicyclohexylcarbodiimide
MH  - Halobacterium
MH  - ion
MH  - Ions
MH  - Light
MH  - membrane
MH  - Membrane Potential
MH  - Onium Compounds
MH  - pH
MH  - Phosphates
MH  - proton
MH  - Proton Pump
MH  - Protons
MH  - Sodium
MH  - synthesis
MH  - Trityl Compounds
RP  - NOT IN FILE
NT  - UI - 81039998LA - engRN - 0 (Onium Compounds)RN - 0 (Phosphates)RN - 0 (Trityl Compounds)RN - 15912-74-0 (triphenylmethylphosphonium)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 7647-14-5 (Sodium Chloride)PT - Journal ArticleDA - 19810116IS - 0006-2960SB - IMCY - UNITED STATES
UR  - PM:7426620
SO  - Biochemistry 1980 Sep 30 ;19(20):4615-4619

385
UI  - 1016
AU  - Mitchell P
TI  - Protonmotive cytochrome system of mitochondria
MH  - Animal
MH  - Biological Transport,Active
MH  - Chemistry
MH  - Cytochrome-c Oxidase
MH  - Cytochromes
MH  - Electron Transport
MH  - Ligands
MH  - metabolism
MH  - Mitochondria
MH  - Models,Biological
MH  - NADH Dehydrogenase
MH  - Oxygen Consumption
MH  - Protons
MH  - Rats
RP  - NOT IN FILE
SO  - Ann N Y Acad Sci 1980  ;341:564-84.():564-584

386
UI  - 903
AU  - Morowitz HJ
TI  - From soup to solid state
MH  - Adenosine Triphosphate
MH  - biosynthesis
MH  - Energy Metabolism
MH  - Human
MH  - metabolism
MH  - Mitochondria
MH  - Oxidation-Reduction
RP  - NOT IN FILE
SO  - Hosp Pract 1980 Jun ;15(6):129, 132

387
UI  - 120
AU  - Negrin RS
AU  - Foster DL
AU  - Fillingame RH
TI  - Energy-transducing H+-ATPase of Escherichia coli. Reconstitution of proton translocation activity of the intrinsic membrane sector
AB  - The intrinsic membrane sector (Fo) of the H+-ATPase complex of Escherichia coli has been purified, incorporated into liposomes, and its proton-translocating activity reconstituted. The Fo sector was prepared by treating a purified, particulate, F1FO-ATPase preparation with EDTA to solubilize the F1-ATPase. The resulting particulate Fo fraction was incorporated into liposomes of E. coli phospholipids by sonication. Proton efflux from these liposomes was measured with a pH electrode after imposition of a membrane potential. The kinetics of proton efflux fits that predicted by the Goldman-flux equation. The rate of proton efflux was increased maximally more than 100-fold on incorporation of the Fo sector into the liposomes. The rate of H+ efflux varied directly with the amount of Fo material added during reconstitution. Dicyclohexylcarbodiimide blocked Fo-mediated H+ efflux. Inhibition was shown to be due to reaction of dicyclohexylcarbodiimide with a specific proteolipid subunit of Fo. The preparation of Fo used in these studies contained the three proteins that had previously been identified as likely subunits of Fo (Foster, D. L., and Fillingame, R. H. (1979) J. Biol. Chem. 254, 8230-8236). It remains to be determined whether all three components are required for reconstitution of proton translocation activity
RP  - NOT IN FILE
NT  - UI - 80204324LA - engRN - 0 (Liposomes)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19800828IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:6445905
SO  - J Biol Chem 1980 Jun 25 ;255(12):5643-5648

388
UI  - 784
AU  - Ohta S
AU  - Tsubo M
AU  - Oshima T
AU  - Yoshida M
AU  - Kagawa Y
TI  - Nucleotide binding to isolated alpha and beta subunits of proton translocating adenosine triphosphatase studied with circular dichroism
AB  - The catalytic and allosteric sites of proton translocating adenosine triphosphatase (ATPase) were studied by measuring the binding of nucleotides to the ATPase, and its alpha and beta subunits purified from thermophilic bacterium PS3, with a circular dichroic spectrometer. In contrast to mesophilic ATPases, this thermophilic enzmye contained no tightly bound nucleotides, and its subunits were stable after their purification. These properties were advantageous for analyzing both catalytic and allosteric sites. The former site showed rapid and loose binding, but the latter slow (t 1/2 = 1 h, for ADP) and tight binding. When a nucleotide was bound, the beta subunits showed a negative ellipticity at 275 nm corresponding to a tyrosyl residue, while the alpha subunits showed an ellipticity change corresponding to the absorption curve of the bound nucleotide. This difference enabled us to distinguish the binding sites in ATPase. At a low concentration, ADP selectively bound to alpha subunits in the ATPase, while at a high concentration, it bound to both subunits. This finding suggests that the tight binding sites are located in the alpha subunits. Although ADP and ATP bound to both the purified alpha and beta subunits, CTP did not bind to beta but only to alpha subunits, and ITP bound to beta but hardly to alpha. These nucleotide specificities also supported the idea that the catalytic sites are located in the beta subunits and the allosteric sites are located in the alpha subunits
RP  - NOT IN FILE
NT  - UI - 80249459LA - engRN - 0 (Macromolecular Systems)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19801024IS - 0021-924XSB - IMCY - JAPANJC - HIF
UR  - PM:6447145
SO  - J Biochem (Tokyo ) 1980 Jun ;87(6):1609-1617

389
UI  - 785
AU  - Ohta S
AU  - Tsuboi M
AU  - Yoshida M
AU  - Kagawa Y
TI  - Intersubunit interactions in proton-translocating adenosine triphosphatase as revealed by hydrogen-exchange kinetics
AB  - The rates of hydrogen-deuterium exchange in the peptide groups of the alpha and beta subunits and the alpha-beta subunit complex of proton- translocating adenosine triphosphatase from the thermophilic bacterium PS3 were examined. The exchange was found to be much slower in the isolated beta subunit than in the isolated alpha subunit. This has been taken as indicating that the structure of the beta subunit is tighter than that of the alpha subunit. Adenosine 5'-triphosphate (ATP) caused tightening of a relatively tight portion of the alpha subunit and of a relatively loose portion of the beta subunit. When the alpha and beta subunits are brought into contact, tightening of the alpha subunit, but not the beta subunit, occurs. The effect of ATP on the structure of the beta subunit is more pronounced in the presence of the alpha subunit than in its absence. These findings support the idea proposed previously that the alpha subunit has an allosteric site and the beta subunit a catalytic site and that the conformation of the beta subunit is controlled by the alpha subunit
RP  - NOT IN FILE
NT  - UI - 80198253LA - engRN - 0 (Macromolecular Systems)RN - 7782-39-0 (Deuterium)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19800815IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:6445747
SO  - Biochemistry 1980 May 13 ;19(10):2160-2165

390
UI  - 849
AU  - Packham NK
AU  - Greenrod JA
AU  - Jackson JB
TI  - Generation of membrane potential during photosynthetic electron flow in chromatophores from Rhodopseudomonas capsulata
AB  - 1. When cytochrome c2 is available for oxidation by the photosynthetic reaction centre, the decay of the carotenoid absorption band shift generated by a short flash excitation of Rhodopseudomonas capsulata chromatophores is very slow (half-time approximately 10 s). Otherwise the decay is fast (half-time approximately 1 s in the absence and 0.05 s in the presence of 1,10-ortho-phenanthroline) and coincides with the photosynthetic back reaction. 2. In each of these situations the carotenoid shift decay, but not electron transport, may be accelerated by ioniophores. The ionophore concentration dependence suggests that in each case the carotenoid response is due to a delocalised membrane potential which may be dissipated either by the electronic back reaction or by electrophoretic ion flux. 3. At high redox potentials, where cytochrome c2 is unavailable for photooxidation, electron transport is believed to proceed only across part of the membrane dielectric. Under such conditions it is shown that the driving force for carbonyl cyanide trifluoromethoxyphenyl hydrazone-mediated H+ efflux is nevertheless decreased by valinomycin/K+; demonstrating that the [BChl]2 leads to Q electron transfer generates a delocalised membrane potential
RP  - NOT IN FILE
NT  - UI - 80242648LA - engRN - 0 (Bacteriochlorophylls)RN - 0 (Carotenoids)RN - 0 (Phenanthrolines)RN - 2001-95-8 (Valinomycin)RN - 370-86-5 (Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone)RN - 66-71-7 (1,10-phenanthroline)PT - Journal ArticleDA - 19801021IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:7397136
SO  - Biochim Biophys Acta 1980 Aug 5 ;592(1):130-142

391
UI  - 550
AU  - Pedersen PL
AU  - Amzel LM
AU  - Cintron N
AU  - Soper JW
AU  - Hullihen J
AU  - Wehrle J
TI  - The ATP synthesizing system of liver mitochondria
RP  - NOT IN FILE
NT  - UI - 81036440LA - engRN - 0 (Multienzyme Complexes)RN - 0 (Nucleotides)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 2.7 (Phosphotransferases)RN - EC 2.7.4.- (ATP synthetase complex)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticlePT - ReviewID - CA 10951/CA/NCIID - GM 25432/GM/NIGMSDA - 19801218IS - 0065-2598SB - IMCY - UNITED STATESJC - 2LU
UR  - PM:6252767
SO  - Adv Exp Med Biol 1980  ;132():317-344

392
UI  - 953
AU  - Rohde K
AU  - Reich JG
TI  - Theoretical study of an energy metabolizing system satisfying Mitchell's postulates
AB  - An energy metabolizing system was modelled, which carries as characteristic features substrate uptake and product extrusion, respiration and ATP-synthesis, coupled by the protonmotive potential corresponding to Mitchell's chemiosmotic hypothesis of oxidative phosphorylation. The influence of the electric part of the protonmotive potential on the rate laws of the different processes has been taken into account by a phenomenological exponent of the proton concentration, thus fortifying the osmotic concentration part of the protonmotive potential. The steady-state behaviour of the model was investigated. It could be shown that a model of such kind reveals multistationarity, homeostasis and trigger behaviour. Curves of respiration control (respiration velocity over ADP/(ATP + ADP) or ATP/ADP) are in qualitative accordance with experimental facts (state 3 and state 4 of mitochondria) as well as curves describing the effect of uncouplers on the respiratory rate
MH  - Adenosine Triphosphate
MH  - biosynthesis
MH  - drug effects
MH  - Energy Metabolism
MH  - Homeostasis
MH  - metabolism
MH  - Mitochondria
MH  - Models,Biological
MH  - Oxidation-Reduction
MH  - Oxidative Phosphorylation
MH  - pharmacology
MH  - Respiration
MH  - Uncoupling Agents
RP  - NOT IN FILE
SO  - Acta Biol Med Ger 1980  ;39(4):367-380

393
UI  - 20973
AU  - Schneider E
AU  - Altendorf K
TI  - Reconstitution of the purified proton conductor (F0) of the adenosine triphosphatase complex from Escherichia coli
MH  - Adenosine
MH  - ADENOSINE-TRIPHOSPHATASE
MH  - Adenosinetriphosphatase
MH  - ATPase
MH  - COMPLEX
MH  - Dicyclohexylcarbodiimide
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - proton
MH  - reconstitution
MH  - Valinomycin
RP  - NOT IN FILE
NT  - UI - 81004373LA - engRN - 2001-95-8 (Valinomycin)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19801124IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:6447620
SO  - FEBS Lett 1980 Jul 28 ;116(2):173-176

394
UI  - 485
AU  - Senior AE
AU  - Richardson LV
AU  - Baker K
AU  - Wise JG
TI  - Tight divalent cation-binding sites of soluble adenosine triphosphatase (F1) from beef heart mitochondria and Escherichia coli
RP  - NOT IN FILE
NT  - UI - 80227912LA - engRN - 148-24-3 (Oxyquinoline)RN - 7439-95-4 (Magnesium)RN - 7439-96-5 (Manganese)RN - 7440-48-4 (Cobalt)RN - 7440-66-6 (Zinc)RN - 84-88-8 (8-hydroxyquinoline-5-sulfonic acid)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19800928IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:6446563
SO  - J Biol Chem 1980 Aug 10 ;255(15):7211-7217

395
UI  - 829
AU  - Shoshan V
AU  - Strotmann H
TI  - The effect of phosphate on light-induced exchange of ADP at the tight nucleotide binding site of CF1
AB  - The effects of phosphate on the energy-dependent exchange of the tightly bound adenine nucleotides in chloroplast thylakoids has been studied. 1. Phosphate was found to have a dual effect on ADP binding; at low light intensities Pi enhanced the rate of binding and the total amount of tightly bound ADP, while at saturating light intensities, it decreased the level of bound nucleotides. Half-maximal stimulation or inhibition of ADP binding occurred at about 30 microM phosphate. 2. Phosphate inhibition of ADP binding at saturating light intensities is prevented in the presence of an ADP-regenerating system. Pi had no effect on the amount of ATP bound to chloroplast coupling factor 1 (CF1), which was lower than the amount of ADP bound. 3. Arsenate acted similarly to phosphate at low light intensities but not at saturating light intensities. 4. At low light intensities the interaction of phosphate with the membrane-bound CF1 increased the binding affinity for ADP about 8-fold. 5. Kinetic analysis of the ADP binding showed that phosphate increased the rate constant for ADP binding to the adenine nucleotide-depleted form of CF1
RP  - NOT IN FILE
NT  - UI - 80094555LA - engRN - 0 (Arsenates)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 2.7.1.1 (Hexokinase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19800327IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:6444303
SO  - J Biol Chem 1980 Feb 10 ;255(3):996-999

396
UI  - 21116
AU  - Sorgato MC
AU  - Branca D
AU  - Ferguson SJ
TI  - The rate of ATP synthesis by submitochondrial particles can be independent of the magnitude of the protonmotive force
AB  - The problem of whether the rate of ATP synthesis is proportional to the magnitude of the protonmotive force has been studied in submitochondrial particles. It was found that the rate of ATP synthesis can decrease at constant protonmotive force and is more closely related to the rate of substrate oxidation
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthesis
MH  - CONSTANT
MH  - England
MH  - Malonates
MH  - proton
MH  - Protons
MH  - Submitochondrial Particles
MH  - synthesis
RP  - NOT IN FILE
NT  - UI - 81133473LA - engRN - 0 (Malonates)RN - 0 (Protons)RN - 56-65-5 (Adenosine Triphosphate)PT - Journal ArticleDA - 19810413IS - 0264-6021SB - IMCY - ENGLAND
UR  - PM:6258563
SO  - Biochem J 1980 Jun 15 ;188(3):945-948

397
UI  - 397
AU  - Vasilyeva EA
AU  - Fitin AF
AU  - Minkov IB
AU  - Vinogradov AD
TI  - Kinetics of interaction of adenosine diphosphate and adenosine triphosphate with adenosine triphosphatase of bovine heart submitochondrial particles
AB  - The short preincubation of submitochondrial particles with low concentrations of ADP in the presence of Mg2+ results in a complete loss of their ATPase and inosine triphosphatase activities. Other nucleoside diphosphates (IDP and GDP) do not affect the ATPase activity. The ADP-inhibited ATPase can be activated in a time-dependent manner by treatment of submitochondrial particles with the enzyme converting ADP into ATP (phosphoenolpyruvate plus pyruvate kinase). The activaton is a first-order reaction with rate constant 0.2 min-1 at 25 degrees C. The rate constant of activation is increased in the presence of ATP up to 2 min-1, and this increase shows saturation kinetics with Km value equal to that for ATPase reaction itself (10(-4) M at 25 degrees C at pH 8.0). The experimental results obtained are consistent with the model where two alternative pathways of ADP dissociation from the inhibitory site of ATPase exist; one is spontaneous dissociation and the second is ATP-dependent dissociation through the formation of the ternary complex between ADP, the enzyme and ATP. ADP-induced inactivation and ATP-dependent activation of ATPase activity of submitochondrial particles is accompanied by the same directed change of their ability to catalyse the ATP-dependent reverse electron transport from succinate to NAD+. The possible implication of the model suggested is discussed in terms of functional role of the inhibitory high-affinity binding site for ADP in the mitochondrial ATPase
RP  - NOT IN FILE
NT  - UI - 81133454LA - engRN - 0 (Macromolecular Systems)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19810413IS - 0264-6021SB - IMCY - ENGLANDJC - 9YO
UR  - PM:6451217
SO  - Biochem J 1980 Jun 15 ;188(3):807-815

398
UI  - 19819
AU  - Vinkler C
AU  - Avron M
AU  - Boyer PD
TI  - Effects of permeant buffers on the initial time course of photophosphorylation and postillumination phosphorylation
AB  - Permeant buffers (pyridine, imidazole, or phosphate) caused similar increases in the time required for onset of ATP synthesis in the light in the presence of valinomycin and K+; and in the illumination time required for postillumination phosphorylation with or without or without valinomycin and K+. Based on prior evidence, the minimum illumination time required for postillumination phosphorylation in thylakoid membranes is taken as a measure of the time required for formation of a transmembrane pH gradient sufficient to drive ATP synthesis. Our results are consistent with the view that, following illumination, as the transient transmembrane electric gradient decays, the establishment and maintenance of a pH gradient serves for energy transfer from the photosystems to the ATP synthase complex
MH  - A
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - Buffers
MH  - COMPLEX
MH  - Energy Transfer
MH  - Light
MH  - membrane
MH  - Membranes
MH  - pH
MH  - Phosphates
MH  - Phosphorylation
MH  - Photophosphorylation
MH  - SYNTHASE
MH  - synthesis
MH  - thylakoid
MH  - thylakoid membrane
MH  - Time
MH  - TRANSFER
MH  - Valinomycin
RP  - NOT IN FILE
NT  - UI - 80137442LA - engRN - 0 (Buffers)RN - 0 (Imidazoles)RN - 0 (Phosphates)RN - 0 (Pyridines)RN - 2001-95-8 (Valinomycin)PT - Journal ArticleDA - 19800523IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:7358668
SO  - J Biol Chem 1980 Mar 25 ;255(6):2263-2266

399
UI  - 872
AU  - von Jagow G
AU  - Engel WD
TI  - Structure and function of the energy-converting system of mitochondria
MH  - Adenine Nucleotide Translocase
MH  - Adenosine Triphosphate
MH  - Adipose Tissue
MH  - Animal
MH  - biosynthesis
MH  - Cattle
MH  - Electron Transport
MH  - Energy Metabolism
MH  - enzymology
MH  - In Vitro
MH  - metabolism
MH  - Mitochondria
MH  - Oxidation-Reduction
MH  - Phosphates
MH  - Thermodynamics
MH  - ultrastructure
RP  - NOT IN FILE
SO  - Angew Chem Int Ed Engl 1980  ;19(9):659-675

400
UI  - 20974
AU  - Wachter E
AU  - Schmid R
AU  - Deckers G
AU  - Altendorf K
TI  - Amino acid replacement in dicyclohexylcarbodiimide-reactive proteins from mutant strains of Escherichia coli defective in the energy- transducing ATPase complex
MH  - ACID
MH  - Adenosinetriphosphatase
MH  - ATPase
MH  - Bacterial Proteins
MH  - COMPLEX
MH  - Dicyclohexylcarbodiimide
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - mutant
MH  - protein
MH  - Proteins
MH  - Proteolipids
RP  - NOT IN FILE
NT  - UI - 80225057LA - engRN - 0 (Bacterial Proteins)RN - 0 (Carbodiimides)RN - 0 (Proteolipids)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19800926IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:6446460
SO  - FEBS Lett 1980 May 5 ;113(2):265-270

401
UI  - 18969
AU  - Wagner R
AU  - Junge W
TI  - Two different types of conformational changes of membrane-bound CF1 as revealed by the triplet probe eosin
MH  - atp
MH  - conformation
MH  - conformational change
MH  - indicator
RP  - NOT IN FILE
SO  - FEBS Lett 1980  ;114():327-333

402
UI  - 649
AU  - Webb MR
AU  - Grubmeyer C
AU  - Penefsky HS
AU  - Trentham DR
TI  - The stereochemical course of phosphoric residue transfer catalyzed by beef heart mitochondrial ATPase
AB  - The stereochemical course of phosphoric residue transfer has been determined for beef heart mitochondrial ATPase. When aden 5'-(3- thiotriphosphate), stereospecifically labeled with 18O in the gamma position, was hydrolyzed in [17O]water in the presence of the ATPase, the product inorganic [16O, 17O, 18O]thiophosphate was chiral. The configuration of the product showed that the hydrolysis had proceeded with inversion at the gamma-phosphorus atom. This result suggests that there is a direct, in-line transfer of the phosphoric residue between ADP and water and that there is no phosphoenzyme intermediate
RP  - NOT IN FILE
NT  - UI - 81069837LA - engRN - 0 (Oxygen Isotopes)RN - 0 (Thionucleotides)RN - 35094-46-3 (adenosine 5'-O-(3-thiotriphosphate))RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleID - AM 23030/AM/NIADDKID - GM21737/GM/NIGMSDA - 19810219IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:6449510
SO  - J Biol Chem 1980 Dec 25 ;255(24):11637-11639

403
UI  - 20927
AU  - al Shawi MK
AU  - Brand MD
TI  - Steady-state H+/O stoichiometry of liver mitochondria
AB  - We have measured the H+/O stoichiometry of rat liver mitochondria respiring in a steady-state, using a novel method. This involves measuring the initial rate of H+ back-flow into mitochondria after respiratory inhibition, with the assumption that this is equal to the steady-state H+-ejection rate. Division by the steady-state O2- consumption rate yields the H+/O ratio. The H+/O values obtained were: 8.3 +/- 1.0 (mean +/- S.E.M.) for 3-hydroxybutyrate: 8.2 +/- 0.7 for glutamate plus malate; 6.0 +/- 0.2 for succinate; 4.1 +/- 0.3 for ascorbate/tetramethylphenylenediamine and 3.0 +/- 0.1 for ascorbate/ferrocyanide. These values correspond to H+/O stoichiometries for electron flow to oxygen from NAD+-linked substrates, succinate and cytochrome c of 8, 6 and 2 (charge/O ratio = 4) respectively
MH  - A
MH  - Antimycin A
MH  - Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone
MH  - cytochrome
MH  - Cytochrome c
MH  - electron
MH  - England
MH  - H+
MH  - Liver
MH  - method
MH  - Mitochondria
MH  - Oxygen
MH  - Potassium
MH  - proton
MH  - Protons
MH  - succinate
RP  - NOT IN FILE
NT  - UI - 82205964LA - engRN - 0 (Protons)RN - 151-50-8 (Potassium Cyanide)RN - 370-86-5 (Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone)RN - 642-15-9 (Antimycin A)PT - Journal ArticleDA - 19820719IS - 0264-6021SB - IMCY - ENGLAND
UR  - PM:6282251
SO  - Biochem J 1981 Dec 15 ;200(3):539-546

404
UI  - 21225
AU  - Arents JC
AU  - van Dekken H
AU  - Hellingwerf KJ
AU  - Westerhoff HV
TI  - Linear relations between proton current and pH gradient in bacteriorhodopsin liposomes
AB  - The dependence of proton movement across the membrane of bacteriorhodopsin liposomes on the pH gradient was investigated. Under the appropriate experimental conditions, endogenous proton (or hydroxyl) leakage, proton movement catalyzed by protonophore or nigericin, and light-driven proton translocation depend linearly on the pH gradient. This justifies the use of linear proton flux vs. protonmotive force relations in a recent mosaic thermodynamic description of ion translocation in bacteriorhodopsin liposomes [Westerhoff, H. V., Scholte, B. J., & Hellingwerf, K. J. (1979) Biochim. Biophys. Acta 547, 544-560]. Since bacteriorhodopsin liposomes are a model system for all biological energy transducing systems in which proton pumps are involved, these findings also explain linear relations between proton flux and protonmotive force observed in and postulated for those systems. In cases where the membrane potential is not clamped at a low value, an initial phase of rapid proton movement occurs, followed by a phase of slower proton movement. The rate of proton movement during the slow phase is again linear with the pH gradient. Such a linear relation is not observed for the fast phase. Since the rapid proton movement phase is also observed in liposomes without bacteriorhodopsin, it is not due (only) to dissociation of scalar protons from bacteriorhodopsin. We suggest that during the initial phase of proton movement, the proton flux is not yet electrically compensated by the fluxes of other ions
MH  - A
MH  - Bacteriorhodopsin
MH  - carotenoid
MH  - Carotenoids
MH  - DEPENDENCE
MH  - ion
MH  - Ionophores
MH  - Ions
MH  - liposome
MH  - Liposomes
MH  - membrane
MH  - Membrane Potential
MH  - model
MH  - Movement
MH  - pH
MH  - proton
MH  - Proton Pump
MH  - protonophore
MH  - Protons
MH  - SYSTEM
MH  - SYSTEMS
MH  - translocation
RP  - NOT IN FILE
NT  - UI - 82046492LA - engRN - 0 (Ionophores)RN - 0 (Liposomes)RN - 0 (Protons)RN - 36-88-4 (Carotenoids)RN - 53026-44-1 (Bacteriorhodopsins)PT - Journal ArticleDA - 19820120IS - 0006-2960SB - IMCY - UNITED STATES
UR  - PM:6271177
SO  - Biochemistry 1981 Sep 1 ;20(18):5114-5123

405
UI  - 971
AU  - Berry MN
TI  - An electrochemical interpretation of metabolism
MH  - Adenosine Triphosphate
MH  - Animal
MH  - Chloroplasts
MH  - Cytoplasm
MH  - Electrochemistry
MH  - Electrophysiology
MH  - Intracellular Membranes
MH  - metabolism
MH  - Mitochondria
MH  - Models,Biological
MH  - Nad
MH  - Oxidation-Reduction
MH  - Oxidative Phosphorylation
MH  - Oxidoreductases
MH  - Phosphorylation
MH  - Phosphotransferases
MH  - physiology
MH  - Protons
MH  - Thermodynamics
RP  - NOT IN FILE
SO  - FEBS Lett 1981 Nov 16 ;134(2):133-138

406
UI  - 9961
AU  - Bowyer JR
AU  - Crofts AR
TI  - On the mechanism of photosynthetic electron transfer in Rhodopseudomonas capsulata and Rhodopseudomonas sphaeroides
MH  - Antimycin A
MH  - Cytochrome b
MH  - Cytochrome c
MH  - Cytochromes
MH  - electron
MH  - mechanism
MH  - Thiazoles
MH  - TRANSFER
MH  - Ubiquinone
RP  - NOT IN FILE
NT  - UI - 82024040LA - engRN - 0 (5-n-undecyl-6-hydroxy-4,7-dioxobenzothiazole)RN - 0 (Cytochromes)RN - 0 (Herbicides, Triazine)RN - 0 (Thiazoles)RN - 1339-63-5 (Ubiquinone)RN - 642-15-9 (Antimycin A)RN - 834-12-8 (ametryne)RN - 9007-43-6 (Cytochrome c)RN - 9035-37-4 (Cytochrome b)PT - Journal ArticleID - GM 26305-01/GM/NIGMSDA - 19811215IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:6269602
SO  - Biochim Biophys Acta 1981 Jul ;636(2):218-233

407
UI  - 21172
AU  - Brusilow WS
AU  - Gunsalus RP
AU  - Hardeman EC
AU  - Decker KP
AU  - Simoni RD
TI  - In vitro synthesis of the F0 and F1 components of the proton translocating ATPase of Escherichia coli
AB  - Specialized lambda transducing phage DNA containing the unc region of the Escherichia coli chromosome was used as template to direct an in vitro transcription/translation system. The results demonstrated synthesis of seven of the eight polypeptides of the proton translocating ATPase of this organism. The three polypeptides a, b, and c, constituting the F0 portion of the complex, were resolved by sodium dodecyl sulfatepolyacrylamide gel analysis and have apparent molecular weights (Mr = 24,000, 18,000, and 8,000-9,000) similar to the corresponding proteins produced in vivo. In addition, the alpha, beta, delta, and epsilon polypeptides of the F1 portion of the ATPase were also detected and their molecular weights correspond to the in vivo peptides. A 4.3-kilobase HindIII-generated lambda unc DNA fragment was cloned onto plasmid vectors and was demonstrated to contain the genes for the three F0 and two of the F1 (alpha, delta) subunits. In addition, the polypeptides synthesized in vitro were precipitable with antibody prepared against purified F1
MH  - A
MH  - Adenine Nucleotide Translocase
MH  - Adenosinetriphosphatase
MH  - alpha
MH  - analysis
MH  - ATPase
MH  - BETA
MH  - COMPLEX
MH  - delta
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - F1
MH  - In Vitro
MH  - protein
MH  - Proteins
MH  - proton
MH  - Proton-Translocating ATPases
MH  - Sodium
MH  - SUBUNIT
MH  - synthesis
MH  - SYSTEM
RP  - NOT IN FILE
NT  - UI - 81142308LA - engRN - 0 (Antibodies)RN - 0 (Plasmids)RN - EC 2.7.7 (Nucleotidyltransferases)RN - EC 2.7.7.- (Adenine Nucleotide Translocase)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleID - GM 18539/GM/NIGMSID - GM07598/GM/NIGMSID - GM07781/GM/NIGMSDA - 19810526IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:6259150
SO  - J Biol Chem 1981 Apr 10 ;256(7):3141-3144

408
UI  - 848
AU  - Clark AJ
AU  - Jackson JB
TI  - The measurement of membrane potential during photosynthesis and during respiration in intact cells of Rhodopseudomonas capsulata by both electrochromism and by permeant ion redistribution
AB  - 1. The membrane potential in intact cells of Rhodopseudomonas capsulata during photosynthesis and during dark respiration has been measured by two independent methods. 2. The light-induced and O2-induced shifts in the carotenoid absorption spectrum were measured in the intact cells. The shift was calibrated with K+-diffusion potentials in chromatophores derived from those cells. The light-induced and O2-induced membrane potentials were -290 mV and -230 mV respectively. 3. The energized uptake of butyltriphenylphosphonium ions was measured in the same batch of cells. The light-induced and O2-induced membrane potentials calculated from the Nernst equation were -160 mV and -120 mV respectively. 4. It is concluded that the two kinds of probe measure the electric potentials across different domains of the cytoplasmic membrane, but it is difficult to reconcile the existence of such domains with simple electrical analogues of the membrane and aqueous phases
RP  - NOT IN FILE
NT  - UI - 82182116LA - engRN - 0 (Ions)RN - 0 (Organophosphorus Compounds)RN - 22949-84-4 (butyltriphenylphosphonium)RN - 370-86-5 (Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone)PT - Journal ArticleDA - 19820621IS - 0264-6021SB - IMCY - ENGLANDJC - 9YO
UR  - PM:7340838
SO  - Biochem J 1981 Nov 15 ;200(2):389-397

409
UI  - 491
AU  - Cox GB
AU  - Downie JA
AU  - Langman L
AU  - Senior AE
AU  - Ash G
AU  - Fayle DR
AU  - Gibson F
TI  - Assembly of the adenosine triphosphatase complex in Escherichia coli: assembly of F0 is dependent on the formation of specific F1 subunits
AB  - A strain of Escherichia coli (AN1007) carrying the polar uncD436 allele which affects the operon coding for the F1-F0 adenosine triphosphatase (ATPase) complex was isolated and characterized. The uncD436 allele affected the two genes most distal to the operon promoter, i.e., uncD and uncC. Although the genes coding for the F0 portion of the ATPase complex were not affected in strains carrying this mutant allele, the lack of reconstitution of washed membranes by normal F1 ATPase suggested that a functional F0 might not be formed. This conclusion was supported by the observation that the 18,000-molecular-weight F0 subunit, coded for by the uncF gene, was absent from the membranes. Plasmid pAN36 (uncD+C+), when inserted into a strain carrying the uncD436 allele, resulted in the incorporation of the 18,000-molecular- weight F0 subunit into the membrane. A further series of experiments with Mu-induced polarity mutants, with and without plasmid pAN36, showed that the formation of both the alpha- and beta-subunits of F1 ATPase was an essential prerequisite to the incorporation into the membrane of the 18,000-molecular-weight F0 subunit and to the formation of a functional F0. Examination of the polypeptide composition of membranes from various unc mutants allowed a sequence for the normal assembly of the F1-F0 ATPase complex to be proposed
RP  - NOT IN FILE
NT  - UI - 82030567LA - engRN - 0 (Macromolecular Systems)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19811221IS - 0021-9193SB - IMCY - UNITED STATESJC - HH3
UR  - PM:6457026
SO  - J Bacteriol 1981 Oct ;148(1):30-42

410
UI  - 19878
AU  - Cross RL
TI  - The mechanism and regulation of ATP synthesis by F1-ATPases
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - COUPLING FACTOR
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - H(+)-Transporting ATP Synthase
MH  - Macromolecular Systems
MH  - mechanism
MH  - Oxidative Phosphorylation
MH  - Oxidative Phosphorylation Coupling Factors
MH  - Phosphates
MH  - Phosphorylation
MH  - regulation
MH  - review
MH  - SYNTHASE
MH  - synthesis
RP  - NOT IN FILE
NT  - UI - 81280523LA - engRN - 0 (Macromolecular Systems)RN - 0 (Oxidative Phosphorylation Coupling Factors)RN - 0 (Phosphates)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewID - GM 23152/GM/NIGMSDA - 19811014IS - 0066-4154SB - IMCY - UNITED STATESJC - 6DJ
UR  - PM:6455964
SO  - Annu Rev Biochem 1981  ;50():681-714

411
UI  - 21208
AU  - Drachev LA
AU  - Kaulen AD
AU  - Skulachev VP
AU  - Khitrina LV
AU  - Chekulaeva LN
TI  - [Phases of photoelectrical response of bacteriorhodopsin]
AB  - The generation of electrical potential difference by bacteriorhodopsin sheets incorporated into collodium film impregnated with a lecithine solution in decane was studied. The electrical response to a short light flash inducing a single turnover of bacteriorhodopsin consists of four phases. Phase I of photoresponse (tau less than 0.2 mks) has a small amplitude and is directed oppositely to the electrical response to continuous light. The negative phase is followed by positive phases, i.e. microsecond (II) and millisecond (III) ones, either of which can be decomposed at least into two exponents. Phase IV of the electrical response is a passive discharge of capacity of the "bacteriorhodopsin sheets--collodium film" system. Although phases II and III correspond to the generation and disappearance of the photochemical cycle intermediate with an absorption maximum at 412 nm, the parameters and activation energy of these two processes do not show complete coincidence. The effects of gramicidin A and the trifluoromethoxycarbonylcyanidephenylhydrazone uncoupler on the photoelectrical response were analyzed. The data obtained are discussed in terms of an equivalent electrical scheme, according to which the bacteriorhodopsin sheets form closed vesicles upon being incorporated into collodium film
MH  - A
MH  - absorption
MH  - ACTIVATION
MH  - Bacteriorhodopsin
MH  - carotenoid
MH  - Carotenoids
MH  - flash
MH  - intermediate
MH  - Light
MH  - microsecond
MH  - Phosphatidylcholines
MH  - SYSTEM
MH  - vesicles
RP  - NOT IN FILE
NT  - UI - 81256760LA - rusRN - 0 (Phosphatidylcholines)RN - 36-88-4 (Carotenoids)RN - 53026-44-1 (Bacteriorhodopsins)PT - Journal ArticleDA - 19811029IS - 0320-9725SB - IMCY - USSR
UR  - PM:7260203
SO  - Biokhimiia 1981 Jun ;46(6):998-1005

412
UI  - 21206
AU  - Drachev LA
AU  - Kalamkarov GR
AU  - Kaulen AD
AU  - Ostrovsky MA
AU  - Skulachev VP
TI  - Fast stages of photoelectric processes in biological membranes. II. Visual rhodopsin
AB  - The functioning of visual rhodopsin as a photoelectric generator has been demonstrated with a direct method. Photoreceptor discs were incorporated into a phospholipid-impregnated collodion film. Illumination of the resulting system with continuous light was found to induce formation of an electric potential (the disc-free side positive) that was measured with two electrodes separated by the film. A photopotential exceeding 40 mV was shown. It dissipated before the light source was switched off. A 15 ns 530-nm laser flash induced the formation of a photopotential of up to 35 mV whose appearance was preceded with a small oppositely directed electrogenic phase. This "negative" photoresponse took less than 200 ns. The "positive" photoresponse was composed of at least two phases (t 1/2 about 500 microseconds and several milliseconds). The latter was shown to correlate with formation of metarhodopsin II. A 347-nm laser flash added after a 530-nm flash resulted in a photoelectric effect similar to that initiated by 530-nm flash but of opposite direction. The 347-nm response was completely abolished by hydroxylamine preventing the accumulation of metarhodopsin II. The response at 530 nm proved to be hydroxylamine-resistant. Both the amplitude and the decay time of the flash-induced potential were maximal in the response to the first flash, each subsequent flash being less effective than the preceding one. Flashes were found to cause acceleration of the photopotential decay. The latter effect proved to be due to a increase of membrane conductance that developed faster than in 50 ms. Addition of 11-cis retinal after illumination improved the amplitude of the photoresponse but not the conductance. The light-induced increase in conductance was insensitive to hydroxylamine. It is suggested that a function of visual rhodopsin consists of generating a potential difference across the photoreceptor disc membrane which responds with a increase in membrane permeability to a rise of the membrane potential. A possible role of an electric break-down of the membrane, induced by the rhodopsin-generated local or partially delocalized electric field has been discussed
MH  - A
MH  - conductance
MH  - Electrodes
MH  - electrogenic
MH  - FIELD
MH  - flash
MH  - function
MH  - Light
MH  - membrane
MH  - Membrane Potential
MH  - Membranes
MH  - method
MH  - microsecond
MH  - Permeability
MH  - Phospholipids
MH  - pigments
MH  - retinal
MH  - SYSTEM
MH  - Time
RP  - NOT IN FILE
NT  - UI - 82027166LA - engRN - 0 (Phospholipids)RN - 0 (Retinal Pigments)RN - 9004-70-0 (Collodion)RN - 9009-81-8 (Rhodopsin)PT - Journal ArticleDA - 19811215IS - 0014-2956SB - IMCY - GERMANY, WEST
UR  - PM:7285901
SO  - Eur J Biochem 1981 Jul ;117(3):471-481

413
UI  - 21209
AU  - Drachev LA
AU  - Kaulen AD
AU  - Skulachev VP
AU  - Khitrina LV
AU  - Chekulaeva LN
TI  - [Some features of photochemical conversions of bacteriorhodopsin at low pH values]
AB  - Association of bacteriorhodopsin sheets and proteoliposomes with collodium film impregnated with a lecithine solution in decane was carried out. The generation of electrical potential difference in response to a light flash within the pH range of 0-6 was studied. The amplitude of microsecond (II) and millisecond (III) phases of the photoelectrical response was shown to decline with a decrease in pH as in the case when the inhibition by acid is due to protonation of a group with a pK of 3,7. The photoelectrical response of the "acid" form of bacteriorhodopsin with a maximun at 605 nm is represented by a negative phase having a small amplitude. This phase is coupled with the formation of the photochemical cycle bathointermediate and is sharply increased in an acid medium simultaneously with the generation of bacteriorhodopsin form with an absorption maximum at 565 nm. The effects of KCl, phenyldicarbaundecaborane and fluoride on the spectral and photoelectrical responses of bacteriorhodopsin are described
MH  - A
MH  - absorption
MH  - ACID
MH  - Bacteriorhodopsin
MH  - carotenoid
MH  - Carotenoids
MH  - flash
MH  - Light
MH  - microsecond
MH  - pH
MH  - Proteolipids
MH  - proteoliposome
MH  - protonation
RP  - NOT IN FILE
NT  - UI - 82046921LA - rusRN - 0 (Proteolipids)RN - 0 (proteoliposomes)RN - 36-88-4 (Carotenoids)RN - 53026-44-1 (Bacteriorhodopsins)PT - Journal ArticleDA - 19820120IS - 0320-9725SB - IMCY - USSR
UR  - PM:7295814
SO  - Biokhimiia 1981 May ;46(5):897-903

414
UI  - 21207
AU  - Drachev LA
AU  - Kaulen AD
AU  - Khitrina LV
AU  - Skulachev VP
TI  - Fast stages of photoelectric processes in biological membranes. I. Bacteriorhodopsin
AB  - Bacteriorhodopsin-containing fragments of Halobacterium halobium membrane (bacteriorhodopsin sheets) were incorporated into a lecithin- impregnated collodion film, and fast stages of flash-induced electrogenesis were measured by two electrodes separated by this film. It is found that a single turnover of bacteriorhodopsin results in an electrogenic response composed of three main stages of the following tau: the first less than 200 ns, the second 15 - 70 microseconds and the third 10 ms. The second and third phases are of the same direction as an electric response to continuous illumination, whereas the first one is oppositely directed. The microseconds and ms stages were shown to correlate, in the first approximation, with formation and decomposition of the bacteriorhodopsin intermediate absorbing with 412 nm, respectively. Both the second and third phases of the photoelectric response are sums of at least two exponents. The third stage is specifically inhibition by La3+ ions which are also shown to decrease the rate of regeneration of the original bacteriorhodopsin absorbing at 570 nm from the intermediate absorbing at 412 nm. Acidification of the medium induces parallel inhibition of the second and third phases and of formation of the intermediate absorbing at 412 nm as if protonation of a group with pK = 3.6 were responsible for this inhibition. The first (opposite) phase survives acidification. It even increases at pH lower than 1.5. At such a low pH, one can show a good correlation of decays of photopotential and of a bacteriorhodopsin bathointermediate. The decays are biphasic (tau 1 = 200 microseconds and tau 2 = 2 ms), formation of both the photopotential and the bathointermediate being faster than 200 ns. At higher pH, when a three-phase photoelectric response is revealed, decay of the formed electric potential difference gives the average tau value of about 1 s. It can be accelerated by compounds that increase ionic conductance of biomembranes. At pH below 4, fluoride is found to completely inhibit the second and third phases, so that only the first phase is observed. The results are discussed in terms of a scheme postulating that the first electrogenic phase is a result of translocation of the protonated Schiff base inside the membrane due to a light-induced conformation change in retinal or protein. The second and third phases are explained by H+ transfer from the Schiff base to the outer membrane surface and from inner (cytoplasmic) surface of membrane to the Schiff base, respectively
MH  - A
MH  - Bacteriorhodopsin
MH  - BASE
MH  - carotenoid
MH  - Carotenoids
MH  - conductance
MH  - conformation
MH  - Electrodes
MH  - electrogenic
MH  - H+
MH  - Halobacterium
MH  - intermediate
MH  - ion
MH  - Ions
MH  - membrane
MH  - Membranes
MH  - microsecond
MH  - pH
MH  - protein
MH  - protonation
MH  - retinal
MH  - Schiff base
MH  - Schiff-base
MH  - SURFACE
MH  - TRANSFER
MH  - translocation
RP  - NOT IN FILE
NT  - UI - 82027165LA - engRN - 0 (Fluorides)RN - 36-88-4 (Carotenoids)RN - 53026-44-1 (Bacteriorhodopsins)RN - 7439-91-0 (Lanthanum)RN - 9004-70-0 (Collodion)PT - Journal ArticleDA - 19811215IS - 0014-2956SB - IMCY - GERMANY, WEST
UR  - PM:7285900
SO  - Eur J Biochem 1981 Jul ;117(3):461-470

415
UI  - 964
AU  - Ernster L
AU  - Schatz G
TI  - Mitochondria: a historical review
MH  - Adenosine Triphosphate
MH  - Animal
MH  - biosynthesis
MH  - Cell Fractionation
MH  - DNA,Mitochondrial
MH  - Electron Transport
MH  - Energy Metabolism
MH  - genetics
MH  - Human
MH  - Intracellular Membranes
MH  - Ions
MH  - metabolism
MH  - Mitochondria
MH  - Morphogenesis
MH  - Nad
MH  - Oxidation-Reduction
MH  - Oxidative Phosphorylation
MH  - Oxygen Consumption
MH  - physiology
MH  - Proteins
MH  - Submitochondrial Particles
MH  - ultrastructure
RP  - NOT IN FILE
SO  - J Cell Biol 1981 Dec ;91(3 Pt 2):227s-255s

416
UI  - 782
AU  - Esch FS
AU  - Bohlen P
AU  - Otsuka AS
AU  - Yoshida M
AU  - Allison WS
TI  - Inactivation of the bovine mitochondrial F1-ATPase with dicyclohexyl[14C]carbodiimide leads to the modification of a specific glutamic acid residue in the beta subunit
AB  - When the bovine mitochondrial F1-ATPase is inactivated with dicyclohexyl[14C]carbodiimide and then gel-filtered, from 2 to 3 g atoms of 14C are incorporated/mol of enzyme. Prior inactivation of the enzyme by the modification of an essential tyrosine residue with 4- chloro-7-nitrobenzofurazan, a reaction that can be reversed by thiols, does not affect the irreversible inactivation of the ATPase by dicyclohexyl[14C]carbodiimide. During the large scale modification of the F1-ATPase by dicyclohexyl[14C]carbodiimide which led to 70% inactivation, 1.9 g atoms of 14C were incorporated/mol of enzyme. Isolation of the alpha, beta, and gamma subunits from this large scale inactivation revealed that the gram atoms of 14C bound per mol of each of the subunits was: alpha, 0.04; beta, 0.56; and gamma, 0.04. The majority of the radioactivity in a cyanogen bromide digest of the 14C- labeled beta subunit was isolated in a fragment that has the following amino acid sequence: Glu-Leu-Ile-Asn-Asn-Val-Ala-Lys-Ala-His-Gly-Gly- Tyr-Ser-Val-Phe-Ala-Gly-Val-Gly -Glu-Arg-Thr-Arg-Glu-Gly-Asn-Asp-Leu- Tyr-Glu*-His-Met; where Glu* represents the N gamma-glutamyl derivative of dicyclohexyl[14C]urea
RP  - NOT IN FILE
NT  - UI - 81264353LA - engRN - 0 (Amino Acids)RN - 0 (Carbodiimides)RN - 0 (Carbon Radioisotopes)RN - 0 (Glutamates)RN - 0 (Macromolecular Systems)RN - 0 (Oxidative Phosphorylation Coupling Factors)RN - 0 (Peptide Fragments)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-86-0 (Glutamic Acid)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM-16974/GM/NIGMSDA - 19811029IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:6114957
SO  - J Biol Chem 1981 Sep 10 ;256(17):9084-9089

417
UI  - 660
AU  - Frangione B
AU  - Rosenwasser E
AU  - Penefsky HS
AU  - Pullman ME
TI  - Amino acid sequence of the protein inhibitor of mitochondrial adenosine triphosphatase
AB  - The complete amino acid sequence of the mitochondrial ATPase inhibitor peptide was determined. The molecule contains 84 residues of which 40 are charged amino acids that occur in clusters along the chain. A section of the chain, located at the COOH-terminal end, contains several duplicated regions, the most prominent of which are pentapeptides. This section of the chain also contains all of the five histidines present in the molecule. Some of the physicochemical properties of the protein and an improved purification procedure are described
RP  - NOT IN FILE
NT  - UI - 82150878LA - engRN - 0 (ATPase inhibitory protein)RN - 0 (Proteins)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleID - AM 01431/AM/NIADDKID - GM 26924/GM/NIGMSDA - 19820521IS - 0027-8424SB - IMCY - UNITED STATESJC - PV3
UR  - PM:6461003
SO  - Proc Natl Acad Sci U S A 1981 Dec ;78(12):7403-7407

418
UI  - 977
AU  - Friedl P
AU  - Schairer HU
TI  - The isolated F0 of Escherichia coli aTP-synthase is reconstitutively active in H+-conduction and ATP-dependent energy-transduction
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - Cell-Free System
MH  - Electric Conductivity
MH  - Energy Metabolism
MH  - enzymology
MH  - Escherichia coli
MH  - Macromolecular Systems
MH  - Membrane Potentials
MH  - metabolism
MH  - Multienzyme Complexes
MH  - Phospholipids
MH  - Phosphotransferases
MH  - Protons
MH  - Structure-Activity Relationship
RP  - NOT IN FILE
SO  - FEBS Lett 1981 Jun 15 ;128(2):261-264

419
UI  - 593
AU  - Galante YM
AU  - Wong SY
AU  - Hatefi Y
TI  - Mitochondrial adenosinetriphosphatase inhibitor protein: reversible interaction with complex V (ATP synthetase complex)
AB  - Mitochondrial ATPase inhibitor protein (IF1) reacts reversibly with complex V and inhibits up to 90% of its ATPase activity. Both the rate and extent of inhibition are pH and temperature dependent and increase as the pH is lowered from pH 8 tp 6.7 (the lowest pH examined) or as the temperature is increased from 4 to 36 degrees C. Nucleotide triphosphates plus Mg2+ ions are required for inhibition of complex V ATPase activity by IF1. In the presence of Mg2+ ions, the effectiveness order of nucleotides is ATP greater than ITP greater than GTP greater than UTP. Highly purified complex V, which requires added phospholipids for expressing ATPase and ATP-Pi exchange activities, cannot be inhibited by IF1 plust ATP-Mg2+ unless phospholipids are also added. This indicates that the active state of the enzyme is necessary for the IF1 effect to be manifested, because F1-ATPase, which does not contain nor require phospholipids for catalyzing ATP hydrolysis, can be inhibited by IF1 plus ATP-Mg2+ in the absence of added phospholipids. The IF1-inhibited complex V, but not IF1-inhibited F1-ATPase, can be reactivated by incubation at pH greater than 7.0 in the absence of ATP- Mg2+. The reactivation rate is pH dependent and is influenced by temperature and enzyme concentration. Complex V preparations contain small and variable amounts of IF1. This endogenous IF1 behaves the same as added IF1 with respect to conditions described above for inhibition and reactivation and can result in 25-50% inhibition in different complex V preparations. However, complex V lacking endogenous IF1 can be reconstituted from F0, F1, oligomycin sensitivity conferring protein, and phospholipids. Inhibition of this reconstituted preparation in the presence of ATP-Mg2+ depends entirely on addition of IF1. In general, the ATP-Pi exchange activity of complex V is more sensitive to the chemical inhibitors of F1-AtPase tha its ATPase activity. This is not so, however, for IF1. Under conditions that IF1 caused approximately 75% inhibition of ATPase activity of complex V, no more than 10% of the ATP-Pi exchange activity was inhibited
RP  - NOT IN FILE
NT  - UI - 81208147LA - engRN - 0 (ATPase inhibitory protein)RN - 0 (Multienzyme Complexes)RN - 0 (Plant Proteins)RN - 0 (Ribonucleotides)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 2.7 (Phosphotransferases)RN - EC 2.7.4.- (ATP synthetase complex)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - AM 08126/AM/NIADDKDA - 19810827IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:6263316
SO  - Biochemistry 1981 Apr 28 ;20(9):2671-2678

420
UI  - 590
AU  - Galante YM
AU  - Wong SY
AU  - Hatefi Y
TI  - Resolution and reconstitution of complex V of the mitochondrial oxidative phosphorylation system: properties and composition of the membrane sector
RP  - NOT IN FILE
NT  - UI - 82066858LA - engRN - 0 (Bromides)RN - 0 (Carrier Proteins)RN - 0 (Membrane Proteins)RN - 0 (Multienzyme Complexes)RN - 0 (Oxidative Phosphorylation Coupling Factors)RN - 0 (complex V (mitochondrial oxidative phosphorylation system))RN - 0 (oligomycin sensitivity-conferring protein)RN - 1404-59-7 (Rutamycin)RN - 57-13-6 (Urea)RN - 7440-23-5 (Sodium)RN - 7647-15-6 (sodium bromide)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - AM 08126/AM/NIADDKDA - 19820109IS - 0003-9861SB - IMCY - UNITED STATESJC - 6SK
UR  - PM:6458242
SO  - Arch Biochem Biophys 1981 Oct 15 ;211(2):643-651

421
UI  - 381
AU  - Gay NJ
AU  - Walker JE
TI  - The atp operon: nucleotide sequence of the region encoding the alpha- subunit of Escherichia coli ATP-synthase
AB  - Part of the atp (or unc) operon encoding the alpha, beta, gamma, delta, and epsilon subunits of Escherichia coli ATP-synthase has been cloned into the plasmid pACYC 184. The DNA coding for the largest of these proteins, the alphas subunit, has been sequenced by cloning into the bacteriophage M13 and sequencing with dideoxy nucleotide chain terminators. It comprises 1539 nucleotides corresponding to a protein of 513 amino acids
RP  - NOT IN FILE
NT  - UI - 82059532LA - engRN - 0 (DNA, Bacterial)RN - 0 (Multienzyme Complexes)RN - 0 (Plasmids)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 2.7 (Phosphotransferases)RN - EC 2.7.4.- (ATP synthetase complex)RN - EC 3.1.21 (DNA Restriction Enzymes)RN - EC 3.1.21.- (Deoxyribonuclease EcoRI)RN - EC 3.1.21.- (endodeoxyribonuclease HpaI)PT - Journal ArticleDA - 19820109IS - 0305-1048SB - IMCY - ENGLANDJC - O8L
UR  - PM:6272228
SO  - Nucleic Acids Res 1981 May 11 ;9(9):2187-2194

422
UI  - 380
AU  - Gay NJ
AU  - Walker JE
TI  - The atp operon: nucleotide sequence of the promoter and the genes for the membrane proteins, and the delta subunit of Escherichia coli ATP- synthase
RP  - NOT IN FILE
NT  - UI - 82059437LA - engRN - 0 (Bacterial Proteins)RN - 0 (Codon)RN - 0 (Membrane Proteins)RN - 0 (Multienzyme Complexes)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 2.7 (Phosphotransferases)RN - EC 2.7.4.- (ATP synthetase complex)PT - Journal ArticleDA - 19820109IS - 0305-1048SB - IMCY - ENGLANDJC - O8L
UR  - PM:6272190
SO  - Nucleic Acids Res 1981 Aug 25 ;9(16):3919-3926

423
UI  - 999
AU  - Gingold MP
AU  - Rigaud JL
AU  - Champeil P
TI  - Fluorescence energy transfer between ATPase monomers in sarcoplasmic reticulum reconstituted vesicles
MH  - Adenosinetriphosphatase
MH  - Energy Transfer
MH  - enzymology
MH  - In Vitro
MH  - metabolism
MH  - Sarcoplasmic Reticulum
MH  - Spectrometry,Fluorescence
RP  - NOT IN FILE
SO  - Biochimie 1981 Nov ;63(11-12):923-925

424
UI  - 592
AU  - Godinot C
AU  - Gautheron DC
AU  - Galante Y
AU  - Hatefi Y
TI  - Labeling of thiols involved in the activity of complex V of the mitochondrial oxidative phosphorylation system
RP  - NOT IN FILE
NT  - UI - 81215658LA - engRN - 0 (Carrier Proteins)RN - 0 (Chloromercuribenzoates)RN - 0 (Membrane Proteins)RN - 0 (Multienzyme Complexes)RN - 0 (Nicotinic Acids)RN - 0 (Oxidative Phosphorylation Coupling Factors)RN - 0 (Phosphates)RN - 0 (Sulfhydryl Reagents)RN - 0 (complex V (mitochondrial oxidative phosphorylation system))RN - 15658-35-2 (6,6'-dithiodinicotinic acid)RN - 56-65-5 (Adenosine Triphosphate)RN - 59-85-8 (p-Chloromercuribenzoic Acid)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleID - AM 08126/AM/NIADDKDA - 19810827IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:6453870
SO  - J Biol Chem 1981 Jul 10 ;256(13):6776-6782

425
UI  - 1182
AU  - Graber P
TI  - Energization of thylakoid membranes by different pulse methods and their relevance to initial ATP synthesis: the reversibility of the DTT-modified ATPase
MH  - atp
MH  - ATP synthesis
MH  - ATPase
MH  - membrane
MH  - Membranes
MH  - method
MH  - Methods
MH  - synthesis
MH  - thylakoid
MH  - thylakoid membrane
RP  - ON REQUEST (03/18/92)
SO  - Dev Biochem 1981  ;20():119-128

426
UI  - 1187
AU  - Graber P
AU  - Schlodder E
TI  - Regulation of the rate of ATP synthesis/hydrolysis by .DELTA.pH and .DELTA..psi
MH  - atp
MH  - regulation
T2  - Photosynth., Proc. Int. Congr., 5th, Meeting Date 1980, Volume 2, 867-79. Edited by: Akoyunoglou, George. Balaban Int. Sci. Serv.: Philadelphia, Pa
Y2  - -32676  
RP  - ON REQUEST (03/18/92)
SO  -  1981  ;():

427
UI  - 1190
AU  - Graber P
AU  - Burmeister M
AU  - Hortsch M
TI  - Regulation of the membrane permeability of spinach chloroplasts by binding of adenine nucleotides
MH  - Adenine Nucleotides
MH  - ADENINE-NUCLEOTIDES
MH  - BINDING
MH  - chloroplast
MH  - Chloroplasts
MH  - membrane
MH  - Nucleotides
MH  - Permeability
MH  - regulation
MH  - Spinach
MH  - SPINACH-CHLOROPLASTS
RP  - ON REQUEST (03/18/92)
SO  - FEBS Lett 1981  ;136():25-31

428
UI  - 21022
AU  - Grber P
AU  - Burmeister M
AU  - Hortsch M
TI  - Regulation of the Membrane Permeability of spinach chloroplasts by binding of Adenine Nucleotides
MH  - Adenine Nucleotides
MH  - ADENINE-NUCLEOTIDES
MH  - atp
MH  - BINDING
MH  - chloroplast
MH  - Chloroplasts
MH  - membrane
MH  - Nucleotides
MH  - Permeability
MH  - pmf
MH  - regulation
MH  - slip
MH  - Spinach
MH  - SPINACH-CHLOROPLASTS
RP  - IN FILE
SO  - FEBS Lett 1981  ;136():25-31

429
UI  - 1432
AU  - Grber P
AU  - Trissl H
TI  - On the rise time and polarity of the photovoltage generated by light gradients in chloroplast suspensions
MH  - A
MH  - absorption
MH  - action spectra
MH  - blebs
MH  - Chlorophyll
MH  - chloroplast
MH  - Chloroplasts
MH  - DCMU
MH  - flash
MH  - Kinetics
MH  - Light
MH  - light-gradient
MH  - M
MH  - photoelectric measurements
MH  - PMS
MH  - RISE TIME
MH  - spectra
MH  - Sucrose
MH  - Time
RP  - IN FILE
NT  - From measurements of the time course of the photovoltage from a chloroplast suspension (1M NaCl; excitation laser flash 530 nm), the half rise time of the photovoltage was calculated ad .ltoreq. 5 ns. The photovoltage was sensitive to DCMU and PMS. With chloroplasts suspended in 2.3 M sucrose, the polarity and kinetics of the signals depended on the excitation wavelength, whereas with swollen chloroplasts (blebs), the polarities were always pos. The complete action spectrum of the photovoltage (0.4 M sucrose) showed that pos. signals matched the blue and red absorption peaks of chlorophyll, whereas neg. signals were obsd. at 510-560 nm and 700-740 nm. Both wavelength regions of positive polarity overlap with regions of negative polarity
AV  - Hawi / light-gradient
SO  - FEBS Lett 1981  ;123():95-99

430
UI  - 647
AU  - Grubmeyer C
AU  - Penefsky HS
TI  - Cooperatively between catalytic sites in the mechanism of action of beef heart mitochondrial adenosine triphosphatase
AB  - Occupancy of only one of two hydrolytic sites on beef heart mitochondrial ATPase (F1) by the radioactive ATP analog, 2',3'-O-(2,4,6- trinitrophenyl) adenosine 5'-[gamma-32P]-triphosphate (TNP-[gamma- 32P]ATP) is associated with a low rate of hydrolysis of the substrate even under conditions otherwise favoring catalysis. Addition of excess nonradioactive TNP-ATP, in concentrations sufficient to fill catalytic Site 2 on the enzyme (Grubmeyer, C., and Penefsky, H. S. (1981) J. Biol. Chem. 256, 3718-3727), accelerates the rate of hydrolysis of the radioactive substrate 15- to 20-fold. Since the excess nonradioactive substrate serves as an effective isotope trap, the involvement of medium TNP-[gamma-32P]-ATP as an intermediate is ruled out. These observations constitute direct evidence for catalytic cooperativity between active sites on F1. It is proposed that the use of high binding affinity substrates or substrate analogs, combined with the isotope trap technique, offers a new approach to the detection and study of catalytic site cooperativity in enzymes. The hydrolyzable nucleotides GTP, ITP, and ATP are excellent promoters of the hydrolysis of previously bound TNP-[gamma-32P]ATP whereas addition of nonhydrolyzable nucleotides such as TNP-ADP, ADP, and adenylyl imidodiphosphate result in a lower rate and extent of hydrolysis. AMP is without effect. Studies of the hydrolysis of [gamma-32P]ATP and TNP-[gamma-32P]ITP, under appropriate conditions, also provide evidence consistent with promoted catalysis. Based upon these findings, a model is presented for the mechanism of action of F1 in which site-site cooperativity reflects promoter-dependent hydrolysis of bound substrate
RP  - NOT IN FILE
NT  - UI - 81168136LA - engRN - 0 (Oxidative Phosphorylation Coupling Factors)RN - 0 (Trinitrobenzenes)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM 21737/GM/NIGMSDA - 19810623IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:6452455
SO  - J Biol Chem 1981 Apr 25 ;256(8):3728-3734

431
UI  - 648
AU  - Grubmeyer C
AU  - Penefsky HS
TI  - The presence of two hydrolytic sites on beef heart mitochondrial adenosine triphosphatase
AB  - The ribose-modified nucleotides 2',3'-O-(2,4,6-trinitrophenyl) adenosine 5'-triphosphate (TNP-ATP) and TNP-ADP were used to probe the catalytic sites on soluble beef heart mitochondrial adenosine triphosphatase (F1). Both compounds were potent competitive inhibitors of ATP hydrolysis catalyzed by F1, Ki = 5.5 and 10 nM, respectively, and by submitochondrial particles, Ki (TNP-ATP) = 21 nM. Both compounds also were potent competitive inhibitors of ATP synthesis during oxidative phosphorylation (Ki = 1300 nM). Both analogs inhibited the 32Pi-ATP exchange reaction and the ATP-dependent reduction of NAD+ by succinate, catalyzed by submitochondrial particles. TNP-ATP and TNP-ADP were bound by F1. The presence of two binding sites on the enzyme for TNP-adenine nucleotides was determined by titrations of difference absorbance spectra, of the increase in fluorescence of the analog which occurred upon interaction with protein, and by titrations with the centrifuge column method using 32P-labeled TNP-adenine nucleotides. The first binding site bound the analogs with an affinity too high to be measured. The Kd for analog binding by the second site was 20 to 80 nM. In the presence of Mg2+, the 2 sites were filled with the TNP-ATP at a rate too rapid to be resolved by the procedure used. TNP-[gamma-32P]ATP was hydrolyzed by F1, Km = 0.2 microM, Vmax = 1.1 mol of 32Pi formed/mol of F1/s. It was shown, using the isotope trap technique as well as the inhibitor efrapeptin, that the 2 binding sites for TNP-ATP on F1 are hydrolytic sites
RP  - NOT IN FILE
NT  - UI - 81168135LA - engRN - 0 (Adenine Nucleotides)RN - 0 (Oxidative Phosphorylation Coupling Factors)RN - 0 (Trinitrobenzenes)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM 21737/GM/NIGMSDA - 19810623IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:6452454
SO  - J Biol Chem 1981 Apr 25 ;256(8):3718-3727

432
UI  - 21104
AU  - Guffanti AA
AU  - Bornstein RF
AU  - Krulwich TA
TI  - Oxidative phosphorylation by membrane vesicles from Bacillus alcalophilus
AB  - ADP and Pi-loaded membrane vesicles from t-malate-grown Bacillus alcalophilus synthesized ATP upon energization with ascorbate/N,N,N',N'- tetra-methyl-P-phenylenediamine. ATP synthesis occurred over a range of external pH from 6.0 to 11.0, under conditions in which the total protonmotive force delta-mu-H+ was as low as -30 mV. The phosphate potentials (delta Gp) were calculated to be 11 and 12 kcal/mol at pH 10.5 and 9.0, respectively, whereas the delta-mu-H+ values in vesicles at these two pH values were quite different (-40 +/- 20 mV at pH 10.5 and -125 +/- 20 mV at pH 9.0). ATP synthesis was inhibited by KCN, gramicidin, and by N,N1-dicyclohexylcarbodiimide. Inward translocation of protons, concomitant with ATP synthesis, was demonstrated using direct pH monitoring and fluorescence methods. No dependence upon the presence of Na+ or K+ was found. Thus, ATP synthesis in B. alcalophilus appears to involve a proton-translocating ATPase which functions at low delta-mu-H+
MH  - A
MH  - ADP
MH  - atp
MH  - ATP synthesis
MH  - ATPase
MH  - Bacillus
MH  - delta
MH  - DEPENDENCE
MH  - Dicyclohexylcarbodiimide
MH  - fluorescence
MH  - function
MH  - membrane
MH  - membrane vesicles
MH  - method
MH  - Methods
MH  - Oxidative Phosphorylation
MH  - pH
MH  - Phosphorylation
MH  - Potassium
MH  - proton
MH  - Protons
MH  - synthesis
MH  - translocation
MH  - vesicles
RP  - NOT IN FILE
NT  - UI - 81208250LA - engRN - 1405-97-6 (Gramicidin)RN - 151-50-8 (Potassium Cyanide)RN - 538-75-0 (Dicyclohexylcarbodiimide)PT - Journal ArticleID - 5 KO4 GM 00020/GM/NIGMSDA - 19810820IS - 0006-3002SB - IMCY - NETHERLANDS
UR  - PM:6165388
SO  - Biochim Biophys Acta 1981 May 13 ;635(3):619-630

433
UI  - 21103
AU  - Guffanti AA
AU  - Blumenfeld H
AU  - Krulwich TA
TI  - ATP synthesis by an uncoupler-resistant mutant of Bacillus megaterium
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthesis
MH  - Bacillus
MH  - Carbonyl Cyanide m-Chlorophenyl Hydrazone
MH  - mutant
MH  - Nitriles
MH  - Potassium
MH  - synthesis
MH  - Valinomycin
RP  - NOT IN FILE
NT  - UI - 81264247LA - engRN - 0 (Nitriles)RN - 2001-95-8 (Valinomycin)RN - 555-60-2 (Carbonyl Cyanide m-Chlorophenyl Hydrazone)RN - 56-65-5 (Adenosine Triphosphate)RN - 7440-09-7 (Potassium)PT - Journal ArticleID - 5K04GM00020/GM/NIGMSDA - 19811025IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:6790540
SO  - J Biol Chem 1981 Aug 25 ;256(16):8416-8421

434
UI  - 615
AU  - Jackson JB
AU  - Venturoli G
AU  - Baccarini-Melandri A
AU  - Melandri BA
TI  - Photosynthetic control and estimation of the optimal ATP: electron stoichiometry during flash activation of chromatophores from Rhodopseudomonas capsulata
AB  - (1) When chromatophores from Rhodopseudomonas capsulata Ala pho+ are exposed to a train of high-frequency, saturating flashes the kinetics of the reaction centre bacteriochlorophyll absorption change enter a pseudo steady-state in which the extent of oxidation during the flashes is equal to the extent of reduction in between the flashes. The level of the pseudo steady-state is lowered by the presence of a phosphate acceptor system, raised by further addition of oligomycin, lowered by a combination of nigericin and valinomycin and raised by antimycin A. (2) In the pseudo steady-state, the extent of reaction centre bacteriochlorophyll oxidation taking place during the flash may be estimated by subtraction from the total concentration of reaction centre bacteriochlorophyll. This value is equated with the amount of electrons transported through the photosynthetic chain. Comparison with the measured ATP yield per flash in the pseudo steady-state permits calculation of the ATP: two electron ratio. The value of the ratio is 1.1 for flash frequencies between 3 and 12.5 Hz and declines at lower and higher frequencies. The ATP: two electron ratio is approximately halved in the presence of antimycin A. (3) An alternative estimate of the ATP: two electron ratio, based on the assumption that high- frequency flashes approximate to the condition of continuous illumination, was approx. 0.8
RP  - NOT IN FILE
NT  - UI - 82024013LA - engRN - 0 (Bacteriochlorophylls)RN - 56-65-5 (Adenosine Triphosphate)RN - 642-15-9 (Antimycin A)PT - Journal ArticleDA - 19811215IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:7284340
SO  - Biochim Biophys Acta 1981 Jun 12 ;636(1):1-8

435
UI  - 20916
AU  - Kanazawa H
AU  - Mabuchi K
AU  - Kayano T
AU  - Noumi T
AU  - Sekiya T
AU  - Futai M
TI  - Nucleotide sequence of the genes for F0 components of the proton- translocating ATPase from Escherichia coli: prediction of the primary structure of F0 subunits
MH  - Adenosinetriphosphatase
MH  - ATPase
MH  - Enzymes
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - Macromolecular Systems
MH  - membrane
MH  - Membrane Proteins
MH  - protein
MH  - Proteins
MH  - proton
MH  - SUBUNIT
MH  - SYSTEM
MH  - SYSTEMS
RP  - NOT IN FILE
NT  - UI - 82134799LA - engRN - 0 (Codon)RN - 0 (Macromolecular Systems)RN - 0 (Membrane Proteins)RN - 0 (Plasmids)RN - EC 3.1.21 (DNA Restriction Enzymes)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19820412IS - 0006-291XSB - IMCY - UNITED STATES
UR  - PM:6277311
SO  - Biochem Biophys Res Commun 1981 Nov 30 ;103(2):613-620

436
UI  - 198
AU  - Kanazawa H
AU  - Mabuchi K
AU  - Kayano T
AU  - Tamura F
AU  - Futai M
TI  - Nucleotide sequence of genes coding for dicyclohexylcarbodiimide- binding protein and the alpha subunit of proton-translocating ATPase of Escherichia coli
MH  - Adenosinetriphosphatase
MH  - ALPHA-SUBUNIT
MH  - ATPase
MH  - BINDING
MH  - Carrier Proteins
MH  - Dicyclohexylcarbodiimide
MH  - Enzymes
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - Macromolecular Systems
MH  - protein
MH  - Proteins
MH  - SUBUNIT
MH  - SYSTEM
MH  - SYSTEMS
RP  - NOT IN FILE
NT  - UI - 81255758LA - engRN - 0 (Carrier Proteins)RN - 0 (DNA, Recombinant)RN - 0 (Macromolecular Systems)RN - 0 (Plasmids)RN - 0 (dicyclohexylcarbodiimide-binding protein)RN - EC 3.1.21 (DNA Restriction Enzymes)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19810922IS - 0006-291XSB - IMCY - UNITED STATES
UR  - PM:6266400
SO  - Biochem Biophys Res Commun 1981 May 15 ;100(1):219-225

437
UI  - 194
AU  - Kanazawa H
AU  - Kayano T
AU  - Mabuchi K
AU  - Futai M
TI  - Nucleotide sequence of the genes coding for alpha, beta and gamma subunits of the proton-translocating ATPase of Escherichia coli
MH  - Adenosinetriphosphatase
MH  - ATPase
MH  - Enzymes
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - membrane
MH  - Membrane Proteins
MH  - protein
MH  - Proteins
MH  - SUBUNIT
RP  - NOT IN FILE
NT  - UI - 82134798LA - engRN - 0 (DNA, Bacterial)RN - 0 (Membrane Proteins)RN - 0 (Plasmids)RN - EC 3.1.21 (DNA Restriction Enzymes)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19820412IS - 0006-291XSB - IMCY - UNITED STATES
UR  - PM:6277310
SO  - Biochem Biophys Res Commun 1981 Nov 30 ;103(2):604-612

438
UI  - 20917
AU  - Kanazawa H
AU  - Futai M
TI  - [Proton-translocating ATPase of Escherichia coli (author's transl)]
MH  - Adenosinetriphosphatase
MH  - ATPase
MH  - Bacterial Proteins
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - protein
MH  - Proteins
MH  - proton
MH  - Protons
MH  - review
RP  - NOT IN FILE
NT  - UI - 82127997LA - jpnRN - 0 (Bacterial Proteins)RN - 0 (DNA, Bacterial)RN - 0 (Protons)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticlePT - ReviewDA - 19820420IS - 0039-9450SB - IMCY - JAPAN
UR  - PM:6276919
SO  - Tanpakushitsu Kakusan Koso 1981 Oct ;26(13):1999-2013

439
UI  - 9909
AU  - Lemasters JJ
AU  - Billica WH
TI  - Non-equilibrium thermodynamics of oxidative phosphorylation by inverted inner membrane vesicles of rat liver mitochondria.
AB  - The relationship of the phosphate potential (delta GP) to the free energy released by the oxidation of NADH (redox potential or delta GR) was studied in suspensions of inverted inner membrane vesicles prepared from rat liver mitochondria. At delta GR values less negative than -52.2 kcal/mol, delta GP was a linear function of delta GR during oxidative phosphorylation at static head. At more negative delta GR, delta GP no longer increased but remained, more or less, at a constant value. At all values of delta GR, delta GP increased as Pi decreased. At high Pi, ATP/ADP was relatively independent of Pi, but at low Pi there was a strong interdependence of ATP/ADP and Pi. The experimental data were analyzed in terms of the theory of non-equilibrium thermodynamics. The degree of coupling, q, averaged 0.8 as estimated from the dependence of respiratory rate on delta GP. From measurements of -delta GR/delta GP at static head and from the estimates of q, an average value of four was calculated for Z, the phenomenological stoichiometry. The results support a 4-proton model of chemiosmotic coupling in which proton stoichiometries are 4H+/site, 3H+/ATPase, and 1H+/translocation of ATP for ADP and Pi. The results further indicate that the site by site reactions of oxidative phosphorylation operate close to thermodynamic equilibrium. This implies that ATP/site ratios are proportional to the redox potentials across each site at static head. Based on the oxidation-reduction potentials of NADH, ubiquinone, and cytochrome c, it follows that the ideal ATP/site ratios of mitochondrial oxidative phosphorylation are 1, 1/2, and 1 1/2, respectively, for sites 1, 2, and 3.
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - ADP
MH  - Animal
MH  - atp
MH  - Carbonyl Cyanide m-Chlorophenyl Hydrazone
MH  - Cytochrome c
MH  - DEPENDENCE
MH  - drug effects
MH  - Intracellular Membranes
MH  - Kinetics
MH  - Liver
MH  - Mathematics
MH  - membrane vesicles
MH  - metabolism
MH  - Mitochondria
MH  - Mitochondria,Liver
MH  - model
MH  - Oligomycins
MH  - Oxidation-Reduction
MH  - Oxidative Phosphorylation
MH  - Oxygen Consumption
MH  - pharmacology
MH  - Phosphates
MH  - Phosphorylation
MH  - proton
MH  - Rats
MH  - Support,Non-U.S.Gov't
MH  - Support,U.S.Gov't,P.H.S.
MH  - Thermodynamics
MH  - Ubiquinone
RP  - NOT IN FILE
SO  - J Biol Chem 1981 Dec 25 ;256(24):12949-12957

440
UI  - 20918
AU  - Mabuchi K
AU  - Kanazawa H
AU  - Kayano T
AU  - Futai M
TI  - Nucleotide sequence of the gene coding for the delta subunit of proton translocating ATPase of Escherichia coli
MH  - Adenosinetriphosphatase
MH  - ATPase
MH  - coupling
MH  - COUPLING FACTOR
MH  - DELTA-SUBUNIT
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - Macromolecular Systems
MH  - Oxidative Phosphorylation
MH  - Oxidative Phosphorylation Coupling Factors
MH  - Phosphorylation
MH  - proton
MH  - SUBUNIT
MH  - SYSTEM
MH  - SYSTEMS
RP  - NOT IN FILE
NT  - UI - 82068433LA - engRN - 0 (DNA, Bacterial)RN - 0 (Macromolecular Systems)RN - 0 (Oxidative Phosphorylation Coupling Factors)RN - 0 (Plasmids)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19820120IS - 0006-291XSB - IMCY - UNITED STATES
UR  - PM:6458296
SO  - Biochem Biophys Res Commun 1981 Sep 16 ;102(1):172-179

441
UI  - 1060
AU  - Masamoto K
AU  - Matsuura K
AU  - Itoh S
AU  - Nishimura M
TI  - Surface potential dependence of the distribution of charged dye molecules onto photosynthetic membranes
MH  - DEPENDENCE
MH  - Membranes
RP  - NOT IN FILE
SO  - J Biochem (Tokyo) 1981  ;89():397-405

442
UI  - 1059
AU  - Masamoto K
AU  - Matsuura K
AU  - Itoh S
AU  - Nishimura M
TI  - Membrane-potential- and surface-potential-induced absorbance changes of merocyanine dyes added to chromatophores from Rhodopseudomonas sphaeroides
MH  - rhodopseudomonas
RP  - NOT IN FILE
SO  - Biochim Biophys Acta 1981  ;638():108-115

443
UI  - 21151
AU  - Melandri BA
AU  - Melandri AB
AU  - Venturoli G
TI  - Functional interaction between photosynthetic electron transport and ATP synthesis as revealed by inhibition studies of flash induced phosphorylation in bacterial chromatophores
MH  - atp
MH  - ATP synthesis
MH  - Bacterial Chromatophores
MH  - chromatophore
MH  - chromatophores
MH  - electron
MH  - Electron Transport
MH  - flash
MH  - Phosphorylation
MH  - synthesis
MH  - transport
RP  - ON REQUEST (03/18/92)
SO  - Dev Bioenerg Biomembr 1981  ;5():381-388

444
UI  - 20972
AU  - Paradies HH
AU  - Mertens G
AU  - Schmid R
AU  - Schneider E
AU  - Altendorf K
TI  - Molecular properties of the ATP synthetase from Escherichia coli
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - atp
MH  - COMPLEX
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - Multienzyme Complexes
MH  - Phosphotransferases
RP  - NOT IN FILE
NT  - UI - 81183968LA - engRN - 0 (Multienzyme Complexes)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 2.7 (Phosphotransferases)RN - EC 2.7.4.- (ATP Synthetase Complexes)PT - Journal ArticleDA - 19810613IS - 0006-291XSB - IMCY - UNITED STATES
UR  - PM:6261744
SO  - Biochem Biophys Res Commun 1981 Feb 12 ;98(3):595-606

445
UI  - 549
AU  - Pedersen PL
AU  - Hullihen J
AU  - Wehrle JP
TI  - Proton adenosine triphosphatase complex of rat liver. The effect of trypsin on the F1 and F0 moieties of the enzyme
RP  - NOT IN FILE
NT  - UI - 81094135LA - engRN - 0 (Macromolecular Systems)RN - 0 (Oligomycins)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 57-13-6 (Urea)RN - EC 3.4.21.4 (Trypsin)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - CA10951/CA/NCIDA - 19810327IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:6450210
SO  - J Biol Chem 1981 Feb 10 ;256(3):1362-1369

446
UI  - 21291
AU  - Prince RC
AU  - Linkletter SJG
AU  - Dutton PL
TI  - The thermodynamic properties of some commonly used oxidation- reduction mediators, inhibitors and dyes, as determined by polarography
MH  - data
MH  - DYE
MH  - dyes
MH  - electron
MH  - indicator
MH  - inhibitor
MH  - inhibitors
MH  - method
RP  - IN FILE
NT  - K 3.3
SO  - Biochim Biophys Acta 1981  ;635():132-148

447
UI  - 591
AU  - Robbins BA
AU  - Wong SY
AU  - Hatefi Y
AU  - Galante YM
TI  - Studies on the immunological properties of complex V (mitochondrial ATP synthetase complex)
RP  - NOT IN FILE
NT  - UI - 82066745LA - engRN - 0 (Antigen-Antibody Complex)RN - 0 (IgG)RN - 0 (Immune Sera)RN - 0 (Multienzyme Complexes)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 2.7 (Phosphotransferases)RN - EC 2.7.4.- (ATP synthetase complex)PT - Journal ArticleID - AM 08126/AM/NIADDKDA - 19820109IS - 0003-9861SB - IMCY - UNITED STATESJC - 6SK
UR  - PM:6272636
SO  - Arch Biochem Biophys 1981 Sep ;210(2):489-497

448
UI  - 379
AU  - Saraste M
AU  - Gay NJ
AU  - Eberle A
AU  - Runswick MJ
AU  - Walker JE
TI  - The atp operon: nucleotide sequence of the genes for the gamma, beta, and epsilon subunits of Escherichia coli ATP synthase
AB  - The nucleotide sequence of the promoter distal region of the atp (or unc) operon of Escherichia coli has been determined. It encodes the gamma, beta and epsilon subunits of the ATP-synthase complex and includes a noncoding sequence in which transcription of the operon probably terminates. This work completes the nucleotide sequence of the operon which contains nine genes: eight encode structural proteins of the ATP-synthase complex; a ninth, the first in the operon, may be a pilot for assembly. The genes for the alpha and beta subunits have evolved from a common ancestor
RP  - NOT IN FILE
NT  - UI - 82059507LA - engRN - 0 (DNA, Bacterial)RN - 0 (Multienzyme Complexes)RN - 0 (Plasmids)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 2.7 (Phosphotransferases)RN - EC 2.7.4.- (ATP synthetase complex)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19820128IS - 0305-1048SB - IMCY - ENGLANDJC - O8L
UR  - PM:6272217
SO  - Nucleic Acids Res 1981 Oct 24 ;9(20):5287-5296

449
UI  - 1186
AU  - Schatz GH
AU  - Schlodder E
AU  - Graber P
TI  - Kinetics of ATP synthesis and AdN exchange in chloroplasts measured by rapid mixing acid-base transitions
MH  - atp
MH  - ATP synthesis
MH  - chloroplast
MH  - Chloroplasts
MH  - Kinetics
MH  - synthesis
T2  - Photosynth., Proc. Int. Congr., 5th, Meeting Date 1980, Volume 2, 945-54. Edited by: Akoyunoglou, George. Balaban Int. Sci. Serv.: Philadelphia, Pa
Y2  - -32676  
RP  - ON REQUEST (03/18/92)
SO  -  1981  ;():

450
UI  - 548
AU  - Schwerzmann K
AU  - Pedersen PL
TI  - Proton--adenosinetriphosphatase complex of rat liver mitochondria: effect of energy state on its interaction with the adenosinetriphosphatase inhibitory peptide
RP  - NOT IN FILE
NT  - UI - 82069018LA - engRN - 0 (ATPase inhibitory protein)RN - 0 (Oxidative Phosphorylation Coupling Factors)RN - 0 (Proteins)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - 1-F05 TWO 2755/TW/FICID - CA 10951/CA/NCIDA - 19820212IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:6458327
SO  - Biochemistry 1981 Oct 27 ;20(22):6305-6311

451
UI  - 483
AU  - Senior AE
TI  - Divalent metals in beef heart mitochondrial adenosine triphosphatase. Demonstration of the metals in membrane-bound enzyme and studies of the interconversion of the "1-Mg" and "2-Mg" forms of the enzyme
AB  - Tight divalent metal binding sites in the beef heart mitochondrial adenosine triphosphatase were studied using the procedure of reconstitution of soluble F1 with F1-depleted membranes (SU particles). Soluble F1 has been shown previously to contain two tight-binding site for Mg. Both of these sites were present on membrane-bound enzyme. Co and Mn, substituted at the second of the two Mg-binding sites on soluble F1, became incorporated with F1 into membrane-bound enzyme. Use of radioactive Co and Mn showed that they behaved differently during short bursts of succinate oxidation or ATP hydrolysis. Co remained stably bound, whereas Mn was released to the extent of 55-80%. The results extend previous work to show that the membrane proton-ATPase is an Mg-metalloenzyme containing a structural Mg site and a second Mg site possibly involved in catalysis. The conversion of 2-Mg F1 to 1-Mg F1 during purification and storage is shown to be due to use of ammonium sulfate precipitation, and the dependence of reuptake of Mg (1- Mg F1 leads to 2-Mg F1) on nucleotides is described
RP  - NOT IN FILE
NT  - UI - 81191858LA - engRN - 7439-95-4 (Magnesium)RN - 7439-96-5 (Manganese)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM-25349/GM/NIGMSDA - 19810720IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:6453121
SO  - J Biol Chem 1981 May 25 ;256(10):4763-4767

452
UI  - 98
AU  - Skerrett KJ
AU  - Wise JG
AU  - Latchney LR
AU  - Senior AE
TI  - Trypsin cleavage of the alpha-subunit of beef heart F1-ATPase abolishes ATP synthesis and ATP-driven energy-transduction capabilities
AB  - Previous work has shown that mild trypsin treatment eliminates energy- transduction capability and tight (non-exchangeable)nucleotide binding in beef heart mitochondrial F1-ATPase (Leimgruber, R.M. and Senior, A.E. (1976) J. Biol. Chem. 251, 7103-7109). The structural change brought about by trypsin was, however, too subtle to be identified by one-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis, and was not defined. In this work we have applied two- dimensional electrophoresis (isoelectric focussing then sodium dodecyl sulfate polyacrylamide gradient electrophoresis) to the problem, and have determined that the alpha-subunit of F1 is altered by the mild trypsin treatment, whereas no change was detected in beta-, gamma-, delta- or epsilon-subunits. Binding of ADP to the trypsin-treated F1 was compared to binding to control enzyme over a range of 0-40 muM ADP in a 30 min incubation period. There was no difference between the two enzymes, KADPd in Mg2+ -containing buffer was about 2 muM in each. Since the tight (nonexchangeable)sites are abolished in trypsin-treated F1, this shows that tight exchangeable ADP-binding sites are different from the tight nonexchangeable ADP-binding sites. There was no effect of trypsin cleavage of the alpha-subunit on beta-subunit conformation as judged by aurovertin fluorescence studies. The cleavage of the alpha- subunit which occurred was judged to occur very close to the C- or N- terminus of the subunit and constitutes therefore a small and specific chemical modification which abolishes overall function in F1 but leaves partial functions intact
RP  - NOT IN FILE
NT  - UI - 82046623LA - engRN - 0 (Aurovertins)RN - 0 (Macromolecular Systems)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.4.21.4 (Trypsin)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM-25349/GM/NIGMSDA - 19820128IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:6457639
SO  - Biochim Biophys Acta 1981 Nov 12 ;638(1):120-124

453
UI  - 199
AU  - Tamura F
AU  - Kanzawa H
AU  - Tsuchiya T
AU  - Futai M
TI  - Structural gene coding for the dicyclohexylcarbodiimide-binding protein of the protein-translocating ATPase from Escherichia coli. Locus of the gene in the F1--F0 gene cluster
MH  - Adenosinetriphosphatase
MH  - ATPase
MH  - Carrier Proteins
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - membrane
MH  - Membrane Proteins
MH  - protein
MH  - Proteins
RP  - NOT IN FILE
NT  - UI - 81237097LA - engRN - 0 (Carrier Proteins)RN - 0 (Membrane Proteins)RN - 0 (Plasmids)RN - 0 (dicyclohexylcarbodiimide-binding protein)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19810922IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:6454589
SO  - FEBS Lett 1981 May 5 ;127(1):48-52

454
UI  - 1183
AU  - Tiemann R
AU  - Renger G
AU  - Graber P
TI  - Electron and proton transport in inside-out thylakoids
MH  - electron
MH  - proton
MH  - thylakoid
MH  - thylakoids
MH  - transport
T2  - Photosynth., Proc. Int. Congr., 5th, Meeting Date 1980, Volume 3, 85-95. Edited by: Akoyunoglou, George. Balaban Int. Sci. Serv.: Philadelphia, Pa
Y2  - -32676  
RP  - ON REQUEST (03/18/92)
SO  -  1981  ;():

455
UI  - 18970
AU  - Wagner R
AU  - Carrillo N
AU  - Junge W
AU  - Vallejos RH
TI  - Heat-activated conformational changes of isolated coupling factor of photophosphorylation CF1. Studies via triplet lifetime of bound eosin-isothiocyanate
MH  - atp
MH  - conformation
MH  - conformational change
MH  - COUPLING FACTOR
MH  - indicator
MH  - Photophosphorylation
RP  - NOT IN FILE
SO  - FEBS Lett 1981  ;136():208-212

456
UI  - 21226
AU  - Westerhoff HV
AU  - Hellingwerf KJ
AU  - Arents JC
AU  - Scholte BJ
AU  - Van Dam K
TI  - Mosaic nonequilibrium thermodynamics describes biological energy transduction
AB  - A procedure, called "mosaic nonequilibrium thermodynamics," for describing ion movement and energy transduction in biological membranes is tested in a model system: bacteriorhodopsin liposomes. The important steps in the theoretical derivations are summarized; one of the experimental tests of the postulated fundamental flow-force relationships is shown. Furthermore, how the quantitative method, even if used only qualitatively, facilitates analysis and understanding of experimental results (in this case, the effect of medium composition on the development of pH gradient and membrane potential in the bacteriorhodopsin liposomes) is shown. The main advantage of this method lies in its quantitative description of the effect of variation of system parameters on the performance of, in this case, the reconstituted proton pump bacteriorhodopsin. As an example, the method is shown to explain quantitatively the dependence of the steady-state pH gradient on the light intensity. Even in more refined analyses of experiments, the quantitative theoretical description is in full accordance with the experimental results; this is illustrated by considering the effect of valinomycin on the dependence of the initial rate of proton uptake into bacteriorhodopsin liposomes on light intensity. It is concluded that mosaic nonequilibrium thermodynamics describes ion movement and energy transduction in the model system of bacteriorhodopsin liposomes and, therefore, may be applied to any other biological system performing such processes
MH  - A
MH  - analysis
MH  - Bacteriorhodopsin
MH  - carotenoid
MH  - Carotenoids
MH  - DEPENDENCE
MH  - development
MH  - ion
MH  - Light
MH  - liposome
MH  - Liposomes
MH  - membrane
MH  - Membrane Potential
MH  - Membranes
MH  - method
MH  - model
MH  - Movement
MH  - pH
MH  - Phosphatidylcholines
MH  - proton
MH  - Proton Pump
MH  - Protons
MH  - SYSTEM
MH  - Thermodynamics
MH  - Valinomycin
MH  - Water
RP  - NOT IN FILE
NT  - UI - 81273096LA - engRN - 0 (Liposomes)RN - 0 (Phosphatidylcholines)RN - 0 (Protons)RN - 2001-95-8 (Valinomycin)RN - 36-88-4 (Carotenoids)RN - 53026-44-1 (Bacteriorhodopsins)RN - 7732-18-5 (Water)PT - Journal ArticleDA - 19811029IS - 0027-8424SB - IMCY - UNITED STATES
UR  - PM:6267598
SO  - Proc Natl Acad Sci U S A 1981 Jun ;78(6):3554-3558

457
UI  - 9924
AU  - Westerhoff HV
AU  - Simonetti AL
AU  - Van DK
TI  - The hypothesis of localized chemiosmosis is unsatisfactory.
AB  - The hypothesis of 'localized' chemiosmosis have been put to an experimental test in the system of oxidative phosphorylation by rat liver mitochondria. We find that the variation of the ratio of phosphate potential to delta muH with delta muH does not depend on how delta muH is varied. This is in conflict with hypotheses of localized chemiosmosis. Of all coupling hypotheses, only the parallel-coupling hypothesis can explain the observations, unless variation of the H+/ATP stoichiometry of the ATPase proton pump is accepted. In the latter case. 'macroscopic' chemiosmosis can explain the observations equally well as hypotheses of localized chemiosmosis. It is concluded that either variation of H+/ATP stoichiometries must be accepted, or that the parallel-coupling hypothesis should be reformulated so that it becomes open to experimental tests.
MH  - Animal
MH  - ATPase
MH  - In Vitro
MH  - Intracellular Membranes
MH  - Liver
MH  - Membrane Potentials
MH  - metabolism
MH  - Mitochondria
MH  - Mitochondria,Liver
MH  - Models,Biological
MH  - Osmosis
MH  - Oxidative Phosphorylation
MH  - Phosphates
MH  - Phosphorylation
MH  - proton
MH  - Proton Pump
MH  - Protons
MH  - Rats
MH  - Support,Non-U.S.Gov't
RP  - NOT IN FILE
SO  - Biochem J 1981 Nov 15 ;200(2):193-202

458
UI  - 482
AU  - Wise JG
AU  - Latchney LR
AU  - Senior AE
TI  - The defective proton-ATPase of uncA mutants of Escherichia coli. Studies of nucleotide binding sites, bound aurovertin fluorescence, and labeling of essential residues of the purified F1-ATPase
RP  - NOT IN FILE
NT  - UI - 82030801LA - engRN - 0 (Aurovertins)RN - 0 (Oxidative Phosphorylation Coupling Factors)RN - 0 (Pyrans)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM-25349/GM/NIGMSDA - 19811215IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:6457039
SO  - J Biol Chem 1981 Oct 25 ;256(20):10383-10389

459
UI  - 19815
AU  - Amory A
AU  - Goffeau A
AU  - McIntosh DB
AU  - Boyer PD
TI  - Exchange of oxygen between phosphate and water catalyzed by the plasma membrane ATPase from the yeast Schizosaccharomyces pombe
AB  - The ATPase of the plasma membrane isolated from the yeast Schizosaccharomyces pombe catalyses a medium Pi in equilibrium H2O exchange in the presence of Mg2+ and in the absence of ATP and ADP. (formula, see text) The Pi in the E.Pi species tumbles in the active site so that each of its oxygens has an equal probability of exchange with water. The partition coefficient (Pc = k2/k2 + k-1) is 0.45. The total rate of oxygen exchange, Vex, representing the rate of incorporation of water oxygens occurring during hydrolysis of E--P into E.Pi (Vex = k-2[E--P]) is dependent on the [Pi] with an apparent Km of 177 mM, reflecting the very low affinity of the enzyme for Pi. The maximal exchange rate is 6.7 micrograms atoms of oxygen X min-1 X mg-1 of protein. The individual kinetic constants are evaluated: k2 = 3.4 X 10(3) min-1, k-2 = 5.50 X 10(5) min-1 and k-1 = 4.11 X 10(3) min-1. Under conditions of uncoupled transport, the hydrolysis of E--P is exergonic as [E.Pi]/[E--P] = k-2/k2 = 164. During hydrolysis of ATP, the rate of medium Pi in equilibrium H2O exchange activity as well as the extent of phosphorylation of the enzyme from Pi are markedly stimulated: 7.9 and 5.3 times, respectively, whereas the Pc is not modified. These data are most simply interpretated by the existence of two isomeric forms of the enzyme; one is specific for binding ATP and the other for binding Pi. The Pc for intermediate Pi in equilibrium H2O exchange, when the E--P species is formed from cleavage of [gamma- 18O]ATP, is the same as for medium exchange, indicating that the same exchange pathway operates under both conditions. Varying the [ATP] had very little effect on the Pc, indicating little or no cooperativity between different catalytic sites under the conditions used in this study
MH  - A
MH  - ACTIVE
MH  - Adenosinetriphosphatase
MH  - ADP
MH  - atp
MH  - ATPase
MH  - BINDING
MH  - Hydrolysis
MH  - membrane
MH  - Oxygen
MH  - Oxygen Isotopes
MH  - Phosphates
MH  - Phosphorylation
MH  - Time
MH  - transport
MH  - Water
RP  - NOT IN FILE
NT  - UI - 83030765LA - engRN - 0 (Oxygen Isotopes)RN - 0 (Phosphates)RN - 7732-18-5 (Water)RN - 7782-44-7 (Oxygen)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19821218IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:6215401
SO  - J Biol Chem 1982 Nov 10 ;257(21):12509-12516

460
UI  - 546
AU  - Amzel LM
AU  - Narayanan P
AU  - Pedersen PL
AU  - Sygusch J
TI  - The three-dimensional structure of F1-ATPases
RP  - NOT IN FILE
NT  - UI - 83176701LA - engRN - 0 (Macromolecular Systems)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - CA 10951/CA/NCIID - GM 25432/GM/NIGMSID - GM 27420/GM/NIGMSDA - 19830505IS - 0077-8923SB - IMCY - UNITED STATESJC - 5NM
UR  - PM:6220637
SO  - Ann N Y Acad Sci 1982  ;402():21-27

461
UI  - 543
AU  - Amzel LM
AU  - McKinney M
AU  - Narayanan P
AU  - Pedersen PL
TI  - Structure of the mitochondrial F1 ATPase at 9-A resolution
AB  - The soluble portion (F1 ATPase) of the mitochondrial ATP-synthesizing system is a multisubunit enzyme of molecular weight 380,000. It is composed of five different subunits, alpha, beta, gamma, and epsilon. The subunit stoichiometry is not known but there are strong suggestions that it is alpha 3 beta 3 gamma delta epsilon. We have determined the three-dimensional structure of the F1 ATPase of rat liver mitochondria to 9-A resolution by using x-ray diffraction techniques. The molecule appears to be formed by two equivalent halves, each formed by three regions of approximately equal size. These regions form a distorted hexagonal or octahedral arrangement. None of the regions form closed symmetrical trimers in the complex. It is proposed that, if the subunit stoichiometry is alpha 3 beta 3 gamma delta epsilon, the major subunits exist in at least two different environments in the complex. In this arrangement, the different copies of the major subunits are functionally not equivalent. This observation appears to offer a natural explanation of the complicated binding and labeling data of F1 ATPases
RP  - NOT IN FILE
NT  - UI - 83299887LA - engRN - 0 (Macromolecular Systems)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - CA 19051/CA/NCIID - GM 25432/GM/NIGMSID - GM 27420/GM/NIGMSDA - 19831028IS - 0027-8424SB - IMCY - UNITED STATESJC - PV3
UR  - PM:6225115
SO  - Proc Natl Acad Sci U S A 1982 Oct ;79(19):5852-5856

462
UI  - 21268
AU  - Antonenko Y
AU  - Yaguzhinsky LS
TI  - Generation of potential in lipid bilayer membranes as a result of proton-transfer reactions in the unstirred layers
AB  - The addition of an uncoupler in the presence of a concentration gradient of weak acids or bases (sodium acetate and ammonium chloride) leads to the generation of a potential on lipid bilayer membranes (LBM) which is positive in sign on the side of the membrane with a high concentration of sodium acetate and negative on the side with a high concentration of ammonium chloride. It is shown that the potential was caused by the pH gradient in the unstirred layers. These effects can be understood in terms of the previously described [Science, 182, 1258 (1973)] model for the transfer of weak acids and bases through LBM. This system described may be useful for quantitation of permeabilities for weak acids and bases through bilayer membranes
MH  - A
MH  - acetate
MH  - Acetic Acids
MH  - ACID
MH  - BASE
MH  - Lipid Bilayers
MH  - membrane
MH  - Membranes
MH  - model
MH  - Permeability
MH  - pH
MH  - proton
MH  - Proton transfer
MH  - Proton-transfer
MH  - Protons
MH  - Sodium
MH  - SYSTEM
MH  - TRANSFER
RP  - NOT IN FILE
NT  - UI - 83135696LA - engRN - 0 (Acetic Acids)RN - 0 (Lipid Bilayers)RN - 0 (Protons)RN - 12125-02-9 (Ammonium Chloride)RN - 64-19-7 (Acetic Acid)PT - Journal ArticleDA - 19830407IS - 0145-479XSB - IMCY - UNITED STATES
UR  - PM:6298198
SO  - J Bioenerg Biomembr 1982 Dec ;14(5-6):457-465

463
UI  - 889
AU  - Binder A
TI  - Respiration and photosynthesis in energy-transducing membranes of cyanobacteria
MH  - Adenosine Triphosphate
MH  - biosynthesis
MH  - Cyanobacteria
MH  - Cytochrome-c Oxidase
MH  - Electron Transport
MH  - Energy Metabolism
MH  - Membranes
MH  - metabolism
MH  - Oxygen Consumption
MH  - Photosynthesis
MH  - Protons
RP  - NOT IN FILE
SO  - J Bioenerg Biomembr 1982 Dec ;14(5-6):271-286

464
UI  - 19818
AU  - Boyer PD
AU  - Kohlbrenner WE
AU  - McIntosh DB
AU  - Smith LT
AU  - O'Neal CC
TI  - ATP and ADP modulations of catalysis by F1 and Ca2+, Mg2+-ATPases
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - ADP
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Ca(2+)-Transporting ATPase
MH  - Catalysis
MH  - COMPLEX
MH  - F1
MH  - H(+)-Transporting ATP Synthase
MH  - Multienzyme Complexes
MH  - Phosphotransferases
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 83176744LA - engRN - 0 (Multienzyme Complexes)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 2.7 (Phosphotransferases)RN - EC 2.7.4.- (ATP synthetase complex)RN - EC 3.6.1.- (Ca(2+) Mg(2+)-ATPase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)RN - EC 3.6.1.38 (Ca(2+)-Transporting ATPase)PT - Journal ArticleID - GM11094/GM/NIGMSDA - 19830505IS - 0077-8923SB - IMCY - UNITED STATESJC - 5NM
UR  - PM:6132584
SO  - Ann N Y Acad Sci 1982  ;402():65-83

465
UI  - 998
AU  - Champeil P
AU  - Rigaud JL
AU  - Gingold MP
TI  - Fluorescence energy transfer between ATPase monomers in sarcoplasmic reticulum reconstituted vesicles, in the presence of low concentrations of a nonionic detergent
AB  - We measured fluorescence energy transfer between ATPase monomers labeled with either a donor or an acceptor fluorophore, in order to test the hypothesis that the presumably oligomeric sarcoplasmic reticulum ATPase would dissociate into monomers within the membrane on the addition of low nonsolubilizing amounts of a nonionic detergent, C12E18. We found that at low nonsolubilizing detergent concentrations below the critical micellar concentration (c.m.c.), there was indeed direct interaction between the detergent and the protein; however, only solubilizing detergent concentrations reduced fluorescence transfer to a minimum. Considering the current literature, fluorescence energy transfer proved insufficient, at the present time, to confirm or reject the hypothesis
MH  - Adenosinetriphosphatase
MH  - Animal
MH  - Detergents
MH  - Energy Transfer
MH  - enzymology
MH  - Fluorescent Dyes
MH  - Kinetics
MH  - metabolism
MH  - pharmacology
MH  - Polyethylene Glycols
MH  - Sarcoplasmic Reticulum
MH  - Spectrometry,Fluorescence
MH  - Surface-Active Agents
MH  - Time
RP  - NOT IN FILE
SO  - Z Naturforsch [C] 1982 May ;37(5-6):513-516

466
UI  - 545
AU  - Cintron NM
AU  - Hullihen J
AU  - Schwerzmann K
AU  - Pedersen PL
TI  - Proton-adenosinetriphosphatase complex of rat liver mitochondria: effect of its inhibitory peptide on adenosine 5'-triphosphate hydrolytic and functional activities of the enzyme
RP  - NOT IN FILE
NT  - UI - 82206687LA - engRN - 0 (ATPase inhibitory protein)RN - 0 (Proteins)RN - 53-84-9 (NAD)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - 1-FO5-TWO 2755/TW/FICID - CA 10951/CA/NCIID - GM 00184/GM/NIGMSDA - 19820807IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:6211190
SO  - Biochemistry 1982 Apr 13 ;21(8):1878-1885

467
UI  - 19877
AU  - Cross RL
AU  - Nalin CM
TI  - Adenine nucleotide binding sites on beef heart F1-ATPase. Evidence for three exchangeable sites that are distinct from three noncatalytic sites
MH  - Adenine Nucleotides
MH  - ADENINE-NUCLEOTIDES
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - atp
MH  - ATP synthase
MH  - BINDING
MH  - Binding Sites
MH  - COUPLING FACTOR
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - H(+)-Transporting ATP Synthase
MH  - NONCATALYTIC SITES
MH  - nucleotide binding
MH  - nucleotide binding sites
MH  - NUCLEOTIDE-BINDING-SITES
MH  - Nucleotides
MH  - Oxidative Phosphorylation
MH  - Oxidative Phosphorylation Coupling Factors
MH  - Phosphorylation
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 82142412LA - engRN - 0 (Adenine Nucleotides)RN - 0 (Oxidative Phosphorylation Coupling Factors)RN - 25612-73-1 (Adenylyl Imidodiphosphate)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM 23152/GM/NIGMSDA - 19820527IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:6460765
SO  - J Biol Chem 1982 Mar 25 ;257(6):2874-2881

468
UI  - 645
AU  - Cross RL
AU  - Grubmeyer C
AU  - Penefsky HS
TI  - Mechanism of ATP hydrolysis by beef heart mitochondrial ATPase. Rate enhancements resulting from cooperative interactions between multiple catalytic sites
RP  - NOT IN FILE
NT  - UI - 83007313LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM 21731/GM/NIGMSID - GM 23152/GM/NIGMSDA - 19821203IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:6214558
SO  - J Biol Chem 1982 Oct 25 ;257(20):12101-12105

469
UI  - 614
AU  - De Santis A
AU  - Bertoli E
AU  - Di Gioia A
AU  - Melandri BA
AU  - Baccarini MA
TI  - The reconstitution of oxidative phosphorylation in mitochondria isolated from a ubiquinone-deficient mutant of Saccharomyces cerevisiae
AB  - Mitochondria, isolated from the ubiquinone-deficient nuclear mutant of Saccharomyces cerevisiae E3-24, are practically unable to oxidize exogenous substrates. Respiratory activity, coupled to ATP synthesis, can, however, be reconstituted by the simple addition of ethanolic solutions of ubiquinones. A minimal length of the isoprenoid side chain (greater than or equal to 3) was required for the restoration. Saturation of the reconstitution required a large amount of exogeneous ubiquinone, in excess over the normal content present in the mitochondria of the wild type strain. A similar pattern of reconstituted activities could be also obtained using sonicated inverted particles. Mitochondria and sonicated particles are also able to carry out a dye-mediated electron flow coupled to ATP synthesis in the absence of added ubiquinone, using ascorbate or succinate as electron donor. This demonstrates that the energy conserving mechanism at the third coupling site of the respiratory chain is fully independent of the presence of the large mobile pool of ubiquinone in the membrane
RP  - NOT IN FILE
NT  - UI - 82239310LA - engRN - 1339-63-5 (Ubiquinone)RN - 56-65-5 (Adenosine Triphosphate)PT - Journal ArticleDA - 19820910IS - 0145-479XSB - IMCY - UNITED STATESJC - HIO
UR  - PM:7047520
SO  - J Bioenerg Biomembr 1982 Jun ;14(3):159-169

470
UI  - 899
AU  - Ferguson SJ
AU  - Sorgato MC
TI  - Proton electrochemical gradients and energy-transduction processes
MH  - Adenosine Triphosphate
MH  - Animal
MH  - Bacteria
MH  - biosynthesis
MH  - Carotenoids
MH  - Chromaffin Granules
MH  - Electron Transport
MH  - Energy Metabolism
MH  - Formates
MH  - Fumarates
MH  - Hydrogen
MH  - Iron
MH  - Membrane Potentials
MH  - metabolism
MH  - Methanol
MH  - Mitochondria
MH  - Oxidation-Reduction
MH  - Oxygen
MH  - Oxygen Consumption
MH  - Potassium
MH  - Protons
MH  - Solutions
MH  - Submitochondrial Particles
MH  - Support,Non-U.S.Gov't
MH  - Thermodynamics
RP  - NOT IN FILE
SO  - Annu Rev Biochem 1982  ;51:185-217.():185-217

471
UI  - 57
AU  - Foster DL
AU  - Fillingame RH
TI  - Stoichiometry of subunits in the H+-ATPase complex of Escherichia coli
AB  - The H+-ATPase (F1F0) of Escherichia coli was purified from cells labeled with either [35S]sulfate or [U-14C-D] glucose, and the molar ratio of subunits in the complex determined. The molar ratio was calculated from the radioactivity incorporated into each subunit, using either the subunit sulfur content or subunit molecular weight. These labeling experiments confirm an alpha 3 beta 3 gamma 1 delta 1 epsilon 1 ratio of subunits in F1, and indicate a chi 1 psi 2 omega 10 ratio of subunits in F0. The chi, psi, and omega designations used here refer to the subunits of F0 in order of decreasing molecular weight. Staining with Coomassie brilliant blue gave a reliable indication of the molar ratio of subunits in F1, but very erroneous values for each of the subunits of F0. We attempted to estimate the ratio of subunits in the native membrane, since the stoichiometry determined for the purified complex could be an anomaly of purification. These estimates were made after labeling cells with [35S]sulfate during amplification of the ATPase genes carried on a lambda transducing phage. The subunit ratios in the native membrane were reasonably close to those obtained with purified F1F0. We conclude that the stoichiometry determined reflects the composition of F1F0 in the native membrane. The most surprising conclusion from this study is that there are 10 +/- 1 omega ("proteolipid") subunits in each F1F0 complex. This is considerably more than had been assumed previously
RP  - NOT IN FILE
NT  - UI - 82120168LA - engRN - 0 (Macromolecular Systems)RN - 0 (Membrane Proteins)RN - 0 (Multienzyme Complexes)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - 5 T32 GM07215/GM/NIGMSID - GM-23105/GM/NIGMSDA - 19820412IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:6460031
SO  - J Biol Chem 1982 Feb 25 ;257(4):2009-2015

472
UI  - 1185
AU  - Graber P
TI  - Phosphorylation in chloroplasts: ATP synthesis driven by .DELTA..psi. and by .DELTA.pH of artificial or light-generated origin
MH  - atp
MH  - ATP synthesis
MH  - chloroplast
MH  - Chloroplasts
MH  - Phosphorylation
MH  - synthesis
RP  - ON REQUEST (03/18/92)
SO  - Curr Top Membr Transp 1982  ;16():215-245

473
UI  - 1181
AU  - Graber P
AU  - Roegner M
AU  - Buchwald HE
AU  - Samoray D
AU  - Hauska G
TI  - Field-driven ATP synthesis by the chloroplast coupling factor complex reconstituted into liposomes
MH  - atp
MH  - ATP synthesis
MH  - chloroplast
MH  - COMPLEX
MH  - coupling
MH  - COUPLING FACTOR
MH  - liposome
MH  - Liposomes
MH  - synthesis
RP  - ON REQUEST (03/18/92)
SO  - FEBS Lett 1982  ;145():35-40

474
UI  - 8505
AU  - Grber P
AU  - Rgner D
AU  - Buchwald HE
AU  - Samoray D
AU  - Hauska G
TI  - Field-driven ATP synthesis by the chloroplast coupling factor complex reconstituted into liposomes
MH  - atp
MH  - ATP synthesis
MH  - chloroplast
MH  - COMPLEX
MH  - coupling
MH  - COUPLING FACTOR
MH  - liposome
MH  - Liposomes
MH  - synthesis
RP  - IN FILE
SO  - FEBS Lett 1982  ;145():39-40

475
UI  - 8150
AU  - Grber P
TI  - Phosphorylation in Chloroplasts: ATP Synthesis Driven by delta psi and by delta pH of Artificial or Light-Generated Origin
MH  - atp
MH  - ATP synthesis
MH  - chloroplast
MH  - Chloroplasts
MH  - DELTA-PH
MH  - membrane
MH  - pH
MH  - Phosphorylation
MH  - PSI
MH  - review
MH  - synthesis
MH  - transport
T2  - Current Topics in Membrane and Transport
Y2  - -32676  
PB  - New York: Academic Press Inc.
RP  - IN FILE
NT  - Ju Rev
SO  -  1982  ;16(120):215-215

476
UI  - 19816
AU  - Gresser MJ
AU  - Myers JA
AU  - Boyer PD
TI  - Catalytic site cooperativity of beef heart mitochondrial F1 adenosine triphosphatase. Correlations of initial velocity, bound intermediate, and oxygen exchange measurements with an alternating three-site model
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - ADENOSINE-TRIPHOSPHATASE
MH  - atp
MH  - ATP synthase
MH  - F1
MH  - H(+)-Transporting ATP Synthase
MH  - model
MH  - Oxygen
MH  - Pyruvate Kinase
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 83007301LA - engRN - 58-64-0 (Adenosine Diphosphate)RN - 7782-44-7 (Oxygen)RN - EC 2.7.1.40 (Pyruvate Kinase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM 11094/GM/NIGMSDA - 19821203IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:6214554
SO  - J Biol Chem 1982 Oct 25 ;257(20):12030-12038

477
UI  - 646
AU  - Grubmeyer C
AU  - Cross RL
AU  - Penefsky HS
TI  - Mechanism of ATP hydrolysis by beef heart mitochondrial ATPase. Rate constants for elementary steps in catalysis at a single site
RP  - NOT IN FILE
NT  - UI - 83007312LA - engRN - 0 (Phosphates)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM 21731/GM/NIGMSID - GM 23152/GM/NIGMSDA - 19821203IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:6214557
SO  - J Biol Chem 1982 Oct 25 ;257(20):12092-12100

478
UI  - 589
AU  - Hatefi Y
AU  - Yagi T
AU  - Phelps DC
AU  - Wong SY
AU  - Vik SB
AU  - Galante YM
TI  - Substrate binding affinity changes in mitochondrial energy-linked reactions
AB  - The effects of uncouplers and valinomycin plus nigericin (in the presence of K+) were studied on the apparent Km for substrates and apparent Vmax of the following energy-linked reactions catalyzed by submitochondrial particles: oxidative phosphorylation, NTP-33Pi exchange, ATP-driven electron transfer from succinate to NAD, and respiration-driven transhydrogenation from NADH to 3-acetylpyridine adenine dinucleotide phosphate. In all cases, partially uncoupling (up to 90%) concentrations of uncouplers of valinomycin plus nigericin were found to decrease apparent Vmax and to increase apparent Km. Results plotted as ln (Vmax/Km) versus the concentration of uncouplers or ionophores showed a linear decrease of the former as a function of increasing perturbant concentration (i.e., decreasing free energy). Because Vmax/Km may be considered as a measure of the apparent first- order rate constant for enzyme-substrate interaction and reflects the affinity between enzyme and substrate to form a complex, the results are consistent with the interpretation that membrane energization leads to a change in enzyme conformation with the resultant increase in enzyme-substrate affinity and facilitation of the reaction rate under consideration. The significance of these findings with respect to the mechanism of action of the energy-transducing systems studied is discussed
RP  - NOT IN FILE
NT  - UI - 82197576LA - engRN - 0 (Nucleotides)RN - 0 (Uncoupling Agents)RN - 2001-95-8 (Valinomycin)RN - 28380-24-7 (Nigericin)RN - 53-84-9 (NAD)RN - 56-65-5 (Adenosine Triphosphate)PT - Journal ArticleID - AM 08126/AM/NIADDKID - GM 24887/GM/NIGMSDA - 19820719IS - 0027-8424SB - IMCY - UNITED STATESJC - PV3
UR  - PM:6952227
SO  - Proc Natl Acad Sci U S A 1982 Mar ;79(6):1756-1760

479
UI  - 10374
AU  - Junge W
TI  - Electrogenic Reactions and Proton Pumping in Green Plant Photosynthesis
MH  - bioenergetics
MH  - Photosynthesis
MH  - plant
MH  - proton
MH  - review
MH  - wox
RP  - IN FILE
NT  - K 4.9.1
SO  - Curr Top Membr Transp 1982  ;16():431-463

480
UI  - 9960
AU  - Kakitani T
AU  - Honig B
AU  - Crofts AR
TI  - Theoretical studies of the electrochromic response of carotenoids in photosynthetic membranes
AB  - Molecular orbital calculations are carried out on a number of carotenoids in the presence of an external charge and a constant electric field. The external charge is used to represent the strong permanent field that is believed to polarize carotenoids in photosynthetic membranes and thus to account for their linear response to the transmembrane potential. Our calculations show that the in vitro leads to in vivo spectral shifts of carotenoids (approximately 25 nm) can be produced by a charge in close proximity to the molecule. The interaction of the induced dipole moment with a constant field accounts for the observed magnitude of the electrochromic response in photosynthetic bacteria. The existence of a second pool of carotenoids that shows a significant (approximately 20 nm) wavelength shift but no electrochromic response can be explained by an external charge positioned near the center of the molecule that affects its absorption maximum while inducing essentially no dipole moment. The spectral shift for this pool is due to the induction of higher multipoles. These also account for discrepancies that arise when one attempts to account quantitatively for available experimental results on carotenoid band shifts in terms of classical electrochromic theory
MH  - Bacteria
MH  - Bacteriorhodopsin
MH  - Carotenoids
MH  - FIELD
MH  - In Vitro
MH  - Membranes
RP  - NOT IN FILE
NT  - UI - 82257647LA - engRN - 0 (Carotenoids)RN - 53026-44-1 (Bacteriorhodopsin)PT - Journal ArticleID - RO1GM 26305/GM/NIGMSDA - 19821021IS - 0006-3495SB - IMCY - UNITED STATESJC - A5S
UR  - PM:7104451
SO  - Biophys J 1982 Jul ;39(1):57-63

481
UI  - 20912
AU  - Kanazawa H
AU  - Mabuchi K
AU  - Futai M
TI  - Nucleotide sequence of the promoter region of the gene cluster for proton-translocating ATPase from Escherichia coli and identification of the active promotor
MH  - ACTIVE
MH  - Adenosinetriphosphatase
MH  - ATPase
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
RP  - NOT IN FILE
NT  - UI - 83022404LA - engRN - 0 (DNA, Bacterial)RN - 0 (Plasmids)RN - EC 2.7.7.6 (DNA-Directed RNA Polymerase)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19821202IS - 0006-291XSB - IMCY - UNITED STATES
UR  - PM:6215041
SO  - Biochem Biophys Res Commun 1982 Jul 30 ;107(2):568-575

482
UI  - 20913
AU  - Kanazawa H
AU  - Kayano T
AU  - Kiyasu T
AU  - Futai M
TI  - Nucleotide sequence of the genes for beta and epsilon subunits of proton-translocating ATPase from Escherichia coli
MH  - Adenosinetriphosphatase
MH  - ATPase
MH  - Enzymes
MH  - EPSILON-SUBUNIT
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - Macromolecular Systems
MH  - membrane
MH  - Membrane Proteins
MH  - protein
MH  - Proteins
MH  - SUBUNIT
MH  - SYSTEM
MH  - SYSTEMS
RP  - NOT IN FILE
NT  - UI - 82256510LA - engRN - 0 (Codon)RN - 0 (DNA, Bacterial)RN - 0 (DNA, Recombinant)RN - 0 (Macromolecular Systems)RN - 0 (Membrane Proteins)RN - 0 (Plasmids)RN - EC 3.1.21 (DNA Restriction Enzymes)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19820910IS - 0006-291XSB - IMCY - UNITED STATES
UR  - PM:6285901
SO  - Biochem Biophys Res Commun 1982 Apr 29 ;105(4):1257-1264

483
UI  - 20915
AU  - Kanazawa H
AU  - Futai M
TI  - Structure and function of H+-ATPase: what we have learned from Escherichia coli H+-ATPase
AB  - We have identified a transducing phage lambda asn5 carrying a set of structure genes coding for F1F0. New transducing phages and plasmids carrying a part of the DNA fragment in lambda asn5 were isolated and assayed by genetic complementation with mutants of F1F0. After analysis of DNA from these phages and plasmids, we mapped the genes for F1F0 within a physically defined segment of DNA of 4.5 megadaltons. The nucleotide sequence of the DNA segment was determined, and the primary amino acid sequences of all the subunits were determined. We discuss the homology of the sequence with those of other proteins capable of nucleotide binding. The secondary structures of the subunits were deduced, and a Rossman fold was found in the beta-subunit. The b- and delta-subunits had unique secondary structures. The roles of the subunits of F1 were studied by analysis of isolated subunits and mutationally altered subunits. Conformational changes of the alpha- and beta-subunits and transmission of conformational change between the two subunits were observed. Intracistronic mapping of mutations was achieved
MH  - A
MH  - ACID
MH  - Adenosinetriphosphatase
MH  - Amino Acid Sequence
MH  - analysis
MH  - ATPase
MH  - BETA-SUBUNIT
MH  - BINDING
MH  - conformational change
MH  - DELTA-SUBUNIT
MH  - Enzymes
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1
MH  - function
MH  - H+-ATPase
MH  - Macromolecular Systems
MH  - mutant
MH  - nucleotide binding
MH  - protein
MH  - Proteins
MH  - secondary
MH  - SUBUNIT
MH  - SYSTEM
MH  - SYSTEMS
RP  - NOT IN FILE
NT  - UI - 83176724LA - engRN - 0 (Macromolecular Systems)RN - EC 3.1.21 (DNA Restriction Enzymes)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19830505IS - 0077-8923SB - IMCY - UNITED STATES
UR  - PM:6301339
SO  - Ann N Y Acad Sci 1982  ;402():45-64

484
UI  - 9905
AU  - Kashket ER
TI  - Stoichiometry of the H+-ATPase of growing and resting, aerobic Escherichia coli.
AB  - The H+/ATP stoichiometry of the proton-translocating ATPase was investigated in growing and nongrowing, respiring cells of Escherichia coli. The protonmotive force, delta p, was determined by measuring the transmembrane chemical gradient of protons, delta pH, from the cellular accumulation of benzoate anions, and the electrical gradient, delta psi, from the accumulation of the lipophilic cation tetraphenylphosphonium (TPP+). The accumulation of lactose was also used to calculate the delta p in this lactose operon constitutive beta-galactosidase negative mutant. The phosphorylation potential, delta GP', was determined by measuring the cellular concentration of ATP, ADP, and inorganic phosphate. According to chemiosmotic principles, at steady state the phosphorylation potential is in thermodynamic equilibrium with the protonmotive force, and thus the ratio delta p/delta GP' can be used to determine the H+/ATP ratio. Respiring E. coli cells, in mid-exponential phase of growth or incubated in buffer, at external pHs from 6.25 to 8.25 had a constant delta GP' of about 500 mV. The H+/ATP ratio was found to be 3 when the delta p value derived from lactose accumulation levels was used. However, when the delta p values derived from delta pH and delta psi were used in the calculations, the H+/ATP ratio varied from about 2.5 at external pH 6.25 to about 4 at pH 8.25. Arguments are presented for the hypothesis that the delta psi values obtained from the TPP+ measurements are likely to be inaccurate and that a value of 3 H+/ATP, independent of the external pH, is likely to be the valid stoichiometry.
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - ADP
MH  - Anions
MH  - atp
MH  - ATPase
MH  - Cells
MH  - Chemistry
MH  - DELTA-PH
MH  - enzymology
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - H+-ATPase
MH  - Hydrogen-Ion Concentration
MH  - INORGANIC-PHOSPHATE
MH  - Ion Channels
MH  - Lactose
MH  - metabolism
MH  - Phosphates
MH  - Phosphorylation
MH  - proton
MH  - Protons
MH  - PSI
MH  - Support,U.S.Gov't,Non-P.H.S.
RP  - NOT IN FILE
SO  - Biochemistry 1982 Oct 26 ;21(22):5534-5538

485
UI  - 21204
AU  - Khitrina LV
AU  - Drachev LA
AU  - Kaulen AD
AU  - Chekulaeva LN
TI  - [Inhibition of bacteriorhodopsin by formalin and lanthanum]
AB  - It was shown that the penetrating ions (PCB-) method can be used for quantitative estimation of the proton translocation function. Using this method it was found that the maximal inhibition (up to 5.5-6 times) can be achieved by 1% formalin treatment for 0.5 hrs in 0.5 M phosphate buffer, pH 7.8, 70 degrees. Using three methods, the kinetics of the obtained preparation were compared to those of the previously known inhibitor, La3+. The changes observed were as follows: inhibition of intermediate M412 decay in membrane suspension, decrease in the amplitude of the electric response millisecond phase in the purple membrane--collodium film system and deceleration of the overall photochemical cycle turnover in this system. The latter was registered by the rate of resporation of the electric response amplitude to the second light flash in the presence of uncoupling agents. The possible role of crosslinks during protein treatment with formalin is discussed
MH  - Bacteriorhodopsin
MH  - buffer
MH  - carotenoid
MH  - Carotenoids
MH  - flash
MH  - function
MH  - inhibitor
MH  - intermediate
MH  - ion
MH  - Ions
MH  - Kinetics
MH  - Light
MH  - M
MH  - membrane
MH  - method
MH  - Methods
MH  - pH
MH  - protein
MH  - proton
MH  - SYSTEM
MH  - Time
MH  - translocation
MH  - Uncoupling Agents
RP  - NOT IN FILE
NT  - UI - 83101559LA - rusRN - 36-88-4 (Carotenoids)RN - 50-00-0 (Formaldehyde)RN - 53026-44-1 (Bacteriorhodopsins)RN - 7439-91-0 (Lanthanum)RN - 9004-70-0 (Collodion)PT - Journal ArticleDA - 19830311IS - 0320-9725SB - IMCY - USSR
UR  - PM:7150667
SO  - Biokhimiia 1982 Nov ;47(11):1763-1772

486
UI  - 779
AU  - Kinosita K
AU  - Ikegami A
AU  - Yoshida M
AU  - Kagawa Y
TI  - Nanosecond fluorometric investigation of hydrodynamic properties of adenosine triphosphatase from thermophilic bacterium PS3
AB  - The soluble portion (TF1) of proton-translocating ATPase from thermophilic bacterium PS3 was labeled with a fluorescent dye N-(1- pyrene)maleimide. The decay of fluorescence anisotropy of the adduct showed that TF1 in aqueous solution was characterized by a volume of equivalent sphere of 1,120 nm3. This value is 2.4 times the volume calculated from the molecular weight and partial specific volume, indicating a non-spherical shape and/or extensive hydration. A prolate ellipsoid with an axial ratio of 2 to 3 is suggested as a first approximation of the shape of hydrated TF1. The presence or absence of ATP, ADP, or Mg2+ did not alter the volume of the equivalent sphere appreciably; the probable conformational change of TF1 induced by these ligands does not lead to a gross alteration of its hydrodynamic properties
RP  - NOT IN FILE
NT  - UI - 83135679LA - engRN - 7732-18-5 (Water)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19830407IS - 0021-924XSB - IMCY - JAPANJC - HIF
UR  - PM:6219102
SO  - J Biochem (Tokyo ) 1982 Dec ;92(6):2043-2046

487
UI  - 19817
AU  - Kohlbrenner WE
AU  - Boyer PD
TI  - Catalytic properties of beef heart mitochondrial ATPase modified with 7- chloro-4-nitrobenzo-2-oxa-1,3-diazole. Evidence for catalytic site cooperativity during ATP synthesis
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - atp
MH  - ATP synthesis
MH  - ATPase
MH  - HEART MITOCHONDRIAL ATPASE
MH  - synthesis
RP  - NOT IN FILE
NT  - UI - 82142503LA - engRN - 0 (Oxadiazoles)RN - 10199-89-0 (4-Chloro-7-nitrobenzofurazan)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleID - GM 11094/GM/NIGMSDA - 19820527IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:6460767
SO  - J Biol Chem 1982 Apr 10 ;257(7):3441-3446

488
UI  - 21102
AU  - Krulwich TA
AU  - Guffanti AA
TI  - ATP synthesis at low proton-motive forces
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - atp
MH  - ATP synthesis
MH  - Proton-Motive Force
MH  - synthesis
RP  - NOT IN FILE
NT  - UI - 83176697LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleID - GM28454/GM/NIGMSDA - 19830505IS - 0077-8923SB - IMCY - UNITED STATES
UR  - PM:6220636
SO  - Ann N Y Acad Sci 1982  ;402():167-168

489
UI  - 900
AU  - Lehninger AL
TI  - Proton and electric charge translocation in mitochondrial energy transduction
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - Adult
MH  - Animal
MH  - Calorimetry
MH  - Electron Transport
MH  - Energy Metabolism
MH  - Human
MH  - Hydrogen-Ion Concentration
MH  - metabolism
MH  - Mitochondria
MH  - Oxidation-Reduction
MH  - Oxidative Phosphorylation
MH  - Oxygen Consumption
RP  - NOT IN FILE
SO  - Adv Exp Med Biol 1982  ;148:171-86.():171-186

490
UI  - 9912
AU  - Maloney PC
AU  - Hansen FC
TI  - Stoichiometry of proton movements coupled to ATP synthesis driven by a pH gradient in Streptococcus lactis.
AB  - An electrochemical potential difference for H+ was established across the plasma membrane of the anaerobe Streptococcus lactis by addition of sulfuric acid to cells suspended in potassium phosphate at pH 8 along with valinomycin or permeant anions. Subsequent acidification of the cell was measured by the distribution of salicyclic acid. A comparison between cells treated or untreated with the inhibitor N,N'-dicyclohexylcarbodiimide was used to reveal that portion of net proton entry attributable to a direct coupling between H+ inflow and synthesis of ATP catalyzed by the reversible proton-translocating ATPase of this microorganism. When the imposed electrochemical proton gradient was below 180-190 mV, proton entry was at the rate expected of passive flux, for both control cells and cells treated with the ATPase inhibitor, However, at higher driving force acidification of control cells was markedly accelerated, coincident with ATP synthesis, while acidification of cells treated with the inhibitor continued at the rate characteristic of passive inflow. This observed threshold (180-190 mV) was identified as the reversal potential for this H+ "pump". Parallel measurements showed that the free energy of hydrolysis for ATP in these washed cells was 8.4 kcal/mole (370mV). The comparison between the reversal (threshold) potential and the free energy of hydrolysis for ATP indicates a stoichiometry of 2 H+/ATP for the coupling of proton movements to ATP formation in bacteria.
MH  - Adenosine Triphosphate
MH  - Anions
MH  - atp
MH  - ATP synthesis
MH  - ATPase
MH  - Bacteria
MH  - biosynthesis
MH  - Cells
MH  - Dicyclohexylcarbodiimide
MH  - H+
MH  - Hydrogen-Ion Concentration
MH  - Hydrolysis
MH  - Lactococcus lactis
MH  - Membrane Potentials
MH  - metabolism
MH  - Movement
MH  - pharmacology
MH  - Potassium
MH  - proton
MH  - Protons
MH  - Support,U.S.Gov't,P.H.S.
MH  - Thiocyanates
MH  - Valinomycin
RP  - NOT IN FILE
SO  - J Membr Biol 1982  ;66(1):63-75

491
UI  - 20911
AU  - Matsuoka I
AU  - Takeda K
AU  - Futai M
AU  - Tonomura Y
TI  - Reactions of a fluorescent ATP analog, 2'-(5-dimethyl-aminonaphthalene- 1-sulfonyl) amino-2'-deoxyATP, with E. coli F1-ATPase and its subunits: the roles of the high affinity binding site in the alpha subunit and the low affinity binding site in the beta subunit
AB  - We performed kinetic studies on the reactions of a fluorescent ATP analog, 2'-(5-dimethyl-aminonaphthalene-1-sulfonyl) amino-2'-deoxyATP (DNS-ATP), with E. coli F1-ATPase (EF1) and its subunits, to clarify the role of each subunit in the ATPase reaction. The following results were obtained. 1. One mol of EF1, which contains nonexchangeable 2 mol ATP and 0.5 mol ADP, binds 3 mol of DNS-ATP. The apparent dissociation constant, in the presence of Mg2+, was 0.23 microM. Upon binding, the fluorescence intensity of DNS-ATP at 520 nm increased exponentially with t1/2 of 35 s, and reached 3.5 times the original fluorescence level. Following the fluorescence increase, DNS-ATP was hydrolyzed, and the fluorescence intensity maintained its enhanced level. 2. The addition of an excess of ATP over the EF1-DNS-nucleotide complex, in the presence of Mg2+, decreased the fluorescence intensity rapidly, indicating the acceleration of DNS-nucleotide release from EF1. ADP and GTP also decreased the fluorescence intensity. 3. DCCD markedly inhibited the accelerating effect of ATP on DNS-nucleotide release from EF1 and the EF1-DNS-ATPase or -ATPase activity in a steady state. On the other hand, DCCD only slightly inhibited the fluorescence increase of DNS-ATP, due to its binding to EF1, and the rate of single cleavage of 1 mol of DNS-ATP per mol of alpha subunit of EF1. 4. In the presence of Mg2+, 0.65-0.82 mol of DNS-ATP binds to 1 mol of the isolated alpha subunit of EF1 with an apparent dissociation constant of 0.06-0.07 microM. Upon binding, the fluorescence intensity of DNS-ATP at 520 nm increased 1.55 fold very rapidly (t1/2 less than 1 s). No hydrolysis of DNS-ATP was observed upon the addition of the isolated alpha subunit. The fluorescence intensity of DNS-ATP was unaffected by the addition of the isolated beta subunit. DNS-ATP was also unhydrolyzed by the isolated beta subunit. 5. EF1-ATPase was reconstituted from alpha, beta, and gamma subunits in the presence of Mg2+ and ATP. The kinetic properties of the fluorescence change of DNS-ATP in the reaction with the reconstituted EF1-ATPase were quite similar to those of native EF1. Most of our findings are consistent with a simple mechanism that the high affinity catalytic site and low affinity regulatory site exist in the alpha subunit and beta subunit, respectively. However, the findings mentioned in (4) suggest that the binding of the alpha and beta subunit, which is mediated by the gamma subunit, induces conformational change(s) in the ATP binding site located probably in the alpha subunit, and that the conformational change(s) is essential to exert the full hydrolyzing activity
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - ADP
MH  - ALPHA-SUBUNIT
MH  - atp
MH  - ATPase
MH  - BETA-SUBUNIT
MH  - BINDING
MH  - COLI F1 ATPASE
MH  - COMPLEX
MH  - CONSTANT
MH  - Dicyclohexylcarbodiimide
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - fluorescence
MH  - Hydrolysis
MH  - Macromolecular Systems
MH  - mechanism
MH  - Site
MH  - SUBUNIT
MH  - SYSTEM
MH  - SYSTEMS
MH  - Time
RP  - NOT IN FILE
NT  - UI - 83108771LA - engRN - 0 (Macromolecular Systems)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - 80214-45-5 (dansylamino deoxy-ATP)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19830311IS - 0021-924XSB - IMCY - JAPAN
UR  - PM:6218158
SO  - J Biochem (Tokyo ) 1982 Nov ;92(5):1383-1398

492
UI  - 1014
AU  - Mitchell P
TI  - Osmoenzymology: the study of molecular machines
MH  - Adenosinetriphosphatase
MH  - Mathematics
MH  - Membrane Proteins
MH  - metabolism
MH  - Models,Biological
MH  - Osmolar Concentration
MH  - Oxidoreductases
MH  - Thermodynamics
RP  - NOT IN FILE
SO  - Prog Clin Biol Res 1982  ;102 Pt B:399-408.():399-408

493
UI  - 1015
AU  - Mitchell P
AU  - Koppenol WH
TI  - Chemiosmotic ATPase mechanisms
MH  - Adenosinetriphosphatase
MH  - Animal
MH  - Biological Transport,Active
MH  - Cell Membrane
MH  - enzymology
MH  - H(+)-Transporting ATP Synthase
MH  - Kinetics
MH  - metabolism
MH  - Models,Biological
MH  - Osmolar Concentration
MH  - Thermodynamics
RP  - NOT IN FILE
SO  - Ann N Y Acad Sci 1982  ;402:584-601.():584-601

494
UI  - 19876
AU  - Nalin CM
AU  - Cross RL
TI  - Adenine nucleotide binding sites on beef heart F1-ATPase. Specificity of cooperative interactions between catalytic sites
AB  - Cooperative interactions between nucleotide binding sites on beef heart mitochondrial F1-ATPase have been studied by measuring substrate- promoted release of 5'adenylyl-beta,gamma-imidodiphosphate (AMP-PNP) from a single high affinity site. The site is initially loaded by incubating F1 with an equimolar amount of the nonhydrolyzable ATP analog. When unbound [3H]AMP-PNP is removed and the complex diluted to a concentration below the Kd, release of ligand shows an apparent absolute requirement for medium ADP. Release is biphasic with the extent of release during the initial rapid phase dependent on the concentration of medium ADP. Although phosphate alone has no effect, it enhances the rapid phase of ADP-promoted release over 2-fold with a half-maximal effect at 60 micrometers P1. The binding of efrapeptin (A23871) to the F1.AMP-PNP complex completely prevents ADP-promoted dissociation. Although AMP-PNP release also occurs in the presence of medium ATP, the F1.AMP-PNP complex does not dissociate if an ATP- regenerating system of sufficient capacity to prevent accumulation of medium ADP is added. Consistent with an inability of nucleoside triphosphate to promote release is the failure of medium, nonradioactive AMP-PNP to affect retention of the 3H-labeled ligand. The stability of F1.AMP-PNP complex in the absence of medium nucleotide and the highly specific ability of ADP plus P1 to promote rapid release of the ATP analog are interpreted as support for an ATP synthesis mechanism that requires substrate binding at one catalytic site for product release from an adjacent interacting site
MH  - A
MH  - Adenine Nucleotides
MH  - ADENINE-NUCLEOTIDES
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - ADP
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - BINDING
MH  - Binding Sites
MH  - COMPLEX
MH  - COUPLING FACTOR
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - H(+)-Transporting ATP Synthase
MH  - mechanism
MH  - MITOCHONDRIAL F1-ATPASE
MH  - nucleotide binding
MH  - nucleotide binding sites
MH  - NUCLEOTIDE-BINDING-SITES
MH  - Nucleotides
MH  - Oxidative Phosphorylation
MH  - Oxidative Phosphorylation Coupling Factors
MH  - Phosphates
MH  - Phosphorylation
MH  - Pyruvate Kinase
MH  - SYNTHASE
MH  - synthesis
RP  - NOT IN FILE
NT  - UI - 82214011LA - engRN - 0 (Adenine Nucleotides)RN - 0 (Oxidative Phosphorylation Coupling Factors)RN - 0 (Phosphates)RN - 25612-73-1 (Adenylyl Imidodiphosphate)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 2.7.1.40 (Pyruvate Kinase)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM 23152/GM/NIGMSDA - 19820826IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:6211449
SO  - J Biol Chem 1982 Jul 25 ;257(14):8055-8060

495
UI  - 1299
AU  - Nishimura M
AU  - Matsuura K
AU  - Masamoto K
TI  - Energy conversion and membrane potential in biological membranes
MH  - Membranes
RP  - NOT IN FILE
SO  - Kagaku no Ryoiki 1982  ;36():155-165

496
UI  - 9913
AU  - Ogawa S
AU  - Lee TM
TI  - Proton stoichiometry of adenosine 5'-triphosphate synthesis in rat liver mitochondria studied by phosphorus-31 nuclear magnetic resonance.
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - Animal
MH  - biosynthesis
MH  - Electrochemistry
MH  - Liver
MH  - Membrane Potentials
MH  - metabolism
MH  - Mitochondria
MH  - Mitochondria,Liver
MH  - Nuclear Magnetic Resonance
MH  - Phosphorylation
MH  - Potassium
MH  - proton
MH  - Protons
MH  - Rats
MH  - Thermodynamics
RP  - NOT IN FILE
SO  - Biochemistry 1982 Aug 31 ;21(18):4467-4473

497
UI  - 21205
AU  - Ovchinnikov YA
AU  - Abdulaev NG
AU  - Dergachev AE
AU  - Drachev AL
AU  - Drachev LA
AU  - Kaulen AD
AU  - Khitrina LV
AU  - Lazarova ZP
AU  - Skulachev VP
TI  - Photoelectric and spectral responses of bacteriorhodopsin modified by carbodiimide and amine derivatives
MH  - Bacteriorhodopsin
MH  - carotenoid
MH  - Carotenoids
RP  - NOT IN FILE
NT  - UI - 83053395LA - engRN - 0 (Amines)RN - 0 (Carbodiimides)RN - 36-88-4 (Carotenoids)RN - 53026-44-1 (Bacteriorhodopsins)PT - Journal ArticleDA - 19830119IS - 0014-2956SB - IMCY - GERMANY, WEST
UR  - PM:7140772
SO  - Eur J Biochem 1982 Oct ;127(2):325-332

498
UI  - 547
AU  - Pedersen PL
TI  - H+-ATPases in biological systems: an overview of their function, structure, mechanism, and regulatory properties
RP  - NOT IN FILE
NT  - UI - 83176691LA - engRN - 0 (Affinity Labels)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19830505IS - 0077-8923SB - IMCY - UNITED STATESJC - 5NM
UR  - PM:6220632
SO  - Ann N Y Acad Sci 1982  ;402():1-20

499
UI  - 676
AU  - Schafer G
AU  - Weber J
TI  - Interaction of high-affinity nucleotide binding sites in mitochondrial ATP synthesis and hydrolysis
AB  - The present study contributes to the problem of the dynamic structure of mitochondrial F1-ATPase and the functional interrelation of so- called tight nucleotide binding sites. Nucleotide analogs are used as a tool to differentiate two distinct functional states of the membrane- bound enzyme, proposed to reflect corresponding conformational states; they reveal F1-ATPase as a "dual-state" enzyme: ATP-synthetase, and ATP- hydrolase. The analogs used are 3'-naphthoyl esters of AD(T)P, and 2'(3')-O-trinitrophenyl ethers of AD(T)P. Both types of analogs act inversely to each other with respect to their relative effects on oxidative phosphorylation and on ATPase in submitochondrial vesicles. The respective ratios of Ki versus both processes are 250/1 compared to 1/170. It is also shown that in the presence of the inhibitory 3'- esters oxidative phosphorylation deviates from linear kinetics and that these inhibitors induce a lag time of oxidative phosphorylation depending on the initial pattern of nucleotides available to energized submitochondrial vesicles. The duration of the lag time coincides with the time course of displacement of the analog from a tight binding site. The conclusions of the study are: (a) the catalytic sites of F1- ATP-synthetase are not operating independently from each other; they rather interact in a cooperative manner; (b) F1-ATPase as a "dual- state" enzyme exhibits highly selective responses to tight binding of nucleotides or analogs in its "energized" (membrane-bound) state versus its "nonenergized" state, respectively
RP  - NOT IN FILE
NT  - UI - 83135698LA - engRN - 0 (Nucleotides)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19830407IS - 0145-479XSB - IMCY - UNITED STATESJC - HIO
UR  - PM:6219105
SO  - J Bioenerg Biomembr 1982 Dec ;14(5-6):479-498

500
UI  - 1188
AU  - Schlodder E
AU  - Graber P
AU  - Witt HT
TI  - Mechanism of phosphorylation in chloroplasts
MH  - chloroplast
MH  - Chloroplasts
MH  - mechanism
MH  - Phosphorylation
RP  - ON REQUEST (03/18/92)
SO  - Top Photosynth 1982  ;4():105-175

501
UI  - 20970
AU  - Schneider E
AU  - Altendorf K
TI  - ATP synthetase (F1F0) of Escherichia coli K-12. High-yield preparation of functional F0 by hydrophobic affinity chromatography
AB  - 1. The purified ATP synthetase complex (F1F0) from Escherichia coli was adsorbed to immobilized poly-(L-lysine)-deoxycholic acid. About 0.7 mg F1F0 were bound per ml of settled gel. The hydrophilic F1 part was dissociated from the complex by treatment with 7 M urea. F0 was eluted in high yield either with deoxycholate (6 mM) or taurodeoxycholate (10 mM). About 14% of the total protein bound to the column was eluted as F0, which corresponds to 64% of the total F0 in the F1F0 complex. 2. The purified F0 preparation obtained was composed of three different kinds of subunits with apparent molecular weights of 24000 (a), 19000 (b) and 8300 (c), respectively as determined by sodium dodecyl sulfate gel electrophoresis. 3. After incorporation into liposomes and the generation of a potassium diffusion potential by valinomycin, the F0 preparation mediated H+ translocation. This H+ uptake is inhibited by either dicyclohexylcarbodiimide or purified F1 ATPase. 4. Incubation of F0-containing liposomes with F1 led to the reconstitution of an ATP- driven quenching of acridine-dye fluorescence. The quenching was abolished by uncoupler and prevented by dicyclohexylcarbodiimide
MH  - A
MH  - ACID
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - atp
MH  - ATPase
MH  - COMPLEX
MH  - Dicyclohexylcarbodiimide
MH  - Diffusion
MH  - diffusion potential
MH  - Electrophoresis
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - fluorescence
MH  - H+
MH  - Liposomes
MH  - M
MH  - Multienzyme Complexes
MH  - Phosphotransferases
MH  - Potassium
MH  - protein
MH  - proton
MH  - Protons
MH  - reconstitution
MH  - Sodium
MH  - SUBUNIT
MH  - translocation
MH  - Valinomycin
RP  - NOT IN FILE
NT  - UI - 83027288LA - engRN - 0 (Liposomes)RN - 0 (Multienzyme Complexes)RN - 0 (Protons)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 2.7 (Phosphotransferases)RN - EC 2.7.4.- (ATP Synthetase Complexes)PT - Journal ArticleDA - 19821216IS - 0014-2956SB - IMCY - GERMANY, WEST
UR  - PM:6290212
SO  - Eur J Biochem 1982 Aug ;126(1):149-153

502
UI  - 544
AU  - Schwerzmann K
AU  - Hullihen J
AU  - Pedersen PL
TI  - Proton adenosine triphosphatase complex of rat liver mitochondria. Interaction with the ATPase inhibitor peptide covalently labeled with N- hydroxysuccinimidyl-p-azidobenzoate
RP  - NOT IN FILE
NT  - UI - 82265597LA - engRN - 0 (ATPase inhibitory protein)RN - 0 (Azides)RN - 0 (Cross-Linking Reagents)RN - 0 (Proteins)RN - 53053-08-0 (hydroxysuccinimidyl-4-azidobenzoate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - 1-F05 TWO 2755/TW/FICID - CA 10951/CA/NCIDA - 19821012IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:6213609
SO  - J Biol Chem 1982 Aug 25 ;257(16):9555-9560

503
UI  - 886
AU  - Skulachev VP
TI  - The localized delta muH+ problem. The possible role of the local electric field in ATP synthesis
MH  - Adenosine Triphosphate
MH  - biosynthesis
MH  - Electrochemistry
MH  - Energy Metabolism
MH  - Hydrogen
MH  - Kinetics
MH  - Oxidation-Reduction
RP  - NOT IN FILE
SO  - FEBS Lett 1982 Sep 6 ;146(1):1-4

504
UI  - 21115
AU  - Sorgato MC
AU  - Galiazzo F
AU  - Panato L
AU  - Ferguson SJ
TI  - Estimation of H+-translation stoicheiometry of mitochondrial ATPase by comparison of proton-motive forces with clamped phosphorylation potentials in submitochondrial particles
AB  - The proton-motive forces generated in submitochondrial particles by both hydrolysis of ATP and oxidation of succinate have been measured by flow dialysis and compared with the ambient phosphorylation potentials. It is concluded that three H+ are translocated for each ATP molecule hydrolysed or synthesised. By utilising rat liver mitochondria respiring with beta-hydroxybutyrate as a new system for regeneration of ATP from ADP and Pi, phosphorylation potentials were clamped at a range of values by using mixtures of particles and mitochondria in various ratios. As the rate of ATP hydrolysis by the particles was lowered, the proton-motive force decreased only slightly except at the very lowest rates, these results paralleling earlier studies on the relation between rate of respiration-driven proton translocation and proton- motive force
MH  - A
MH  - Adenosinetriphosphatase
MH  - ADP
MH  - atp
MH  - ATPase
MH  - Dialysis
MH  - H+
MH  - Hydrolysis
MH  - Liver
MH  - Mitochondria
MH  - Phosphorylation
MH  - proton
MH  - Proton-Motive Force
MH  - Proton-Translocating ATPases
MH  - Submitochondrial Particles
MH  - succinate
MH  - SYSTEM
MH  - translocation
RP  - NOT IN FILE
NT  - UI - 83049037LA - engRN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19830119IS - 0006-3002SB - IMCY - NETHERLANDS
UR  - PM:6215943
SO  - Biochim Biophys Acta 1982 Oct 18 ;682(1):184-188

505
UI  - 20971
AU  - Steffens K
AU  - Kiltz HH
AU  - Schneider E
AU  - Schmid R
AU  - Altendorf K
TI  - ATP-synthetase complex (F1F0) from Escherichia coli. Purification and characterization of subunits A and B of the F0 part
MH  - A
MH  - ACID
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Amino Acids
MH  - atp
MH  - Bacterial Proteins
MH  - COMPLEX
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - Multienzyme Complexes
MH  - Phosphotransferases
MH  - protein
MH  - Proteins
MH  - purification
MH  - Sodium
MH  - SUBUNIT
RP  - NOT IN FILE
NT  - UI - 82262052LA - engRN - 0 (Amino Acids)RN - 0 (Bacterial Proteins)RN - 0 (Multienzyme Complexes)RN - 151-21-3 (Sodium Dodecyl Sulfate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 2.7 (Phosphotransferases)RN - EC 2.7.4.- (ATP Synthetase Complexes)PT - Journal ArticleDA - 19821021IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:6286344
SO  - FEBS Lett 1982 Jun 1 ;142(1):151-154

506
UI  - 20914
AU  - Takeda K
AU  - Hirano M
AU  - Kanazawa H
AU  - Nukiwa N
AU  - Kagawa Y
AU  - Futai M
TI  - Hybrid ATPase's formed from subunits of coupling factor F1's of Escherichia coli and thermophilic bacterium PS3
AB  - ATPase complexes were reconstituted from homologous and heterologous combinations of alpha, beta, and gamma subunits of coupling factor ATPase TF1 of thermophilic bacterium PS3 and EF1 of Escherichia coli. TF1 and alpha beta gamma complex reconstituted from TF1 subunits were thermostable and activated by methanol, sodium dodecyl sulfate and anions and they were halophilic, whereas EF1 and its three-subunit complex did not show these properties. The hybrid ATPase alpha T beta T gamma E (complex of the alpha and beta subunits of TF1 and the gamma subunit of EF1) showed closely similar properties to TF1 except for thermostability, while alpha E beta E gamma T (alpha and beta from EF1 and gamma from TF1) had similar properties to EF1. These results suggest that alpha and/or beta is required for the properties of F1. The complex alpha E beta T gamma E showed similar properties to EF1 except for its optimum pH: this complex had a broad pH optimum at about pH 7, whereas EF1 had an optimum at pH 8.5. No hybrid complexes were thermostable, suggesting that all three subunits of TF1 are required for heat stability. These hybrids showed higher halophilicity than EF1, although they were less halophilic than TF1. The hybrid enzymes studied here are the first thermophilic-mesophilic hybrid enzymes obtained
MH  - A
MH  - Adenosinetriphosphatase
MH  - Anions
MH  - ATPase
MH  - Bacteria
MH  - BETA-SUBUNIT
MH  - COMPLEX
MH  - coupling
MH  - COUPLING FACTOR
MH  - Enzymes
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1
MH  - Heat
MH  - Methanol
MH  - pH
MH  - PS3
MH  - Sodium
MH  - SUBUNIT
RP  - NOT IN FILE
NT  - UI - 82167350LA - engRN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19820614IS - 0021-924XSB - IMCY - JAPAN
UR  - PM:6461646
SO  - J Biochem (Tokyo ) 1982 Feb ;91(2):695-701

507
UI  - 677
AU  - Tiedge H
AU  - Lucken U
AU  - Weber J
AU  - Schafer G
TI  - High-affinity binding of ADP and of ADP analogues to mitochondrial F1- ATPase
AB  - Nucleotide-depleted F1-ATPase was prepared from beef heart mitochondria. By use of fluorescence techniques and isotope binding analyses, we investigated the occupation of the high-affinity binding sites on F1 by ADP and the ADP analogues 3'-O-(1-naphthoyl)adenosine diphosphate (N-ADP) and 3'-O-[1-(5-dimethylamino)-naphthoyl]adenosine diphosphate (DMAN-ADP). F1-ATPase was found to exhibit three binding sites for ADP (Kd = 50 nM for one site; Kd = 3 microM for the remaining two sites), two binding sites for N-ADP (Kd = 20 - 50 nM for both of the sites), and three binding sites for DMAN-ADP (Kd = 50 nM for all of the sites). Since the adenine nucleotides under consideration are bound to the same class of sites, the binding data can be explained best on the basis of the hypothesis that the binding process is anticooperative with ADP, whereas the analogues are able to overcome anticooperativity partially (N-ADP) or completely (DMAN-ADP). This binding model is consistent with the view that the exchangeable tight sites are involved directly in the catalytical process of ATP-synthesis in oxidative phosphorylation
RP  - NOT IN FILE
NT  - UI - 83053390LA - engRN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19830119IS - 0014-2956SB - IMCY - GERMANY, WESTJC - EMZ
UR  - PM:6216106
SO  - Eur J Biochem 1982 Oct ;127(2):291-299

508
UI  - 395
AU  - Vasilyeva EA
AU  - Minkov IB
AU  - Fitin AF
AU  - Vinogradov AD
TI  - Kinetic mechanism of mitochondrial adenosine triphosphatase. ADP- specific inhibition as revealed by the steady-state kinetics
AB  - 1. A substantial increase of the initial rate of ATP hydrolysis was observed after preincubation of bovine heart submitochondrial particles with phosphoenolpyruvate and pyruvate kinase. 2. The activation was accompanied by an increase of Vmax, without change of Km for ATP. 3. The activated particles catalysed the biphasic hydrolysis of ATP in the presence of an ATP-regenerating system; the initial rapid phase was followed by a second, slower, phase in a time-dependent fashion. 4. The higher the ATP concentration used as a substrate, the higher is the rate of transition between these two phases. 5. The particles catalysed the hydrolysis of ITP with a lag phase; after preincubation with phosphoenolpyruvate and pyruvate kinase, ITP was hydrolysed at a constant rate. 6. Qualitatively the same phenomena were observed when soluble mitochondrial ATPase (F1-ATPase) prepared by the conventional method in the presence of ATP was used as nucleotide triphosphatase. 7. A kinetic scheme is proposed, in which the intermediate active enzyme- product complex (E.ADP) formed during ATP hydrolysis is in slow equilibrium with the inactive E*.ADP complex forming as a result of dislocation of ADP from the active site of ATPase to the other site, which is not in rapid equilibrium with the surrounding medium
RP  - NOT IN FILE
NT  - UI - 82206080LA - engRN - 132-06-9 (Inosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 73-89-2 (Phosphoenolpyruvate)RN - EC 2.7.1.40 (Pyruvate Kinase)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19820708IS - 0264-6021SB - IMCY - ENGLANDJC - 9YO
UR  - PM:6211173
SO  - Biochem J 1982 Jan 15 ;202(1):9-14

509
UI  - 396
AU  - Vasilyeva EA
AU  - Minkov IB
AU  - Fitin AF
AU  - Vinogradov AD
TI  - Kinetic mechanism of mitochondrial adenosine triphosphatase. Inhibition by azide and activation by sulphite
AB  - 1. The initial rapid phase of ATP hydrolysis by bovine heart submitochondrial particles or by soluble F1-ATPase is insensitive to anion activation (sulphite) or inhibition (azide). 2. The second slow phase of ATP hydrolysis is hyperbolically inhibited by azide (Ki approximately 10(-5) M); the inosine triphosphatase activity of submitochondrial particles or F1-ATPase is insensitive to azide or sulphite. 3. The rate of interconversion between rapid azide- insensitive and slow azide-sensitive phases of ATP hydrolysis does not depend on azide concentration, but strongly depends on ATP concentration. 4. Sulphite prevents the interconversion of the rapid initial phase of the reaction into the slower second phase, and also prevents and slowly reverses the inhibition by azide. 5. The presence of sulphite in the mixture when ADP reacts with ATPase of submitochondrial particles changes the pattern of the following activation process. 6. Azide blocks the activation of ATP-inhibited ATPase of submitochondrial particles by phosphoenolpyruvate and pyruvate kinase. 7. The results obtained suggest that the inhibiting effect of azide on mitochondrial ATPase is due to stabilization of inactive E*.ADP complex formed during ATP hydrolysis; the activation of ATPase by sulphite is also realized through the equilibrium between intermediate active E.ADP complex and inactive E*.ADP complex
RP  - NOT IN FILE
NT  - UI - 82206049LA - engRN - 0 (Azides)RN - 0 (Sulfites)RN - 26628-22-8 (Sodium Azide)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 7631-90-5 (sodium bisulfite)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19820708IS - 0264-6021SB - IMCY - ENGLANDJC - 9YO
UR  - PM:6211171
SO  - Biochem J 1982 Jan 15 ;202(1):15-23

510
UI  - 781
AU  - Wakabayashi T
AU  - Tomioka A
AU  - Tokunaga M
AU  - Yoshida M
AU  - Kagawa Y
AU  - Usukura J
AU  - Yamada E
TI  - Image analysis of electron micrographs of ATPase (Coupling Factor TF1) from thermophilic bacteria and luminal epithelium of mouse urinary bladder
AB  - Electron micrographs of two dimensional array of H+-ATPase (Coupling Factor TF1 from the thermophilic bacteria) and luminal epithelial protein of urinary bladder were obtained using negative staining method and freeze-fracture method, respectively. Images which showed good optical diffraction pattern were digitized by a computer-linked flat- bed two dimensional microdensitometer and processed digitally. The results of translational computer noise filtering showed that the outline of TF1 molecules looks like a hexagon or an asterisk in the presence of sodium azide which is a specific inhibitor of TF1 or AMPPNP which is an unsplitable analogue of ATP, respectively. The luminal epithelial protein also looks like a hexagon. Rotational harmonic analysis was carried out to examine the rotational symmetry of the filtered images of TF1. It was found that 2-fold or 6-fold symmetry is dominant in the presence of sodium azide or AMPPNP, respectively
RP  - NOT IN FILE
NT  - UI - 83303149LA - engRN - 0 (Azides)RN - 25612-73-1 (Adenylyl Imidodiphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19831021IS - 0385-0005SB - IMCY - JAPANJC - VZM
UR  - PM:6225225
SO  - Tokai J Exp Clin Med 1982  ;7 Suppl():15-22

511
UI  - 376
AU  - Walker JE
AU  - Saraste M
AU  - Gay NJ
TI  - E. coli F1-ATPase interacts with a membrane protein component of a proton channel
AB  - The ATP synthases of bacteria, mitochondria and chloroplasts, which use the energy of a transmembrane proton gradient to power the synthesis of ATP, consist of an integral membrane component F0--thought to contain a proton channel--and a catalytic component, F1. To help investigate the way F0 and F1 are coupled, we have sequenced the b-subunit of the Escherichia coli F0, which seems to be the counterpart of a thermophilic bacteria F0 subunit thought to be essential for F1 binding. We report here that its sequence is remarkable, being hydrophobic around the N-terminus and highly charged in the remainder. We propose that the N-terminal segment lies in the membrane and the rest outside. The extramembranous section contains two adjacent stretches of 31 amino acids where the sequence is very similar: in the second of these stretches there is further internal homology. These duplicated stretches of the polypeptide probably fold into two alpha- helices which have many common features able to make contact with F1 subunits. Thus protein b occupies a central position in the enzyme, where it may be involved in proton translocation. It is possibly also important in biosynthetic assembly
RP  - NOT IN FILE
NT  - UI - 82272344LA - engRN - 0 (Ion Channels)RN - 0 (Membrane Proteins)RN - 0 (Multienzyme Complexes)RN - 1333-74-0 (Hydrogen)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 2.7 (Phosphotransferases)RN - EC 2.7.4.- (ATP synthetase complex)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19821012IS - 0028-0836SB - IMCY - ENGLANDJC - NSC
UR  - PM:6180323
SO  - Nature 1982 Aug 26 ;298(5877):867-869

512
UI  - 377
AU  - Walker JE
AU  - Eberle A
AU  - Gay NJ
AU  - Runswick MJ
AU  - Saraste M
TI  - Conservation of structure in proton-translocating ATPases of Escherichia coli and mitochondria
RP  - NOT IN FILE
NT  - UI - 83054312LA - engRN - 0 (Macromolecular Systems)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19830107IS - 0300-5127SB - IMCY - ENGLANDJC - E48
UR  - PM:6216131
SO  - Biochem Soc Trans 1982 Aug ;10(4):203-206

513
UI  - 375
AU  - Walker JE
AU  - Runswick MJ
AU  - Saraste M
TI  - Subunit equivalence in Escherichia coli and bovine heart mitochondrial F1F0 ATPases
RP  - NOT IN FILE
NT  - UI - 83053989LA - engRN - 0 (Macromolecular Systems)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19830107IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:6216120
SO  - FEBS Lett 1982 Sep 20 ;146(2):393-396

514
UI  - 378
AU  - Walker JE
AU  - Saraste M
AU  - Runswick MJ
AU  - Gay NJ
TI  - Distantly related sequences in the alpha- and beta-subunits of ATP synthase, myosin, kinases and other ATP-requiring enzymes and a common nucleotide binding fold
AB  - The alpha- and beta-subunits of membrane-bound ATP synthase complex bind ATP and ADP: beta contributes to catalytic sites, and alpha may be involved in regulation of ATP synthase activity. The sequences of beta- subunits are highly conserved in Escherichia coli and bovine mitochondria. Also alpha and beta are weakly homologous to each other throughout most of their amino acid sequences, suggesting that they have common functions in catalysis. Related sequences in both alpha and beta and in other enzymes that bind ATP or ADP in catalysis, notably myosin, phosphofructokinase, and adenylate kinase, help to identify regions contributing to an adenine nucleotide binding fold in both ATP synthase subunits
RP  - NOT IN FILE
NT  - UI - 84236073LA - engRN - 0 (Macromolecular Systems)RN - 0 (Multienzyme Complexes)RN - 0 (Myosin)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 2.7 (Phosphotransferases)RN - EC 2.7.1.11 (6-Phosphofructokinase)RN - EC 2.7.4.- (ATP synthetase complex)RN - EC 2.7.4.3 (Adenylate Kinase)RN - EC 2.7.7.- (Adenine Nucleotide Translocase)PT - Journal ArticleDA - 19840727IS - 0261-4189SB - IMCY - ENGLANDJC - EMB
UR  - PM:6329717
SO  - EMBO J 1982  ;1(8):945-951

515
UI  - 588
AU  - Wong SY
AU  - Galante YM
AU  - Hatefi Y
TI  - Equilibrium binding of 125I-labeled adenosinetriphosphatase inhibitor protein to complex V of the mitochondrial oxidative phosphorylation system
RP  - NOT IN FILE
NT  - UI - 83101213LA - engRN - 0 (ATPase inhibitory protein)RN - 0 (Bicarbonates)RN - 0 (Buffers)RN - 0 (Iodine Radioisotopes)RN - 0 (Proteins)RN - 0 (complex V (mitochondrial oxidative phosphorylation system))RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - AM 08126/AM/NIADDKDA - 19830311IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:6217833
SO  - Biochemistry 1982 Nov 9 ;21(23):5781-5787

516
UI  - 780
AU  - Yoshida M
AU  - Allison WS
AU  - Esch FS
AU  - Futai M
TI  - The specificity of carboxyl group modification during the inactivation of the Escherichia coli F1-ATPase with dicyclohexyl[14C]carbodiimide
RP  - NOT IN FILE
NT  - UI - 82265650LA - engRN - 0 (Amino Acids)RN - 0 (Carbodiimides)RN - 0 (Carbon Radioisotopes)RN - 0 (Membrane Proteins)RN - 0 (Peptide Fragments)RN - 506-68-3 (Cyanogen Bromide)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - EC 3.4.21.4 (Trypsin)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM-16974/GM/NIGMSDA - 19821021IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:6213615
SO  - J Biol Chem 1982 Sep 10 ;257(17):10033-10037

517
UI  - 542
AU  - Amzel LM
AU  - Pedersen PL
TI  - Proton atpases: structure and mechanism
RP  - NOT IN FILE
NT  - UI - 83307251LA - engRN - 0 (Macromolecular Systems)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewID - CA 10951/CA/NCIID - GM 25432/GM/NIGMSDA - 19831021IS - 0066-4154SB - IMCY - UNITED STATESJC - 6DJ
UR  - PM:6225377
SO  - Annu Rev Biochem 1983  ;52():801-824

518
UI  - 21242
AU  - Andersen OS
TI  - Ion movement through gramicidin A channels. Studies on the diffusion- controlled association step
AB  - The permeability characteristics of gramicidin A channels are generally considered to reflect accurately the intrinsic properties of the channels themselves; i.e., the aqueous convergence regions are assumed to be negligible barriers for ion movement through the channels. The validity of this assumption has been examined by an analysis of gramicidin A single-channel current-voltage characteristics up to very high potentials (500 mV). At low permeant ion concentrations the currents approach a voltage-independent limiting value, whose magnitude is proportional to the permeant ion concentration. The magnitude of this current is decreased by experimental maneuvers that decrease the aqueous diffusion coefficient of the ions. It is concluded that the magnitude of this limiting current is determined by the diffusive ion movement through the aqueous convergence regions up to the channel entrance. It is further shown that the small-signal (ohmic) permeability properties also reflect the existence of the aqueous diffusion limitation. These results have considerable consequences for the construction of kinetic models for ion movement through gramicidin A channels. It is shown that the simple two-site-three-barrier model commonly used to interpret gramicidin A permeability data may lead to erroneous conclusions, as biionic potentials will be concentration dependent even when the channel is occupied by at most one ion. The aqueous diffusion limitation must be considered explicitly in the analysis of gramicidin A permeability characteristics. Some implications for understanding the properties of ion-conducting channels in biological membranes will be considered
MH  - A
MH  - analysis
MH  - Diffusion
MH  - ion
MH  - Ion Channels
MH  - Ions
MH  - membrane
MH  - Membranes
MH  - model
MH  - Movement
MH  - Permeability
MH  - Sucrose
RP  - NOT IN FILE
NT  - UI - 83179177LA - engRN - 0 (Glycerides)RN - 0 (Ion Channels)RN - 1405-97-6 (Gramicidin)RN - 25496-72-4 (monoolein)RN - 57-50-1 (Sucrose)RN - 7440-46-2 (Cesium)PT - Journal ArticleID - GM 21342/GM/NIGMSDA - 19830623IS - 0006-3495SB - IMCY - UNITED STATES
UR  - PM:6188502
SO  - Biophys J 1983 Feb ;41(2):147-165

519
UI  - 21267
AU  - Antonenko YN
AU  - Yaguzhinsky LS
TI  - A new method of the measurement of the electrically neutral fluxes of cations through lipid bilayer membranes induced by Men+/nH+-exchangers
AB  - Electrically neutral ionophores (nigericin, monencin) incorporated into a planar bilayer lipid membrane (BLM) bring about hydrogen ion gradient formation in the unstirred layers of BLM if a metal ion gradient on the membrane is prepared. Under these conditions a diffusion potential of a hydrogen ion is generated after addition of a protonophore. Cation selectivity of nigericin, monencin and A23187 has been studied by means of electrical potential measurements in the presence of a protonophore and Men+/nH+-exchangers mentioned above. The data on cation selectivity are in a good agreement with the well known results of the direct measurements of metal ion fluxes. This shows that the effect of generation of the potential on BLM in the presence of a protonophore and a Men+/nH+-exchanger can be used for the estimation of electrically neutral ion fluxes through BLM
MH  - A
MH  - Antibiotics
MH  - Cations
MH  - Diffusion
MH  - diffusion potential
MH  - Hydrogen
MH  - ion
MH  - Ionophores
MH  - Lipid Bilayers
MH  - membrane
MH  - Membranes
MH  - method
MH  - protonophore
RP  - NOT IN FILE
NT  - UI - 84029200LA - engRN - 0 (Antibiotics)RN - 0 (Cations)RN - 0 (Ionophores)RN - 0 (Lipid Bilayers)RN - 17090-79-8 (Monensin)RN - 28380-24-7 (Nigericin)RN - 52665-69-7 (Calcimycin)PT - Journal ArticleDA - 19831217IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:6414844
SO  - FEBS Lett 1983 Oct 31 ;163(1):42-45

520
UI  - 847
AU  - Clark AJ
AU  - Cotton NP
AU  - Jackson JB
TI  - The influence of the ionic conductance on the relation between electron transport and proton-motive force in intact cells of Rhodopseudomonas capsulata
AB  - 1. The dependence of membrane potential (delta psi) on the rate of respiration in darkened intact cell suspensions of Rhodopseudomonas capsulata was distinctly non-linear: severe inhibition of respiration with either rotenone or KCN led to only a small drop in delta psi. 2. In the presence of 0.3 microMs carbonylcyanide p- trifluoromethoxyphenylhydrazone [CF3OPhzC(CN)2] the dependence of delta psi on respiratory rate became linear. Consequently, and particularly at lower concentrations of CF3OPhzC(CN)2, there was a pronounced, synergistic depression of the respiratory delta psi with CF3OPhzC(CN)2 and either rotenone or KCN. 3. Antimycin A, at a concentration which strongly inhibited the photosynthetic electron transport chain, only slightly lowered the light-induced delta psi in anaerobic cell suspensions. Antimycin and CF3OPhzC(CN)2 synergistically lowered delta psi generated by illumination. 4. The light-induced delta psi in anaerobic cells was only about 1.5-times larger than the respiratory- induced delta psi in darkened cells. Nevertheless it required approximately 16-times more CF3OPhzC(CN)2 to collapse the photosynthetic delta psi than the respiratory delta psi. 5. These results are discussed with reference to the ionic current/delta psi relation described in [J.B. Jackson (1982) FEBS Lett. 139, 139-143]. The unifying feature is that the intrinsic conductance of the cell membrane is strongly dependent on delta psi but the conductance due to CF3OPhzC(CN)2 is independent of delta psi
RP  - NOT IN FILE
NT  - UI - 83131702LA - engRN - 0 (Protons)RN - 370-86-5 (Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone)RN - 642-15-9 (Antimycin A)PT - Journal ArticleDA - 19830415IS - 0014-2956SB - IMCY - GERMANY, WESTJC - EMZ
UR  - PM:6297908
SO  - Eur J Biochem 1983 Feb 15 ;130(3):575-580

521
UI  - 846
AU  - Cotton NP
AU  - Clark AJ
AU  - Jackson JB
TI  - Interaction between the respiratory and photosynthetic electron transport chains of intact cells of Rhodopseudomonas capsulata mediated by membrane potential
AB  - 1. The inhibition of respiration by light and the relief from this inhibition by uncoupling agents have been studied in intact cells of Rhodopseudomonas capsulata. 2. The same concentration range of carbonylcyanide p-trifluoromethoxyphenylhydrazone [CF3OPhzC(CN)2] prevented light-inhibition of respiration and depressed the membrane potential (delta psi) in illuminated, aerobic bacterial suspensions. 3. For a wide range of CF3OPhzC(CN)2 concentration the dependence of the respiration rate on delta psi in darkened cells coincided with that in illuminated cells. 4. When present separately, antimycin A and low concentrations of CF3OPhzC(CN)2 had little effect on light-inhibition of respiration or on delta psi in illuminated, aerobic cells, but added together they gave relief from light-inhibition and they lowered delta psi. 5. In the absence of CF3OPhzC(CN)2 even very low light intensities were sufficient to inhibit respiration and increase delta psi. In the presence of CF3OPhzC(CN)2 much higher light intensities were necessary. 6. The results are interpreted as evidence that light-inhibition of respiration is mediated by delta psi. The modification of the response by uncouplers, inhibitors and with light intensity is critically determined by the ionic conductance properties of the membrane in accordance with the findings in [Clark, A.J., Cotton, N.P.J., and Jackson, J.B. (1983) Eur. J. Biochem. 130, 575-580]
RP  - NOT IN FILE
NT  - UI - 83131703LA - engRN - 370-86-5 (Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone)RN - 642-15-9 (Antimycin A)PT - Journal ArticleDA - 19830415IS - 0014-2956SB - IMCY - GERMANY, WESTJC - EMZ
UR  - PM:6825711
SO  - Eur J Biochem 1983 Feb 15 ;130(3):581-587

522
UI  - 21263
AU  - Crofts AR
AU  - Wraight CA
TI  - The electrochemical domain of photosynthesis. 
MH  - bc1
MH  - Cytochrome b
MH  - Cytochrome c1
MH  - Photosynthesis
RP  - NOT IN FILE
SO  - Biochim Biophys Acta 1983  ;726():149-185

523
UI  - 56
AU  - Fillingame RH
AU  - Mosher ME
AU  - Negrin RS
AU  - Peters LK
TI  - H+-ATPase of Escherichia coli uncB402 mutation leads to loss of chi subunit of subunit of F0 sector
AB  - The uncB402 mutation in Escherichia coli results in formation of an H+- ATPase complex that is defective in energy-transducing capacity. The mutation, originally described by Butlin et al. (Butlin, J.D., Cox, G.B., and Gibson, F. (1973) Biochim. Biophys. Acta 292, 366-375), alters the F0 sector of the H+-ATPase complex. Here, we show that uncB402 is an amber-suppressible, chain-terminating mutation that results in loss of the chi subunit from F0. This was demonstrated in crude membrane fractions after overproduction of the ATPase complex by heat induction of a lambda transducing phage carrying the unc operon of uncB402. The lambda-uncB402 DNA was used as a template in an in vitro transcription-translation system. A synthesis product that may correspond to the truncated form of the chi subunit was observed. Despite the absence of chi, the F1-ATPase was still bound to the membrane, although more weakly than in wild type. The omega subunit of F0 ("dicyclohexylcarbodiimide-binding protein") shows normal reactivity with dicyclohexylcarbodiimide, indicating that at least this portion of F0 integrates properly in the membrane in the absence of the chi subunit. The F0 of uncB402 was not functional in H+ translocation activity. This was shown by direct H+ flux measurements with crude membrane vesicles that were treated with guanidine to disrupt the binding of F1 to F0. Secondly, a method was developed for isolation of F0 from F1-depleted membranes. The F0 from uncB402 was shown to have less than 5% the proton-translocase activity of wild type F0 when reconstituted into liposomes. Although the uncB402 mutant shows these defects, the question of whether the chi subunit plays a direct role in F1-binding or H+ translocation remains open, since the loss of chi may lead to subtle changes in the assembly of the other F0 subunits. Analysis of other mutants should permit a more definitive assignment of function
RP  - NOT IN FILE
NT  - UI - 83082929LA - engRN - 0 (Macromolecular Systems)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - 5 T32 GM07215/GM/NIGMSID - GM-23105/GM/NIGMSDA - 19830214IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:6217206
SO  - J Biol Chem 1983 Jan 10 ;258(1):604-609

524
UI  - 478
AU  - Fimmel AL
AU  - Jans DA
AU  - Langman L
AU  - James LB
AU  - Ash GR
AU  - Downie JA
AU  - Senior AE
AU  - Gibson F
AU  - Cox GB
TI  - The F1F0-ATPase of Escherichia coli. Substitution of proline by leucine at position 64 in the c-subunit causes loss of oxidative phosphorylation
AB  - The uncE410 allele differs from the normal uncE gene in that C leads to T base changes occur at nucleotides 190 and 191, resulting in proline at position 64 in the c-subunit of the F1F0-ATPase being replaced by leucine. Two partial-revertant strains were isolated in which alanine- 20 of the c-subunit was replaced by proline, owing to a G leads to C base change at nucleotide 58. These c-subunits, coded for by the uncE501 and uncE502 alleles, therefore contained two amino acid changes, namely proline-64 leads to leucine, and alanine-20 leads to proline. Membranes prepared from the partial-revertant strains lacked ATP-dependent atebrin-fluorescence-quenching activity but were able to carry out oxidative phosphorylation. The ATPase activity of the F1- ATPase was inhibited when bound to membranes from strains carrying the uncE410, uncE501 and uncE502 alleles. It is concluded that a bend in the helix axis in one of the arms of the c-subunit hairpin structure is required for integration of the c-subunit into a functional F1F0-ATPase
RP  - NOT IN FILE
NT  - UI - 83308564LA - engRN - 147-85-3 (Proline)RN - 7005-03-0 (Leucine)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19831008IS - 0264-6021SB - IMCY - ENGLANDJC - 9YO
UR  - PM:6193778
SO  - Biochem J 1983 Aug 1 ;213(2):451-458

525
UI  - 21387
AU  - Friedl P
AU  - Hoppe J
AU  - Gunsalus RP
AU  - Michelsen O
AU  - von Meyenburg K
AU  - Schairer HU
TI  - Membrane integration and function of the three F0 subunits of the ATP synthase of Escherichia coli K12
AB  - Integration into the cytoplasmic membrane and function of the three F0 subunits, a, b and c, of the membrane-bound ATP synthase of Escherichia coli K12 were analysed in situations where synthesis of only one or two types of subunits was possible. This was achieved by combined use of atp mutations and plasmids carrying and expressing one or two of the atp genes coding for ATP synthase subunits. AU three F0 subunits were found to be required for the establishment of efficient H+ conduction. Subunits a and b individually as well as together were found to bind F1 ATPase to the membrane while subunit c did not. The ATPase activity bound to either of these single subunits, or in pairwise combinations, was not inhibited by N,N'-dicyclohexylcarbodiimide. Also ATP-dependent H+ translocation was not catalysed unless all three F0 subunits were present in the membrane. The integration into the membrane of the subunits a and b was independent of the presence of other ATP synthase subunits
MH  - A
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Bacterial Proton-Translocating ATPases
MH  - Cell Membrane
MH  - Cell Membrane Permeability
MH  - enzymology
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - function
MH  - genetics
MH  - H+
MH  - membrane
MH  - metabolism
MH  - mutagenesis
MH  - Mutation
MH  - P
MH  - Plasmids
MH  - Protons
MH  - SUBUNIT
MH  - Support,Non-U.S.Gov't
MH  - SYNTHASE
MH  - synthesis
MH  - translocation
RP  - NOT IN FILE
NT  - Department of Stoffwechselregulation, GBF-Gesellschaft fur Biotechnologische Forschung mbH, Braunschweig-Stockheim, FRGFAU - Friedl, P
SO  - EMBO J 1983  ;2(1):99-103

526
UI  - 20908
AU  - Futai M
AU  - Kanazawa H
TI  - Structure and function of proton-translocating adenosine triphosphatase (F0F1): biochemical and molecular biological approaches
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - ADENOSINE-TRIPHOSPHATASE
MH  - ATPase
MH  - F0F1
MH  - function
MH  - review
RP  - NOT IN FILE
NT  - UI - 84039286LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - 9007-49-2 (DNA)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticlePT - ReviewDA - 19831220IS - 0146-0749SB - IMCY - UNITED STATES
UR  - PM:6226867
SO  - Microbiol Rev 1983 Sep ;47(3):285-312

527
UI  - 373
AU  - Gay NJ
AU  - Walker JE
TI  - Homology between human bladder carcinoma oncogene product and mitochondrial ATP-synthase
AB  - More than 10 different dominant transforming genes (oncogenes) have been identified in human tumours. A human bladder carcinoma oncogene, closely related in sequence to retroviral transforming genes, is split into four exons; the first encodes the N-terminal 37 residues of p21, a protein of unknown function. The oncogene is activated by a single point mutation (guanine to thymine) resulting in the change glycine to valine at position 12 of p21 (refs 3, 4). We report here that the amino acid sequence surrounding this residue is highly homologous to the beta- subunit of mitochondrial and bacterial ATP-synthase in the region of the polypeptide that is believed to contribute to nucleotide binding. Thus, p21 may form part of an enzyme that uses purine nucleotides in catalysis. This is consistent with the finding that an equivalent murine oncogene product binds GTP
RP  - NOT IN FILE
NT  - UI - 83115270LA - engRN - 0 (Multienzyme Complexes)RN - EC 2.7 (Phosphotransferases)RN - EC 2.7.4.- (ATP synthetase complex)PT - Journal ArticleDA - 19830317IS - 0028-0836SB - IMCY - ENGLANDJC - NSC
UR  - PM:6296696
SO  - Nature 1983 Jan 20 ;301(5897):262-264

528
UI  - 21101
AU  - Guffanti AA
AU  - Fuchs RT
AU  - Krulwich TA
TI  - Oxidative phosphorylation by isolated membrane vesicles from Bacillus megaterium and its uncoupler-resistant mutant derivative
AB  - ATP synthesis was studied in ADP + Pi-loaded, right-side-out membrane vesicles from Bacillus megaterium and its uncoupler-resistant mutant strain, C8. Upon energization with ascorbate/phenazine methosulfate, more ATP synthesis was observed in C8 vesicles than in those from the wild type. ATP synthesis by C8 vesicles was more resistant to low levels (0.5-1.0 microM) of carbonyl cyanide m-chlorophenylhydrazone than was synthesis by wild type vesicles, whereas synthesis by both preparations was completely inhibited by N,N'-dicyclohexylcarbodiimide. Upon energization by a valinomycin-induced potassium diffusion potential, vesicles from the wild type strain synthesized more ATP than vesicles from C8, but that synthesis was still lower than observed with electron donors. The results indicate that the characteristic bioenergetic properties exhibited by whole cells of C8 are retained in a vesicle system and thus cannot be attributed to a cytoplasmic, substrate level activity. Interestingly, lipophilic cations that were efficacious in measuring the transmembrane electrical potential of whole cells appeared to accurately measure artificially generated potentials across vesicle membranes, but were not taken up upon addition of ascorbate/phenazine methosulfate
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - ADP
MH  - atp
MH  - ATP synthesis
MH  - Bacillus
MH  - Carbonyl Cyanide m-Chlorophenyl Hydrazone
MH  - Cations
MH  - Cells
MH  - Diffusion
MH  - diffusion potential
MH  - electron
MH  - membrane
MH  - membrane vesicles
MH  - Membranes
MH  - mutant
MH  - Oxidative Phosphorylation
MH  - Phosphorylation
MH  - Potassium
MH  - synthesis
MH  - SYSTEM
MH  - vesicles
RP  - NOT IN FILE
NT  - UI - 83082889LA - engRN - 555-60-2 (Carbonyl Cyanide m-Chlorophenyl Hydrazone)RN - 56-65-5 (Adenosine Triphosphate)PT - Journal ArticleID - GM28454/GM/NIGMSDA - 19830214IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:6401293
SO  - J Biol Chem 1983 Jan 10 ;258(1):35-37

529
UI  - 54
AU  - Hermolin J
AU  - Gallant J
AU  - Fillingame RH
TI  - Topology, organization, and function of the psi subunit in the F0 sector of the H+-ATPase of Escherichia coli
AB  - The F1F0 H+-ATPase in membranes of Escherichia coli was amplified by heat induction of a lysogenic lambda-unc+ transducing phage. Inverted membrane vesicles were stripped of the F1 sector of the ATPase complex by washing with EDTA. The stripped membranes were treated with dithiobis(succimidylpropionate) to cross-link subunits of the F0 sector of the ATPase complex. After electrophoresis under nonreducing conditions in one dimension, cross-linked subunits were identified by off-diagonal electrophoresis in a second dimension following cleavage of the cross-linked products with beta-mercaptoethanol. A psi-psi dimer was the major cross-linked product identified. In addition, a chi-psi product and chi-psi2 product were identified. These results support the proposed chi-psi2 stoichiometry of subunits in F0. When the F1-stripped membranes were treated with trypsin, the psi subunit was rapidly degraded, whereas psi was protected from degradation when F1 was bound to the membrane. Trypsin-treated, stripped membranes, lacking an intact psi subunit, did not bind the F1 portion of the ATPase with high affinity. However, these trypsin-treated stripped membranes remained as permeable to protons as untreated stripped membranes, and the H+ conductivity was blocked by dicyclohexylcarbodiimide. These results indicate that the portion of the psi subunit exposed on the cytoplasmic face of the inner membrane is involved in the binding of the F1 portion of the ATPase, but is not necessary for H+ conduction mediated by the F0 sector of the complex
RP  - NOT IN FILE
NT  - UI - 84061911LA - engRN - 0 (Macromolecular Systems)RN - EC 3.4.21.4 (Trypsin)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM-23105/GM/NIGMSDA - 19840127IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:6227625
SO  - J Biol Chem 1983 Dec 10 ;258(23):14550-14555

530
UI  - 371
AU  - Hollemans M
AU  - Runswick MJ
AU  - Fearnley IM
AU  - Walker JE
TI  - The sites of labeling of the beta-subunit of bovine mitochondrial F1- ATPase with 8-azido-ATP
AB  - Reaction of the photoaffinity label 8-azido-[2-3H] ATP with bovine heart mitochondrial F1-ATPase abolishes its enzyme activity; inhibition is prevented by ATP (Wagenvoord, R.J., van der Kraan, I., and Kemp, A. (1977) Biochim. Biophys. Acta 460, 17-24). More than 65% of the radioactivity is associated with the beta-subunit and about 25% with the alpha-chains. Radioactivity in the beta-subunit is localized in two specific regions. One corresponds to residues 1-12 (Runswick, M.J., and Walker, J.E. (1983) J. Biol. Chem. 258, 3081-3089), a region which is nonessential for catalysis. Radioactivity in the second region is localized predominantly on three amino acids, lysine 301, isoleucine 304, and tyrosine 311. It seems likely that these residues are found in the vicinity of the ATP binding site of F1-ATPase
RP  - NOT IN FILE
NT  - UI - 83265727LA - engRN - 0 (Azides)RN - 0 (Macromolecular Systems)RN - 53696-59-6 (8-azidoadenosine 5'-triphosphate)RN - 55520-40-6 (Tyrosine)RN - 56-65-5 (Adenosine Triphosphate)RN - 56-87-1 (Lysine)RN - 7004-09-3 (Isoleucine)RN - EC 3.4.21.4 (Trypsin)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19830909IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:6223927
SO  - J Biol Chem 1983 Aug 10 ;258(15):9307-9313

531
UI  - 8283
AU  - Hong YQ
AU  - Junge W
TI  - Localized or delocalized protons in photophosphorylation? On the accessibility of the thylakoid lumen for ions and buffers
MH  - bioenergetics
MH  - buffer
MH  - Buffers
MH  - indicator
MH  - ion
MH  - Ions
MH  - localized coupling
MH  - NEUTRALRED
MH  - Photophosphorylation
MH  - proton
MH  - Protons
MH  - SURFACE
MH  - thylakoid
RP  - NOT IN FILE
SO  - Biochim Biophys Acta 1983  ;722():197-208

532
UI  - 21388
AU  - Hoppe J
AU  - Friedl P
AU  - Schairer HU
AU  - Sebald W
AU  - von Meyenburg K
AU  - Jorgensen BB
TI  - The topology of the proton translocating F0 component of the ATP synthase from E. coli K12: studies with proteases
AB  - The accessibility of the three F0 subunits a, b and c from the Escherichia coli K12 ATP synthase to various proteases was studied in F1-depleted inverted membrane vesicles. Subunit b was very sensitive to all applied proteases. Chymotrypsin produced a defined fragment of mol. wt. 15,000 which remained tightly bound to the membrane. The cleavage site was located at the C-terminal region of subunit b. Larger amounts of proteases were necessary to attack subunit a (mol. wt. 30,000). There was no detectable cleavage of subunit c. It is suggested that the major hydrophilic part of subunit b extends from the membrane into the cytoplasm and is in contact with the F1 sector. The F1 sector was found to afford some protection against proteolysis of the b subunit in vitro and in vivo. Protease digestion had no influence on the electro-impelled H+ conduction via F0 but ATP-dependent H+ translocation could not be reconstituted upon binding of F1. A possible role for subunit b as a linker between catalytic events on the F1 component and the proton pathway across the membrane is discussed
MH  - A
MH  - atp
MH  - ATP synthase
MH  - Bacterial Proton-Translocating ATPases
MH  - BINDING
MH  - catalytic
MH  - Chymotrypsin
MH  - Cytoplasm
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - F1
MH  - H+
MH  - In Vitro
MH  - membrane
MH  - membrane vesicles
MH  - metabolism
MH  - proton
MH  - Protons
MH  - Site
MH  - Subtilisin
MH  - SUBUNIT
MH  - Support,Non-U.S.Gov't
MH  - SYNTHASE
MH  - translocation
MH  - Trypsin
MH  - vesicles
RP  - NOT IN FILE
NT  - Department of Stoffwechselregulation, GBF-Gesellschaft fur Biotechnologische Forschung mbH, Braunschweig-Stockheim, FRGFAU - Hoppe, J
SO  - EMBO J 1983  ;2(1):105-110

533
UI  - 490
AU  - Jans DA
AU  - Fimmel AL
AU  - Langman L
AU  - James LB
AU  - Downie JA
AU  - Senior AE
AU  - Ash GR
AU  - Gibson F
AU  - Cox GB
TI  - Mutations in the uncE gene affecting assembly of the c-subunit of the adenosine triphosphatase of Escherichia coli
AB  - The amino acid substitutions in the mutant c-subunits of Escherichia coli F1F0-ATPase coded for by the uncE429, uncE408 and uncE463 alleles affect the incorporation of these proteins into the cell membrane. The DNA sequence of the uncE429 allele differed from normal in that a G leads to A base change occurred at nucleotide 68 of the uncE gene, resulting in glycine being replaced by aspartic acid at position 23 in the c-subunit. The uncE408 and uncE463 mutant DNA sequences were identical and differed from normal in that a C leads to T base change occurred at nucleotide 91 of the uncE gene, resulting in leucine being replaced by phenylalanine at position 31 in the c-subunit. An increased gene dosage of the uncE408 or uncE463 alleles resulted in the incorporation into the membranes of the mutant c-subunits. The results are discussed in terms of the 'Helical Hairpin Hypothesis' of Engelman & Steitz [(1981) Cell 23,411-422]
RP  - NOT IN FILE
NT  - UI - 83282613LA - engRN - 0 (Carrier Proteins)RN - 0 (DNA, Bacterial)RN - 0 (Plasmids)RN - 0 (dicyclohexylcarbodiimide-binding protein)RN - EC 3.1.21 (DNA Restriction Enzymes)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19830920IS - 0264-6021SB - IMCY - ENGLANDJC - 9YO
UR  - PM:6309138
SO  - Biochem J 1983 Jun 1 ;211(3):717-726

534
UI  - 20909
AU  - Kanazawa H
AU  - Noumi T
AU  - Futai M
AU  - Nitta T
TI  - Escherichia coli mutants defective in the gamma subunit of proton- translocating ATPase: intracistronic mapping of the defective site and the biochemical properties of the mutants
AB  - Various hybrid plasmids carrying a portion of the gene for the gamma subunit of the H+-ATPase of Escherichia coli complemented five mutants defective in the enzyme in a genetic test, indicating that the mutants are defective in the gamma subunit. Since the nucleotide sequence of genomic DNA carried on the plasmids is known, the defective site(s) of the mutants could be located within the gene for the gamma subunit as follows: KF10 and NR70, KF1, and KF12 and KF13 have a mutation causing a defect(s) in amino acid residues 1 to 82, 83 to 167, and 168 to 287, respectively, of the gamma subunit. The biochemical properties of all these mutants except NR70 were analyzed in terms of proton permeability of the membranes and assembly of F1. Results suggested that KF1 and KF10 have defective F1 without at least the alpha and beta subunits on their membranes, whereas KF12 and KF13 have F1's of rather similar structure to that of the wild type. Attempts were made to purify F1 oF KF12 as a single complex. Although the F1 complex dissociated during purification, active alpha and beta subunits of KF12 were partially purified. On the basis of these biochemical and genetic results, it is suggested that structural alterations in the primary sequence of the gamma subunit corresponding to residues 1 to 167 cause more extensive defects in the assembly of F1 than alteration in the sequence of residues 168 to 287
MH  - A
MH  - ACID
MH  - ACTIVE
MH  - Adenosinetriphosphatase
MH  - ATPase
MH  - BETA-SUBUNIT
MH  - COMPLEX
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1
MH  - H+-ATPase
MH  - membrane
MH  - Membranes
MH  - mutant
MH  - Permeability
MH  - proton
MH  - Protons
MH  - purification
MH  - RESIDUE
MH  - Site
MH  - SUBUNIT
RP  - NOT IN FILE
NT  - UI - 83229687LA - engRN - 0 (Protons)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19830708IS - 0003-9861SB - IMCY - UNITED STATES
UR  - PM:6190439
SO  - Arch Biochem Biophys 1983 Jun ;223(2):521-532

535
UI  - 20907
AU  - Kanazawa H
AU  - Noumi T
AU  - Oka N
AU  - Futai M
TI  - Intracistronic mapping of the defective site and the biochemical properties of beta subunit mutants of Escherichia coli H+-ATPase: correlation of structural domains with functions of the beta subunit
AB  - Sixteen mutants of Escherichia coli defective in H+-ATPase (proton- translocating ATPase) were tested for their ability to recombine with hybrid plasmids carrying various portions of the beta subunit cistron. Twelve mutations were mapped within the carboxyl half of the cistron corresponding to amino acid residues 279 to 459 (domain II), while four mutations were mapped within residues 17 to 278 (domain I). The biochemical properties of these mutants were analyzed in terms of the proton permeability of their membranes and the assembly properties of their F1F0 complex. The mutants were classified according to the properties into three types, I, II, and III. In 12 mutants of type I, proton conduction in membrane vesicles was blocked and little F1 was released from the membranes under conditions in which F1 could be released from wild-type membranes, suggesting that assembly of the F1F0 complex is structurally and functionally defective. F1 was partially purified with very low recovery from one of the type I mutants, KF16. ATPase activity was reconstituted from this F1 with the beta subunit of the wild type, confirming the genetic results. Only one mutant, KF38, was classified as type II. Its membranes were partially leaky to protons and its F1 was releasable, suggesting that the interaction of its F1 and F0 was unstable. Type III mutants, KF11 and KF43, had an F1F0 complex with very low activity, in which the structure of F1 was relatively similar to that of the wild type. F1 was purified as a single complex from KF43 in this study and from KF11 previously (H. Kanazawa, Y. Horiuchi, M. Takagi, Y. Ishino, and M. Futai (1980) J. Biochem. 88, 695-703). Reconstitution experiments in vitro showed that the F1's of both mutants were defective in the beta subunit. The properties of the altered F1 of KF43 differed from those of F1 of KF11, suggesting that the mutation sites of KF43 and KF11 were different. From the results of mapping mutation sites and the biochemical properties of the mutants, the correlation of structural domains with function of the beta subunit is discussed. Most type I and type II mutations except that of KF39 were mapped in domain II, while the type III mutations were mapped in domain I, suggesting that domain II is more important than domain I for the function of the beta subunit, especially in terms of proper assembly of the F1F0 complex
MH  - A
MH  - ACID
MH  - ATPase
MH  - BETA-SUBUNIT
MH  - COMPLEX
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - F1
MH  - function
MH  - H+-ATPase
MH  - In Vitro
MH  - M
MH  - membrane
MH  - membrane vesicles
MH  - Membranes
MH  - mutant
MH  - Permeability
MH  - proton
MH  - Protons
MH  - reconstitution
MH  - RESIDUE
MH  - Site
MH  - SUBUNIT
RP  - NOT IN FILE
NT  - UI - 84126786LA - engRN - 0 (Enzyme Precursors)RN - 0 (Protons)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19840306IS - 0003-9861SB - IMCY - UNITED STATES
UR  - PM:6320730
SO  - Arch Biochem Biophys 1983 Dec ;227(2):596-608

536
UI  - 9906
AU  - Kashket ER
TI  - Stoichiometry of the H+-ATPase of Escherichia coli cells during anaerobic growth.
AB  - The H+/ATP stoichiometry of the H+-ATPase was investigated in Escherichia coli cells growing under anaerobic conditions at pH 6 and 7. The protonmotive force was determined from the intracellular accumulation of benzoate and tetraphenylphosphonium ions, as well as the accumulation of lactose in this lac operon inducible, but beta-galactosidase negative strain. The phosphorylation potential was calculated from the cellular concentrations of ATP, ADP and inorganic phosphate. By comparing the phosphorylation potential and the proton motive force under these steady state conditions, the H+/ATP stoichiometry was determined to be 3, similar to the value previously found in the same cells growing under aerobic conditions.
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - ADP
MH  - Anaerobiosis
MH  - atp
MH  - Benzoates
MH  - Cells
MH  - enzymology
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - H+-ATPase
MH  - Hydrogen-Ion Concentration
MH  - INORGANIC-PHOSPHATE
MH  - Ions
MH  - Lactose
MH  - metabolism
MH  - Onium Compounds
MH  - Phosphorylation
MH  - proton
MH  - Support,U.S.Gov't,Non-P.H.S.
RP  - NOT IN FILE
SO  - FEBS Lett 1983 Apr 18 ;154(2):343-346

537
UI  - 21171
AU  - Klionsky DJ
AU  - Brusilow WS
AU  - Simoni RD
TI  - Assembly of a functional F0 of the proton-translocating ATPase of Escherichia coli
AB  - We have investigated both structural and functional assembly of the F0 portion of the Escherichia coli proton-translocating ATPase in vivo. Fractionation of E. coli minicells containing plasmids which code for parts of the unc operon shows that each of the F0 peptides a, b, and c insert into the cytoplasmic membrane independent of each other and without the polypeptides which form the F1 portion of the complex alpha, beta, gamma, delta, and epsilon. Assays of membrane energization indicate that, while formation of a functional proton channel requires the presence of all three F0 polypeptides a, b and c, they are not sufficient. Synthesis of both the alpha and beta subunits of the F1 are required for formation of a functional proton channel
MH  - A
MH  - Adenosinetriphosphatase
MH  - alpha
MH  - ATPase
MH  - BETA
MH  - BETA-SUBUNIT
MH  - COMPLEX
MH  - delta
MH  - Enzymes
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - F1
MH  - Macromolecular Systems
MH  - membrane
MH  - proton
MH  - Proton-Translocating ATPases
MH  - SUBUNIT
MH  - synthesis
MH  - SYSTEM
MH  - SYSTEMS
RP  - NOT IN FILE
NT  - UI - 83290854LA - engRN - 0 (Macromolecular Systems)RN - 0 (Plasmids)RN - EC 3.1.21 (DNA Restriction Enzymes)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleID - GM07276/GM/NIGMSID - GM07598/GM/NIGMSID - GM18539/GM/NIGMSDA - 19831008IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:6309770
SO  - J Biol Chem 1983 Aug 25 ;258(16):10136-10143

538
UI  - 19813
AU  - Kohlbrenner WE
AU  - Boyer PD
TI  - Probes of catalytic site cooperativity during catalysis by the chloroplast adenosine triphosphate and the adenosine triphosphate synthase
AB  - During net nucleoside triphosphate synthesis by chloroplast ATP synthase the extent of water oxygen incorporation into each nucleoside triphosphate released increases with decrease in ADP, GDP or IDP concentration. Likewise, during net ATP hydrolysis by the Mg2+- activated chloroplast ATPase, the extent of water oxygen incorporation into each Pi released increases as the ATP, GTP, or ITP concentration is decreased. However, the concentration ranges in which substrate modulation occurs differs with each nucleotide. Modulation of oxygen exchange during synthesis and hydrolysis of adenine nucleotides, as measured by variation in the extent of water oxygen incorporation into products, occurs below 250 microM. In contrast, guanosine and inosine nucleotides alter the extent of exchange at higher and much wider concentration ranges. Activation of the chloroplast ATPase by either heat or trypsin results in similar catalytic behavior as monitored by ATP modulation of oxygen exchanges during hydrolysis in the presence of Mg2+. More exchange capacity is evident with octylglucoside-activated enzyme at all ATP concentrations. High levels of tentoxin were also found to alter the catalytic exchange parameters resulting in continued water oxygen exchange into Pi released during hydrolysis at high ATP concentrations. Little or no oxygen exchange accompanies ATP hydrolysis in the presence of Ca2+. The [18O]Pi species formed from highly gamma- 18O-labeled ATP at lower ATP concentrations gives a distribution as expected if only one catalytic pathway is operative at a given ATP concentration. This and other results support the concept of catalytic cooperativity between alternating sites as explanation for the modulation of oxygen exchange by nucleotide concentration
MH  - A
MH  - ACTIVATION
MH  - Adenine Nucleotides
MH  - ADENINE-NUCLEOTIDES
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - adenosine triphosphate synthase
MH  - ADP
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Calcium
MH  - Catalysis
MH  - chloroplast
MH  - COMPLEX
MH  - Glucosides
MH  - H(+)-Transporting ATP Synthase
MH  - Heat
MH  - Hydrolysis
MH  - Magnesium
MH  - Multienzyme Complexes
MH  - Nucleotides
MH  - Oxygen
MH  - Phosphotransferases
MH  - SYNTHASE
MH  - synthesis
MH  - Trypsin
MH  - Water
RP  - NOT IN FILE
NT  - UI - 83291035LA - engRN - 0 (Glucosides)RN - 0 (Multienzyme Complexes)RN - 0 (Peptides, Cyclic)RN - 28540-82-1 (tentoxin)RN - 29836-26-8 (octyl-beta-D-glucoside)RN - 56-65-5 (Adenosine Triphosphate)RN - 7439-95-4 (Magnesium)RN - 7440-70-2 (Calcium)RN - EC 2.7 (Phosphotransferases)RN - EC 2.7.4.- (ATP synthetase complex)RN - EC 3.4.21.4 (Trypsin)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM 11094/GM/NIGMSDA - 19831028IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:6309819
SO  - J Biol Chem 1983 Sep 25 ;258(18):10881-10886

539
UI  - 970
AU  - Mathis P
TI  - Photosynthesis: transduction of light energy into chemical energy
MH  - Adenosine Triphosphate
MH  - Bacteria
MH  - biosynthesis
MH  - Carbon Dioxide
MH  - Cell Membrane
MH  - Electron Transport
MH  - Light
MH  - metabolism
MH  - Models,Biological
MH  - NADH,NADPH Oxidoreductases
MH  - Oxidation-Reduction
MH  - Photosynthesis
MH  - Quinones
MH  - Rhodobacter sphaeroides
MH  - Thermodynamics
RP  - NOT IN FILE
SO  - Symp Soc Exp Biol 1983  ;36:223-48.():223-248

540
UI  - 21114
AU  - McCarthy JE
AU  - Ferguson SJ
TI  - The effects of partial uncoupling upon the kinetics of ATP synthesis by vesicles from Paracoccus denitrificans and by bovine heart submitochondrial particles. Implications for the mechanism of the proton-translocating ATP synthase
AB  - 1. Reduction in the magnitude of the respiration-dependent protonmotive force (proton electrochemical gradient in mV) of vesicles from Paracoccus denitrificans, and of submitochondrial particles, has been found to be paralleled small increases in S50% values for both ADP and Pi. For example, reduction of the protonmotive force of P. denitrificans vesicles from 145 mV to 110 mV was accompanied by an increase of S50% (ADP) from 8 microM to 18 microM, and an increase of S50% (Pi) from 0.33 mM to 1.4 mM. This result was obtained with partial uncoupling quantities of both carbonyl-cyanide p- trifluoromethoxyphenylhydrazone and of the synergistic combination of nigericin plus valinomycin in the presence of K+. In view of the similar effects of these two different methods of uncoupling it is concluded that the changes in S50% were a consequence of the diminished protonmotive force acting on the ATP synthase rather than of a secondary, direct interaction of the uncouplers with the enzyme. Changes in S50% rather than Km are described because under several sets of conditions double-reciprocal plots were nonlinear. 2. For equivalent attenuations in the rate of ATP synthesis by submitochondrial particles, 2,4-dinitrophenol caused much larger increases in S50% (ATP) than did carbonylcyanide p-trifluoromethoxyphenylhydrazone. Therefore it is concluded that the effect of 2,4-dinitrophenol was primarily a consequence of its previously recognized direct interaction with the F1 segment of the mitochondrial ATPase. The concentration range of 2,4- dinitrophenol that raised S50% (ADP) is similar to that which weakens the binding of ADP to a particular type of site on the purified F1 sector of ATP synthase. This correlation is consistent with such a site having a catalytic role during ATP synthesis. 3. A titration of the rate of ATP synthesis by vesicles of P. denitrificans with increasing quantities of carbonylcyanide p-trifluoromethoxyphenylhydrazone showed that the initial titres of the uncoupler caused large decreases in the rate of ATP synthesis for relatively small attenuations in the protonmotive force. Thus the initial 20 mV drop in the protonmotive force was accompanied by a reduction of more than 65% in the rate of ATP synthesis. Over the lowest range of values of protonmotive force that drove detectable rates of ATP synthesis however, the dependence of the rate was a less steep function of the protonmotive force. A plot of the logarithm of the rate of ATP synthesis against protonmotive force reveals a biphasic relationship. There does not appear to be a 'threshold' value of the protonmotive force below which ATP synthesis is blocked by kinetic factors. 4. The relationships of the protonmotive force with S50% values and with the rate of ATP synthesis (at near saturating concentrations of ADP and Pi) are discussed in relation to possible mechanisms for the coupling of proton translocation to ATP synthesis
MH  - A
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - ADP
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - ATPase
MH  - BINDING
MH  - COMPLEX
MH  - coupling
MH  - DEPENDENCE
MH  - F1
MH  - function
MH  - Kinetics
MH  - mechanism
MH  - MECHANISMS
MH  - method
MH  - Methods
MH  - Multienzyme Complexes
MH  - P
MH  - Phosphotransferases
MH  - proton
MH  - Protons
MH  - secondary
MH  - Site
MH  - Submitochondrial Particles
MH  - SYNTHASE
MH  - synthesis
MH  - translocation
MH  - Uncoupling Agents
MH  - Valinomycin
MH  - vesicles
RP  - NOT IN FILE
NT  - UI - 83182477LA - engRN - 0 (Multienzyme Complexes)RN - 0 (Protons)RN - 0 (Uncoupling Agents)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 2.7 (Phosphotransferases)RN - EC 2.7.4.- (ATP Synthetase Complexes)PT - Journal ArticleDA - 19830610IS - 0014-2956SB - IMCY - GERMANY, WEST
UR  - PM:6301834
SO  - Eur J Biochem 1983 May 2 ;132(2):425-431

541
UI  - 19814
AU  - McIntosh DB
AU  - Boyer PD
TI  - Adenosine 5'-triphosphate modulation of catalytic intermediates of calcium ion activated adenosinetriphosphatase of sarcoplasmic reticulum subsequent to enzyme phosphorylation
MH  - Adenine Nucleotides
MH  - ADENINE-NUCLEOTIDES
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - ATPase
MH  - Ca(2+)-Transporting ATPase
MH  - Calcium
MH  - ion
MH  - Nucleotides
MH  - Oxygen
MH  - Oxygen Isotopes
MH  - Phosphorylation
MH  - Sarcoplasmic Reticulum
RP  - NOT IN FILE
NT  - UI - 83257206LA - engRN - 0 (Adenine Nucleotides)RN - 0 (Oxygen Isotopes)RN - 0 (Ribonucleotides)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.38 (Ca(2+)-Transporting ATPase)PT - Journal ArticleDA - 19830923IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:6223659
SO  - Biochemistry 1983 Jun 7 ;22(12):2867-2875

542
UI  - 55
AU  - Mosher ME
AU  - Peters LK
AU  - Fillingame RH
TI  - Use of lambda unc transducing bacteriophages in genetic and biochemical characterization of H+-ATPase mutants of Escherichia coli
AB  - The eight subunits of the H+-ATPase of Escherichia coli are coded by the genes of the unc operon, which maps between bglB and asnA. A collection of unc mutations were transferred via P1 transduction into a strain in which lambda cI857 S7 was inserted into bglB. The lambda phage was induced, and asnA+ transducing phage that carried unc were selected. Transducing phage carrying mutations in the uncA, B, D, E, and F genes were used for complementation analysis with a collection of unc mutants, including mutants which had been reported previously but not genetically characterized. Some mutations gave a simple complementation pattern, indicating a single defective gene, whereas other mutations gave more complex patterns. Two mutants (uncE105 and uncE107) altered in the proteolipid (omega) subunit of F0 were not complemented by any of the lambda unc phage, even though both mutants had a fully functional F1 ATPase and therefore normal A and D genes. Hence, only limited conclusions can be drawn from genetic complementation alone, since it cannot distinguish normal from abnormal genes in certain classes of unc mutants. The lambda unc phage proved to be essential in characterizing several mutants defective in F0-mediated H+ translocation. The unc gene products were overproduced by heat induction of the lysogenized lambda unc phage to determine whether all the F0 subunits were in the membrane. Two mutants that gave a simple complementation pattern, indicative of one defective gene, did not assemble a three-subunit F0. The uncB108 mutant was shown to lack the chi subunit of F0 but to retain psi and omega. Trace amounts of an altered omega subunit and normal amounts of chi and psi were found in the uncE106 mutant. A substitution of aspartate for glycine at residue 58 of the protein was determined by DNA sequence analysis of the uncE gene cloned from the lambda uncE106 phage DNA. One of the omega- defective, noncomplementing mutants (uncE107) was shown to retain all three F0 subunits. The uncE gene from this mutant was also sequenced to confirm an asparagine-for-aspartate substitution at position 61 (the dicyclohexylcarbodiimide-binding site) of the omega subunit
RP  - NOT IN FILE
NT  - UI - 84061611LA - engRN - 0 (Proteolipids)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - 5 T32 GM07215/GM/NIGMSID - GM-23105/GM/NIGMSDA - 19840127IS - 0021-9193SB - IMCY - UNITED STATESJC - HH3
UR  - PM:6227607
SO  - J Bacteriol 1983 Dec ;156(3):1078-1092

543
UI  - 19811
AU  - Myers JA
AU  - Boyer PD
TI  - Catalytic properties of the ATPase on submitochondrial particles after exchange of tightly bound nucleotides under different steady state conditions
AB  - Energized submitochondrial particles were subjected to high or low [3H]ATP/[3H]ADP ratios, maintained during steady state by a pyruvate kinase or hexokinase regenerating system, respectively. Under both steady state conditions, about 1.4 mol [3H]nucleotide/mol ATPase was retained but considerably more [3H]ATP was retained with the high [3H]ATP/[3H]ADP ratio. The ATPase activity and the oxygen exchange of these differentially labeled SMP were the same, suggesting a lack of control function of non-catalytic tightly bound nucleotides
MH  - A
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - ATPase
MH  - BOUND NUCLEOTIDES
MH  - Hexokinase
MH  - Nucleotides
MH  - Oxygen
MH  - Pyruvate Kinase
MH  - Submitochondrial Particles
RP  - NOT IN FILE
NT  - UI - 84029157LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleID - GM 11904/GM/NIGMSID - GM 17285/GM/NIGMSDA - 19831217IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:6226536
SO  - FEBS Lett 1983 Oct 17 ;162(2):277-281

544
UI  - 644
AU  - Penefsky HS
TI  - On the mechanism of ATP synthesis in oxidative phosphorylation
RP  - NOT IN FILE
NT  - UI - 86020393LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19851121IS - 0028-7113SB - IMCY - UNITED STATESJC - W8Y
UR  - PM:6242323
SO  - Trans N Y Acad Sci 1983  ;41():139-146

545
UI  - 477
AU  - Perlin DS
AU  - Cox DN
AU  - Senior AE
TI  - Integration of F1 and the membrane sector of the proton-ATPase of Escherichia coli. Role of subunit "b" (uncF protein)
AB  - Membranes derived from the Escherichia coli strain AN1460 which carries the multicopy plasmid pAN45 (unc+) (Downie, J. A., Langman, L., Cox, G. B., Yanofsky, C., and Gibson, (1980) J. Bacteriol. 143, 8-17) were enriched 5- to 10-fold in proton-ATPase activity. Incubation of F1- depleted AN1460 membranes with trypsin abolished F1-binding ability but did not inhibit proton transport through the membrane sector (F0). Sodium dodecyl sulfate-gel electrophoresis indicated that subunit "b" (uncF protein) of F0 was cleaved by trypsin and prebound F1 protected against the trypsin effect. Subunits "a" (uncB protein) and "c" (uncE protein) were unaffected by the trypsin treatment. A water-soluble fragment (Mr = 14,800) was liberated after trypsin treatment and appeared to arise from subunit b. Studies of enzyme hybridization and of F1 binding to membranes derived from strains containing mutations in uncB, F, and E genes supported the suggestion that subunit b is involved in F1 binding to the F0. Also, extraction of membranes with KSCN increased the relative proportion of subunit b in the membrane and this coincided with a parallel increase in trypsin-sensitive F1-binding ability. It is proposed that subunit b is involved in binding of F1 to the F0; this agrees with the presumed role of the protein as deduced from predictions of its secondary and tertiary structure (Walker, J. E., Saraste, M., and Gay, N. J. (1982) Nature (Lond.) 298, 867-869; Senior, A. E. (1983) Biochim. Biophys. Acta, in press)
RP  - NOT IN FILE
NT  - UI - 83290900LA - engRN - 0 (Bacterial Proteins)RN - 0 (Macromolecular Systems)RN - EC 3.4.21.4 (Trypsin)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM 29805/GM/NIGMSDA - 19831008IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:6193110
SO  - J Biol Chem 1983 Aug 25 ;258(16):9793-9800

546
UI  - 21170
AU  - Porter AC
AU  - Brusilow WS
AU  - Simoni RD
TI  - Promoter for the unc operon of Escherichia coli
AB  - Fragments of DNA carrying possible promoters for the unc operon of Escherichia coli were cloned into a promoter detection plasmid (pRZ5255). Similar fragments were transcribed in vitro to produce transcripts whose sizes were used to determine the approximate start site for transcription. One strong promoter and at least two very much weaker ones were detected by these methods. The exact position of the strongest promoter, presumed to be the true unc promoter, was determined by S1 nuclease mapping and shown to lie 73 base pairs upstream from the open reading frame that precedes uncB. It therefore appears that this reading frame (uncI) is part of the unc operon. S1 mapping also revealed the presence of a third weak promoter 25 base pairs upstream of uncI. All of the weak promoters occur between the proposed unc promoter and uncB, but their role in vivo, if any, is unclear
MH  - A
MH  - Adenosinetriphosphatase
MH  - ATPase
MH  - BASE
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - In Vitro
MH  - method
MH  - Methods
MH  - Proton-Translocating ATPases
MH  - Site
RP  - NOT IN FILE
NT  - UI - 83290727LA - engRN - 0 (Plasmids)RN - 0 (RNA, Bacterial)RN - 0 (RNA, Messenger)RN - EC 3.1.- (Endonucleases)RN - EC 3.1.30.1 (Aspergillus Nuclease S1)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleID - GM07598/GM/NIGMSID - GM18539/GM/NIGMSDA - 19831021IS - 0021-9193SB - IMCY - UNITED STATES
UR  - PM:6193096
SO  - J Bacteriol 1983 Sep ;155(3):1271-1278

547
UI  - 19875
AU  - Power J
AU  - Cross RL
AU  - Harris DA
TI  - Interaction of F1-ATPase, from ox heart mitochondria with its naturally occurring inhibitor protein. Studies using radio-iodinated inhibitor protein
AB  - The ox heart mitochondrial inhibitor protein may be iodinated with up to 0.8 mol 125I per mol inhibitor with no loss of inhibitory activity, with no change in binding affinity to submitochondrial particles, and without alteration in the response of membrane-bound inhibitor to energisation. Tryptic peptide maps reveal a single labelled peptide, consistent with modification of the single tyrosine residue of the protein. A single type of high-affinity binding site (Kd=96 . 10 (-9)M) for the inhibitor protein has been measured in submitochondrial particles. The concentration of this site is proportional to the amount of membrane-bound F1, and there appears to be one such site per F1 molecule. The ATp hydrolytic activity of submitochondrial particles is inversely proportional to the occupancy of the high-affinity binding site for the inhibitor protein. No evidence is found for a non- inhibitory binding site on the membrane or on other mitochondrial proteins. In intact mitochondria from bovine heart, the inhibitor protein is present in an approx. 1:1 ratio with F1. Submitochondrial particles prepared by sonication of these mitochondria with MgATP contain about 0.75 mol inhibitor protein per mol F1, and show about 25% of the ATPase activity of inhibitor-free submitochondrial particles. Additional inhibitor protein can be bound to these particles to a level of 0.2 mol/mol F1, with consequent loss of ATPase activity. If MgATP is omitted from the medium, or inhibitors of ATP hydrolysis are present, the rate of combination between F1 and its inhibitor protein is very much reduced. The equilibrium level of binding is, however, unaltered. These results suggest the presence of a single, high-affinity, inhibitory binding site for inhibitor protein on membrane-bound F1. The energisation of coupled submitochondrial particles by succinate oxidation or by ATP hydrolysis results in both the dissociation of inhibitor protein into solution, and the activation of ATP hydrolysis. At least 80% of the membrane-bound F1-inhibitor complex responds to this energisation by participating in a new equilibrium between bound and free inhibitor protein. This finding suggests that a delocalised energy pool is important in promoting inhibitor protein release from F1. Dissipation of the electrochemical gradient by uncouplers, or the binding of oligomycin or efrapetin effectively blocks energised release of the inhibitor protein. Conversely, the addition of aurovertin or adenosine 5'--[beta, lambda--imido]triphosphate enhances energy-driven release. The mode of action of various inhibitors on binding and energised release of the protein inhibitor is discussed
MH  - A
MH  - ACTIVATION
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BINDING
MH  - COMPLEX
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - H(+)-Transporting ATP Synthase
MH  - Hydrolysis
MH  - INHIBITOR PROTEIN
MH  - membrane
MH  - Mitochondria
MH  - Peptide Fragments
MH  - Proteins
MH  - Submitochondrial Particles
MH  - succinate
MH  - SYNTHASE
MH  - Trypsin
RP  - NOT IN FILE
NT  - UI - 83257230LA - engRN - 0 (ATPase inhibitory protein)RN - 0 (Iodine Radioisotopes)RN - 0 (Peptide Fragments)RN - 0 (Proteins)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.4.21.4 (Trypsin)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM23152/GM/NIGMSDA - 19830909IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:6223660
SO  - Biochim Biophys Acta 1983 Jul 29 ;724(1):128-141

548
UI  - 372
AU  - Runswick MJ
AU  - Walker JE
TI  - The amino acid sequence of the beta-subunit of ATP synthase from bovine heart mitochondria
AB  - The amino acid sequence of the beta-subunit of bovine heart mitochondrial ATP synthase has been determined by protein sequence analysis. The polypeptide chain of 478 amino acids is blocked at its NH2 terminal. Comparison of this sequence with sequences of the corresponding proteins from Escherichia coli (Saraste, M., Gay, N.J., Eberle, A., Runswick, M.J., and Walker, J.E. (1981) Nucleic Acids Res. 9, 5287-5296) and maize and spinach chloroplasts Krebbers, E.T., Larrinua, I. M., McIntosh, L., and Bogorad, L. (1982) Nucleic Acids Res. 10, 4985-5002; Kurawski, G., Bottomley, W., and Whitfield, P.R. (1982) Proc. Natl. Acad. Sci. U.S.A. 79, 6260-6264) shows that the protein is highly conserved. 70% of residues are identical in E. coli and beef mitochondria. This contrasts with some of the other subunits in the enzyme complex which are much less conserved
RP  - NOT IN FILE
NT  - UI - 83135760LA - engRN - 0 (Macromolecular Systems)RN - 0 (Multienzyme Complexes)RN - 0 (Peptide Fragments)RN - EC 2.7 (Phosphotransferases)RN - EC 2.7.4.- (ATP synthetase complex)PT - Journal ArticleDA - 19830407IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:6298222
SO  - J Biol Chem 1983 Mar 10 ;258(5):3081-3089

549
UI  - 19624
AU  - Saishu T
AU  - Kagawa Y
AU  - Shimizu R
TI  - Resistance of thermophilic ATPase (TF1) to specific F1-atpase inhibitors including local anesthetics
AB  - F1-ATPase obtained from mesophilic organisms is inhibited by specific inhibitors, such as aurovertin, efrapeptin, quercetin and several local anesthetics. This property has been explained by the common structure at the catalytic center of F1. However thermophilic F1 (TF1), which has the same primary structure at the center as other F1's, was shown to be resistant to these F1-specific inhibitors. Thus, the inhibitory mechanism may be explained not by the common structure at the catalytic site, but by some conformational changes of the flexible mesophilic F1 molecules or the absence of an inhibitor binding site in thermophilic F1
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - aurovertin
MH  - Bacterial Proteins
MH  - BINDING
MH  - BINDING SITE
MH  - catalytic
MH  - conformational change
MH  - conformational changes
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - H(+)-Transporting ATP Synthase
MH  - inhibitor
MH  - inhibitors
MH  - mechanism
MH  - primary
MH  - protein
MH  - Proteins
MH  - RESISTANCE
MH  - Site
MH  - structure
MH  - SYNTHASE
MH  - THERMOPHILIC
MH  - united states
RP  - NOT IN FILE
NT  - UI - 83204160LA - engRN - 0 (Anesthetics, Local)RN - 0 (Bacterial Proteins)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19830623IS - 0006-291XSB - IMCY - UNITED STATESJC - 9Y8
UR  - PM:6221726
SO  - Biochem Biophys Res Commun 1983 May 16 ;112(3):822-826

550
UI  - 20910
AU  - Senda M
AU  - Kanazawa H
AU  - Tsuchiya T
AU  - Futai M
TI  - Conformational change of the alpha subunit of Escherichia coli F1 ATPase: ATP changes the trypsin sensitivity of the subunit
AB  - Conformational change in the alpha subunit of Escherichia coli proton- translocating ATPase was studied using trypsin. The subunit was cleaved with a small amount of trypsin (1 microgram/mg subunit) to peptides of less than 8000 daltons. On the other hand, the subunit was cleaved to two main polypeptides (30,000 and 25,000 daltons) in the presence of sufficient ATP (1 mM-0.5 microM) to saturate the high-affinity site of the subunit. Analysis of digests of the subunit combined with fluorescent maleimide suggested that the subunit was digested in the middle of the polypeptide chain in the presence of the nucleotide. ADP and adenylyl imidodiphosphate had the same effect as ATP. These results suggest that the conformation of the subunit changed to form two trypsin-resistant domains upon binding of ATP to the high-affinity site
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - ADP
MH  - ALPHA-SUBUNIT
MH  - analysis
MH  - atp
MH  - ATPase
MH  - Bacterial Proteins
MH  - BINDING
MH  - COLI F1 ATPASE
MH  - conformation
MH  - conformational change
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - protein
MH  - Proteins
MH  - proton
MH  - Site
MH  - SUBUNIT
MH  - Trypsin
RP  - NOT IN FILE
NT  - UI - 83125602LA - engRN - 0 (Bacterial Proteins)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.4.21.4 (Trypsin)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19830324IS - 0003-9861SB - IMCY - UNITED STATES
UR  - PM:6218786
SO  - Arch Biochem Biophys 1983 Feb 1 ;220(2):398-404

551
UI  - 481
AU  - Senior AE
AU  - Wise JG
TI  - The proton-ATPase of bacteria and mitochondria
RP  - NOT IN FILE
NT  - UI - 83241666LA - engRN - 0 (DNA, Bacterial)RN - 0 (Macromolecular Systems)RN - 0 (Protons)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewID - GM 25349/GM/NIGMSDA - 19830826IS - 0022-2631SB - IMCY - UNITED STATESJC - J4E
UR  - PM:6191035
SO  - J Membr Biol 1983  ;73(2):105-124

552
UI  - 480
AU  - Senior AE
AU  - Langman L
AU  - Cox GB
AU  - Gibson F
TI  - Oxidative phosphorylation in Escherichia coli. Characterization of mutant strains in which F1-ATPase contains abnormal beta-subunits
AB  - To facilitate study of the role of the beta-subunit in the membrane- bound proton-translocating ATPase of Escherichia coli, we identified mutant strains from which an F1-ATPase containing abnormal beta- subunits can be purified. Seventeen strains of E. coli, characterized by genetic complementation tests as carrying mutations in the uncD gene (which codes for the beta-subunit), were studied. The majority of these strains (11) were judged to be not useful, as their membranes lacked ATPase activity, and were either proton-permeable as prepared or remained proton-impermeable after washing with buffer of low ionic strength. A further two strains were of a type not hitherto reported, in that their membranes had ATPase activity, were proton-impermeable as prepared, and were not rendered proton-permeable by washing in buffer of low ionic strength. Presumably in these two strains F1-ATPase is not released in soluble form by this procedure. F1-ATPase of normal molecular size were purified from strains AN1340 (uncD478), AN937 (uncD430), AN938 (uncD431) and AN1543 (uncD484). F1-ATPase from strain AN1340 (uncD478) had 15% of normal specific Mg-dependent ATPase activity and 22% of normal ATP-synthesis activity. The F1-ATPase preparations from strains AN937, AN938 and AN1543 had respectively 1.7%, 1.8% and 0.2% of normal specific Mg-dependent ATPase activity, and each of these preparations had very low ATP-synthesis activity. The yield of F1-ATPase from the four strains described was almost twice that obtained from a normal haploid strain. The kinetics of Ca- dependent ATPase activity were unusual in each of the four F1-ATPase preparations. It is likely that these four mutant uncD F1-ATPase preparations will prove valuable for further experimental study of the F1-ATPase catalytic mechanism
RP  - NOT IN FILE
NT  - UI - 83230713LA - engRN - 0 (Membrane Proteins)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19830729IS - 0264-6021SB - IMCY - ENGLANDJC - 9YO
UR  - PM:6222731
SO  - Biochem J 1983 Feb 15 ;210(2):395-403

553
UI  - 479
AU  - Senior AE
TI  - Secondary and tertiary structure of membrane proteins involved in proton translocation
RP  - NOT IN FILE
NT  - UI - 83257236LA - engRN - 0 (Bacterial Proteins)RN - 0 (Membrane Proteins)RN - 53026-44-1 (Bacteriorhodopsin)RN - EC 1.9.3.1 (Cytochrome-c Oxidase)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewDA - 19830909IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:6307357
SO  - Biochim Biophys Acta 1983 Jul 15 ;726(2):81-95

554
UI  - 9919
AU  - Sholtz KF
AU  - Gorskaya IA
AU  - Kotelnikova AV
TI  - The stoichiometry of proton translocation through H+-ATPase of rat-liver mitochondria.
AB  - The ratio of the number of protons transported directly by H+-ATPase of intact mitochondria and that of the hydrolyzed ATP (the proton translocation quotient) is determined. A special kinetic method which makes possible determination of ATPase and H+-translocase activities in the same experiment has been used to estimate the proton translocation quotient for mitochondrial H+-ATPase. The quotient is found to be, on average, equal to 3.31 mol/mol. It is shown that the protonophore 3,5-di-tert-butyl-4-hydroxybenzylidenemalononitrile can greatly decrease the proton translocation quotient. This supports our assumption concerning a labile coupling between H+-translocase and ATPase in mitochondria [I. A. Gorskaya, K. F. Sholtz, S. A. Moreva, A. V. Kotelnikova (1979) Biochemistry (Engl. Transl. Biokhimiya) 44, 765-770]. A decrease in the translocation quotient in the presence of the protonophore is likely to be due to the ability of this uncoupler to take back the translocated protons into the mitochondrial matrix before their release into the medium. An electrostatic model of the molecular mechanism of H+-translocase and ATPase coupling in the H+-ATPase complex is discussed. The model is in agreement with the results obtained: transport of more than two protons per each hydrolyzed ATP molecule, and variable efficiency of the process.
MH  - Adenosinetriphosphatase
MH  - Animal
MH  - atp
MH  - ATPase
MH  - Biochemistry
MH  - Biological Transport
MH  - COMPLEX
MH  - drug effects
MH  - Enzyme Activation
MH  - enzymology
MH  - H+-ATPase
MH  - Hydrogen-Ion Concentration
MH  - Ionophores
MH  - mechanism
MH  - metabolism
MH  - Mitochondria
MH  - Mitochondria,Liver
MH  - model
MH  - Models,Chemical
MH  - Nitriles
MH  - pharmacology
MH  - proton
MH  - Protons
MH  - Rats
MH  - transport
MH  - Valinomycin
RP  - NOT IN FILE
SO  - Eur J Biochem 1983 Oct 17 ;136(1):129-134

555
UI  - 19812
AU  - Smith LT
AU  - Rosen G
AU  - Boyer PD
TI  - Properties of ATP tightly bound to catalytic sites of chloroplast ATP synthase
AB  - Under steady state photophosphorylating conditions, each ATP synthase complex from spinach thylakoids contains, at a catalytic site, about one tightly bound ATP molecule that is rapidly labeled from medium 32Pi. The level of this bound [32P]ATP is markedly reduced upon de- energization of the spinach thylakoids. The reduction is biphasic, a rapid phase in which the [32P] ATP/synthase complex drops about 2-fold within 10 s, followed by a slow phase, kobs = 0.01/min. A decrease in the concentration of medium 32Pi to well below its apparent Km for photophosphorylation is required to decrease the amount of tightly bound ATP/synthase found just after de-energization and before the rapid phase of bound ATP disappearance. The [32P]ATP that remains bound after the rapid phase appears to be mostly at a catalytic site as demonstrated by a continued exchange of the oxygens of the bound ATP with water oxygens. This bound [32P]ATP does not exchange with medium Pi and is not removed by the presence of unlabeled ATP. The levels of tightly bound ADP and ATP arising from medium ADP were measured by a novel method based on use of [beta-32P]ADP. After photophosphorylation and within minutes after the rapid phase of bound ATP loss, the measured ratio of bound ADP to ATP was about 1.4 and the sum of bound ADP plus ATP was about 1/synthase. This ratio is smaller than that found about 1 h after de-energization. Hence, while ATP bound at catalytic sites disappears, bound ADP appears. The results suggest that during and after de-energization the bound ATP disappears from the catalytic site by hydrolysis to bound ADP and Pi with subsequent preferential release of Pi. These and related observations can be accommodated by the binding change mechanism for ATP synthase with participation of alternating catalytic sites and are consistent with a deactivated state arising from occupancy of one catalytic site on the synthase complex by an inhibitory ADP without presence of Pi
MH  - A
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - ADP
MH  - atp
MH  - ATP synthase
MH  - BINDING
MH  - CHANGE MECHANISM
MH  - chloroplast
MH  - COMPLEX
MH  - Hydrolysis
MH  - mechanism
MH  - method
MH  - Multienzyme Complexes
MH  - Oxygen
MH  - Phosphates
MH  - Phosphotransferases
MH  - Photophosphorylation
MH  - Spinach
MH  - SYNTHASE
MH  - thylakoid
MH  - Water
RP  - NOT IN FILE
NT  - UI - 83291036LA - engRN - 0 (Multienzyme Complexes)RN - 0 (Phosphates)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 2.7 (Phosphotransferases)RN - EC 2.7.4.- (ATP synthetase complex)PT - Journal ArticleDA - 19831028IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:6309820
SO  - J Biol Chem 1983 Sep 25 ;258(18):10887-10894

556
UI  - 877
AU  - Tanford C
TI  - Mechanism of free energy coupling in active transport
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - Animal
MH  - Biological Transport,Active
MH  - Energy Metabolism
MH  - Ion Channels
MH  - Kinetics
MH  - metabolism
MH  - Models,Biological
MH  - Thermodynamics
RP  - NOT IN FILE
SO  - Annu Rev Biochem 1983  ;52:379-409.():379-409

557
UI  - 374
AU  - Walker JE
AU  - Gay NJ
TI  - Analysis of Escherichia coli ATP synthase subunits by DNA and protein sequencing
RP  - NOT IN FILE
NT  - UI - 84092750LA - engRN - 0 (DNA, Bacterial)RN - 0 (Multienzyme Complexes)RN - 0 (Plasmids)RN - EC 2.7 (Phosphotransferases)RN - EC 2.7.4.- (ATP synthetase complex)PT - Journal ArticleDA - 19840214IS - 0076-6879SB - IMCY - UNITED STATESJC - MVA
UR  - PM:6318027
SO  - Methods Enzymol 1983  ;97():195-218

558
UI  - 476
AU  - Wise JG
AU  - Duncan TM
AU  - Latchney LR
AU  - Cox DN
AU  - Senior AE
TI  - Properties of F1-ATPase from the uncD412 mutant of Escherichia coli
AB  - Properties of purified F1-ATPase from Escherichia coli mutant strain AN484 (uncD412) have been studied in an attempt to understand why the amino acid substitution in the beta-subunit of this enzyme causes a tenfold reduction from normal MgATP hydrolysis rate. In most properties that were studied, uncD412 F1-ATPase resembled normal E. coli F1- ATPase. Both enzymes were found to contain a total of six adenine- nucleotide-binding sites, of which three were found to be non- exchangeable and three were exchangeable (catalytic) sites. Binding of the non-hydrolysable substrate analogue adenosine 5'-[beta gamma- imido]triphosphate (p[NH]ppA) to the three exchangeable sites showed apparent negative co-operativity. The binding affinities for p[NH]ppA, and also ADP, at the exchangeable sites were similar in the two enzymes. Both enzymes were inhibited by efrapeptin, aurovertin and p[NH]ppA, and were inactivated by dicyclohexylcarbodi-imide, 4-chloro-7- nitrobenzofurazan and p-fluorosulphonyl-benzoyl-5'-adenosine. Km values for CaATP and MgATP were similar in the two enzymes. uncD412 F1-ATPase was abnormally unstable at high pH, and dissociated into subunits readily with consequent loss of activity. The reason for the impairment of catalysis in uncD412 F1-ATPase cannot be stated with certainty from these studies. However we discuss the possibility that the mutation interrupts subunit interaction, thereby causing a partial impairment in the site-site co-operativity which is required for 'promotion' of catalysis in this enzyme
RP  - NOT IN FILE
NT  - UI - 84079710LA - engRN - 0 (5'-(4-fluorosulfonylbenzoyl)adenosine)RN - 0 (Adenine Nucleotides)RN - 0 (Aurovertins)RN - 10199-89-0 (4-Chloro-7-nitrobenzofurazan)RN - 25612-73-1 (Adenylyl Imidodiphosphate)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-61-7 (Adenosine)RN - 65256-31-7 (aurovertin D)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM-25349/GM/NIGMSDA - 19840107IS - 0264-6021SB - IMCY - ENGLANDJC - 9YO
UR  - PM:6228224
SO  - Biochem J 1983 Nov 1 ;215(2):343-350

559
UI  - 778
AU  - Yoshida M
TI  - The synthesis of enzyme-bound ATP by the F1-ATPase from the thermophilic bacterium PS3 in 50% dimethylsulfoxide
AB  - Purified TF1 (F1-ATPase from a thermophilic bacterium PS3) synthesizes enzyme-bound ATP from medium Pi and enzyme-bound ADP in the presence of 50% dimethylsulfoxide (DMSO). Once ATP was formed on the enzyme, it was not released even after removal of DMSO and Pi from the solution. The half maximal concentration of medium Pi for ATP synthesis was 1mM. The pH optimum for enzyme-bound ATP formation was about 6.5. Under the optimum conditions, a yield of up to 0.8 mol of ATP/mol of TF1 was obtained
RP  - NOT IN FILE
NT  - UI - 83308745LA - engRN - 0 (Phosphates)RN - 0 (Solvents)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 67-68-5 (Dimethyl Sulfoxide)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19831028IS - 0006-291XSB - IMCY - UNITED STATESJC - 9Y8
UR  - PM:6225432
SO  - Biochem Biophys Res Commun 1983 Aug 12 ;114(3):907-912

560
UI  - 777
AU  - Yoshida M
AU  - Allison WS
TI  - Modulation by ADP and Mg2+ of the inactivation of the F1-ATPase from the thermophilic bacterium, PS3, with dicyclohexylcarbodiimide
AB  - The soluble F1-ATPase from the thermophilic bacterium PS3 (TF1) contains no endogenous adenine nucleotides and contains about 0.2 g ions of Mg2+/mol which resists removal by repeated centrifugation- elution on columns of Sephadex G-50. The isolated enzyme will not bind additional Mg2+ added in the absence of adenine nucleotides nor is the rate of inactivation of the isolated enzyme by dicyclohexylcarbodiimide (DCCD) affected by the addition of Mg2+. When ADP is added to isolated TF1, a 1:1 TF1 X ADP complex is formed which is stable to repeated gel permeation on columns of Sephadex G-50 subjected to centrifugation- elution. On formation of the 1:1 TF1 X ADP complex, the rate of inactivation of the enzyme by DCCD is accelerated 6-fold. The rate of inactivation of the 1:1 TF1 X ADP complex by DCCD is not further stimulated in the presence of 2 mM ADP which indicates that the binding of ADP to a single site in the enzyme is sufficient to promote maximal stimulation of the inactivation. Addition of Mg2+ to the 1:1 TF1 X ADP complex results in the binding of about 1 g ion of Mg2+/mol of enzyme. The 1:1:1 TF1 X ADP X Mg2+ complex thus formed is sluggishly inactivated by DCCD. When the Mg2+ is removed from the TF1 X ADP X Mg2+ complex by treatment with trans-1,2-diaminocyclohexane-N,N,N',N'- tetraacetic acid, the rate of inactivation of the enzyme by DCCD is accelerated 4-fold. Other divalent metal ions protect the 1:1 TF1 X ADP complex against inactivation by DCCD. Of these, Mn2+, Zn2+, Co2+, and Cd2+, which are about as equally effective as Mg2+ as cofactors for the hydrolytic reaction when present at 0.2 mM, offer about equal protection of the complex against inactivation by DCCD also when present at 0.2 mM. These results indicate that the binding site for ADP in the 1:1 TF1 X ADP complex is a catalytic site. TF1, inactivated by 92% with DCCD, has the same capacity to bind ADP as the active enzyme, forming a tight 1:1 TF1 X ADP complex which is stable to repeated centrifugation-elution on columns of Sephadex G-50. The 1:1 TF1 X ADP complex retains its capacity to bind Mg2+ to form the 1:1:1 TF1 X ADP X Mg2+ complex after it is inactivated by 88% with DCCD
RP  - NOT IN FILE
NT  - UI - 84061889LA - engRN - 0 (Carbodiimides)RN - 0 (Cations, Divalent)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 58-64-0 (Adenosine Diphosphate)RN - 7439-95-4 (Magnesium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM-16974/GM/NIGMSDA - 19840127IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:6227624
SO  - J Biol Chem 1983 Dec 10 ;258(23):14407-14412

561
UI  - 775
AU  - Andrews WW
AU  - Yoshida M
AU  - Hill FC
AU  - Allison WS
TI  - Identification of an essential lysine residue in the beta subunit of the F1-ATPase from the thermophilic bacterium, PS3, using 7-chloro-4- nitro[14C]benzofurazan
AB  - When the F1-ATPase from the thermophilic bacterium, PS3, was inactivated by 90% with 7-chloro-4-nitro[14C]benzofurazan ([14C]Nbf-Cl) at pH 7.3 and then gel-filtered, 1.25 mols of [14C]Nbf-O-Tyr and less than 0.1 mol of Nbf-N-Lys were formed per mol of enzyme. After adjusting the pH of the gel-filtered, modified enzyme to 9.0 and incubating it for 14 hrs. at 23 degrees C to promote O----N migration, 0.68 mol of Nbf-N-Lys were formed per mol of enzyme while about 16% of the original activity reappeared. Isolation of the subunits after the O- ---N migration showed that 90% of the incorporated 14C was present in the beta subunit, which contained 0.21 mols of [14C]Nbf-N-Lys per mol. A tryptic peptide which contained the majority of the 14C incorporated into the beta subunit was isolated and subjected to automatic amino acid sequence analysis contained 38 residues. The amino acid sequence immediately around the lysine residue labeled with [14C]Nbf-, K*, was found to be: ...I-G-L-F-G-G-A-G-V-G-K*-T-V-L-I-G...
RP  - NOT IN FILE
NT  - UI - 85022562LA - engRN - 0 (Carbon Radioisotopes)RN - 0 (Macromolecular Systems)RN - 0 (Oxadiazoles)RN - 0 (Peptide Fragments)RN - 10199-89-0 (4-Chloro-7-nitrobenzofurazan)RN - 56-87-1 (Lysine)RN - EC 3.4.21.4 (Trypsin)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM-16974/GM/NIGMSDA - 19841109IS - 0006-291XSB - IMCY - UNITED STATESJC - 9Y8
UR  - PM:6237650
SO  - Biochem Biophys Res Commun 1984 Sep 28 ;123(3):1040-1046

562
UI  - 21202
AU  - Bayramashvili DI
AU  - Drachev AL
AU  - Drachev LA
AU  - Kaulen AD
AU  - Kudelin AB
AU  - Martynov VI
AU  - Skulachev VP
TI  - Proteinase-treated photoreceptor discs. Photoelectric activity of the partially-digested rhodopsin and membrane orientation
AB  - Photoreceptor discs from rod outer segments of cattle retina were treated with (a) papain, (b) thermolysin or (c) trypsin, the procedures resulting in the cleavage of the rhodopsin polypeptide chain between (a) 323 and 324, 236 and 237, 241 and 242, (b) 327 and 328, 240 and 241, or (c) 339 and 340 amino acid residues, respectively. In all the cases, partially digested rhodopsins proved to be competent in generating photoelectric potential and increasing membrane conductance of the discs adsorbed onto phospholipid-impregnated collodion film. The kinetics of generation and dissipation of photopotential as well as of formation of metarhodopsin II and of the light-induced rhodopsin protonation were found to be the same in the partially digested preparations and in the intact one. Incubation of papain-treated or thermolysin-treated discs at pH 6.0 induced formation of inside-out vesicles which, when incorporated into the collodion film, generated an oppositely directed photopotential. Treatment of such vesicles with papain gave rise to further cleavages of the polypeptide localized between 30 and 31, 186 and 187 amino acid residues. One more proteinase- sensitive site, localized between 104 and 105 residues, has been discovered in the inside-out vesicles treated with thermolysin. This fact consistent with the scheme of the 'seven column' arrangement of the visual rhodopsin [Ovchinnikov, Yu. A. (1982) FEBS Lett. 148, 179- 191]. Rhodopsin, when treated with papain on both sides, was deprived of sixty amino acid residues being split in two sites in the middle part of the polypeptide, but was still active as a photoelectric energy transducer. The main specific feature inherent in the photoelectric response of the papain-treated or thermolysin-treated rhodopsin and absent from the native protein is that the former survives addition of long trains of saturating flashes when the response of the intact preparation becomes negligible. This effect was shown to be due to conversion of partially digested rhodopsin to a photolytic product that at room temperature lived for minutes even in the presence of NH2OH. A 532-nm laser flash effectively converted this product back to rhodopsin
MH  - A
MH  - ACID
MH  - ACTIVE
MH  - Cattle
MH  - conductance
MH  - flash
MH  - inside-out
MH  - Kinetics
MH  - membrane
MH  - pH
MH  - pigments
MH  - protein
MH  - protonation
MH  - RESIDUE
MH  - retinal
MH  - Site
MH  - Temperature
MH  - Trypsin
MH  - vesicles
RP  - NOT IN FILE
NT  - UI - 84285369LA - engRN - 0 (Hydroxylamines)RN - 0 (Retinal Pigments)RN - 7803-49-8 (Hydroxylamine)RN - 9009-81-8 (Rhodopsin)RN - EC 3.4.22.2 (Papain)RN - EC 3.4.24.27 (Thermolysin)PT - Journal ArticleDA - 19841003IS - 0014-2956SB - IMCY - GERMANY, WEST
UR  - PM:6468381
SO  - Eur J Biochem 1984 Aug 1 ;142(3):583-590

563
UI  - 892
AU  - Campo ML
AU  - Zhang CJ
AU  - Tedeschi H
TI  - Energy transduction, ion transport and protonmotive force: are they linked?
MH  - Adenosine Triphosphate
MH  - Animal
MH  - Arylsulfonates
MH  - biosynthesis
MH  - Electrophysiology
MH  - Energy Metabolism
MH  - Ion Channels
MH  - Membrane Potentials
MH  - metabolism
MH  - Mice
MH  - Mitochondria
MH  - Models,Biological
MH  - pharmacology
MH  - physiology
MH  - Potassium
MH  - Protons
MH  - Succinates
MH  - Succinic Acid
RP  - NOT IN FILE
SO  - Biochem Soc Trans 1984 Jun ;12(3):384-386

564
UI  - 845
AU  - Cotton NP
AU  - Clark AJ
AU  - Jackson JB
TI  - Changes in membrane ionic conductance, but not changes in slip, can account for the non-linear dependence of the electrochemical proton gradient upon the electron-transport rate in chromatophores
AB  - Decrease in the rate of cyclic electron transport (JE) measured from the absorbance changes associated with reaction centre bacteriochlorophyll led to a less than proportionate decrease in the membrane potential (delta psi) measured by electrochromism. In principle this result can be explained either by a delta psi-dependent slip in the H+/e- coupling ratio (nE) or by a delta psi-dependent change in the membrane ionic conductance. Simultaneous measurement of the membrane ionic current (JDIS) did not reveal any significant changes in the H+/e- ratio (JDIS/JE) and showed that conductance changes (JDIS/delta psi) account quantitatively for the curved dependence of delta psi on JE. Simultaneous recordings of JDIS and the extravesicular pH from cresol-red absorbance changes, suggest that protons are the main current-carrying species across the chromatophore membrane at high values of delta psi in the presence and absence of Fo- ATPase inhibitor. At reduced delta psi the flux of other ions outweighs the hydrogen ion current
RP  - NOT IN FILE
NT  - UI - 84261530LA - engPT - Journal ArticleDA - 19840904IS - 0014-2956SB - IMCY - GERMANY, WESTJC - EMZ
UR  - PM:6745265
SO  - Eur J Biochem 1984 Jul 2 ;142(1):193-198

565
UI  - 19874
AU  - Cross RL
AU  - Cunningham D
AU  - Tamura JK
TI  - Binding change mechanism for ATP synthesis by oxidative phosphorylation and photophosphorylation
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - BINDING
MH  - CHANGE MECHANISM
MH  - H(+)-Transporting ATP Synthase
MH  - mechanism
MH  - Oxidative Phosphorylation
MH  - Phosphorylation
MH  - Photophosphorylation
MH  - SYNTHASE
MH  - synthesis
RP  - NOT IN FILE
NT  - UI - 85050366LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM 23152/GM/NIGMSDA - 19850114IS - 0070-2137SB - IMCY - UNITED STATESJC - DWM
UR  - PM:6238809
SO  - Curr Top Cell Regul 1984  ;24():335-344

566
UI  - 997
AU  - Damiano E
AU  - Bassilana M
AU  - Rigaud JL
AU  - Leblanc G
TI  - Use of the pH sensitive fluorescence probe pyranine to monitor internal pH changes in Escherichia coli membrane vesicles
AB  - Measurements of the fluorescent properties of 8-hydroxy-1,3,6-pyrenetrisulfonate (pyranine) enclosed within the internal space of Escherichia coli membrane vesicles enable recordings and quantitative analysis of: (i) changes in intravesicular pH taking place during oxidation of electron donors by the membrane respiratory chain; (ii) transient alkalization of the internal aqueous space resulting from the creation of outwardly directed acetate diffusion gradients across the vesicular membrane. Quantitation of the fluorescence variations recorded during the creation of transmembrane acetate gradients shows a close correspondence between the measured shifts in internal pH value and those expected from the amplitude of the imposed acetate gradients
MH  - Acetates
MH  - Arylsulfonates
MH  - Cell Membrane
MH  - Cell-Free System
MH  - diagnostic use
MH  - Diffusion
MH  - Electron Transport
MH  - Escherichia coli
MH  - Hydrogen-Ion Concentration
MH  - Lactates
MH  - metabolism
MH  - Oxidation-Reduction
MH  - physiology
RP  - NOT IN FILE
SO  - FEBS Lett 1984 Jan 23 ;166(1):120-124

567
UI  - 21297
AU  - Drachev AL
AU  - Kaulen AD
AU  - Skulachev VI
TI  - Correlation of photochemical cycle, H+ release and uptake, and electric events in bacteriorhodopsin
MH  - H+
MH  - Bacteriorhodopsin
RP  - IN FILE
SO  - FEBS Lett 1984  ;178():331-336

568
UI  - 21203
AU  - Drachev AL
AU  - Drachev LA
AU  - Kaulen AD
AU  - Khitrina LV
TI  - The action of lanthanum ions and formaldehyde on the proton-pumping function of bacteriorhodopsin
AB  - The photochemical cycle and the proton-pumping function of bacteriorhodopsin modified with lanthanum and formaldehyde has been studied. In both preparations, the M412 leads to BR570 transition time has been found to increase considerably. The deceleration of the photochemical cycle has been shown to be accompanied by inhibition of the millisecond phase of the photoelectrical response of bacteriorhodopsin membranes associated with phospholipid-impregnated collodion film. Photoelectrogenic activity measured with permeable ion probe in proteoliposomes was also inhibited. Effects of lanthanum were reversed by EDTA. Formation of M412 was slightly accelerated and the microsecond electrogenic phase was not affected by lanthanum and by formaldehyde. It is concluded that lanthanum, but not formaldehyde, can be used as a specific reversible inhibitor of the second half of the bacteriorhodopsin photocycle and of the associated H+ uptake on the cytoplasmic side of the halobacterial membrane. Possible mechanisms of these effects are discussed
MH  - A
MH  - Anions
MH  - Bacteriorhodopsin
MH  - carotenoid
MH  - Carotenoids
MH  - electrogenic
MH  - function
MH  - H+
MH  - inhibitor
MH  - ion
MH  - Ions
MH  - mechanism
MH  - MECHANISMS
MH  - membrane
MH  - Membranes
MH  - microsecond
MH  - proteoliposome
MH  - proton
MH  - Protons
MH  - Time
RP  - NOT IN FILE
NT  - UI - 84131996LA - engRN - 0 (Anions)RN - 0 (Protons)RN - 36-88-4 (Carotenoids)RN - 50-00-0 (Formaldehyde)RN - 53026-44-1 (Bacteriorhodopsins)RN - 7439-91-0 (Lanthanum)PT - Journal ArticleDA - 19840411IS - 0014-2956SB - IMCY - GERMANY, WEST
UR  - PM:6321172
SO  - Eur J Biochem 1984 Jan 16 ;138(2):349-356

569
UI  - 21113
AU  - Ferguson SJ
AU  - Parsonage D
TI  - Analysis of relationships between the protonmotive force and rates and extents of ATP synthesis
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - analysis
MH  - atp
MH  - ATP synthesis
MH  - COMPLEX
MH  - England
MH  - Multienzyme Complexes
MH  - Nad
MH  - Phosphotransferases
MH  - proton
MH  - Protons
MH  - synthesis
RP  - NOT IN FILE
NT  - UI - 84236959LA - engRN - 0 (Multienzyme Complexes)RN - 0 (Protons)RN - 53-84-9 (NAD)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 2.7 (Phosphotransferases)RN - EC 2.7.4.- (ATP Synthetase Complexes)PT - Journal ArticleDA - 19840730IS - 0300-5127SB - IMCY - ENGLAND
UR  - PM:6329846
SO  - Biochem Soc Trans 1984 Jun ;12(3):416-419

570
UI  - 53
AU  - Fillingame RH
AU  - Peters LK
AU  - White LK
AU  - Mosher ME
AU  - Paule CR
TI  - Mutations altering aspartyl-61 of the omega subunit (uncE protein) of Escherichia coli H+ -ATPase differ in effect on coupled ATP hydrolysis
AB  - Mutations in the H+-translocating ATPase complex (F1F0) of Escherichia coli have been described in which aspartyl-61 of the omega subunit ( uncE protein) is substituted by either glycine ( uncE105 ) or asparagine ( uncE107 ). Either substitution blocks the H+-translocation activity of the F0 sector of the complex. Here we report a difference in the effects of the two substitutions on the coupled ATPase activity of F1 bound to F0. Wild-type F1 was bound to the F0 of either mutant with affinities comparable to wild-type. The ATPase activity of F1 bound to uncE107 F0 was inhibited by 50%, whereas that bound to uncE105 F0 was not inhibited. Complementation studies with a pBR322-derived plasmid that carried the E gene of the unc operon only indicated that a single mutation in the host strain was responsible for the respective phenotypes. In mutants complemented by the uncE + plasmid, restoration of wild-type biochemical properties was only partial and may be attributed to a mixing of wild-type and mutant omega subunits in a hybrid F0 complex. The activity of membrane-bound F1 was less inhibited in the uncE +/ uncE107 hybrid. Paradoxically, complementation of uncE105 by the uncE + plasmid resulted in substantial inhibition of the activity of membrane-bound F1. The results indicate that a glycine- versus-asparagine substitution for aspartyl-61 must lead to altered conformations of omega and that these differences in conformation are important in the coupling between the F0 and F1 sectors of the complex
RP  - NOT IN FILE
NT  - UI - 84212269LA - engRN - 0 (Macromolecular Systems)RN - 0 (Plasmids)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.1.21 (DNA Restriction Enzymes)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - 5 T32 GM07215/GM/NIGMSID - GM23105/GM/NIGMSDA - 19840713IS - 0021-9193SB - IMCY - UNITED STATESJC - HH3
UR  - PM:6327626
SO  - J Bacteriol 1984 Jun ;158(3):1078-1083

571
UI  - 9900
AU  - Freedman JA
AU  - Lemasters JJ
TI  - Thermodynamics of reverse electron transfer across site 1: ATP/2e- is greater than one.
AB  - ATP-driven, rotenone-sensitive, reverse electron transfer from succinate to acetoacetate was measured in rat liver mitochondria in the presence of cyanide. In the approach to equilibrium, the absolute ratio of the free energy change of electron transfer to that of ATP hydrolysis exceeded 1, tending towards about 1 1/4. The data support an H+/2e- stoichiometry of 5 for Site 1 as predicted by a thirteen-proton model of chemiosmotic coupling.
MH  - acetoacetate
MH  - Acetoacetates
MH  - Adenosine Triphosphate
MH  - Animal
MH  - atp
MH  - Cyanides
MH  - electron
MH  - Electron Transport
MH  - Hydrolysis
MH  - Liver
MH  - Mathematics
MH  - metabolism
MH  - Mitochondria
MH  - Mitochondria,Liver
MH  - model
MH  - pharmacology
MH  - Rats
MH  - Rotenone
MH  - succinate
MH  - Succinates
MH  - Support,Non-U.S.Gov't
MH  - Support,U.S.Gov't,P.H.S.
MH  - Thermodynamics
MH  - TRANSFER
RP  - NOT IN FILE
SO  - Biochem Biophys Res Commun 1984 Nov 30 ;125(1):8-13

572
UI  - 20902
AU  - Futai M
AU  - Kanazawa H
TI  - [H+-ATPase--its genetic studies]
MH  - Adenosinetriphosphatase
MH  - ATPase
MH  - proton
MH  - Protons
MH  - review
RP  - NOT IN FILE
NT  - UI - 85064624LA - jpnRN - 0 (DNA, Bacterial)RN - 0 (Protons)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticlePT - ReviewDA - 19850114IS - 0039-9450SB - IMCY - JAPAN
UR  - PM:6095366
SO  - Tanpakushitsu Kakusan Koso 1984 Aug ;29(8):635-660

573
UI  - 21099
AU  - Garcia ML
AU  - Kitada M
AU  - Eisenstein HC
AU  - Krulwich TA
TI  - Voltage-dependent proton fluxes in liposomes
AB  - Liposomes containing buffered KCl were prepared from bacterial lipids, were diluted into K+-free media and were treated with valinomycin to induce the formation of a diffusion potential (delta psi). Upon formation of such a potential, substantial proton influx was observed, as assayed by the quenching of 9-aminoacridine fluorescence. Complete reversal of fluorescence quenching occurred when the potential was collapsed by addition of KCl or when methylamine was added. Studies of proton influx as a function of the theoretical magnitude of the delta psi indicated that the phenomenon occurred only above a delta psi of about -60 mV. Establishment of a Na+ diffusion potential also resulted in proton influx. Treatment of K+-loaded liposomes with N,N'- dicyclohexylcarbodiimide did not reduce the delta psi-dependent proton influx. Moreover, proton influx could be demonstrated upon imposition of a diffusion potential in liposomes prepared from a synthetic lipid. The proton fluxes associated with generation of a diffusion potential in liposomes may complicate studies of reconstituted systems in which proton translocation should occur, and may affect the magnitude of the electrochemical proton gradient that is operant under some conditions
MH  - A
MH  - delta
MH  - Dicyclohexylcarbodiimide
MH  - Diffusion
MH  - diffusion potential
MH  - fluorescence
MH  - function
MH  - Lipids
MH  - liposome
MH  - Liposomes
MH  - Potassium
MH  - Potassium Chloride
MH  - proton
MH  - PSI
MH  - SYSTEM
MH  - SYSTEMS
MH  - translocation
MH  - Valinomycin
RP  - NOT IN FILE
NT  - UI - 84257649LA - engRN - 0 (Liposomes)RN - 2001-95-8 (Valinomycin)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 7447-40-7 (Potassium Chloride)RN - 90-45-9 (Aminacrine)PT - Journal ArticleID - GM28454/GM/NIGMSDA - 19840912IS - 0006-3002SB - IMCY - NETHERLANDS
UR  - PM:6743646
SO  - Biochim Biophys Acta 1984 Jul 27 ;766(1):109-115

574
UI  - 9901
AU  - Gober JW
AU  - Kashket ER
TI  - H+/ATP stoichiometry of cowpea Rhizobium sp. strain 32H1 cells grown under nitrogen-fixing and nitrogen-nonfixing conditions.
AB  - The obligate aerobe Cowpea Rhizobium sp. strain 32H1 in axenic culture is able to fix N2 when grown under 0.2% O2 but not when grown under 21% O2. It was, therefore, of interest to investigate ATP synthesis in these cells grown under the two conditions. When respiring in buffers having pHs ranging from 6 to 8.5, cells grown under either O2 tension maintained an intracellular pH more alkaline than the exterior. The transmembrane chemical gradient of H+ (delta pH) was essentially the same under both conditions of growth, decreasing from ca. 90 mV at medium pH 6 to ca. 30 mV at pH 8.5. However, the transmembrane electrical gradient (delta psi) was significantly higher in cells grown under 21% O2 (150 to 166 mV) than in cells grown under 0.2% O2, the latter being 16 mV at pH 6 and increasing to 88 mV at pH 8.5. Therefore, the proton motive force of 21% O2-grown cells ranged from 237 mV at external pH 6 to 185 mV at pH 8.5, compared with a proton motive force of 114 to 121 mV in the 0.2% O2-grown cells. The cells grown in 0.2% O2 had the same proton motive force whether tested at 21 or at 0.2% O2. The phosphorylation potential, calculated from the intracellular ATP, ADP, and Pi concentrations, was 424 mV in the 21% O2-grown cells and 436 mV in the 0.2% O2-grown cells. Thus, the 21% O2-grown cells translocated 1.8 to 2.3 H+/ATP synthesized by the H+-ATPase, whereas the H+/ATP ratio for 0.2% O2-grown cells was 3.7 to 3.8.
MH  - Adenosine Triphosphate
MH  - ADP
MH  - Aerobiosis
MH  - Anaerobiosis
MH  - atp
MH  - ATP synthesis
MH  - Buffers
MH  - Cells
MH  - DELTA-PH
MH  - H(+)-Transporting ATP Synthase
MH  - H+
MH  - H+-ATPase
MH  - Hydrogen-Ion Concentration
MH  - Kinetics
MH  - metabolism
MH  - Nitrogen Fixation
MH  - Oxygen Consumption
MH  - Phosphorylation
MH  - proton
MH  - PSI
MH  - Rhizobium
MH  - Support,U.S.Gov't,Non-P.H.S.
RP  - NOT IN FILE
SO  - J Bacteriol 1984 Oct ;160(1):216-221

575
UI  - 1175
AU  - Graber P
AU  - Bauermeister H
TI  - Electric measurements of the kinetics of photosynthetic events
MH  - Kinetics
T2  - Adv. Photosynth. Res., Proc. Int. Congr. Photosynth., 6th, Meeting Date 1983, Volume 2, 333-6. Edited by: Sybesma, C. Nijhoff: The Hague, Neth
Y2  - -32676  
RP  - ON REQUEST (03/18/92)
SO  -  1984  ;():

576
UI  - 1170
AU  - Graber P
AU  - Schlodder E
AU  - Witt HT
TI  - Mechanism of the regulation of ATP synthesis/hydrolysis in chloroplasts and ATP synthesis in model systems
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - chloroplast
MH  - Chloroplasts
MH  - COMPLEX
MH  - coupling
MH  - F0
MH  - F1
MH  - H+-ATPase
MH  - mechanism
MH  - model
MH  - regulation
MH  - SYNTHASE
MH  - synthesis
MH  - SYSTEM
MH  - SYSTEMS
T2  - H+-ATPase (ATP Synthase): Struct., Funct., Biog., F0 F1 Complex Coupling Membr., Int. Workshop, 431-40. Edited by: Papa, S. Adriatica Ed.: Bari, Italy
Y2  - -32676  
RP  - ON REQUEST (03/18/92)
SO  -  1984  ;():

577
UI  - 1169
AU  - Graber P
TI  - Electrical and electrochemical aspects of the primary processes of photosynthesis
MH  - FIELD
MH  - Photosynthesis
T2  - Charge Field Eff. Biosyst., [Pap. - Int. Symp. Bioelectrochem. Bioenerg.], Meeting Date 1983, 227-42. Edited by: Allen, Milton Joel; Usherwood, Peter Norman Russell. Abacus Press: Tunbridge Wells, UK
Y2  - -32676  
RP  - ON REQUEST (03/18/92)
SO  -  1984  ;():

578
UI  - 1167
AU  - Graber P
AU  - Renger G
TI  - Functional and structural organization of primary photosynthetic processes in plants
MH  - plant
MH  - Plants
RP  - ON REQUEST (03/18/92)
SO  - Wiss Z Humboldt-Univ Berlin,Math -Naturwiss Reihe 1984  ;33():280-284

579
UI  - 1171
AU  - Graber P
AU  - Junesch U
AU  - Schatz GH
TI  - Kinetics of proton-transport-coupled ATP synthesis in chloroplasts. Activation of the ATPase by an artificially generated .DELTA.pH and .DELTA..psi
MH  - ACTIVATION
MH  - atp
MH  - ATP synthesis
MH  - ATPase
MH  - chloroplast
MH  - Chloroplasts
MH  - Kinetics
MH  - synthesis
RP  - ON REQUEST (03/18/92)
SO  - Ber Bunsen-Ges Phys Chem 1984  ;88():599-608

580
UI  - 1174
AU  - Graber P
AU  - Roegner M
AU  - Samoray D
AU  - Hauska G
TI  - ATP synthesis catalyzed by reconstituted CF0F1 liposomes driven by an artificially-generated .DELTA..psi. and .DELTA.pH
MH  - atp
MH  - ATP synthesis
MH  - CF0F1
MH  - liposome
MH  - Liposomes
MH  - RECONSTITUTED CF0F1
MH  - synthesis
T2  - Adv. Photosynth. Res., Proc. Int. Congr. Photosynth., 6th, Meeting Date 1983, Volume 2, 427-30. Edited by: Sybesma, C. Nijhoff: The Hague, Neth
Y2  - -32676  
RP  - ON REQUEST (03/18/92)
SO  -  1984  ;():

581
UI  - 8506
AU  - Grber P
AU  - Junesch U
AU  - Schatz GH
TI  - Kinetics of Proton-Transport-Coupled ATP Synthesis in Chloroplasts. Activation of the ATPase by an Artificially Generated delta pH and delta Psi
MH  - ACTIVATION
MH  - atp
MH  - ATP synthesis
MH  - ATPase
MH  - bioenergetics
MH  - chloroplast
MH  - Chloroplasts
MH  - delta
MH  - DELTA-PH
MH  - Kinetics
MH  - pH
MH  - PSI
MH  - regulation
MH  - synthesis
RP  - IN FILE
NT  - Ju, ATP 9
SO  - BBG 1984  ;88():599-608

582
UI  - 21098
AU  - Guffanti AA
AU  - Mann M
AU  - Sherman TL
AU  - Krulwich TA
TI  - Patterns of electrochemical proton gradient formation by membrane vesicles from an obligately acidophilic bacterium
AB  - Isolated membrane vesicles from the obligately acidophilic bacterium Bacillus acidocaldarius generated an electrochemical gradient of protons (delta mu- H+) upon energization with ascorbate-phenazine methosulfate at pH 6.0 or 3.0. At pH 6.0, there was little or no transmembrane pH gradient (delta pH), but a transmembrane electrical potential (delta psi) of ca. -77 mV, positive out, was observed. At pH 3.0, a delta pH equivalent to - 100 mV, acid out, and a delta psi of - 73 mV, positive out, were observed upon energization. The total magnitude of the delta mu- H+ was higher than that of whole cells at acid pH, but the very large delta pHs and the reversed delta psi s, i.e., inside positive, that are typical of acidophile cells were not observed in the vesicles. The vesicles exhibited energy-dependent accumulation of alpha-aminoisobutyric acid that was inhibited by both nigericin and valinomycin (plus K+) at pH 3.0 but was inhibited little by nigericin at pH 6.0
MH  - A
MH  - ACID
MH  - Bacillus
MH  - Bacteria
MH  - Cells
MH  - delta
MH  - DELTA-PH
MH  - H+
MH  - membrane
MH  - membrane vesicles
MH  - pH
MH  - proton
MH  - Protons
MH  - PSI
MH  - Valinomycin
MH  - vesicles
RP  - NOT IN FILE
NT  - UI - 84264321LA - engRN - 0 (Aminoisobutyric Acids)RN - 62-57-7 (2-aminoisobutyric acid)PT - Journal ArticleDA - 19840917IS - 0021-9193SB - IMCY - UNITED STATES
UR  - PM:6746570
SO  - J Bacteriol 1984 Aug ;159(2):448-452

583
UI  - 891
AU  - Guffanti AA
AU  - Krulwich TA
TI  - Bioenergetic problems of alkalophilic bacteria
MH  - Adenosine Triphosphate
MH  - Bacteria
MH  - biosynthesis
MH  - Energy Metabolism
MH  - Hydrogen-Ion Concentration
MH  - metabolism
MH  - Models,Biological
MH  - Oxidative Phosphorylation
MH  - physiology
MH  - Potassium
MH  - Support,U.S.Gov't,P.H.S.
RP  - NOT IN FILE
SO  - Biochem Soc Trans 1984 Jun ;12(3):411-412

584
UI  - 21100
AU  - Guffanti AA
AU  - Fuchs RT
AU  - Schneier M
AU  - Chiu E
AU  - Krulwich TA
TI  - A transmembrane electrical potential generated by respiration is not equivalent to a diffusion potential of the same magnitude for ATP synthesis by Bacillus firmus RAB
AB  - ATP synthesis by starved whole cells of alkalophilic Bacillus firmus RAB was energized by addition of DL-malate or by imposition of a valinomycin-mediated K+ diffusion potential. At pH 9.0, the transmembrane electrical potentials produced by these two means were similar in magnitude, at close to -170 mV. While N,N'- dicyclohexylcarbodiimide-sensitive ATP synthesis occurred upon the addition of DL-malate, no ATP was synthesized in response to a diffusion potential. In contrast, Na+-dependent accumulation of alpha- aminoisobutyric acid was energized equally well by DL-malate or a diffusion potential at pH 9.0. Even at pH 7.0, DL-malate was more efficacious than a diffusion potential in energizing ATP synthesis as assessed by determining the phosphorylation potentials generated at transmembrane electrical potential values of different magnitudes. Both modes of energization did, however, result in N,N'- dicyclohexylcarbodiimide-sensitive ATP synthesis at pH 7.0. The results of these studies are consistent with a model of energy coupling in which one pathway of protons between the respiratory proton pumps and the H+-translocating ATPase is direct or localized. Whereas an artificially imposed electrochemical gradient of protons can energize ATP synthesis under certain experimental conditions, in the proton-poor milieu that is optimal for growth of alkalophilic bacilli, the direct proton pathway may be crucial
MH  - A
MH  - ACID
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - alpha
MH  - atp
MH  - ATP synthesis
MH  - ATPase
MH  - Bacillus
MH  - Cells
MH  - coupling
MH  - Dicyclohexylcarbodiimide
MH  - Diffusion
MH  - diffusion potential
MH  - model
MH  - pH
MH  - Phosphorylation
MH  - proton
MH  - Proton Pump
MH  - Protons
MH  - Respiration
MH  - synthesis
MH  - Valinomycin
RP  - NOT IN FILE
NT  - UI - 84135793LA - engRN - 0 (Malates)RN - 2001-95-8 (Valinomycin)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - 6915-15-7 (malic acid)PT - Journal ArticleID - GM28454/GM/NIGMSDA - 19840424IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:6699003
SO  - J Biol Chem 1984 Mar 10 ;259(5):2971-2975

585
UI  - 20903
AU  - Hirano M
AU  - Takeda K
AU  - Kanazawa H
AU  - Futai M
TI  - Detection of ATP-dependent conformational change in the F1 portion and beta subunit of Escherichia coli H+-ATPase using 8-anilinonaphthalene-1- sulfonate
AB  - The bindings of ATP to Escherichia coli coupling factor ATPase (F1) and its isolated beta subunit were studied with 8-anilinonaphthalene-1- sulfonate (ANS). The fluorescence of ANS increased upon addition of F1 or the beta subunit, and this fluorescence was quenched on addition of ATP. Thus, ANS bound to the hydrophobic region of F1 or the beta subunit, and the binding of ATP was detected as a quenching of ANS fluorescence. The quenching of the fluorescence suggested that F1 or the beta subunit underwent a conformational change on binding to ATP. With and without ATP, 5.0 and 3.6 mol of ANS, respectively, bound to 1 mol of F1. On the other hand, with or without ATP about 1 mol of ANS bound to beta. The fluorescence quenching was specific for ATP and was not observed with GTP or CTP. It was dependent on pH, being higher at acidic pH, but it was not enhanced by MgCl2. The dissociation constants of F1 and the beta subunit for ATP were estimated to be 10(-4)-10(-5) M. The significance of these binding sites is discussed in relation to the mechanism of the ATPase
MH  - A
MH  - ANS
MH  - atp
MH  - ATPase
MH  - BETA-SUBUNIT
MH  - BINDING
MH  - Binding Sites
MH  - conformational change
MH  - CONSTANT
MH  - coupling
MH  - COUPLING FACTOR
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1
MH  - fluorescence
MH  - H+-ATPase
MH  - M
MH  - Macromolecular Systems
MH  - mechanism
MH  - pH
MH  - Site
MH  - SUBUNIT
MH  - SYSTEM
MH  - SYSTEMS
RP  - NOT IN FILE
NT  - UI - 84203529LA - engRN - 0 (Macromolecular Systems)RN - 0 (Mesylates)RN - 103-06-0 (anilinomethanesulfonate)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19840720IS - 0006-2960SB - IMCY - UNITED STATES
UR  - PM:6326806
SO  - Biochemistry 1984 Apr 10 ;23(8):1652-1656

586
UI  - 20905
AU  - Hsu SY
AU  - Senda M
AU  - Kanazawa H
AU  - Tsuchiya T
AU  - Futai M
TI  - Comparison of F1's of oxidative phosphorylation from Escherichia coli and Salmonella typhimurium and demonstration of interchangeability of their subunits
AB  - The peripheral membrane portion (SF1) of proton-translocating ATPase of Salmonella typhimurium and its alpha, beta, and gamma subunits were purified and compared with the same portion (EF1) from Escherichia coli. The alpha, beta, and gamma subunits of these F1's were found to be mutually interchangeable, and all possible combinations of the three subunits from EF1 and SF1 showed ATPase activity. Both F1's could bind functionally to the integral membrane part (F0) of either bacterium, suggesting that F0 and F1 are interchangeable in these two bacteria and thus that the two F1's are closely similar at the level of subunit structure. However, SF1 differed from EF1 in some enzymological properties such as its specific activity and susceptibilities to sodium dodecyl sulfate and methanol. The specific ATPase activity of EF1 was more than twice that of SF1, and hybrid enzymes containing the beta subunit of EF1 had higher activity than other hybrids. Amino acid analysis suggested that the primary structures of the alpha subunits of the two F1's are less homologous than those of the beta subunits. Thus, the primary structure of the alpha subunit may be more species specific than that of the beta subunit
MH  - ACID
MH  - ALPHA-SUBUNIT
MH  - Amino Acids
MH  - analysis
MH  - ATPase
MH  - Bacteria
MH  - BETA-SUBUNIT
MH  - Enzymes
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - F1
MH  - Macromolecular Systems
MH  - membrane
MH  - Methanol
MH  - Oxidative Phosphorylation
MH  - Phosphorylation
MH  - Sodium
MH  - SUBUNIT
MH  - SYSTEM
MH  - SYSTEMS
RP  - NOT IN FILE
NT  - UI - 84179076LA - engRN - 0 (Amino Acids)RN - 0 (Macromolecular Systems)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19840607IS - 0006-2960SB - IMCY - UNITED STATES
UR  - PM:6231953
SO  - Biochemistry 1984 Feb 28 ;23(5):988-993

587
UI  - 21042
AU  - Joliot P
AU  - Joliot A
TI  - Electron transfer between the two photosystems I. Flash excitation under oxidizing conditions
MH  - electron
MH  - electron transfer
MH  - Electron-transfer
MH  - flash
MH  - history
MH  - P
MH  - p700
MH  - thylakoid
MH  - TRANSFER
MH  - transport
MH  - wox
RP  - IN FILE
NT  - P 40; K
SO  - Biochim Biophys Acta 1984  ;765():210-218

588
UI  - 1173
AU  - Junesch U
AU  - Graber P
TI  - The rate of ATP-synthesis as function of .DELTA.pH and .DELTA..psi. in preactivated and non-preactivated chloroplasts
MH  - ATP synthesis
MH  - chloroplast
MH  - Chloroplasts
MH  - function
T2  - Adv. Photosynth. Res., Proc. Int. Congr. Photosynth., 6th, Meeting Date 1983, Volume 2, 431-6. Edited by: Sybesma, C. Nijhoff: The Hague, Neth
Y2  - -32676  
RP  - ON REQUEST (03/18/92)
SO  -  1984  ;():

589
UI  - 9869
AU  - Junesch U
AU  - Grber P
TI  - The rate of ATP-synthesis as function of pH and Psi in preactivated and non-preactivated chloroplasts
MH  - atp
MH  - ATP synthesis
MH  - chloroplast
MH  - Chloroplasts
MH  - function
MH  - Phosphorylation
MH  - Photosynthesis
MH  - regulation
T2  - Advances in Photosynthesis Research, Vol. II
A2  - Sybesma C
Y2  - -32676  
PB  - The Hauge/Boston/Lancaster: Nijhoff,M./Junk,W.
RP  - IN FILE
NT  - G
SO  -  1984  ;():II.5.431-II.5.436

590
UI  - 21294
AU  - Junge W
AU  - Hong YQ
AU  - Qian LP
AU  - Viale A
TI  - Cooperative transient trapping of photosystem II protons by the integral membrane portion (CF0) of chloroplast ATP-synthase after mild extraction of the four-subunit catalytic part (CF1)
AB  - Fo: n=6, pK=7.35 for spinach, pK=7.7 for peas.
MH  - trapping
MH  - photosystem II
MH  - Protons
MH  - proton
MH  - membrane
MH  - chloroplast
MH  - ATP synthase
MH  - catalytic
MH  - FO
MH  - Spinach
MH  - pea
RP  - NOT IN FILE
SO  - Proc Natl Acad Sci U S A 1984  ;81():3078-3082

591
UI  - 901
AU  - Kagawa Y
TI  - [ATPase in energy production system]
MH  - Adenosinetriphosphatase
MH  - biosynthesis
MH  - Energy Metabolism
RP  - NOT IN FILE
SO  - Tanpakushitsu Kakusan Koso 1984 Aug ;29(8):597-600

592
UI  - 20904
AU  - Kanazawa H
AU  - Kiyasu T
AU  - Noumi T
AU  - Futai M
TI  - Overproduction of subunit a of the F0 component of proton-translocating ATPase inhibits growth of Escherichia coli cells
AB  - A hybrid plasmid, pKY159, carrying the promoter and the proximal region of the gene cluster for proton-translocating ATPase caused growth inhibition of Escherichia coli cells (K. Yamaguchi and M. Yamaguchi, J. Bacteriol. 153:550-554, 1983). The mechanism of this growth inhibition was studied, especially in terms of the responsible gene(s). Insertion of IS1, IS5, or gamma delta between the promoter and the gene for a possible component of the ATPase of 14,000 daltons (14K protein) released the inhibitory effect by pKY159. Deletion of the gene for subunit a also released the effect. However, deletion in the gene for the 14K protein released the effect only with an additional insertion within the gene. These results suggested that overproduction of subunit a is closely related to growth inhibition, whereas the 14K protein is not
MH  - A
MH  - ATPase
MH  - Cells
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - M
MH  - mechanism
MH  - protein
MH  - SUBUNIT
RP  - NOT IN FILE
NT  - UI - 84185438LA - engRN - 0 (DNA Transposable Elements)RN - 0 (Plasmids)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19840607IS - 0021-9193SB - IMCY - UNITED STATES
UR  - PM:6325392
SO  - J Bacteriol 1984 Apr ;158(1):300-306

593
UI  - 20906
AU  - Kanazawa H
AU  - Noumi T
AU  - Matsuoka I
AU  - Hirata T
AU  - Futai M
TI  - F1ATPase of Escherichia coli: a mutation (uncA401) located in the middle of the alpha subunit affects the conformation essential for F1 activity
AB  - F1ATPase from the Escherichia coli mutant of H+-ATPase, AN120 (uncA401), has less than 1% of the wild type activity and has been shown to be defective in the alpha subunit by in vitro reconstitution experiments. In the present study, the mutation site was located within a domain of the subunit by recombinant DNA technology. For this, a series of recombinant plasmids carrying various portions of the alpha subunit gene were constructed and used for genetic recombination with AN120. Analysis of the recombinants indicated that the mutation site could be located between amino acid residues 370 and 387. The biochemical properties of the mutant F1 were analyzed further using the fluorescent ATP analog DNS-ATP (2'-(5-dimethylaminonaphthalene-1- sulfonyl)-amino-2'-deoxy ATP). The single turnover process of E. coli F1ATPase proposed by Matsuoka et al. [(1982) J. Biochem. 92, 1383- 1398.] was compared in the mutant and wild type F1's. Mutant F1 bound DNS-ATP and hydrolyzed it as efficiently as wild type F1. Results showed that binding of ATP to a low affinity site, possibly in the beta subunit, caused decrease of fluorescence of DNS-ATP in the wild type F1 and that this effect of ATP binding was inhibited by DCCD (dicyclohexyl carbodiimide). However, this effect was not inhibited by DCCD in the mutant F1, suggesting that in the proposed process some step(s) after ATP binding to the low affinity site differed in the mutant and wild F1's. When Pi was added to F1 bound to DNS-ATP or to aurovertin, a fluorescent probe capable of binding to the beta subunit, the opposite changes of fluorescence of these probes in the mutant and wild type F1's were observed, suggesting that the conformational change induced by phosphate binding was altered in the mutant F1. On the basis of the estimated mutation site and the biochemical properties of the mutant F1, the correlation of the domain of this site in the alpha subunit with the function of F1 ATPase is discussed
MH  - A
MH  - ACID
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - ALPHA-SUBUNIT
MH  - analysis
MH  - atp
MH  - ATPase
MH  - BETA-SUBUNIT
MH  - BINDING
MH  - conformation
MH  - conformational change
MH  - Dicyclohexylcarbodiimide
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - fluorescence
MH  - function
MH  - H+-ATPase
MH  - In Vitro
MH  - Macromolecular Systems
MH  - mutant
MH  - reconstitution
MH  - RESIDUE
MH  - Site
MH  - SUBUNIT
MH  - SYSTEM
MH  - SYSTEMS
RP  - NOT IN FILE
NT  - UI - 84126808LA - engRN - 0 (DNA, Recombinant)RN - 0 (Dansyl Compounds)RN - 0 (Macromolecular Systems)RN - 0 (Plasmids)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - 80214-45-5 (dansylamino deoxy-ATP)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19840314IS - 0003-9861SB - IMCY - UNITED STATES
UR  - PM:6230047
SO  - Arch Biochem Biophys 1984 Jan ;228(1):258-269

594
UI  - 19809
AU  - Kasho VN
AU  - Boyer PD
TI  - Relationships of inosine triphosphate and bicarbonate effects on F1 ATPase to the binding change mechanism
AB  - Two interesting previously reported properties of mitochondrial F1 ATPase have been confirmed and have been examined by 18O exchange measurements to assess if they are consistent with sequential participation of catalytic sites during ATP hydrolysis. These are the ability of
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BINDING
MH  - CHANGE MECHANISM
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - H(+)-Transporting ATP Synthase
MH  - Hydrolysis
MH  - mechanism
MH  - Nucleotides
MH  - Phosphates
MH  - SYNTHASE
MH  - Water
RP  - NOT IN FILE
NT  - UI - 85289195LA - engRN - 0 (Bicarbonates)RN - 0 (Inosine Nucleotides)RN - 0 (Phosphates)RN - 132-06-9 (Inosine Triphosphate)RN - 7732-18-5 (Water)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM-11094/GM/NIGMSDA - 19850930IS - 0145-479XSB - IMCY - UNITED STATESJC - HIO
UR  - PM:6242244
SO  - J Bioenerg Biomembr 1984 Dec ;16(5-6):407-419

595
UI  - 18784
AU  - Kotlyar AB
AU  - Vinogradov AD
TI  - Interaction of the membrane-bound succinate dehydrogenase with substrate and competitive inhibitors
MH  - A
MH  - ACTIVE SITE
MH  - affinity
MH  - Animal
MH  - Binding Sites
MH  - Binding,Competitive
MH  - Catalysis
MH  - COMPLEX
MH  - data
MH  - enzyme
MH  - Enzyme Inhibitors
MH  - Ethylmaleimide
MH  - fumarate
MH  - Fumarates
MH  - inhibitor
MH  - inhibitors
MH  - INTERACTION
MH  - Kinetics
MH  - Malonates
MH  - metabolism
MH  - method
MH  - Methods
MH  - oxaloacetates
MH  - pharmacology
MH  - REACTIVITY
MH  - redox
MH  - REDOX STATE
MH  - REDUCTION
MH  - Site
MH  - succinate
MH  - succinate dehydrogenase
MH  - Succinates
RP  - NOT IN FILE
NT  - The protective effect of dicarboxylates on the active-site- directed inhibition of the membrane-bound succinate dehydrogenase by N- ethylmaleimide, steady-state kinetics methods for Ki and Ks determinations, and equilibrium studies were employed to quantitate the relative affinities of succinate, fumarate, malonate and oxaloacetate to the reduced and oxidized species of the enzyme. A more than 10-fold difference in the relative affinities of the reduced and oxidized succinate dehydrogenase to succinate, fumarate and oxaloacetate is found, whereas the reactivity of the active-site sulphydryl group does not depend on the redox state of the enzyme. The redox-state-dependent changes in the affinity of the membrane-bound succinate dehydrogenase to oxaloacetate can be quantitatively accounted for by a 10-fold increase in the rate of dissociation of the enzyme-inhibitor complex which occurs upon reduction of the enzyme. The data obtained give no support for either the existence of a sulphydryl group other than the active-site one important for the catalysis or for the presence of a separate dicarboxylate-specific regulatory site in the succinate dehydrogenase molecule
SO  - Biochim Biophys Acta 1984  ;784(1):24-34

596
UI  - 9910
AU  - Lemasters JJ
TI  - The ATP-to-oxygen stoichiometries of oxidative phosphorylation by rat liver mitochondria. An analysis of ADP-induced oxygen jumps by linear nonequilibrium thermodynamics.
AB  - Uncertainty exists as to the proton stoichiometries of mitochondrial oxidative phosphorylation and consequently as to the ATP stoichiometries. In rat liver mitochondria, ADP/O ratios were determined from the total and extra oxygen consumed during ADP-stimulated respiration under conditions of quantitative conversion of ADP to ATP. For succinate, glutamate plus malate, 3-hydroxybutyrate, and 2-oxoglutarate, respectively, ADP/total O was 1.71, 2.71, 2.61, and 3.45. ADP/extra O was 2.03, 3.04, 3.23, and 4.15. The results were interpreted in terms of linear nonequilibrium thermodynamics. It was shown that ADP/extra O = Z/q where Z is the phenomenological stoichiometry and q is the degree of coupling. q was determined from the dependence of respiratory rate on delta Gp, the phosphorylation potential, and was about 0.98 for all substrates. The results were consistent with ideal ATP/O stoichiometries of 2 for succinate, 3 for glutamate plus malate, 3 or 3 1/4 for 3-hydroxybutyrate, and 4 for 2-oxoglutarate. Taking into account the oxidation-reduction free-energy changes measured across Sites 1 + 2 at static head (J.J. Lemasters, R. Grunwald, and R.K. Emaus J. Biol. Chem. 259, 3058-3063), an ideal ATP/O stoichiometry of 3 1/4 for 3-hydroxybutyrate is proposed. The lower ATP/O for glutamate plus malate is then accounted for by proton translocation linked to glutamate/aspartate exchange. The data suggest a new 13-proton scheme of chemiosmotic coupling in which proton stoichiometries are 3 for the F1Fo-ATPase, 1 for the exchange of ATP for ADP and Pi, and 5, 4, and 4 for Sites 1, 2, and 3.
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - ADP
MH  - analysis
MH  - Animal
MH  - atp
MH  - DEPENDENCE
MH  - Kinetics
MH  - Liver
MH  - Mathematics
MH  - metabolism
MH  - Mitochondria
MH  - Mitochondria,Liver
MH  - Models,Biological
MH  - Oxidation-Reduction
MH  - Oxidative Phosphorylation
MH  - Oxygen
MH  - Oxygen Consumption
MH  - Phosphorylation
MH  - proton
MH  - Rats
MH  - Respiration
MH  - Software
MH  - succinate
MH  - Support,Non-U.S.Gov't
MH  - Support,U.S.Gov't,P.H.S.
MH  - Thermodynamics
RP  - NOT IN FILE
SO  - J Biol Chem 1984 Nov 10 ;259(21):13123-13130

597
UI  - 196
AU  - Lotscher HR
AU  - deJong C
AU  - Capaldi RA
TI  - Inhibition of the adenosinetriphosphatase activity of Escherichia coli F1 by the water-soluble carbodiimide 1-ethyl-3-[3- (dimethylamino)propyl]carbodiimide is due to modification of several carboxyls in the beta subunit
AB  - Reaction of the ATPase of Escherichia coli (ECF1) with 1-ethyl-3-[3- (dimethylamino)propyl]carbodiimide (EDC) resulted in a time- and concentration-dependent inhibition of ATPase activity. The inactivation was greatly reduced by Mg2+ ions. Close to 13 mol of EDC per mol of ECF1 was incorporated into the enzyme at 95% inhibition of ATPase activity. Two-thirds of the label was found to be associated with subunit beta with a stoichiometry of about 3 mol of EDC per mol of beta. Cleavage of EDC-modified subunit beta with cyanogen bromide and fractionation of the peptides by high-pressure liquid chromatography revealed a short segment of 33 amino acids (CB8, residues 162-194) containing 3 mol of EDC per mol of peptide. In tryptic peptide maps, two EDC-labeled fragments could be identified (T18, residues 166-183, and T20, residues 186-202). The analyses were complicated by significant internal cross-linking within the beta subunit induced by EDC. The results show that EDC modifies multiple sites in a short segment of subunit beta which includes the glutamic acids modified by dicyclohexylcarbodiimide in F1 from both E. coli and PS3. In addition to covalent modification, EDC also promoted the formation of intersubunit cross-links. The predominant cross-linked product was identified as a beta-epsilon complex by antibody binding experiments
RP  - NOT IN FILE
NT  - UI - 85023296LA - engRN - 0 (Amino Acids)RN - 0 (Carbodiimides)RN - 0 (Carbon Radioisotopes)RN - 0 (Macromolecular Systems)RN - 1892-57-5 (Ethyldimethylaminopropyl Carbodiimide)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - HL 24526/HL/NHLBIDA - 19841212IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:6237683
SO  - Biochemistry 1984 Aug 28 ;23(18):4134-4140

598
UI  - 201
AU  - Lotscher HR
AU  - Capaldi RA
TI  - Structural asymmetry of the F1 of Escherichia coli as indicated by reaction with dicyclohexylcarbodiimide
AB  - Dicyclohexylcarbodiimide (DCCD) inhibits the ATPase activity of F1 from Escherichia coli by covalent modification of a single glutamic acid in the beta subunit. 95% inhibition was obtained after incorporation of around 1 mole of DCCD per mole F1, i.e. 1 mole of reagent per 3 beta subunits; and up to 2 moles of DCCD per mole F1 were readily incorporated into the protein. One of the 3 beta subunits per F1 can be crosslinked to the epsilon subunit by 1-ethyl-3- [3(dimethylamino)propyl]carbodiimide (EDC). This beta subunit (beta 1) is here shown to be shielded from reaction with DCCD, presumably by its association with epsilon and also possibly the gamma subunit. Thus the three beta subunits are not equivalent in the enzyme complex
RP  - NOT IN FILE
NT  - UI - 84231364LA - engRN - 0 (Carbodiimides)RN - 0 (Cross-Linking Reagents)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - HL 24526/HL/NHLBIDA - 19840709IS - 0006-291XSB - IMCY - UNITED STATESJC - 9Y8
UR  - PM:6233975
SO  - Biochem Biophys Res Commun 1984 May 31 ;121(1):331-339

599
UI  - 195
AU  - Lotscher HR
AU  - deJong C
AU  - Capaldi RA
TI  - Interconversion of high and low adenosinetriphosphatase activity forms of Escherichia coli F1 by the detergent lauryldimethylamine oxide
AB  - The amphipathic detergent lauryldimethylamine oxide (LDAO) stimulated ATP hydrolytic activity of Escherichia coli membranes and isolated ECF1 and ECF1-F0 ATPase complexes in a concentration-dependent manner. The enzyme was maximally activated 3-fold in membranes and 5-6-fold for isolated ECF1 or the ECF1-F0 complex. The maximal specific activity of activated ECF1 was 140-160 mumol of ATP hydrolyzed min-1 mg-1. The activation by LDAO was reversible. LDAO specifically released subunit delta from ECF1, generating a four subunit enzyme (alpha, beta, gamma, and epsilon subunits). The removal of subunit delta was not responsible for the stimulation of ATPase activity as evidenced by the full activation of the four subunit enzyme by LDAO. Treatment of ECF1 with 1- ethyl-3-[3-(dimethylamino)propyl]carbodiimide generated a beta-epsilon cross-link in high yield [Lotscher, H.R., DeJong, C., & Capaldi, R. A. (1984) Biochemistry (accompanying paper in this issue)]. The formation of this cross-link was greatly reduced in the presence of LDAO, indicating that the detergent perturbated the interaction between epsilon and beta subunits although epsilon was not removed from the ECF1 complex. The results suggest that the interconversion of ECF1 from a low to a high ATPase activity form by LDAO is in major part due to a release of the inhibitory action of subunit epsilon on subunit beta
RP  - NOT IN FILE
NT  - UI - 85023297LA - engRN - 0 (Detergents)RN - 0 (Dimethylamines)RN - 0 (Macromolecular Systems)RN - 0 (Surface-Active Agents)RN - 1643-20-5 (dodecyldimethylamine oxide)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - HL 24526/HL/NHLBIDA - 19841212IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:6237684
SO  - Biochemistry 1984 Aug 28 ;23(18):4140-4143

600
UI  - 197
AU  - Lotscher HR
AU  - deJong C
AU  - Capaldi RA
TI  - Modification of the F0 portion of the H+-translocating adenosinetriphosphatase complex of Escherichia coli by the water- soluble carbodiimide 1-ethyl-3-[3-(dimethylamino)propyl]carbodiimide and effect on the proton channeling function
AB  - 1-Ethyl-3-[3-(dimethylamino)propyl]carbodiimide (EDC), a water-soluble carbodiimide, inhibited ECF1-F0 ATPase activity and proton translocation through F0 when reacted with Escherichia coli membrane vesicles. The site of modification was found to be in subunit c of the F0 portion of the enzyme but did not involve Asp-61, the site labeled by the hydrophobic carbodiimide dicyclohexylcarbodiimide (DCCD). EDC was not covalently incorporated into subunit c in contrast to DCCD. Instead, EDC promoted a cross-link between the C-terminal carboxyl group (Ala-79) and a near-neighbor phosphatidylethanolamine as evidenced by fragmentation of subunit c with cyanogen bromide followed by high-pressure liquid chromatography and thin-layer chromatography
RP  - NOT IN FILE
NT  - UI - 85023295LA - engRN - 0 (Carbodiimides)RN - 0 (Carbon Radioisotopes)RN - 0 (Macromolecular Systems)RN - 0 (Peptide Fragments)RN - 1892-57-5 (Ethyldimethylaminopropyl Carbodiimide)RN - 506-68-3 (Cyanogen Bromide)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - HL 24526/HL/NHLBIDA - 19841212IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:6237682
SO  - Biochemistry 1984 Aug 28 ;23(18):4128-4134

601
UI  - 675
AU  - Lubben M
AU  - Lucken U
AU  - Weber J
AU  - Schafer G
TI  - Azidonaphthoyl-ADP: a specific photolabel for the high-affinity nucleotide-binding sites of F1-ATPase
AB  - 3'-O-[5-azidonaphthoyl]-ADP has been synthesized as a photoreactive analog to 3'-O-naphthoyl(1)-ADP which is known to bind to the high- affinity nucleotide sites of mitochondrial F1-ATPase, considered to be the catalytic sites. The photolabel in the dark acts as a ligand to F1- ATPase and as a competitive inhibitor with Ki = 11 microM. Binding to the enzyme is accompanied by a quench of endogenous protein fluorescence leveling off at an occupancy of 1 mol/mol F1, whereas the total number of reversible sites accessible to the analog is 3 mol/mol F1 as measured by isotope studies. Covalent insertion by near ultraviolet activation of the probe yields labeling of both alpha and beta polypeptides of F1; it is accompanied by corresponding removal of reversible high-affinity sites for ADP or naphthoyl-ADP and by an inhibition of the enzyme; total inactivation occurs at a covalent occupancy of 2 mol/mol F1. This is the maximum number of sites accessible to covalent modification by the label; one reversible site is still available in the totally inactivated enzyme. This observation is discussed in terms of a stochastic model requiring a minimum of two interacting catalytic domains out of three in order to commence catalysis
RP  - NOT IN FILE
NT  - UI - 85003621LA - engRN - 0 (Affinity Labels)RN - 0 (Azides)RN - 0 (Nucleotides)RN - 58-64-0 (Adenosine Diphosphate)RN - 84453-67-8 (3'-O-(5-azidonaphthoyl)adenosine diphosphate)RN - 93835-65-5 (5-azidonaphthoic acid)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19841113IS - 0014-2956SB - IMCY - GERMANY, WESTJC - EMZ
UR  - PM:6236974
SO  - Eur J Biochem 1984 Sep 17 ;143(3):483-490

602
UI  - 585
AU  - Matsuno-Yagi A
AU  - Hatefi Y
TI  - Inhibitory chemical modifications of F1-ATPase: effects on the kinetics of adenosine 5'-triphosphate synthesis and hydrolysis in reconstituted systems
AB  - The purified, soluble F1-ATPase was modified by several covalently reacting inhibitors, either known or considered to bind to the active site bearing beta-subunit, to cause partial inhibition up to 99%. The modified enzyme was then reconstituted in the presence of OSCP (oligomycin sensitivity conferring protein) with submitochondrial particles (SMP) almost completely (greater than 99%) denuded of active F1-ATPase and was assayed for oligomycin-sensitive ATPase and oxidative phosphorylation activities. The inhibitors used were 1-fluoro-2,4- dinitrobenzene (FDNB), N-(ethoxycarbonyl)-2-ethoxy-1,2-dihydroquinoline (EEDQ), 1-cyclohexyl-3-(2-morpholinoethyl)carbodiimide metho-p- toluenesulfonate (CMCD), quinacrine mustard (QM), 5-(dimethylamino)- naphthalene-1-sulfonyl chloride (dansyl-Cl), 5'-[p-(fluoro- sulfonyl)benzoyl]adenosine (FSBA), and N,N'-dicyclohexylcarbodiimide (DCCD). The SMP reconstituted with unmodified F1 exhibited oxidative phosphorylation and oligomycin-sensitive ATPase (in the presence of uncouplers) activities as high as 500 nmol min-1 mg-1 and 8 mumol min-1 mg-1, respectively. The systems reconstituted with F1 modified to cause various degrees of inhibition with FDNB, EEDQ, CMCD, QM, and dansyl-Cl exhibited the same degree of inhibition of oxidative phosphorylation and oligomycin-sensitive ATPase activities as the inhibition of the ATPase activity of the modified F1 before reconstitution. The systems reconstituted with FSBA-modified F1 showed the following relative degrees of inhibition: oxidative phosphorylation greater than oligomycin-sensitive ATPase of particles greater than ATPase of soluble F1. In contrast, the systems reconstituted with DCCD-modified F1 showed much greater inhibition of oligomycin-sensitive ATPase than of oxidative phosphorylation activity.(ABSTRACT TRUNCATED AT 250 WORDS)
RP  - NOT IN FILE
NT  - UI - 84280934LA - engRN - 0 (5'-(4-fluorosulfonylbenzoyl)adenosine)RN - 0 (Affinity Labels)RN - 370-86-5 (Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-61-7 (Adenosine)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - AM08126/AM/NIADDKDA - 19841012IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:6235851
SO  - Biochemistry 1984 Jul 17 ;23(15):3508-3514

603
UI  - 540
AU  - McEnery MW
AU  - Buhle EL
AU  - Aebi U
AU  - Pedersen PL
TI  - Proton ATPase of rat liver mitochondria. Preparation and visualization of a functional complex using the novel zwitterionic detergent 3-[(3- cholamidopropyl)dimethylammonio]-1-propanesulfonate
AB  - The proton ATPase of rat liver mitochondria has been purified by a simple procedure which involves the use of the novel, zwitterionic detergent 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate to solubilize the membrane-bound complex. The purified enzyme has a high, oligomycin-sensitive ATPase activity (11.3 +/- 2.9 mumol/min/mg) in the absence of added phospholipids. It shows, in four different gel electrophoretic systems, the five bands characteristic of the F1 portion of the complex and three additional Coomassie blue-stainable bands which have apparent molecular weights of 28,000, 19,000, and 13,600. A fourth Coomassie blue-stainable component of about 10,000- 12,500 daltons comigrates with the delta subunit, whereas a fifth component, detectable only by absorption at 280 nm, is observed between the dye front and the 10,000-dalton species. The enzyme complex has been reconstituted into liposomal vesicles of asolectin. Under these conditions the enzyme catalyzes an ATP-Pi exchange reaction and is capable of translocating protons in an ATP-dependent manner as assayed by quenching of 9-amino-6-chloro-2-methoxyacridine. Both activities are inhibited by the addition of oligomycin, uncoupler, dicyclohexylcarbodiimide, and cadmium. At high detergent concentration, the complex appears in negative stain electron microscopy in a dispersed state. The tripartite structure is clearly visible in monomeric, dimeric, or trimeric forms of the molecule. At the low detergent concentration, the proton ATPase tends to cluster into densely packed arrays. This represents the first report of the properties of a functionally active proton ATPase solubilized and purified in the presence of a zwitterionic detergent
RP  - NOT IN FILE
NT  - UI - 84162180LA - engRN - 0 (Cholic Acids)RN - 0 (Detergents)RN - 0 (Liposomes)RN - 0 (Oligomycins)RN - 75621-03-3 (3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - CA 10951/CA/NCIID - GM 07445/GM/NIGMSID - GM 31940/GM/NIGMSDA - 19840511IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:6231296
SO  - J Biol Chem 1984 Apr 10 ;259(7):4642-4651

604
UI  - 623
AU  - Melandri BA
AU  - Mehlhorn RJ
AU  - Packer L
TI  - Light-induced proton gradients and internal volumes in chromatophores of Rhodopseudomonas sphaeroides
AB  - To test the predictions of the chemiosmotic hypothesis, it is essential to have sensitive and accurate measures of the aqueous volume and pH within membrane compartments. One unique feature of the present investigation is the application of electron spin resonance probes to determine internal aqueous volume and pH changes in bacterial chromatophores under virtually identical conditions. Volumes of the chromatophores ranged from 6 to 16 microliter/mg bacteriochlorophyll among different preparations, and were sensitive to the osmolarity of the suspending buffer. pH gradients reached two units in illuminated chromatophores as determined with ESR methods, and increased when KCl and valinomycin were added to the assay. Measurements with the fluorescent dye 9-amino-acridine yielded similar pH gradients, provided that an operational vesicle volume, which corrected for the binding of the dye to the membrane, was used in the calculation. The sensitivity of the ESR method allowed the measurement of pH gradients resulting from only a few light flashes. A plot of pH gradients versus number of flashes was linear up to about 30 flashes, and intercepted the origin. This result is consistent with proton release into the bulk aqueous phase after only a single light flash. This ability to measure small pH gradients offers new opportunities for the study of energy-transducing mechanisms
RP  - NOT IN FILE
NT  - UI - 85045543LA - engRN - 0 (Cyclic N-Oxides)RN - 14691-88-4 (tempamine)RN - 2001-95-8 (Valinomycin)RN - 7447-40-7 (Potassium Chloride)PT - Journal ArticleDA - 19841218IS - 0003-9861SB - IMCY - UNITED STATESJC - 6SK
UR  - PM:6093711
SO  - Arch Biochem Biophys 1984 Nov 15 ;235(1):97-105

605
UI  - 9876
AU  - Mills JD
AU  - Mitchell P
TI  - A Dual pH Optimal Model for Activation of the Chloroplast ATPase, CF0-CF1
MH  - atp
MH  - model
MH  - Photosynthesis
MH  - regulation
T2  - Advances in Photosynthesis Research, Vol. II
A2  - Sybesma C
Y2  - -32676  
PB  - The Hauges/Boston/Lancaster: Nijhoff,M./Junk,W.
RP  - IN FILE
NT  - G
SO  -  1984  ;():II.6.523-II.6.526

606
UI  - 20900
AU  - Noumi T
AU  - Futai M
AU  - Kanazawa H
TI  - Replacement of serine 373 by phenylalanine in the alpha subunit of Escherichia coli F1-ATPase results in loss of steady-state catalysis by the enzyme
AB  - The mutant allele (uncA401) of the gene for the alpha subunit of Escherichia coli F1-ATPase was cloned from the total DNA of the mutant AN120 on a hybrid plasmid pAN120. Determination of the DNA sequence of the alpha subunit gene from pAN120 revealed a single base change of cytosine at nucleotide residue 1118 to thymine and indicated that serine 373 was replaced by phenylalanine. It has been reported that the mutant F1 is defective in a step of steady-state catalysis, whereas its single turnover process is normal (Kanazawa, H., Noumi, T., Matsuoka, I., Hirata T., and Futai, M. (1984) Arch. Biochem. Biophys. 228, 258- 269). Thus, we concluded that serine 373 in the alpha subunit is essential for steady-state catalysis by F1-ATPase
MH  - A
MH  - ALPHA-SUBUNIT
MH  - ATPase
MH  - BASE
MH  - Catalysis
MH  - COLI F1 ATPASE
MH  - Enzymes
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - M
MH  - Macromolecular Systems
MH  - mutant
MH  - RESIDUE
MH  - SUBUNIT
MH  - SYSTEM
MH  - SYSTEMS
RP  - NOT IN FILE
NT  - UI - 84289385LA - engRN - 0 (Macromolecular Systems)RN - 56-45-1 (Serine)RN - 63-91-2 (Phenylalanine)RN - EC 3.1.21 (DNA Restriction Enzymes)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19840928IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:6088480
SO  - J Biol Chem 1984 Aug 25 ;259(16):10076-10079

607
UI  - 20901
AU  - Noumi T
AU  - Mosher ME
AU  - Natori S
AU  - Futai M
AU  - Kanazawa H
TI  - A phenylalanine for serine substitution in the beta subunit of Escherichia coli F1-ATPase affects dependence of its activity on divalent cations
AB  - A mutant (KF11) of Escherichia coli H+-translocating ATPase (F1-F0) has a single point mutation in the beta subunit of F1 that has lost 90% of its Mg2+-dependent ATPase activity (Kanazawa, H., Horiuchi, Y., Takagi, M., Ishino, Y., and Futai, M. (1980) J. Biochem. (Tokyo) 88, 695-703). The mutation was mapped at about the 500th nucleotide residue from the 5' end of the beta subunit gene by a genetic recombination test on the physical map of the cistron coding for the beta subunit. The mutant allele of KF11 (uncD11) was cloned on a hybrid plasmid (pKF11) via DNA isolated from a lambda uncD11 transducing phage. Restriction fragments of pKF11 containing the estimated mutation site were subjected to polyacrylamide gel electrophoresis under conditions where strands were separated into single strands. The two strands of a DNA segment, which was shown to carry an altered base, showed anomalous migration compared with those from the wild-type fragment. The results confirmed the result of mapping of the altered site by genetic tests. On the basis of these results, the nucleotide sequence of the mutated gene was determined, and a single base change of the 524th cytosine to thymine resulting in a phenylalanine for serine substitution at residue 174 of the beta subunit was found. This result, together with results on the altered properties of F1 from KF11 reported previously, indicates that residue 174 is essential for the Mg2+-dependent ATPase activity of F1 but not for the Ca2+-dependent ATPase activity
MH  - A
MH  - ATPase
MH  - BASE
MH  - BETA-SUBUNIT
MH  - Cations
MH  - COLI F1 ATPASE
MH  - DEPENDENCE
MH  - Electrophoresis
MH  - Enzymes
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - M
MH  - Macromolecular Systems
MH  - mutant
MH  - point mutation
MH  - RESIDUE
MH  - Site
MH  - SUBUNIT
MH  - SYSTEM
MH  - SYSTEMS
RP  - NOT IN FILE
NT  - UI - 84289384LA - engRN - 0 (Cations, Divalent)RN - 0 (Macromolecular Systems)RN - 56-45-1 (Serine)RN - 63-91-2 (Phenylalanine)RN - EC 3.1.21 (DNA Restriction Enzymes)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleID - GM-23105/GM/NIGMSDA - 19840928IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:6088479
SO  - J Biol Chem 1984 Aug 25 ;259(16):10071-10075

608
UI  - 19810
AU  - O'Neal CC
AU  - Boyer PD
TI  - Assessment of the rate of bound substrate interconversion and of ATP acceleration of product release during catalysis by mitochondrial adenosine triphosphatase
AB  - The oxygen exchange parameters for the hydrolysis of ATP by the F1- ATPase have been determined over a 140,000-fold range of ATP concentrations and a 5,000-fold range of reaction velocity. The average number of water oxygens incorporated into each Pi product ranges from a limit of about 1.02 at saturating ATP concentrations to a limit of about 3.97 at very low ATP concentrations. The latter value represents 400 reversals of hydrolysis of bound ATP prior to Pi dissociation. In accord with the binding change mechanism, this means that ATP binding at one catalytic site increases the off constant of Pi and ADP from another catalytic site by at least 20,000-fold, equivalent to the use of 6 kcal mol-1 of ATP binding energy to promote product release. The estimated rate of reversal of hydrolysis of F1-ATPase-bound ATP to bound ADP + Pi varies only about 5-fold with ATP concentration. The rate is similar that observed previously for reversal of bound ATP hydrolysis or synthesis with the membrane-bound enzyme and is greater than the rate of net ATP formation during oxidative phosphorylation. This adds to evidence that energy input or membrane components are not required for bound ATP synthesis
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - ADENOSINE-TRIPHOSPHATASE
MH  - ADP
MH  - atp
MH  - ATP FORMATION
MH  - ATP synthase
MH  - ATP synthesis
MH  - ATPase
MH  - BINDING
MH  - Catalysis
MH  - CHANGE MECHANISM
MH  - F1
MH  - F1 ATPASE
MH  - H(+)-Transporting ATP Synthase
MH  - Hydrolysis
MH  - mechanism
MH  - membrane
MH  - Oxidative Phosphorylation
MH  - Oxygen
MH  - Oxygen Isotopes
MH  - Phosphorylation
MH  - SYNTHASE
MH  - synthesis
MH  - Water
RP  - NOT IN FILE
NT  - UI - 84185724LA - engRN - 0 (Oxygen Isotopes)RN - 56-65-5 (Adenosine Triphosphate)RN - 73-89-2 (Phosphoenolpyruvate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM 11094/GM/NIGMSDA - 19840614IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:6232276
SO  - J Biol Chem 1984 May 10 ;259(9):5761-5767

609
UI  - 538
AU  - Pedersen PL
AU  - Hullihen J
TI  - Inhibitor peptide of mitochondrial proton adenosine triphosphatase. Neutralization of its inhibitory action by calmodulin
AB  - In the presence of ATP and Mg2+, the homogeneous ATPase peptide inhibitor of rat liver mitochondria markedly inhibits the proton ATPase from this source (Cintron N. M., and Pedersen, P. L. (1979) J. Biol. Chem. 254, 3439-3443). Under these conditions, calmodulin prevents the inhibitor peptide from inhibiting the liver H+-ATPase. About 1.5 mol of calmodulin/mol of inhibitor is necessary to effect a half-maximal response (apparent Km = 0.5 microM calmodulin). The capacity of calmodulin to neutralize the action of the ATPase inhibitor peptide appears highly specific. This effect is not produced by insulin, trypsin inhibitor, lysozyme, ribonuclease, myoglobin, cytochrome c, ovalbumin, or bovine albumin. Only polyglutamate was found to mimic the action of calmodulin. However, when added together with calmodulin, polyglutamate failed to elicit an additive effect indicating that its site of interaction on the ATPase inhibitor peptide differs from that of calmodulin. Calcium is not essential in the assay medium for calmodulin to neutralize the action of the ATPase inhibitor peptide. The neutralization effect produced by calmodulin is also source- independent, with preparations of calmodulin from bovine brain and rat testes being equally competent. Calmodulin has no direct effect on the ATPase activity of the proton ATPase, nor does it affect the capacity of the enzyme to participate in either ATP synthesis or the ATP- dependent transhydrogenase reaction. Moreover, calmodulin fails to reverse inhibition of the H+-ATPase to which ATPase inhibitor peptide is already bound. Overall, these results indicate that calmodulin interacts in a direct and highly specific manner with the "free" ATPase peptide inhibitor of rat liver mitochondria
RP  - NOT IN FILE
NT  - UI - 85079996LA - engRN - 0 (ATPase inhibitory protein)RN - 0 (Calmodulin)RN - 0 (Proteins)RN - 0 (Trypsin Inhibitors)RN - 11061-68-0 (Insulin)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - CA 10951/CA/NCIDA - 19850125IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:6239865
SO  - J Biol Chem 1984 Dec 25 ;259(24):15148-15153

610
UI  - 472
AU  - Perlin DS
AU  - Latchney LR
AU  - Wise JG
AU  - Senior AE
TI  - Specificity of the proton adenosinetriphosphatase of Escherichia coli for adenine, guanine, and inosine nucleotides in catalysis and binding
AB  - Specificity of the Escherichia coli proton ATPase for adenine, guanine, and inosine nucleotides in catalysis and binding was studied. MgADP, CaADP, MgGDP, and MgIDP were each good substrates for oxidative phosphorylation. The corresponding triphosphates were each substrates for hydrolysis and proton pumping. At 1 mM concentration, MgATP, MgGTP, and MgITP drove proton pumping with equal efficiency. At 0.1 mM concentration, MgATP was 4-fold more efficient than MgITP or MgGTP. Nucleotide-depleted soluble F1 could rebind to F1-depleted membranes and block proton conductivity through F0; rebound nucleotide-depleted F1 catalyzed pH gradient formation with MgATP, MgGTP, or MgITP. This showed that the nonexchangeable nucleotide sites on F1 need not be occupied by adenine nucleotide for proton pumping to occur. It was further shown that no nucleotide was tightly bound in the nonexchangeable sites of F1 during proton pumping driven by MgGTP in these reconstituted membranes, whereas adenine nucleotide was tightly bound when MgATP was the substrate. Nucleotide-depleted soluble F1 bound maximally 5.9 ATP, 3.2 GTP, and 3.6 ITP of which half the ATP and almost all of the GTP and ITP exchanged over a period of 30-240 min with medium ADP or ATP. Also, half of the bound ATP exchanged with medium GTP or ITP. These data showed that inosine and guanine nucleotides do not bind to soluble F1 in nonexchangeable fashion, in contrast to adenine nucleotides. Purified alpha-subunit from F1 bound ATP at a single site but showed no binding of GTP nor ITP, supporting previous suggestions that the non-exchangeable sites in intact F1 are on alpha-subunits
RP  - NOT IN FILE
NT  - UI - 85047212LA - engRN - 0 (Ribonucleotides)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM-25349/GM/NIGMSID - GM-29805/GM/NIGMSDA - 19850114IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:6238624
SO  - Biochemistry 1984 Oct 9 ;23(21):4998-5003

611
UI  - 908
AU  - Robinson JD
TI  - The chemiosmotic hypothesis of energy coupling and the path of scientific opportunity
MH  - Adenosine Triphosphate
MH  - biosynthesis
MH  - Electron Transport
MH  - Energy Metabolism
MH  - Hydrogen
MH  - metabolism
MH  - Models,Chemical
MH  - Osmosis
MH  - Potassium
MH  - Sodium
MH  - Support,U.S.Gov't,P.H.S.
RP  - NOT IN FILE
SO  - Perspect Biol Med 1984  ;27(3):367-383

612
UI  - 20968
AU  - Schneider E
AU  - Altendorf K
TI  - Subunit b of the membrane moiety (F0) of ATP synthase (F1F0) from Escherichia coli is indispensable for H+ translocation and binding of the water-soluble F1 moiety
AB  - The ATP synthase complex, designated F1F0, of Escherichia coli is composed of a water-soluble portion (F1; membrane-associated ATPase, EC 3.6.1.3) with ATP-hydrolyzing activity and a membrane-integrated part (F0) with H+-translocating activity. F0 is built up from three kinds of subunits (a, b, and c). We have isolated the F0 portion directly from membranes of an E. coli strain (KY 7485) that overproduces the enzyme several fold. Subunit b was extracted from purified F0 by two methods. One method included prolonged incubation of the F0 complex in the presence of trichloroacetate (2.5 M) and the separation of subunit b and an a-c complex by gel filtration. Alternatively, subunit b was extracted by deoxycholate and separated from the a-c complex by hydrophobic-interaction chromatography. Integrated into liposomes, the a-c complex exhibited neither H+ uptake nor binding of F1. However, a functional F0 complex was reconstituted by adding stoichiometric amounts of subunit b to the a-c complex
MH  - A
MH  - Adenosinetriphosphatase
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BINDING
MH  - COMPLEX
MH  - Dicyclohexylcarbodiimide
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - F1
MH  - H+
MH  - Liposomes
MH  - M
MH  - Macromolecular Systems
MH  - membrane
MH  - Membranes
MH  - method
MH  - Methods
MH  - Multienzyme Complexes
MH  - Phosphotransferases
MH  - SUBUNIT
MH  - SYNTHASE
MH  - SYSTEM
MH  - SYSTEMS
MH  - translocation
RP  - NOT IN FILE
NT  - UI - 85063774LA - engRN - 0 (Macromolecular Systems)RN - 0 (Multienzyme Complexes)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - EC 2.7 (Phosphotransferases)RN - EC 2.7.4.- (ATP Synthetase Complexes)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19850122IS - 0027-8424SB - IMCY - UNITED STATES
UR  - PM:6209711
SO  - Proc Natl Acad Sci U S A 1984 Dec ;81(23):7279-7283

613
UI  - 475
AU  - Senior AE
AU  - Latchney LR
AU  - Ferguson AM
AU  - Wise JG
TI  - Purification of F1-ATPase with impaired catalytic activity from partial revertants of Escherichia coli uncA mutant strains
AB  - It is shown that F1-ATPase preparations having impaired catalytic rates may be purified from partial revertants of uncA mutant strains of Escherichia coli. Recovery of catalytic activity in the partial revertant F1 was accompanied by recovery of alpha in equilibrium beta intersubunit conformational interaction, supporting the hypothesis that such interaction is required for normal catalysis in F1. The specific ATPase activities of the partial revertant F1 preparations were in the range 1-29% of normal, and some of the preparations showed unusual insensitivity to inhibitors. The properties of a new uncA mutant F1 preparation (uncA498) which has approximately half of normal catalytic rate are also briefly described
RP  - NOT IN FILE
NT  - UI - 84126823LA - engRN - 0 (Aurovertins)RN - 0 (Azides)RN - 0 (Macromolecular Systems)RN - 0 (Peptides)RN - 26628-22-8 (Sodium Azide)RN - 56645-91-1 (efrapeptin)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSDA - 19840314IS - 0003-9861SB - IMCY - UNITED STATESJC - 6SK
UR  - PM:6230049
SO  - Arch Biochem Biophys 1984 Jan ;228(1):49-53

614
UI  - 473
AU  - Senior AE
TI  - Disposition of polar and nonpolar residues on outer surfaces of transmembrane helical segments of proteins involved in proton translocation
AB  - "Helical wheel" projections of transmembrane helical segments of membrane proteins involved in proton translocation were constructed. The particular proteins studied were the uncF protein subunit of the Escherichia coli proton-ATPase, the uncE protein subunit of the E. coli proton-ATPase, and cytochrome oxidase subunit III. Clear demarcation of polar and nonpolar regions on surfaces of transmembrane helical segments was seen in the uncF protein and in uncE protein helical segment two, but not in uncE protein helical segment one. The transmembrane segment of cytochrome oxidase subunit III which includes the dicyclohexylcarbodiimide (DCCD)-reactive residue was very similar to E. coli uncE protein helical segment two. The DCCD-reactive residue in both was clearly located on a nonpolar surface
RP  - NOT IN FILE
NT  - UI - 85021432LA - engRN - 0 (Bacterial Proteins)RN - 0 (Membrane Proteins)RN - 0 (Protons)RN - EC 1.9.3.1 (Cytochrome-c Oxidase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM-29805/GM/NIGMSDA - 19841120IS - 0003-9861SB - IMCY - UNITED STATESJC - 6SK
UR  - PM:6091562
SO  - Arch Biochem Biophys 1984 Oct ;234(1):138-143

615
UI  - 9959
AU  - Snozzi M
AU  - Crofts AR
TI  - Electron transport in chromatophores from Rhodopseudomonas sphaeroides GA fused with liposomes
AB  - Chromatophores from Rhodopseudomonas sphaeroides GA were fused with liposomes in order to dilute the components of the cyclic photosynthetic electron-transport chain within the membrane. This dilution led to a decrease in the rate of cytochrome b-561 reduction. The original rates could be restored at potentials around 100 mV (where a large part of the quinone pool is chemically reduced), if ubiquinone was incorporated into the liposomes prior to fusion. Similar dilution effects could be observed in synchronized cultures. The membrane obtained after division contained about twice the amount of phospholipids per reaction center when compared to chromatophores prepared from cells harvested just before division. Chromatophores from synchronized cultures are more uniform with respect to the concentration of the different electron-transport components in the membrane than the membranes from normally grown cells. The kinetic behaviour both of fused chromatophores and of membranes from synchronized cultures are in agreement with a modified Q-cycle model for photosynthetic electron transport in Rps. sphaeroides. The results presented in this paper cannot be explained by postulating the presence of a firmly bound quinone, Qz, in the ubiquinol: cytochrome c2 oxidoreductase, as previously proposed
MH  - Cells
MH  - Cytochrome b
MH  - electron
MH  - Electron Transport
MH  - Liposomes
MH  - Membranes
MH  - model
MH  - Phospholipids
MH  - transport
MH  - Ubiquinone
RP  - NOT IN FILE
NT  - UI - 84280949LA - engRN - 0 (Liposomes)RN - 11130-51-1 (cytochrome b561)RN - 1339-63-5 (Ubiquinone)RN - 9035-37-4 (Cytochrome b)PT - Journal ArticleID - 5 RO1 GM26305/GM/NIGMSDA - 19841009IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:6331848
SO  - Biochim Biophys Acta 1984 Aug 31 ;766(2):451-463

616
UI  - 20969
AU  - Steffens K
AU  - Schneider E
AU  - Herkenhoff B
AU  - Schmid R
AU  - Altendorf K
TI  - Chemical modification of the F0 part of the ATP synthase (F1F0) from Escherichia coli. Effects on proton conduction and F1 binding
AB  - The purified F0 part of the ATP synthase complex from Escherichia coli was incorporated into liposomes and chemically modified by various reagents. The modified F0-liposomes were assayed for H+ uptake and, after reconstitution with F1, for total and dicyclohexylcarbodiimide- sensitive ATPase activity. The water-soluble carbodiimide, 1-ethyl-3-(- 3-dimethylaminopropyl)carbodiimide methiodide, (1.2 mM), inhibited H+ uptake to a great extent. Binding of F1 was almost unaffected, but the hydrolysis of ATP was uncoupled from H+ transport. This is reflected by the inhibition of dicyclohexylcarbodiimide-sensitive ATPase activity. Woodward's reagent K, N-ethyl-5-phenylisoxazolium-3'-sulfonate, inhibited both H+ uptake and total ATPase activity. Modification of arginine residues by phenylglyoxal (20 mM) was followed by inhibition of the F1 binding activity by 80% of the control. H+ translocation was reduced to 70%. Diethylpyrocarbonate (3 mM) exhibited a strong inhibiting effect on H+ uptake but not on F1 binding. Modification of tyrosine (by tetranitromethane) as well as lysine residues (by succinic anhydride) did not affect F0 functions. From the data presented we conclude that carboxyl-groups, different from the dicyclohexylcarbodiimide-binding site, are involved in H+ translocation through F0 and, in part, in the functional binding of F1. Furthermore, for the latter function, also arginine residues seem to be important. The role of histidine residues remains unclear at present
MH  - A
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BINDING
MH  - COMPLEX
MH  - Dicyclohexylcarbodiimide
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - F1
MH  - function
MH  - H+
MH  - Histidine
MH  - Hydrolysis
MH  - Liposomes
MH  - Multienzyme Complexes
MH  - Peptide Fragments
MH  - Phosphotransferases
MH  - proton
MH  - Protons
MH  - reconstitution
MH  - RESIDUE
MH  - Site
MH  - SYNTHASE
MH  - translocation
MH  - transport
RP  - NOT IN FILE
NT  - UI - 84108421LA - engRN - 0 (Liposomes)RN - 0 (Multienzyme Complexes)RN - 0 (Peptide Fragments)RN - 0 (Protons)RN - 1609-47-8 (Diethyl Pyrocarbonate)RN - 55520-40-6 (Tyrosine)RN - 56-87-1 (Lysine)RN - 74-79-3 (Arginine)RN - EC 2.7 (Phosphotransferases)RN - EC 2.7.4.- (ATP Synthetase Complexes)PT - Journal ArticleDA - 19840321IS - 0014-2956SB - IMCY - GERMANY, WEST
UR  - PM:6319139
SO  - Eur J Biochem 1984 Feb 1 ;138(3):617-622

617
UI  - 94
AU  - Tybulewicz VL
AU  - Falk G
AU  - Walker JE
TI  - Rhodopseudomonas blastica atp operon. Nucleotide sequence and transcription
AB  - The nucleotide sequence has been determined of a 12,368 base-pair region of DNA cloned from the non-sulphur photosynthetic bacterium Rhodopseudomonas blastica. It contains a cluster of six genes of which five encode the subunits of F1-ATPase; the sixth codes for an unknown protein. The genes are arranged in the same order as in the Escherichia coli unc operon, except that the unknown gene is placed between those for gamma and beta subunits. Neither the genes for F0 subunits, nor a homologue of the E. coli uncI gene is associated with this locus. The six genes are transcribed from a single promoter and we have designated this region the R. blastica atp operon. The two distal genes, beta and epsilon, may also be transcribed from a second promoter. Initiation and termination points for transcription have been identified by primer extensions and S1 nuclease mapping experiments. Signals involved in initiation of translation (Shine and Dalgarno sequences) and termination of transcription in the photosynthetic bacterium resemble those in E. coli. However, no common features can be identified in these two bacteria between 5' regions adjacent to sites of initiation of transcription. The sequence also contains a gene that encodes a protein homologous to discoidin, a cell surface lectin of Dictyostelium discoideum thought to be involved in cell--cell aggregation. Seven other reading frames have not been identified
RP  - NOT IN FILE
NT  - UI - 85058188LA - engRN - 0 (Codon)RN - 0 (DNA, Bacterial)RN - 0 (RNA, Bacterial)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19841227IS - 0022-2836SB - IMCY - ENGLANDJC - J6V
UR  - PM:6209404
SO  - J Mol Biol 1984 Oct 25 ;179(2):185-214

618
UI  - 887
AU  - Van Dam K
AU  - Woelders H
AU  - Colen A
AU  - Westerhoff HV
TI  - A structural basis for mosaic protonic energy coupling
MH  - Animal
MH  - Energy Metabolism
MH  - Mitochondria
MH  - Models,Biological
MH  - physiology
MH  - Protons
RP  - NOT IN FILE
SO  - Biochem Soc Trans 1984 Jun ;12(3):401-402

619
UI  - 394
AU  - Vinogradov AD
TI  - [Catalytic properties of mitochondrial ATP-synthetase]
RP  - NOT IN FILE
NT  - UI - 85047338LA - rusRN - 0 (Multienzyme Complexes)RN - 0 (Phosphates)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 2.7 (Phosphotransferases)RN - EC 2.7.4.- (ATP synthetase complex)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewDA - 19850103IS - 0320-9725SB - IMCY - USSRJC - A28
UR  - PM:6093895
SO  - Biokhimiia 1984 Aug ;49(8):1220-1238

620
UI  - 95
AU  - Walker JE
AU  - Saraste M
AU  - Gay NJ
TI  - The unc operon. Nucleotide sequence, regulation and structure of ATP- synthase
RP  - NOT IN FILE
NT  - UI - 85000488LA - engRN - 0 (Codon)RN - 0 (DNA, Bacterial)RN - 0 (Macromolecular Systems)RN - 0 (Multienzyme Complexes)RN - 0 (RNA, Bacterial)RN - 0 (RNA, Messenger)RN - EC 2.7 (Phosphotransferases)RN - EC 2.7.4.- (ATP synthetase complex)RN - EC 2.7.7.6 (DNA-Directed RNA Polymerase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewDA - 19841109IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:6206892
SO  - Biochim Biophys Acta 1984 Sep 6 ;768(2):164-200

621
UI  - 370
AU  - Walker JE
AU  - Falk G
AU  - Gay NJ
AU  - Tybulewicz VL
TI  - Genes for bacterial and mitochondrial ATP synthase
RP  - NOT IN FILE
NT  - UI - 84209363LA - engRN - 0 (DNA, Bacterial)RN - 0 (DNA, Mitochondrial)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19840622IS - 0300-5127SB - IMCY - ENGLANDJC - E48
UR  - PM:6233197
SO  - Biochem Soc Trans 1984 Apr ;12(2):234-235

622
UI  - 612
AU  - Westerhoff HV
AU  - Melandri BA
AU  - Venturoli G
AU  - Azzone GF
AU  - Kell DB
TI  - A minimal hypothesis for membrane-linked free-energy transduction. The role of independent, small coupling units
AB  - Experimental data are reviewed that are not in keeping with the scheme of 'delocalized' protonic coupling in membrane-linked free-energy transduction. It turns out that there are three main types of anomalies: (i) rates of electron transfer and of ATP synthesis do not solely depend on their own driving force and on delta mu H, (ii) the ('static head') ratio of delta Gp to delta mu H varies with delta mu H and (iii) inhibition of either some of the electron-transfer chains or some of the H+-ATPases, does not cause an overcapacity in the other, non-inhibited proton pumps. None of the earlier free-energy coupling schemes, alternative to delocalized protonic coupling, can account for these three anomalies. We propose to add a fifth postulate, namely that of the coupling unit, to the four existing postulates of 'delocalized protonic coupling' and show that, with this postulate, protonic coupling can again account for most experimental observations. We also discuss: (i) how experimental data that might seem to be at odds with the 'coupling unit' hypothesis can be accounted for and (ii) the problem of the spatial arrangement of the electrical field in the different free-energy coupling schemes
RP  - NOT IN FILE
NT  - UI - 85072875LA - engRN - 0 (Multienzyme Complexes)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 2.7 (Phosphotransferases)RN - EC 2.7.4.- (ATP synthetase complex)PT - Journal ArticlePT - ReviewDA - 19850221IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:6095906
SO  - Biochim Biophys Acta 1984 Dec 17 ;768(3-4):257-292

623
UI  - 613
AU  - Westerhoff HV
AU  - Melandri BA
AU  - Venturoli G
AU  - Azzone GF
AU  - Kell DB
TI  - Mosaic protonic coupling hypothesis for free energy transduction
RP  - NOT IN FILE
NT  - UI - 84108856LA - engRN - 0 (Ion Channels)RN - 0 (Liposomes)RN - 0 (Protons)RN - 56-65-5 (Adenosine Triphosphate)PT - Journal ArticleDA - 19840302IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:6319178
SO  - FEBS Lett 1984 Jan 2 ;165(1):1-5

624
UI  - 541
AU  - Williams N
AU  - Hullihen JM
AU  - Pedersen PL
TI  - The proton adenosinetriphosphatase complex of rat liver mitochondria. Temperature-dependent dissociation-reassociation of the F1-ATPase subunits
AB  - The soluble F1 moiety of the rat liver mitochondrial proton ATPase dissociates into two easily separable fractions when cold treated and then warmed. One fraction is soluble in potassium phosphate buffer, pH 7.4, whereas the other is insoluble. Neither of these two fractions alone can catalyze ATP hydrolysis under assay conditions optimal for the native F1-ATPase. The insoluble fraction when resolved via sodium dodecyl sulfate--polyacrylamide gel electrophoresis is shown to be composed of only alpha and gamma subunits. When this fraction is chromatographed on Sephadex G-75, it is resolved into an alpha gamma complex and into the alpha subunit alone. The soluble fraction when resolved in the same electrophoretic system is shown to contain the remaining subunits, beta, delta, epsilon, and some gamma. This fraction is resolved into two major components by chromatography on Sepharose CL- 6B, a beta gamma complex and beta subunit alone. The cold-dissociated enzyme can be readily associated when the temperature is raised to 20 degrees C. In the presence of either ATP or MgATP the enzyme completely regains its original ATPase specific activity. In contrast, Mg2+ is only about 15% effective in restoring ATPase activity. The results presented here define conditions for the dissociation and reassociation of the major subunits comprising the F1-ATPase of rat liver and thus provide a unique system among mammalian enzymes for testing the function of individual subunits. In addition, they strongly indicate that neither the alpha nor beta subunits, nor complexes of these subunits with the gamma subunit, are capable of catalyzing ATP hydrolysis.(ABSTRACT TRUNCATED AT 250 WORDS)
RP  - NOT IN FILE
NT  - UI - 84179046LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - 7439-95-4 (Magnesium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - CA 10951/CA/NCIDA - 19840601IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:6231951
SO  - Biochemistry 1984 Feb 14 ;23(4):780-785

625
UI  - 539
AU  - Williams N
AU  - Amzel LM
AU  - Pedersen PL
TI  - Proton ATPase of rat liver mitochondria: a rapid procedure for purification of a stable, reconstitutively active F1 preparation using a modified chloroform method
AB  - A method is described for the purification of rat liver F1-ATPase by a modification of the chloroform extraction procedure originally described by Beechey et al. (Biochem. J. (1975) 148, 533). Purified liver membrane vesicles are extracted with chloroform in the presence of ATP and EDTA. The procedure yields pure F1 in only 2-3 h without the necessity of ion-exchange chromatography. The enzyme exhibits the alpha, beta, gamma, delta, and epsilon bands characteristic of F1- ATPase. It has a high ATPase specific activity, and is reconstitutively active, catalyzing high rates of ATP synthesis. Significantly, it can be readily crystallized. If desired, the enzyme can be passed over a gel filtration column to place it in a stabilizing phosphate-EDTA buffer, lyophilized and stored indefinitely at -20 degrees C
RP  - NOT IN FILE
NT  - UI - 85020704LA - engRN - 67-66-3 (Chloroform)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - CA 10951/CA/NCIID - GM 25432/GM/NIGMSDA - 19841114IS - 0003-2697SB - IMCY - UNITED STATESJC - 4NK
UR  - PM:6237596
SO  - Anal Biochem 1984 Aug 1 ;140(2):581-588

626
UI  - 972
AU  - Wilson JE
TI  - Some thoughts on the evolutionary basis for the prominent role of ATP and ADP in cellular energy metabolism
AB  - The predominance of the adenosine triphosphate/adenosine diphosphate (ATP/ADP) couple in cellular phosphorylation reactions, including those that form the basis for cellular energy metabolism, cannot be explained on thermodynamic grounds since a variety of &quot;high energy phosphate&quot; compounds (including ADP itself) found in the cell would, based on thermodynamic considerations, be at least as effective as ATP in serving as a phosphoryl donor. How then did present-day organisms come to rely on the ATP/ADP couple as the principal mediator of phosphorylation reactions? The early appearance of adenine compounds in the prebiotic environment is suggested by experiments indicating that, relative to other purine or pyridimine compounds, adenine derivatives are preferentially synthesized under simulated prebiotic conditions (Ponnamperuma et al., 1963). In addition to the roles of adenine nucleotides in phosphorylation reactions, other adenine derivatives (e.g. Coenzyme A, flavin adenine dinucleotide, puridine nucleotides) are employed in a variety of metabolic roles. The principal function of the adenine moiety in these latter cases is in the binding of these derivatives to the relevant enzyme. The capability for binding of the adenine moiety appears to have arisen early in evolution and been exploited in a multitude of contexts, a suggestion consistent with observed similarities between the binding sites of several enzymes employing adenine derivatives as substrate. The early availability of suitable adenine compounds in the biosphere and development of complementary binding sites on cellular proteins, coupled with the expected advantages in having a limited number of metabolites as central mediators of endergonic and exergonic metabolism could readily have led to the observed pre-eminence of adenine nucleotides in cellular energy metabolism.(ABSTRACT TRUNCATED AT 250 WORDS)
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Monophosphate
MH  - Adenosine Triphosphate
MH  - Binding Sites
MH  - Brain
MH  - development
MH  - Energy Metabolism
MH  - enzymology
MH  - Evolution
MH  - Hexokinase
MH  - Magnesium
MH  - metabolism
MH  - Phosphorylation
MH  - Proteins
MH  - Pyruvate Kinase
MH  - Support,U.S.Gov't,P.H.S.
MH  - Thermodynamics
RP  - NOT IN FILE
SO  - J Theor Biol 1984 Dec 21 ;111(4):615-623

627
UI  - 474
AU  - Wise JG
AU  - Latchney LR
AU  - Ferguson AM
AU  - Senior AE
TI  - Defective proton ATPase of uncA mutants of Escherichia coli. 5'- Adenylyl imidodiphosphate binding and ATP hydrolysis
AB  - The Escherichia coli uncA gene codes for the alpha-subunit of the F1 sector of the membrane proton ATPase. In this work purified soluble F1 enzymes from three mutant strains ( uncA401 , uncA447 , and uncA453 ) have been compared to F1 from a normal strain in respect to (a) binding of 5'-adenylyl imidodiphosphate (AMPPNP) to native enzyme in both the presence and absence of Mg, (b) high-affinity binding of MgATP to native enzyme, (c) total reloading of MgAMPPNP to nucleotide-depleted F1 preparations, (d, e) ability to hydrolyze MgATP at both high MgATP concentrations (d) (steady-state conditions) and low MgATP concentrations (e) where substrate hydrolysis occurs under nonsteady- state (" unisite ") conditions, and (f) sensitivity of steady-state ATPase activities to inhibitors of normal F1-ATPase activity. uncA mutant F1 showed normal stoichiometry of MgAMPPNP binding to both native (three sites per F1) and nucleotide-depleted preparations (six sites per F1). Native uncA F1 preparations showed lower-than-normal affinity for MgAMPPNP and MgATP at the first site filled. Binding of AMPPNP in the absence of Mg was similar to normal, except that no increase in affinity for AMPPNP was induced by aurovertin. The uncA F1- ATPases had low but real steady-state rates of ATP hydrolysis, which were inhibited by aurovertin but relatively insensitive to inhibition by AMPPNP, efrapeptin, and sodium azide. Non-steady-state ( unisite ) ATP hydrolysis rates catalyzed at low substrate concentrations by uncA F1-ATPases were similar to normal.(ABSTRACT TRUNCATED AT 250 WORDS)
RP  - NOT IN FILE
NT  - UI - 84203498LA - engRN - 25612-73-1 (Adenylyl Imidodiphosphate)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM-25349/GM/NIGMSDA - 19840622IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:6232946
SO  - Biochemistry 1984 Mar 27 ;23(7):1426-1432

628
UI  - 584
AU  - Wong SY
AU  - Matsuno-Yagi A
AU  - Hatefi Y
TI  - Kinetics of ATP hydrolysis by F1-ATPase and the effects of anion activation, removal of tightly bound nucleotides, and partial inhibition of the ATPase by covalent modification
AB  - Eadie-Hofstee plots (v/[S] vs. v) of the kinetics of ATP hydrolysis by purified bovine heart mitochondrial F1-ATPase (MF1) over a substrate (MgATP) concentration range of 1-5000 microM were curvilinear, indicating negative cooperativity with respect to [MgATP] as originally shown by Ebel & Lardy (1975) [Ebel, R. E., & Lardy, H. A. (1975) J. Biol. Chem. 250, 191-196]. The data were computer analyzed for the best fit of the least number of straight lines, each representing a different apparent Km and Vmax. The best fits for MF1 and TF1 from the thermophilic bacterium PS3 were three lines in each case. The upper limits of the apparent Km values for MF1 were of the order of 10(-6), 10(-4), and 10(-3) M, and the corresponding apparent Vmax values (per minute per milligram of protein) were in the range of micromoles or less for the lowest Km line and decamicromoles for the other two. The results for TF1 were very similar. The presence of an activating anion (10 mM KHCO3) in the MF1 assay medium increased the overall Vmax by about 50% and eliminated the high Km but had essentially no effect on the intermediate and low Km's, indicating retention of negative cooperativity in the corresponding substrate concentration range. Kinetic data for MgITP as substrate also yielded two Km values (in the absence of KHCO3) differing by about 10(4)-fold. The relationship between [14C]dicyclohexylcarbodiimide [( 14C]-DCCD) binding to MF1 and activity inhibition was linear up to approximately 1 mol of DCCD bound/mol of MF1. At this point, the degree of inhibition was about 95%.(ABSTRACT TRUNCATED AT 250 WORDS)
RP  - NOT IN FILE
NT  - UI - 85047213LA - engRN - 0 (Anions)RN - 0 (Bicarbonates)RN - 0 (Macromolecular Systems)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - AM 08126/AM/NIADDKDA - 19850114IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:6238625
SO  - Biochemistry 1984 Oct 9 ;23(21):5004-5009

629
UI  - 586
AU  - Yagi T
AU  - Hatefi Y
TI  - Thiols in oxidative phosphorylation: inhibition and energy-potentiated uncoupling by monothiol and dithiol modifiers
AB  - Three apparently different modifications of submitochondrial particles (SMP) or ATP synthase preparations (complex V) inhibit oxidative phosphorylation and ATP-32Pi exchange activities, all of which are reversible by addition of mono- or dithiols. (a) Triphenyltin chloride inhibits ATP synthesis and hydrolysis without uncoupling. The inhibition by triphenyltin chloride is reversible by addition of beta- mercaptoethanol, dithiothreitol, or dihydrolipoamide. (b) Factor B is a water-soluble protein of Mr (11-12) X 10(3), contains a vicinal dithiol, and is required for energy transfer to and from F1-ATPase when tested with SMP-rendered factor B deficient by extraction with ammonia- ethylenediaminetetraacetic acid (EDTA) (AE-SMP). Treatment of factor B with mono- and dithiol modifiers, such as p- (chloromercuri)benzenesulfonate (PCMPS), Cd2+, or diazenedicarboxylic acid bis(dimethylamide) (diamide), inhibits factor B. This inhibition is reversed by addition to modified factor B of appropriate mono- and dithiol compounds. Preparations of AE-SMP are partially F1 deficient and partially uncoupled. The uncoupling can be repaired completely by addition of factor B or low levels of oligomycin, or to a large extent by addition of F1-ATPase + oligomycin sensitivity conferring protein. (c) SMP, AE-SMP, and complex V can be completely uncoupled by treatment at 30 degrees C with phenylarsine oxide, Cd2+, diamide, PCMPS, monobromobimane, and mono- and bifunctional maleimides. The uncoupling by these reagents is potentiated by membrane energization. Uncoupling by diamide is greater than or equal to 80% reversed by dihydrolipoamide or beta-mercaptoethanol, the former being much more potent. Dithiothreitol and dithioerythritol are poorly effective.(ABSTRACT TRUNCATED AT 250 WORDS)
RP  - NOT IN FILE
NT  - UI - 85000436LA - engRN - 0 (Oxidative Phosphorylation Coupling Factors)RN - 0 (Sulfhydryl Compounds)RN - 0 (Sulfhydryl Reagents)RN - 10465-78-8 (Diamide)PT - Journal ArticleID - AM08126/AM/NIADDKDA - 19841102IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:6477876
SO  - Biochemistry 1984 May 22 ;23(11):2449-2455

630
UI  - 587
AU  - Yagi T
AU  - Matsuno-Yagi A
AU  - Vik SB
AU  - Hatefi Y
TI  - Modulation of the kinetics and the steady-state level of intermediates of mitochondrial coupled reactions by inhibitors and uncouplers
AB  - In oxidative phosphorylation and ATP-driven uphill electron transfer from succinate to NAD, double-reciprocal plots of rates vs. substrate concentrations of the energy-driven reactions are a family of parallel lines at several fixed subsaturating concentrations of the substrates or at several moderate concentrations of the inhibitors of the energy- yielding reactions. Thus, as shown elsewhere [Hatefi, Y., Yagi, T., Phelps, D. C., Wong, S.-Y., Vik, S. B., & Galante, Y. M. (1982) Proc. Natl. Acad. Sci. U.S.A. 79, 1756-1760], partial uncoupling decreases the Vappmax and increases the Kappm of the substrates of the energy- driven reactions, resulting in a decrease of Vmax/Km as a function of increased uncoupling. However, partial limitation of the flow rates of the energy-yielding reactions decreases both the Vappmax and the Kappm of the substrates of the energy-driven reactions, resulting in no change in Vmax/Km. This is true as long as the rate limitation is moderate (e.g., less than 60%), under which conditions the steady-state membrane potential (delta psi) remains essentially unchanged. At high inhibition of the energy-yielding reactions, or at moderate inhibition in the presence of low levels of an uncoupler to cause partial uncoupling, then the family of double-reciprocal plots is no longer parallel and tends to converge toward the left. Under these conditions, steady-state delta psi and Vmax/Km also decrease as inhibition is increased. The relationship between the magnitude of steady-state delta psi and the rate of the energy-driven reaction was studied in oxidative phosphorylation, ATP-driven electron transfer from succinate to NAD, and respiration-driven uniport calcium transport by intact mitochondria.(ABSTRACT TRUNCATED AT 250 WORDS)
RP  - NOT IN FILE
NT  - UI - 84179050LA - engRN - 0 (Azides)RN - 0 (Malonates)RN - 0 (Oligomycins)RN - 0 (Uncoupling Agents)RN - 141-82-2 (malonic acid)RN - 53-84-9 (NAD)RN - 555-60-2 (Carbonyl Cyanide m-Chlorophenyl Hydrazone)RN - 56-65-5 (Adenosine Triphosphate)RN - 7440-70-2 (Calcium)RN - 83-79-4 (Rotenone)PT - Journal ArticleID - AM 08126/AM/NIADDKDA - 19840607IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:6712922
SO  - Biochemistry 1984 Feb 28 ;23(5):1029-1036

631
UI  - 776
AU  - Yoshida M
AU  - Kagawa Y
TI  - [Mechanism of action of H+-ATPase(F0F1)]
RP  - NOT IN FILE
NT  - UI - 85064621LA - jpnRN - 0 (Protons)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewDA - 19850114IS - 0039-9450SB - IMCY - JAPANJC - Q7D
UR  - PM:6209745
SO  - Tanpakushitsu Kakusan Koso 1984 Aug ;29(8):601-609

632
UI  - 773
AU  - Bar-Zvi D
AU  - Yoshida M
AU  - Shavit N
TI  - Photoaffinity labeling of the TF1-ATPase from the thermophilic bacterium PS3 with 3'-O-(4-benzoyl)benzoyl ADP
AB  - 3'-O-(4-Benzoyl)benzoyl ADP (BzADP) was used as a photoaffinity label for covalent binding of adenine nucleotide analogs to the nucleotide binding site(s) of the thermophilic bacterium PS3 ATPase (TF1). As with the CF1-ATPase (Bar-Zvi, D. and Shavit, N. (1984) Biochim. Biophys. Acta 765, 340-356) noncovalently bound BzADP is a reversible inhibitor of the TF1-ATPase. BzADP changes the kinetics of ATP hydrolysis from noncooperative to cooperative in the same way as ADP does, but, in contrast to the effect on the CF1-ATPase, it has no effect on the Vmax. In the absence of Mg2+ 1 mol BzADP binds noncovalently to TF1, while with Mg2+ 3 mol are bound. Photoactivation of BzADP results in the covalent binding of the analog to the nucleotide binding site(s) on TF1 and correlates with the inactivation of the ATPase. Complete inactivation of the TF1-ATPase occurs after covalent binding of 2 mol BzADP/mol TF1. Photoinactivation of TF1 by BzADP is prevented if excess of either ADP or ATP is present during irradiation. Analysis by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate of the Bz[3H]ADP-labeled TF1-ATPase shows that all the radioactivity is incorporated into the beta subunit
RP  - NOT IN FILE
NT  - UI - 85199893LA - engRN - 0 (Affinity Labels)RN - 0 (Macromolecular Systems)RN - 58-64-0 (Adenosine Diphosphate)RN - 87550-03-6 (3'-O-(4-benzoyl)benzoyladenosine diphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19850702IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:2859890
SO  - Biochim Biophys Acta 1985 May 31 ;807(3):293-299

633
UI  - 909
AU  - Bogucka K
TI  - [Current theories on the mechanism of energy coupling in mitochondria]
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - Energy Metabolism
MH  - H(+)-Transporting ATP Synthase
MH  - Human
MH  - metabolism
MH  - Mitochondria
MH  - Models,Biological
MH  - Nad
MH  - Oxidation-Reduction
MH  - Oxidative Phosphorylation
MH  - Protons
RP  - NOT IN FILE
SO  - Postepy Biochem 1985  ;31(1):5-28

634
UI  - 955
AU  - Bohnensack R
TI  - Mathematical modeling of mitochondrial energy transduction
AB  - A mathematical model of mitochondrial energy transduction is presented. The model contains rate equations for the main steps of oxidative phosphorylation. It was used to simulate the relations of respiration and ATP formation to extra- and intramitochondrial ATP/ADP ratios under various steady-state conditions. Furthermore, the model equations allowed to compute control coefficients, which quantify the control exerted by different steps on respiration. The distribution of control within mitochondria is demonstrated to depend on the metabolic state of mitochondria and also on the properties of extramitochondrial enzymes involved in ATP turnover. The simulated steady-state data as well as computed control coefficients were found in close agreement with experimental data
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - Animal
MH  - Energy Metabolism
MH  - Kinetics
MH  - Mathematics
MH  - metabolism
MH  - Mitochondria
MH  - Models,Biological
MH  - Oxidation-Reduction
MH  - Oxidative Phosphorylation
MH  - Respiration
RP  - NOT IN FILE
SO  - Biomed Biochim Acta 1985  ;44(6):853-862

635
UI  - 774
AU  - Bullough DA
AU  - Kwan M
AU  - Laikind PK
AU  - Yoshida M
AU  - Allison WS
TI  - The varied responses of different F1-ATPases to chlorpromazine
AB  - The effects of chlorpromazine on various properties of the F1-ATPases from bovine heart mitochondria (MF1), the plasma membranes of Escherichia coli (EF1), and plasma membranes of the thermophilic bacterium PS3 (TF1) have been examined. While chlorpromazine inhibited MF1 with an I0.5 of about 50 microM and EF1 with an I0.5 of about 150 microM at 23 degrees C, the ATPase activity of TF1 was stimulated by chlorpromazine concentrations up to 0.6 mM at this temperature. Maximal activation of about 20% was observed at 0.2 mM chlorpromazine at 23 degrees C. Chlorpromazine concentrations greater than 0.6 mM inhibited TF1 at 23 degrees C. At 37 degrees C the ATPase activity of TF1 was doubled in the presence of 0.5 mM chlorpromazine, the concentration at which maximal stimulation was observed at this temperature. Chlorpromazine inhibited the rate of inactivation of EF1 by dicyclohexylcarbodiimide (DCCD) at 23 degrees C and pH 6.5. Concentrations of chlorpromazine which inhibited the ATPase activity of TF1 at pH 7.0 accelerated the rate of inactivation of the enzyme by DCCD at pH 6.5, while lower concentrations of the phenothiazine, which stimulated the ATPase, had no effect on DCCD inactivation. Chlorpromazine concentrations up to 1.0 mM had no effect on the rate of inactivation of TF1 by DCCD at 37 degrees C and pH 6.5. Chlorpromazine at 0.5 mM accelerated the rate of inactivation of MF1 by 5'-p- fluorosulfonylbenzoyladenosine (FSBA), while it slowed the rate of inactivation of EF1 by FSBA. The inactivation of TF1 by FSBA in the absence of chlorpromazine was complex and was not included in this comparison. Chlorpromazine protected MF1 and EF1 against cold inactivation. Whereas 100 microM chlorpromazine afforded about 90% stabilization of MF1 at 4 degrees C, only about 30% stabilization of EF1 was observed under the same conditions in the presence of 400 microM chlorpromazine. Each of the ATPases was inactivated by the structural analog of chlorpromazine, quinacrine mustard. Whereas 5 mM ATP and 5 mM ADP protected MF1 and TF1 against inactivation by 0.5 mM quinacrine mustard, the rate of inactivation of EF1 by quinacrine mustard was accelerated fourfold by 5 mM ATP and slightly accelerated by 5 mM ADP
RP  - NOT IN FILE
NT  - UI - 85120878LA - engRN - 0 (5'-(4-fluorosulfonylbenzoyl)adenosine)RN - 4213-45-0 (Quinacrine Mustard)RN - 50-53-3 (Chlorpromazine)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 58-61-7 (Adenosine)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM-16974/GM/NIGMSDA - 19850321IS - 0003-9861SB - IMCY - UNITED STATESJC - 6SK
UR  - PM:2857548
SO  - Arch Biochem Biophys 1985 Feb 1 ;236(2):567-575

636
UI  - 21201
AU  - Drachev LA
AU  - Kaulen AD
AU  - Skulachev VP
TI  - [Nature of the electrogenic phases of the bacteriorhodopsin photocycle and the localization of retinal]
MH  - Bacteriorhodopsin
MH  - carotenoid
MH  - Carotenoids
MH  - electrogenic
MH  - proton
MH  - Protons
MH  - Retinaldehyde
RP  - NOT IN FILE
NT  - UI - 85203398LA - rusRN - 0 (Protons)RN - 0 (Retinoids)RN - 0 (Suspensions)RN - 116-31-4 (Retinaldehyde)RN - 36-88-4 (Carotenoids)RN - 53026-44-1 (Bacteriorhodopsins)RN - 9004-70-0 (Collodion)PT - Journal ArticleDA - 19850711IS - 0002-3264SB - IMCY - USSR
UR  - PM:2986925
SO  - Dokl Akad Nauk SSSR 1985 Mar ;281(1):176-180

637
UI  - 470
AU  - Duncan TM
AU  - Senior AE
TI  - The defective proton-ATPase of uncD mutants of Escherichia coli. Two mutations which affect the catalytic mechanism
AB  - The catalytic characteristics of F1-ATPases from uncD412 and uncD484 mutant strains of Escherichia coli were studied in order to understand how these beta-subunit mutations cause defective catalysis. Both mutant enzymes showed reduced affinity for ATP at the first catalytic site. While uncD412 F1 was similar to normal in other aspects of single site catalysis, uncD484 F1 showed a Keq of bound reactants greatly biased toward bound substrate ATP and an abnormally fast rate of Pi release. Impairment of productive catalytic cooperativity was the major cause of the reduced steady state ("multisite") catalytic rate in both mutant enzymes. Addition of excess ATP to saturate second and/or third catalytic sites did promote ATP hydrolysis and product release at the first catalytic site of uncD412 F1, but the multisite turnover rate was significantly slower than normal. In contrast, with uncD484 F1, addition of excess ATP induced rapid release of ATP from the first catalytic site and so productive catalytic cooperativity was almost completely absent. The results show that both mutations affect properties of the catalytic site and catalytic site cooperativity and further that the relatively more severe uncD484 mutation affects a residue which acts as a determinant of the fate of bound substrate ATP during promotion of catalysis. Taken together with previous studies of uncA mutant F1-ATPases (Wise, J. G., Latchney, L. R., Ferguson, A. M., and Senior, A. E. (1984) Biochemistry 23, 1426-1432) the results indicate that catalytic site cooperativity in F1-ATPases involves concerted beta-alpha-beta intersubunit communication between catalytic sites on the beta-subunits
RP  - NOT IN FILE
NT  - UI - 85182608LA - engRN - 0 (Adenine Nucleotides)RN - 0 (Aurovertins)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - 7439-95-4 (Magnesium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - 5T32GM07136/GM/NIGMSID - GM25349/GM/NIGMSDA - 19850531IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:2859284
SO  - J Biol Chem 1985 Apr 25 ;260(8):4901-4907

638
UI  - 368
AU  - Falk G
AU  - Walker JE
TI  - Transcription of Rhodospirillum rubrum atp operon
AB  - The photosynthetic non-sulphur bacterium Rhodospirillum rubrum contains a cluster of five genes encoding the subunits of F1-ATPase [Falk, Hampe & Walker (1985) Biochem. J. 228, 391-407]. Transcription of these genes has been studied by two methods, transcriptional mapping with S1 nuclease and primer extension analysis. Thereby a 5'-end in RNA derived from this region has been demonstrated at a guanine residue 236 bases before the initiation codon of the gene for the delta-subunit, the first in this cluster. DNA sequences on the 5' side of this nucleotide show some similarity to promoters in Escherichia coli, but are not apparently related to sequences upstream of the Rhodopseudomonas blastica atp operon. A 3'-end in RNA derived from this gene cluster has been demonstrated by S1-nuclease mapping. This is found before a run of thymidylate residues in the DNA, on the 3' side of a region of dyad symmetry. In E. coli these features are characteristic of rho- independent transcriptional termination signals. It appears from these studies and from the organization of the genes that the five genes in the atp cluster may be co-transcribed from this promoter and that transcripts terminate at the region of dyad symmetry
RP  - NOT IN FILE
NT  - UI - 86025375LA - engRN - 0 (DNA, Bacterial)RN - 0 (RNA, Messenger)RN - EC 3.1.- (Endonucleases)RN - EC 3.1.30.1 (Aspergillus Nuclease S1)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19851104IS - 0264-6021SB - IMCY - ENGLANDJC - 9YO
UR  - PM:2864916
SO  - Biochem J 1985 Aug 1 ;229(3):663-668

639
UI  - 369
AU  - Falk G
AU  - Hampe A
AU  - Walker JE
TI  - Nucleotide sequence of the Rhodospirillum rubrum atp operon
AB  - The nucleotide sequence was determined of a 8775-base-pair region of DNA cloned from the photosynthetic non-sulphur bacterium Rhodospirillum rubrum. It contains a cluster of five genes encoding F1-ATPase subunits. The genes are arranged in the same order as F1 genes in the Escherichia coli unc operon. However, as in the related organism Rhodopseudomonas blastica, neither genes for components of F0, the membrane sector of ATP synthase, nor a homologue of the E. coli uncI gene are associated with this locus, as they are in E. coli
RP  - NOT IN FILE
NT  - UI - 85251588LA - engRN - 0 (DNA, Bacterial)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19850821IS - 0264-6021SB - IMCY - ENGLANDJC - 9YO
UR  - PM:2861810
SO  - Biochem J 1985 Jun 1 ;228(2):391-407

640
UI  - 19807
AU  - Feldman RI
AU  - Boyer PD
TI  - The role of tightly bound ADP on chloroplast ATPase
AB  - Isolated chloroplast coupling factor 1 ATPase is known to retain about 1 mol of tightly bound ADP/mol of enzyme. Some experimental results have given evidence that the bound ADP is at catalytic sites, but this view has not been supported by observations of a slow replacement of the bound ADP when CaATP or MgATP is added. The experiments reported in this paper show why a slow replacement of ADP bound at a catalytic site can occur. When coupling factor 1, labeled with tightly bound [3H]ADP, is exposed to Mg2+ or Ca2+ prior to the addition of MgATP or CaATP, a pronounced lag in the onset of ATP hydrolysis is observed, and only slow replacement of the [3H]ADP occurs. Mg2+ or Ca2+ can induce inhibition very rapidly, as if an inhibited form of the enzyme results whenever the enzyme with tightly bound ADP encounters Mg2+ or Ca2+ prior to ATP. The inhibited form can be slowly reactivated by incubation with EDTA, although some irreversible loss in activity is encountered. In contrast, when MgATP or CaATP is added to enzyme depleted of Mg2+ and Ca2+ by incubation with EDTA, a rapid onset of ATP hydrolysis occurs and most of the tightly bound [3H]ADP is released within a few seconds, as expected for binding at a catalytic site. The Mg2+-induced inhibition of both the ATPase activity and the lack of replacement of tightly bound [3H] ADP can be largely prevented by incubation with Pi under conditions favoring Pi addition to the site containing the tightly bound ADP. Our and other results can be explained if enzyme catalysis is greatly hindered when MgADP or CaADP without accompanying Pi is tightly bound at one of the three catalytic sites on the enzyme in a high affinity conformation
MH  - A
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - ADP
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BINDING
MH  - Calcium
MH  - Catalysis
MH  - chloroplast
MH  - conformation
MH  - COUPLING FACTOR
MH  - Edetic Acid
MH  - H(+)-Transporting ATP Synthase
MH  - Hydrolysis
MH  - Magnesium
MH  - Phosphates
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 86033746LA - engRN - 0 (Enzyme Reactivators)RN - 0 (Phosphates)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 60-00-4 (Edetic Acid)RN - 7439-95-4 (Magnesium)RN - 7440-70-2 (Calcium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM 11094/GM/NIGMSDA - 19851129IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:2865256
SO  - J Biol Chem 1985 Oct 25 ;260(24):13088-13094

641
UI  - 21136
AU  - Ferguson SJ
TI  - Fully delocalised chemiosmotic or localised proton flow pathways in energy coupling? A scrutiny of experimental evidence 
MH  - proton
MH  - coupling
MH  - A
RP  - NOT IN FILE
SO  - Biochim Biophys Acta 1985  ;811():47-95

642
UI  - 366
AU  - Gay NJ
AU  - Walker JE
TI  - Two genes encoding the bovine mitochondrial ATP synthase proteolipid specify precursors with different import sequences and are expressed in a tissue-specific manner
AB  - Two cDNAs encoding different precursor proteins of the same mature proteolipid subunit of mitochondrial ATP synthase have been cloned from a bovine cDNA library. The hybridisation probe was a mixture of 17-mer oligonucleotides containing 256 discrete sequences. The coding sequences of the two cDNAs differ in 25 silent positions of codons and the 3' non-coding sequences are only weakly related. The precursor sequences, which direct the import of the proteolipid into the mitochondrion, are 61 and 68 amino acids long. They are related to each other in regions which probably are recognition signals for the processing protease. The corresponding genes are expressed differently in various tissues in a way that reflects their embryonic origin
RP  - NOT IN FILE
NT  - UI - 86135991LA - engRN - 0 (Protein Precursors)RN - 0 (Proteolipids)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19860414IS - 0261-4189SB - IMCY - ENGLANDJC - EMB
UR  - PM:2868890
SO  - EMBO J 1985 Dec 16 ;4(13A):3519-3524

643
UI  - 20897
AU  - Gromet-Elhanan Z
AU  - Khananshvili D
AU  - Weiss S
AU  - Kanazawa H
AU  - Futai M
TI  - ATP synthesis and hydrolysis by a hybrid system reconstituted from the beta-subunit of Escherichia coli F1-ATPase and beta-less chromatophores of Rhodospirillum rubrum
AB  - Photophosphorylation and ATPase activities were restored to beta-less Rhodospirillum rubrum chromatophores by their reconstitution with purified beta-subunits of either R. rubrum F1-ATPase (Rr beta) or Escherichia coli F1-ATPase (Ec beta). In the homologous reconstituted system both activities were restored to the same extent, whereas in the hybrid system ATP synthesis was restored to about 10% when the hydrolysis was restored to 200%. This difference in rates of synthesis and hydrolysis was not due to any general uncoupling effect of Ec beta leading to an increased membrane permeability to protons, because with both hybrid and homologous systems an identical light-induced quenching of quinacrine fluorescence was observed. They differed, however, in ATP- driven quenching of quinacrine fluorescence, which was much lower in the hybrid system. These results suggest that the hybrid has a decreased capacity for proton-translocation through the membrane-bound Fo channel during ATP hydrolysis, and probably also during ATP synthesis. The very high ATPase activity of the hybrid system indicates that it might enable the released protons to leak to the outside medium rather than to move inside through the Fo channel. The activities restored by Rr beta and Ec beta exhibit a similar sensitivity to dicyclohexylcarbodiimide, but different sensitivities to oligomycin and to an anti-E. coli F1 (EcF1) antibody. Oligomycin inhibited only the homologous R. rubrum system whereas anti-EcF1 was a much more effective inhibitor of the hybrid system. It is therefore concluded that Rr beta plays a role, that the Ec beta cannot fulfill, in conferring oligomycin sensitivity to the RrFo X F1-ATP synthase-ATPase complex
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthesis
MH  - ATPase
MH  - BETA-SUBUNIT
MH  - chromatophore
MH  - chromatophores
MH  - COLI F1 ATPASE
MH  - COMPLEX
MH  - Dicyclohexylcarbodiimide
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - fluorescence
MH  - Hydrolysis
MH  - membrane
MH  - Oligomycins
MH  - Permeability
MH  - Photophosphorylation
MH  - proton
MH  - Protons
MH  - reconstitution
MH  - Rhodospirillum
MH  - Rhodospirillum rubrum
MH  - synthesis
MH  - SYSTEM
MH  - SYSTEMS
RP  - NOT IN FILE
NT  - UI - 86008351LA - engRN - 0 (Oligomycins)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - 83-89-6 (Quinacrine)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19851114IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:2864345
SO  - J Biol Chem 1985 Oct 15 ;260(23):12635-12640

644
UI  - 21096
AU  - Guffanti AA
AU  - Chiu E
AU  - Krulwich TA
TI  - Failure of an alkalophilic bacterium to synthesize ATP in response to a valinomycin-induced potassium diffusion potential at high pH
AB  - Starved whole cells of the obligately alkalophilic Bacillus firmus RAB synthesize ATP upon addition of L-malate at pH 9.0 as expected of an aerobic organism that grows oxidatively on nonfermentable carbon sources at pH values as high as 11.0. The current study was a detailed examination of the perplexing inability of such cells to exhibit ATP synthesis in response to a valinomycin-mediated potassium diffusion potential at pH 9.0. While there were minor differences in the patterns of generation of the potential and the proton influx that accompanies its generation in the three different buffering systems employed, the magnitude of the transmembrane electro-chemical potential of protons was at least as high as pH 9.0 as at pH 7.0. Nevertheless, a diffusion potential consistently energized ATP synthesis at pH 7.0 but not at 9.0; these findings were independent of the presence or absence of Tris or of Na+. By contrast, the artificial electron donor ascorbate, in the presence of phenazine methosulfate, energized ATP synthesis by the starved whole cells at both pH values. The same phenomenon, i.e., efficacy of a respiration-derived potential but not of a diffusion potential at pH 9.0, was demonstrated in ADP + Pi-loaded membrane vesicles. On the other hand, electrogenic Na+-coupled solute transport could be energized by both ascorbate/phenazine and methosulfate and a diffusion potential in the vesicles at pH 9.0. The results are discussed in connection with models of a localized path of proton flow between proton pumps and the ATP synthase
MH  - A
MH  - ACID
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - ADP
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - Bacillus
MH  - Bacteria
MH  - Cells
MH  - Diffusion
MH  - diffusion potential
MH  - electrogenic
MH  - electron
MH  - membrane
MH  - membrane vesicles
MH  - model
MH  - Onium Compounds
MH  - pH
MH  - Phosphates
MH  - Potassium
MH  - proton
MH  - Proton Pump
MH  - Protons
MH  - SYNTHASE
MH  - synthesis
MH  - SYSTEM
MH  - SYSTEMS
MH  - transport
MH  - Valinomycin
MH  - vesicles
RP  - NOT IN FILE
NT  - UI - 85224230LA - engRN - 0 (Aminoisobutyric Acids)RN - 0 (Onium Compounds)RN - 0 (Organophosphorus Compounds)RN - 0 (Phosphates)RN - 144-90-1 (3-aminoisobutyric acid)RN - 18198-39-5 (tetraphenylphosphonium)RN - 2001-95-8 (Valinomycin)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 7440-09-7 (Potassium)PT - Journal ArticleID - GM28454/GM/NIGMSDA - 19850717IS - 0003-9861SB - IMCY - UNITED STATES
UR  - PM:4004268
SO  - Arch Biochem Biophys 1985 Jun ;239(2):327-333

645
UI  - 21039
AU  - Harold FM
AU  - Kakinuma Y
TI  - Primary and secondary transport of cations in bacteria
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - ATPase
MH  - Bacteria
MH  - Carrier Proteins
MH  - Cations
MH  - Dicyclohexylcarbodiimide
MH  - ion
MH  - Ion Channels
MH  - Potassium
MH  - protein
MH  - Proteins
MH  - proton
MH  - Protons
MH  - review
MH  - secondary
MH  - Sodium
MH  - transport
RP  - NOT IN FILE
NT  - UI - 86128769LA - engRN - 0 (Carrier Proteins)RN - 0 (Cations)RN - 0 (Ion Channels)RN - 0 (Protons)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - 7440-09-7 (Potassium)RN - 7440-23-5 (Sodium)RN - EC 3.6.1.- (sodium-translocating ATPase)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticlePT - ReviewID - AI 03568/AI/NIAIDDA - 19860312IS - 0077-8923SB - IMCY - UNITED STATES
UR  - PM:2418733
SO  - Ann N Y Acad Sci 1985  ;456():375-383

646
UI  - 583
AU  - Hatefi Y
TI  - The mitochondrial electron transport and oxidative phosphorylation system
RP  - NOT IN FILE
NT  - UI - 85277980LA - engRN - 0 (Macromolecular Systems)RN - 0 (Membrane Proteins)RN - 0 (Multienzyme Complexes)RN - EC 1. (Oxidoreductases)RN - EC 1.10.2.2 (Ubiquinol-Cytochrome-c Reductase)RN - EC 1.3.5.1 (succinate dehydrogenase (ubiquinone))RN - EC 1.3.99.1 (Succinate Dehydrogenase)RN - EC 1.6.99. (Quinone Reductases)RN - EC 1.6.99.2 (NAD(P)H Dehydrogenase (Quinone))RN - EC 1.9.3.1 (Cytochrome-c Oxidase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewID - AM08126/AM/NIADDKID - GM24887/GM/NIGMSDA - 19850904IS - 0066-4154SB - IMCY - UNITED STATESJC - 6DJ
UR  - PM:2862839
SO  - Annu Rev Biochem 1985  ;54():1015-1069

647
UI  - 1166
AU  - Junesch U
AU  - Thulke G
AU  - Graber P
TI  - Kinetics of proton-transport-coupled ATP synthesis driven by an artificially generated .DELTA.pH and .DELTA..psi
MH  - atp
MH  - ATP synthesis
MH  - Kinetics
MH  - synthesis
RP  - ON REQUEST (03/18/92)
SO  - NATO ASI Ser ,Ser A 1985  ;91():425-437

648
UI  - 21023
AU  - Junesch U
AU  - Grber P
TI  - The rate of ATP synthesis as a function of .Delta. pH in normal and dithiothreitol-modified chloroplasts
MH  - A
MH  - atp
MH  - ATP synthesis
MH  - chloroplast
MH  - Chloroplasts
MH  - function
MH  - pH
MH  - synthesis
RP  - IN FILE
SO  - Biochim Biophys Acta 1985  ;809():429-434

649
UI  - 21038
AU  - Kakinuma Y
AU  - Harold FM
TI  - ATP-driven exchange of Na+ and K+ ions by Streptococcus faecalis
AB  - We describe the characterization of KtrII, a novel potassium transport system of Streptococcus faecalis, first discovered by H. Kobayashi [1982) J. Bacteriol. 150, 506-511). KtrII requires sodium ions and mediates the stoichiometric exchange of internal Na+ for external K+. Potassium accumulation is not energized by the electrochemical potentials of either H+ or Na+; the energy source is probably ATP. Two lines of evidence indicate that KtrII is a manifestation of the sodium- stimulated ATPase reported earlier (Heefner, D. L., and Harold, F. M. (1982) Proc. Natl. Acad. Sci. U.S.A. 79, 2798-2802). (i) Mutants that lack the ATPase also lack KtrII, and revertants recover both in parallel. (ii) KtrII and the Na+-ATPase are induced in parallel when cells are grown on media rich in sodium, particularly under conditions that limit the generation of a proton potential. KtrII is not induced in response to K+ deprivation. We propose that the Na+-ATPase exchanges Na+ for K+ ions
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - atp
MH  - ATPase
MH  - Cells
MH  - Dicyclohexylcarbodiimide
MH  - H+
MH  - ion
MH  - Ions
MH  - M
MH  - mutant
MH  - Potassium
MH  - proton
MH  - Sodium
MH  - Streptococcus
MH  - SYSTEM
MH  - transport
RP  - NOT IN FILE
NT  - UI - 85130934LA - engRN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - 7440-09-7 (Potassium)RN - 7440-23-5 (Sodium)RN - EC 3.6.1.- (sodium-translocating ATPase)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleID - AI-03568/AI/NIAIDDA - 19850404IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:2857711
SO  - J Biol Chem 1985 Feb 25 ;260(4):2086-2091

650
UI  - 21292
AU  - Kamensky Y
AU  - Konstantinov AA
AU  - Kunz WS
AU  - Surkov S
TI  - Effect of bc1 site electron transfer inhibitors on the absorption spectra of mitochondrial cytochromes b
MH  - absorption
MH  - bc1
MH  - cytochrome
MH  - Cytochromes
MH  - electron
MH  - electron transfer
MH  - Electron-transfer
MH  - inhibitor
MH  - inhibitors
MH  - Site
MH  - spectra
MH  - TRANSFER
RP  - IN FILE
SO  - FEBS Lett 1985  ;181():95-99

651
UI  - 20898
AU  - Kanazawa H
AU  - Hama H
AU  - Rosen BP
AU  - Futai M
TI  - Deletion of seven amino acid residues from the gamma subunit of Escherichia coli H+-ATPase causes total loss of F1 assembly on membranes
AB  - A mutant gene for the gamma subunit of H+-translocating ATPase was cloned from Escherichia coli mutant NR70 isolated by B. P. Rosen [J. Bacteriol. 116, 1124-1129 (1973)]. Determination of its nucleotide sequence revealed a deletion of 21 base pairs between nucleotide residues 64 and 84, resulting in a deletion of seven amino acid residues (LysAlaMetGluMetValAla) from the amino-terminal portion. This deletion resulted in the loss of a hydrophobic domain of the subunit estimated by an analysis of its hydropathic character. Since F1 subunits are reported not to be assembled on the normal F0 portion of NR70, it is concluded that the hydrophobic domain deleted in the mutant subunit is important for assembly of the F1 portion. Introduction of a plasmid pNR70 carrying the mutant allele of NR70 into a wild-type strain gave no recombinants resistant to neomycin. This result suggested that the neomycin-resistant phenotype is not directly related to the defect in the gamma subunit of NR70
MH  - A
MH  - ACID
MH  - analysis
MH  - ATPase
MH  - BASE
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - F1
MH  - H+-ATPase
MH  - membrane
MH  - Membranes
MH  - mutant
MH  - P
MH  - RESIDUE
MH  - SUBUNIT
RP  - NOT IN FILE
NT  - UI - 85305726LA - engRN - 0 (Plasmids)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19851017IS - 0003-9861SB - IMCY - UNITED STATES
UR  - PM:2864018
SO  - Arch Biochem Biophys 1985 Sep ;241(2):364-370

652
UI  - 19808
AU  - Kandpal RP
AU  - Melese T
AU  - Stroop SD
AU  - Boyer PD
TI  - Mitochondrial F1-ATPase will bind and cleave ATP but only slowly release ADP after N,N'-dicyclohexylcarbodiimide or 7-chloro-4- nitrobenzo-2-oxa-1,3-diazole derivatization
AB  - The ATPase from the inner mitochondrial membrane is known to be inhibited by modification of one of the three catalytic subunits with N,N'-dicyclohexylcarbodiimide (DCCD) or 7-chloro-4-nitrobenzo-2-oxa-1,3- diazole. An experimental approach described in this paper shows that most of the residual ATPase activity observed after the usual DCCD or 7- chloro-4-nitrobenzo-2-oxa-1,3-diazole modification is due to the presence of unmodified enzyme, although the large fraction of modified enzyme retains a weak catalytic activity. This weak catalytic activity can be stimulated by methanol or dimethyl sulfoxide. When the modified enzymes are exposed to Mg2+ and [3H]ATP, about equal amounts of [3H]ATP and [3H]ADP appear at catalytic sites. The turnover rate for these enzymes is less than 1/1000 that of the native enzyme when it is calculated from the rate at which the enzyme becomes labeled at the catalytic sites with [3H]ATP and [3H]ADP during steady state hydrolysis. In addition, a higher ATP concentration is required for steady state turnover and, after ATP binding, the principal rate- limiting step is the capacity of the derivatized enzyme to undergo the binding changes necessary for the release of ADP and Pi. When the modified enzymes are not hydrolyzing ATP, they convert to form(s) that show a distinct lag in the replacement of bound nucleotides at catalytic sites. The replacement of bound nucleotides is still promoted by MgATP, even though the enzymes have been converted to sluggish forms. Contrary to a recent suggestion based on the study of the DCCD- modified enzyme (Soong, K.S., and Wang, J.H. (1984) Biochemistry 23, 136-141), our data provide evidence for the existence of catalytic cooperatively between at least two alternating sites in the modified enzyme and are consistent with continued sequential participation of all three sites
MH  - A
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - ADP
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BINDING
MH  - Biochemistry
MH  - BOUND NUCLEOTIDES
MH  - Dicyclohexylcarbodiimide
MH  - Enzymes
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - H(+)-Transporting ATP Synthase
MH  - Hydrolysis
MH  - Magnesium
MH  - membrane
MH  - Methanol
MH  - MITOCHONDRIAL F1-ATPASE
MH  - Nucleotides
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 85182704LA - engRN - 0 (Carbodiimides)RN - 0 (Oxadiazoles)RN - 10199-89-0 (4-Chloro-7-nitrobenzofurazan)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 67-56-1 (Methanol)RN - 67-68-5 (Dimethyl Sulfoxide)RN - 7439-95-4 (Magnesium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM 11094/GM/NIGMSDA - 19850613IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:2859288
SO  - J Biol Chem 1985 May 10 ;260(9):5542-5547

653
UI  - 906
AU  - Kihara H
TI  - [Physiological time and diffusion process]
MH  - Adenosine Triphosphate
MH  - Diffusion
MH  - Energy Metabolism
MH  - Enzyme Activation
MH  - Evolution
MH  - metabolism
MH  - Time
RP  - NOT IN FILE
SO  - Tanpakushitsu Kakusan Koso 1985 Sep ;(28):37-44

654
UI  - 21283
AU  - Kornyshev AA
TI  - Nonlocal Electrostatics of Solvation
MH  - SOLVATION
T2  - The Chemical Physics of Solvation
A2  - Dogonadze RR
A2  - Kalman E
A2  - Kornyshev AA
A2  - Ulstrup J
PB  - Amsterdam: Elsevier
RP  - IN FILE
M2  - Part A
SO  -  1985  ;(3):77-118

655
UI  - 21097
AU  - Krulwich TA
AU  - Agus R
AU  - Schneier M
AU  - Guffanti AA
TI  - Buffering capacity of bacilli that grow at different pH ranges
AB  - Cytoplasmic buffering capacities and buffering by whole cells were examined in six bacterial species: Bacillus acidocaldarius, Bacillus stearothermophilus, Escherichia coli, Bacillus subtilis, Bacillus alcalophilus, and Bacillus firmus RAB. Acid-base titrations were conducted on whole cells and cells permeabilized with Triton X-100 or n- butanol. In all of the species examined, the buffering capacity of intact cells was generally a significant proportion of the total buffering capacity, but the magnitude of the buffering capacity varied from species to species. Over the entire range of pH values from 4 to 9.5, B. subtilis exhibited a cytoplasmic buffering capacity that was much higher than that of B. stearothermophilus, B. acidocaldarius, or E. coli. The latter three species had comparable cytoplasmic buffering capacities at pH 4 to 9.5, as long as optimal conditions for cell permeabilization were employed. All of the nonalkalophiles exhibited a decrease in cytoplasmic buffering capacity as the external pH increased from pH 5 to 7. At alkaline pH values, the two thermophiles in the study had particularly low cytoplasmic buffering capacities, and the two alkalophilic bacteria had appreciably higher cytoplasmic buffering capacities than any of the other species studied. Cytoplasmic buffering capacities as high as 1,100 nmol of H+ per pH unit per mg of protein were observed in alkalophilic B. firmus RAB. Since previous studies have shown that immediate cytoplasmic alkalinization occurs upon loss of the active mechanisms for pH homeostasis in the alkalophiles, the very high buffering capacities apparently offer no global protection of internal pH. Perhaps, the high buffering capacities reflect protective mechanisms for specific macromolecules or process rather than part of the mechanisms for bulk pH homeostasis
MH  - A
MH  - ACTIVE
MH  - Bacillus
MH  - Bacteria
MH  - buffer
MH  - Buffers
MH  - Cells
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - H+
MH  - Homeostasis
MH  - mechanism
MH  - MECHANISMS
MH  - pH
MH  - protein
RP  - NOT IN FILE
NT  - UI - 85182501LA - engRN - 0 (Buffers)PT - Journal ArticleDA - 19850613IS - 0021-9193SB - IMCY - UNITED STATES
UR  - PM:3886633
SO  - J Bacteriol 1985 May ;162(2):768-772

656
UI  - 581
AU  - Matsuno-Yagi A
AU  - Hatefi Y
TI  - Studies on the mechanism of oxidative phosphorylation. Catalytic site cooperativity in ATP synthesis
AB  - Oxidative phosphorylation catalyzed by bovine heart submitochondrial particles appears to exhibit negative cooperativity with respect to [ADP] and positive cooperativity in catalysis. Eadie-Hofstee plots (v/[S]versus v) of the kinetics of oxidative phosphorylation at the variable ADP concentration range of 1-1200 microM were curvilinear and could be analyzed for two apparent KmADP values differing by one order of magnitude, and two apparent Vmax values. The KmADP values with either NADH or succinate as the respiratory substrate were in the ranges of 10 and 100 microM, and the Vmax values in nmol of ATP formed X min-1 (mg of protein)-1 were, respectively, 500 and 1840 when NADH was the oxidizable substrate, and 550 and 100 when succinate was the energy source. Site-site cooperativity of the ATP synthase, which is a central feature of current theories for the mechanism of oxidative phosphorylation, has been well-documented for ATP hydrolysis by isolated F1-ATPase, but never before demonstrated for mitochondrial ATP synthesis
RP  - NOT IN FILE
NT  - UI - 86033934LA - engRN - 0 (Phosphorus Radioisotopes)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)PT - Journal ArticleID - AM08126/AM/NIADDKDA - 19851220IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:4055778
SO  - J Biol Chem 1985 Nov 25 ;260(27):11424-11427

657
UI  - 582
AU  - Matsuno-Yagi A
AU  - Yagi T
AU  - Hatefi Y
TI  - Studies on the mechanism of oxidative phosphorylation: effects of specific F0 modifiers on ligand-induced conformation changes of F1
AB  - Aurovertin is a fluorescent antibiotic that binds to the catalytic beta subunits of the mitochondrial F1-ATPase and inhibits ATP synthesis and hydrolysis. ATP, ADP, and membrane energization in submitochondrial particles (SMP) alter the fluorescence of F1-bound aurovertin. These fluorescence changes are considered to be in response to the conformation changes of F1-ATPase. This paper shows that the ATP- induced fluorescence change of aurovertin bound to SMP or complex V (purified ATP synthase complex F0-F1) is inhibited when these preparations are pretreated with oligomycin or N,N'- dicyclohexylcarbodiimide (DCCD). This inhibition is not seen with isolated F1-ATPase. These and other results have suggested that modifications of the DCCD-binding protein in the membrane sector (F0) of the ATP synthase complex are communicated to F1, thereby altering the binding characteristics of ATP to the beta subunits. By analogy, it is proposed that modifications (e.g., protonation/deprotonation) of the DCCD-binding protein effected by protonic energy alter the conformation of F1 and bring about the substrate/product binding changes that appear to be essential features of the mechanism and regulation of oxidative phosphorylation
RP  - NOT IN FILE
NT  - UI - 86067942LA - engRN - 0 (Aurovertins)RN - 0 (Ligands)RN - 0 (Oligomycins)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - AM08126/AM/NIADDKDA - 19851224IS - 0027-8424SB - IMCY - UNITED STATESJC - PV3
UR  - PM:2866511
SO  - Proc Natl Acad Sci U S A 1985 Nov ;82(22):7550-7554

658
UI  - 19806
AU  - Melese T
AU  - Boyer PD
TI  - Derivatization of the catalytic subunits of the chloroplast ATPase by 2- azido-ATP and dicyclohexylcarbodiimide. Evidence for catalytically induced interchange of the subunits
AB  - Modifications of the catalytic beta subunits of the chloroplast ATPase (CF1-ATPase) are reported which support the proposal that all three subunits participate sequentially during catalysis. The beta subunits of the CF1-ATPase are sufficiently homogeneous to allow detection of their derivatization with dicyclohexylcarbodiimide (DCCD) or the substrate analog 2-azido-ATP by two-dimensional isoelectric focusing. Whether the DCCD reacts with the same beta subunit that tightly binds ATP has not been known. Our results show that when CF1-ATPase is covalently labeled with 2-azido-ATP followed by reaction with DCCD, different beta subunits are labeled. The DCCD labeling does not stop catalytic cooperativity of the CF1-ATPase and allows slow enzyme turnover. When the DCCD-modified enzyme catalyzes 2-azido-ATP cleavage and the enzyme with tightly bound nucleotide is photolyzed, both DCCD- modified and unmodified subunits are randomly labeled by the azido nucleotide. This result is as expected if during the catalytic cycle one beta subunit with unique properties is replaced by another subunit that gains these properties. The participation of all three subunits in the catalytic cycle is suggested by the apparent retention of catalytic cooperativity by the two remaining subunits after one subunit has already catalyzed 2-azido-ATP cleavage and been labeled
MH  - 2-AZIDO-ATP
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Azides
MH  - BETA-SUBUNIT
MH  - Catalysis
MH  - chloroplast
MH  - Dicyclohexylcarbodiimide
MH  - H(+)-Transporting ATP Synthase
MH  - Macromolecular Systems
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 86059402LA - engRN - 0 (Azides)RN - 0 (Carbodiimides)RN - 0 (Macromolecular Systems)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - 72884-75-4 (2-azidoadenosine 5'-triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM 11094/GM/NIGMSDA - 19860116IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:2866185
SO  - J Biol Chem 1985 Dec 15 ;260(29):15398-15401

659
UI  - 1001
AU  - Mitchell P
AU  - Mitchell R
AU  - Moody AJ
AU  - West IC
AU  - Baum H
AU  - Wrigglesworth JM
TI  - Chemiosmotic coupling in cytochrome oxidase. Possible protonmotive O loop and O cycle mechanisms
AB  - Using the principle of specific vectorial ligand conduction, we outline directly coupled protonmotive O loop and O cycle mechanisms of cytochrome oxidase action that are analogous to protonmotive Q loop and Q cycle mechanisms of QH2 dehydrogenase action. We discuss these directly coupled mechanisms in the light of available experimental knowledge, and suggest that they may stimulate useful new research initiatives designed to elucidate the osmochemistry of protonmotive oxygen reduction in cytochrome oxidase
MH  - Cytochrome-c Oxidase
MH  - Hydrogen
MH  - Ion Channels
MH  - Light
MH  - Oxygen
MH  - Protons
RP  - NOT IN FILE
NT  - UI - 85258119LA - engRN - 0 (Ion Channels)RN - 0 (Protons)RN - 11062-77-4 (Superoxides)RN - 7440-50-8 (Copper)RN - 7722-84-1 (Hydrogen Peroxide)RN - 7782-44-7 (Oxygen)RN - EC 1.9.3.1 (Cytochrome-c Oxidase)PT - Journal ArticlePT - ReviewDA - 19850925IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:2410291
SO  - FEBS Lett 1985 Aug 19 ;188(1):1-7

660
UI  - 1013
AU  - Mitchell P
TI  - Molecular mechanics of protonmotive F0F1 ATPases. Rolling well and turnstile hypothesis
AB  - The reversible protonmotive F0F1 ATPases perform the uniquely important function of balancing the forces, and interconverting the potential energies, of phosphoryl transfer and proton translocation. The molecular mechanics of the processes of ligand conduction catalysed by the F0F1 ATPases is therefore especially interesting. This paper summarises the main structural and functional knowledge of the F0F1 ATPases in the light of current mechanistic hypotheses, and suggests a new type of rotating subunit hypothesis, which is related to that recently developed for bacterial flagellar motors
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - H(+)-Transporting ATP Synthase
MH  - Ligands
MH  - Light
MH  - Macromolecular Systems
MH  - metabolism
MH  - Models,Chemical
MH  - Protein Conformation
MH  - Protons
MH  - Support,Non-U.S.Gov't
RP  - NOT IN FILE
SO  - FEBS Lett 1985 Mar 11 ;182(1):1-7

661
UI  - 1012
AU  - Mitchell P
AU  - Mitchell R
AU  - Moody AJ
AU  - West IC
AU  - Baum H
AU  - Wrigglesworth JM
TI  - Chemiosmotic coupling in cytochrome oxidase. Possible protonmotive O loop and O cycle mechanisms
AB  - Using the principle of specific vectorial ligand conduction, we outline directly coupled protonmotive O loop and O cycle mechanisms of cytochrome oxidase action that are analogous to protonmotive Q loop and Q cycle mechanisms of QH2 dehydrogenase action. We discuss these directly coupled mechanisms in the light of available experimental knowledge, and suggest that they may stimulate useful new research initiatives designed to elucidate the osmochemistry of protonmotive oxygen reduction in cytochrome oxidase
MH  - Biological Transport
MH  - Chemistry
MH  - Copper
MH  - Cytochrome-c Oxidase
MH  - Electron Transport
MH  - Hydrogen Peroxide
MH  - Hydrogen-Ion Concentration
MH  - Ion Channels
MH  - Light
MH  - Membrane Potentials
MH  - metabolism
MH  - Osmosis
MH  - Oxidation-Reduction
MH  - Oxygen
MH  - Protons
MH  - Superoxides
MH  - Support,Non-U.S.Gov't
RP  - NOT IN FILE
SO  - FEBS Lett 1985 Aug ;%19;188(1):1-7

662
UI  - 119
AU  - Mosher ME
AU  - White LK
AU  - Hermolin J
AU  - Fillingame RH
TI  - H+-ATPase of Escherichia coli. An uncE mutation impairing coupling between F1 and Fo but not Fo-mediated H+ translocation
AB  - The uncE114 mutation from Escherichia coli strain KI1 (Nieuwenhuis, F. J. R. M., Kanner, B. I., Gutnick, D. L., Postma, P. W., and Van Dam, K. (1973) Biochim. Biophys. Acta 325, 62-71) was characterized after transfer to a new genetic background. A defective H+-ATPase complex is formed in strains carrying the mutation. Based upon the genetic complementation pattern of other unc mutants by a lambda uncE114 transducing phage, and complementation of uncE114 recipients by an uncE+ plasmid (pCP35), the mutation was concluded to lie in the uncE gene. The uncE gene codes for the omega subunit ("dicyclohexylcarbodiimide binding protein") of the H+-ATPase complex. The mutation was defined by sequencing the mutant gene. The G----C transversion found results in a substitution of Glu for Gln at position 42 of the omega subunit in the Fo sector of the H+-ATPase. The substitution did not significantly impair H+ translocation by Fo or affect inhibition of H+ translocation by dicyclohexylcarbodiimide. Wild- type F1 was bound by uncE114 Fo with near normal affinity, but the functional coupling between F1 and Fo was disrupted. The uncoupling was indicated by an H+-leaky membrane, even when saturating levels of wild- type F1 were bound. Disassociation of F1 from Fo under conditions of assay did partially contribute to the H+ leakiness, but the major contributor to the high H+ conductance was Fo with bound F1. The F1 bound to uncE114 membranes exhibited normal ATPase activity, but ATP hydrolysis was uncoupled from H+ translocation and was resistant to inhibition by dicyclohexylcarbodiimide. The F1 isolated from the uncE114 mutant was modified with partial loss of coupling function. However, this modification did not account for the uncoupled properties of the mutant Fo described above, since these properties were retained after reconstitution of mutant membrane (Fo) with wild-type F1
RP  - NOT IN FILE
NT  - UI - 85182594LA - engRN - 0 (Succinates)RN - 110-15-6 (Succinic Acid)RN - 2001-95-8 (Valinomycin)RN - 53-84-9 (NAD)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 7440-09-7 (Potassium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - 5 T32 GM07215/GM/NIGMSID - GM-23105/GM/NIGMSDA - 19850531IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:2859283
SO  - J Biol Chem 1985 Apr 25 ;260(8):4807-4814

663
UI  - 19764
AU  - Nyren P
AU  - Lundin A
TI  - Enzymatic method for continuous monitoring of inorganic pyrophosphate synthesis
AB  - A sensitive method for the analysis of inorganic pyrophosphate (PPi) which utilizes the enzymes ATP sulfurylase and firefly luciferase is described. The assay is based on continuous monitoring of the ATP formed in the ATP sulfurylase reaction using purified firefly luciferase. The assay can be completed in less than 2 s and is not affected by inorganic phosphate. The method has been used for continuous monitoring of formation of PPi in Rhodospirillum rubrum chromatophores. The assay is extremely sensitive, the linear range of the assay being 1 X 10(-9) - 5 X 10(-7) M PPi. It is suitable for routine applications. It is also possible to use the method for determination of low amounts of adenosine 5'-phosphosulfate
MH  - A
MH  - Adenosine
MH  - analysis
MH  - atp
MH  - chromatophore
MH  - chromatophores
MH  - Diphosphates
MH  - Enzymes
MH  - INORGANIC-PHOSPHATE
MH  - method
MH  - Pyrophosphatases
MH  - Rhodospirillum
MH  - Rhodospirillum rubrum
MH  - synthesis
RP  - NOT IN FILE
NT  - UI - 86156700LA - engRN - 0 (Diphosphates)RN - EC 1.13.12.- (Luciferase)RN - EC 2.7.7.4 (Sulfate Adenylyltransferase)RN - EC 3.6.1.- (Pyrophosphatases)RN - EC 3.6.1.1 (inorganic pyrophosphatase)PT - Journal ArticleDA - 19860331IS - 0003-2697SB - IMCY - UNITED STATESJC - 4NK
UR  - PM:3006540
SO  - Anal Biochem 1985 Dec ;151(2):504-509

664
UI  - 393
AU  - Panchenko MV
AU  - Vinogradov AD
TI  - Interaction between the mitochondrial ATP synthetase and ATPase inhibitor protein. Active/inactive slow pH-dependent transitions of the inhibitor protein
AB  - The rate of mitochondrial ATPase inactivation by the naturally occurring inhibitor protein in the presence of saturating ATP and Mg2+ at pH 8.0 depends hyperbolically on the amount of inhibitor added; the upper limit of an apparent first-order constant for the inactivation process is 1.0(-1) at 25 degrees C. A dramatic difference in the inactivation rate is observed when the protein inhibitor is added to the same assay system from either acidic (pH 4.8) or alkaline (pH 8.2) solutions. The slow reversible transition of the inhibitor from its rapidly reacting 'acidic' form to the slow reacting 'alkaline' form occurs when the solution of the protein inhibitor is subjected to a pH- jump from 4.8 to 8.2 (t1/2 approximately 30s at 25 degrees C). The pH- profile of the inhibitor active/inactive equilibrium suggests that a group with pKa approximately 6.5 is involved in the transition. Treatment of the inhibitor protein with a histidine-specific reagent (e.g. diethyl pyrocarbonate) abolishes its inactivating effect on the ATPase activity. It is concluded that the protonation/deprotonation of the inhibitor protein followed by its slow conformational changes is the rate-limiting step in the inhibitor-ATP synthetase interaction
RP  - NOT IN FILE
NT  - UI - 85204352LA - engRN - 0 (ATPase inhibitory protein)RN - 0 (Proteins)RN - 1609-47-8 (Diethyl Pyrocarbonate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19850709IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:2860010
SO  - FEBS Lett 1985 May 20 ;184(2):226-230

665
UI  - 643
AU  - Penefsky HS
TI  - Mechanism of inhibition of mitochondrial adenosine triphosphatase by dicyclohexylcarbodiimide and oligomycin: relationship to ATP synthesis
AB  - Measurement of the rate of [gamma-32P]ATP binding (k1) and release (k- 1) from catalytic sites on submitochondrial particles permitted calculation of the affinity constant in catalytic sites (k1 = K1/k1-1) of 10(12) M-1. This value is the same as that determined previously for the solubilized ATPase (F1) from beef heart mitochondria. Treatment of submitochondrial particles with dicyclohexylcarbodiimide or oligomycin so as to cause about 90% inhibition of ATPase activity was accompanied by a decrease in the binding of [gamma-32P]ATP in high-affinity catalytic sites. Under the conditions of the experiment, it is expected that the inhibitors reacted not with the ATPase itself but with other proteins in the oligomycin-sensitive ATPase complex (F0-F1). It is proposed that dicyclohexylcarbodiimide and oligomycin inhibit ATPase activity by causing a conformational change in the F0 portion of the complex that is transmitted to F1, resulting in an impaired binding of substrate in catalytic sites. These observations of apparent conformational interactions between F0 and F1 on the mitochondrial membrane are relevant to the mechanism of the coupling device that links the energy store to ATP formation in oxidative phosphorylation. It is proposed that a change in the state of ionization of one or more charged amino acid residues in F0 results in a conformational change in F0 which, transmitted to F1, reversibly alters the catalytic sites and facilitates the release of product ATP
RP  - NOT IN FILE
NT  - UI - 85166177LA - engRN - 0 (Carbodiimides)RN - 0 (Oligomycins)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM 21737/GM/NIGMSDA - 19850503IS - 0027-8424SB - IMCY - UNITED STATESJC - PV3
UR  - PM:2858849
SO  - Proc Natl Acad Sci U S A 1985 Mar ;82(6):1589-1593

666
UI  - 642
AU  - Penefsky HS
TI  - Reaction mechanism of the membrane-bound ATPase of submitochondrial particles from beef heart
AB  - Submitochondrial particles from beef heart, washed with dilute solutions of KCl so as to activate the latent, membrane-bound ATPase, F1, may be used to study single site catalysis by the enzyme. [gamma- 32P]ATP, incubated with a molar excess of catalytic sites, a condition which favors binding of substrate in only a single catalytic site on the enzyme, is hydrolyzed via a four-step reaction mechanism. The mechanism includes binding in a high affinity catalytic site, Ka = 10(12)M-1, a hydrolytic step for which the equilibrium constant is near unity, and two product release steps in which Pi dissociates from catalytic sites about 10 times more rapidly than ADP. Catalysis by the membrane-bound ATPase also is characterized by a 10(6)-fold acceleration in the rate of net hydrolysis of [gamma-32P]ATP, bound in the high affinity catalytic site, that occurs when substrate is made available to additional catalytic sites on the enzyme. These aspects of the reaction mechanism of the ATPase of submitochondrial particles closely parallel the reaction mechanism determined for solubilized, homogeneous F1 (Grubmeyer, C., Cross, R. L., and Penefsky, H. S. (1982) J. Biol. Chem. 257, 12092-12100). The finding that removal of the enzyme from the membrane does not significantly alter the properties of single site catalysis lends support to models of ATP synthesis in oxidative phosphorylation, catalyzed by membrane-bound F1, that have been based on the study of the soluble enzyme
RP  - NOT IN FILE
NT  - UI - 86033840LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleID - GM 21731/GM/NIGMSDA - 19851205IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:2932441
SO  - J Biol Chem 1985 Nov 5 ;260(25):13728-13734

667
UI  - 641
AU  - Penefsky HS
TI  - Energy-dependent dissociation of ATP from high affinity catalytic sites of beef heart mitochondrial adenosine triphosphatase
AB  - Incubation of [gamma-32P]ATP with a molar excess of the membrane-bound form of mitochondrial ATPase (F1) results in binding of the bulk of the radioactive nucleotide in high affinity catalytic sites (Ka = 10(12) M- 1). Subsequent initiation of respiration by addition of succinate or NADH is accompanied by a profound decrease in the affinity for ATP. About one-third of the bound radioactive ATP appears to dissociate, that is, the [gamma-32P]ATP becomes accessible to hexokinase. The NADH- stimulated dissociation of [gamma-32P]ATP is energy-dependent since the stimulation is inhibited by uncouplers of oxidative phosphorylation and is prevented by respiratory chain inhibitors. The rate of the energy- dependent dissociation of ATP that occurs in the presence of NADH, ADP, and Pi is commensurate with the measured initial rate of ATP synthesis in NADH-supported oxidative phosphorylation catalyzed by the same submitochondrial particles. Thus, the rate of dissociation of ATP from the high affinity catalytic site of submitochondrial particles meets the criterion of kinetic competency under the conditions of oxidative phosphorylation. These experiments provide evidence in support of the argument that energy conserved during the oxidation of substrates by the respiratory chain can be utilized to reduce the very tight binding of product ATP in high affinity catalytic sites and to promote dissociation of the nucleotide
RP  - NOT IN FILE
NT  - UI - 86033841LA - engRN - 0 (Phosphorus Radioisotopes)RN - 53-84-9 (NAD)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleID - GM 21731/GM/NIGMSDA - 19851205IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:2932442
SO  - J Biol Chem 1985 Nov 5 ;260(25):13735-13741

668
UI  - 469
AU  - Perlin DS
AU  - Latchney LR
AU  - Senior AE
TI  - Inhibition of Escherichia coli H+-ATPase by venturicidin, oligomycin and ossamycin
AB  - The antibiotics venturicidin, oligomycin and ossamycin were investigated as potential inhibitors of the Escherichia coli H+-ATPase. It was found that venturicidin strongly inhibited ATP-driven proton transport and ATP hydrolysis, while oligomycin weakly inhibited these functions. Inhibition of the H+-ATPase by venturicidin and oligomycin was correlated with inhibition of F0-mediate proton transport. Both inhibitors were found to interfere with the covalent reaction between dicyclohexyl[14C]carbodiimide and the F0 subunit c (uncE protein). Ossamycin had no direct inhibitory effect on E. coli F0 or F1; rather, it was found to uncouple ATP hydrolysis from proton transport
RP  - NOT IN FILE
NT  - UI - 85199890LA - engRN - 0 (Aminoglycosides)RN - 0 (Antibiotics)RN - 0 (Lactones)RN - 0 (Oligomycins)RN - 0 (Uncoupling Agents)RN - 0 (Venturicidins)RN - 11015-84-2 (ossamycin)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSID - GM29805/GM/NIGMSDA - 19850702IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:2859888
SO  - Biochim Biophys Acta 1985 May 31 ;807(3):238-244

669
UI  - 471
AU  - Perlin DS
AU  - Senior AE
TI  - Functional effects and cross-reactivity of antibody to purified subunit b (uncF protein) of Escherichia coli proton-ATPase
AB  - Subunit b (uncF protein) of the proton-ATPase (F1F0) of Escherichia coli was purified from membranes of strain AN1460 (unc+). Antibody to purified subunit b was raised in rabbits. It reacted with F1-depleted membranes and blocked F1 binding. Bound antibody had no effect on proton transport through F0. F1-Depleted membranes competed with purified subunit b for antibody in an enzyme-linked immunosorbent assay. F1-Depleted membranes which had been pretreated with trypsin or preincubated with saturating amounts of soluble F1 competed poorly with purified subunit b for antibody. The antibody to subunit b was used to further evaluate the trypsin-cleavage data previously reported [D. S. Perlin, D. N. Cox, and A. E. Senior (1983) J. Biol. Chem. 258, 9793- 9800]. The results indicated that trypsin proteolysis of F1-depleted membranes resulted in the transient appearance of three fragments of subunit b (Mr = 16,400, 15,700, and 15,500) that remained tightly bound to the membrane. A water-soluble fragment (Mr 14,800), previously thought to be derived from subunit b, was not detected by the antibody. The antibody to subunit b did not cross-react with any subunit of mitochondrial, chloroplast, or other bacterial proton-ATPase, or with the proton-ATPase of clathrin-coated vesicles, plant microsomal membranes, or Neurospora crassa plasma membranes
RP  - NOT IN FILE
NT  - UI - 85120882LA - engRN - 0 (Peptide Fragments)RN - 9004-70-0 (Collodion)RN - EC 3.4.21.4 (Trypsin)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM-29805/GM/NIGMSDA - 19850321IS - 0003-9861SB - IMCY - UNITED STATESJC - 6SK
UR  - PM:2857549
SO  - Arch Biochem Biophys 1985 Feb 1 ;236(2):603-611

670
UI  - 880
AU  - Reid D
TI  - ATP-adenosine triphosphate: the answer to power
MH  - Adenosine Triphosphate
MH  - Energy Metabolism
MH  - Human
MH  - metabolism
MH  - Nad
MH  - Oxidative Phosphorylation
RP  - NOT IN FILE
SO  - Can Oper Room Nurs J 1985 Sep ;3(4):37-41

671
UI  - 1168
AU  - Schmidt G
AU  - Graber P
TI  - The rate of ATP synthesis by reconstituted CF0F1 liposomes
MH  - atp
MH  - ATP synthesis
MH  - CF0F1
MH  - liposome
MH  - Liposomes
MH  - RECONSTITUTED CF0F1
MH  - synthesis
RP  - ON REQUEST (03/18/92)
SO  - Biochim Biophys Acta 1985  ;808():46-51

672
UI  - 20967
AU  - Schneider E
AU  - Altendorf K
TI  - All three subunits are required for the reconstitution of an active proton channel (F0) of Escherichia coli ATP synthase (F1F0)
AB  - The membrane-integrated, proton-translocating F0 portion of the ATP synthase (F1F0) from Escherichia coli is built up from three kinds of subunits a, b and c with the proposed stoichiometry of 1:2:10 +/- 1. We have dissociated the F0 complex by treatment with trichloroacetate (3 M) at pH 8.0, in the presence of deoxycholate (1%) and N-tetradecyl-N, N-dimethyl-3-ammonio-1-propanesulfonate (Zwittergent 3-14, 5%). The subunits were separated by gel filtration with trichloroacetate (1 M) included in the elution buffer. The homogeneity of the fractions was checked by rechromatography and SDS-gel electrophoresis. After integration into phospholipid vesicles each subunit alone as well as all possible combinations were tested for H+ translocating activity and binding of F1. A functional H+ channel could only be reconstituted by the combination a1b2c10 which corresponds to that of native F0
MH  - A
MH  - ACTIVE
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BINDING
MH  - buffer
MH  - COLI ATP SYNTHASE
MH  - COMPLEX
MH  - Detergents
MH  - Electrophoresis
MH  - England
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - F1
MH  - H+
MH  - ion
MH  - Ion Channels
MH  - Liposomes
MH  - M
MH  - Macromolecular Systems
MH  - pH
MH  - proton
MH  - Protons
MH  - reconstitution
MH  - SUBUNIT
MH  - SYNTHASE
MH  - SYSTEM
MH  - SYSTEMS
RP  - NOT IN FILE
NT  - UI - 85257483LA - engRN - 0 (Detergents)RN - 0 (Ion Channels)RN - 0 (Liposomes)RN - 0 (Macromolecular Systems)RN - 0 (Protons)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19850904IS - 0261-4189SB - IMCY - ENGLAND
UR  - PM:2410260
SO  - EMBO J 1985 Feb ;4(2):515-518

673
UI  - 20966
AU  - Schneider E
AU  - Altendorf K
TI  - Modification of subunit b of the F0 complex from Escherichia coli ATP synthase by a hydrophobic maleimide and its effects on F0 functions
AB  - Purified F0 from Escherichia coli ATP synthase was labelled with N-(7- dimethylamino-4-methyl-coumarinyl)-maleimide (DACM), a hydrophobic reagent which forms a stable, strongly fluorescent adduct with SH groups. Sodium dodecyl sulfate gel electrophoresis clearly demonstrated that subunit b was exclusively labelled, most likely at Cys-21, the only cysteine residue in E. coli F0. The amount of two molecules of DACM bound per F0, which was calculated from the absorption spectrum at 380 nm, is in full agreement with the postulated stoichiometry of two copies of subunit b/F0 complex. Thus the label provides a useful tool for simply detecting subunit b in protein chemical studies. DACM- labelled F0 was incorporated into liposomes and assayed for H+ translocating activity and its capacity to bind purified F1. Whereas the initial rate of H+ uptake was inhibited about 40% the reconstitution of a dicyclohexylcarbodiimide-sensitive F1F0 ATPase activity was completely unaffected. In a second set of experiments we reconstituted an F0 complex from DACM-labelled purified subunit b and an ac complex. In contrast to the results obtained with intact, DACM- labelled F0, both H+ translocating activity and F1 binding capacity were greatly reduced. Our data indicate that cysteine-21, probably together with other amino acids, is involved in maintaining a proper interaction of the hydrophobic N-terminal region of subunit b with the ac complex. This interplay seems to be a prerequisite for at least the in vitro assembly of a functional F0 complex
MH  - A
MH  - absorption
MH  - ACID
MH  - Amino Acids
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BINDING
MH  - COLI ATP SYNTHASE
MH  - COMPLEX
MH  - Cysteine
MH  - DYE
MH  - dyes
MH  - Electrophoresis
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - F1
MH  - Fluorescent Dyes
MH  - function
MH  - H+
MH  - In Vitro
MH  - Liposomes
MH  - protein
MH  - reconstitution
MH  - RESIDUE
MH  - Sodium
MH  - spectra
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 86055842LA - engRN - 0 (Amino Acids)RN - 0 (Fluorescent Dyes)RN - 0 (Liposomes)RN - 0 (Maleimides)RN - 0 (Sulfhydryl Compounds)RN - 55145-14-7 (N-(7-dimethylamino-4-methylcoumarinyl)maleimide)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19851227IS - 0014-2956SB - IMCY - GERMANY, WEST
UR  - PM:2866095
SO  - Eur J Biochem 1985 Nov 15 ;153(1):105-109

674
UI  - 21111
AU  - Seren S
AU  - Caporin G
AU  - Galiazzo F
AU  - Lippe G
AU  - Ferguson SJ
AU  - Sorgato MC
TI  - Current-voltage relationships for proton flow through the F0 sector of the ATP-synthase, carbonylcyanide-p-trifluoromethoxyphenylhydrazone or leak pathways in submitochondrial particles
AB  - Respiring submitochondrial particles from which the F1 sector of ATP- synthase was displaced generated a membrane potential in the range of 115-140 mV. Addition of oligomycin raised the membrane potential by approximately 40 mV. The lower membrane potential in particles with F1 displaced is attributed to partial dissipation of the proton electrochemical gradient as a consequence of proton flow through the open proton channels provided by the F0 sectors of the ATP-synthase. The characteristics of proton flow through the open F0 channels were studied by varying the rate of electron transport-driven proton translocation which permitted the establishment of a range of steady- state membrane potentials. Open F0 channels appeared to have a gated response to the membrane potential such that they were inoperative when the potential fell below approximately 110 mV. The membrane potential was measured as a function of respiratory rate in intact Mg-ATP submitochondrial particles that had been treated with low concentrations of the protonophore carbonylcyanide-p- trifluoromethoxyphenylhydrazone. In general a linear dependence of membrane potential upon respiratory rate was observed except at the lowest concentrations of protonophore and highest respiratory rates, presumably because the effect of the protonophore was then offset by an increased rate of proton translocation driven by the respiratory chain. The effect of increasing concentrations of carbonylcyanide-p- trifluoromethoxyphenylhydrazone on the membrane potential of respiring submitochondrial particles was studied. It was found that equal amounts of the protonophore lowered the membrane potential to a lesser extent at lower values of the membrane potential. Treatment of Mg-ATP submitochondrial particles with oligomycin slightly increased (by approximately 10 mV) the size of the respiration-dependent membrane potential, but did not alter the profile of membrane potential as a function of succinate oxidation rate. The latter was controlled by titration with malonate. This result indicates that the F0 sector of the ATP-synthase does not significantly contribute to leak pathways in intact submitochondrial particles
MH  - A
MH  - ACID
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone
MH  - DEPENDENCE
MH  - Edetic Acid
MH  - electron
MH  - F0
MH  - F1
MH  - function
MH  - membrane
MH  - Membrane Potential
MH  - Membrane Potentials
MH  - Nitriles
MH  - Oligomycins
MH  - proton
MH  - Proton-Translocating ATPases
MH  - protonophore
MH  - Protons
MH  - Submitochondrial Particles
MH  - succinate
MH  - SYNTHASE
MH  - translocation
RP  - NOT IN FILE
NT  - UI - 86030278LA - engRN - 0 (Nitriles)RN - 0 (Oligomycins)RN - 0 (Protons)RN - 370-86-5 (Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone)RN - 56-65-5 (Adenosine Triphosphate)RN - 60-00-4 (Edetic Acid)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19851125IS - 0014-2956SB - IMCY - GERMANY, WEST
UR  - PM:2865136
SO  - Eur J Biochem 1985 Oct 15 ;152(2):373-379

675
UI  - 959
AU  - Skulachev VP
TI  - [The sodium cycle--a new type of bacterial energetics]
AB  - The literature data and experimental results of the author's laboratory on the role of Na+ in bacterial energetics are reviewed. It was shown that certain bacterial species utilize the transmembrane difference of Na+ electrochemical potentials (delta mu Na+) as a convertible membrane-linked form of energy. The membranes of such bacteria were found to contain delta mu Na+ generators (e. g., decarboxylases of some carboxylic acids of NADH-menaquinone reductase). It was shown that delta mu Na+ formed by these generators may support all the three main types of work of the bacterial cell, i. e., chemical (ATP synthesis), osmotic (substrate accumulation) and mechanical (motility)
MH  - Adenosine Triphosphate
MH  - Bacteria
MH  - Biological Transport
MH  - biosynthesis
MH  - Energy Metabolism
MH  - Hydrogen
MH  - Membranes
MH  - metabolism
MH  - Osmolar Concentration
MH  - Protons
MH  - Sodium
RP  - NOT IN FILE
SO  - Biokhimiia 1985 Feb ;50(2):179-183

676
UI  - 468
AU  - Smith RA
AU  - Latchney LR
AU  - Senior AE
TI  - Tight divalent metal binding to Escherichia coli F1- adenosinetriphosphatase. Complete substitution of intrinsic magnesium by manganese or cobalt and studies of metal binding sites
AB  - Tight divalent metal binding sites in Escherichia coli F1- adenosinetriphosphatase (F1-ATPase) were studied. Native enzyme contained two Mg per F1, confirming previous results. All of the Mg may be replaced by Co or Mn using a dissociation-repolymerization procedure. The substituted enzymes are homogeneous and contain two Mn per F1 or two Co per F1. They are fully active as ATPases, they rebind to F1-depleted membranes, and they catalyze ATP-driven proton pumping. N,N'-Dicyclohexylcarbodiimide-(DCCD) inactivated F1 retains all the intrinsic tightly bound Mg. Evidence is presented that DCCD affects at least two beta subunits in E. coli F1, and therefore, the tightly bound metals appear not to be bound at the DCCD-reactive glutamate residue on the beta subunit. However, the nature of the tightly bound metal (Mg, Mn, or Co) as well as the presence of added (2 mM) MgSO4, MnSO4, or CoSO4 affected the rate of DCCD inactivation, showing that metal binding changes the beta-subunit conformation. Isolated F1 alpha subunit bound Mg, Mn, or Co stoichiometrically and independently of ATP binding. Isolated F1 beta subunit bound only small amounts of Mg, and no Co or Mn. Therefore, it is possible, although not conclusively shown, that the alpha subunit is the site of tight metal binding in the intact F1
RP  - NOT IN FILE
NT  - UI - 86026307LA - engRN - 0 (Macromolecular Systems)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 7439-95-4 (Magnesium)RN - 7439-96-5 (Manganese)RN - 7440-48-4 (Cobalt)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSDA - 19851205IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:2864956
SO  - Biochemistry 1985 Jul 30 ;24(16):4490-4494

677
UI  - 21112
AU  - Sutton R
AU  - Ferguson SJ
TI  - Identification of an essential beta chain lysine residue from bovine heart mitochondrial ATPase specifically modified with nitrobenzofurazan
AB  - A tetrapetide containing an essential lysine residue chemically modified with the nitrobenzofurazan group has been purified from bovine heart mitochondrial ATPase. The composition of the peptide indicates that this lysine is residue 401 in the sequence of a beta chain. The modification was achieved by incubation at pH 9 of ATPase that had been previously labelled on a single essential tyrosine residue by reaction of the enzyme with 4-chloro-7-nitrobenzofurazan. The specific transfer of the nitrobenzofurazan group from the tyrosine residue to a particular lysine residue is consistent with the previously demonstrated intramolecular character of this transfer reaction
MH  - A
MH  - ATPase
MH  - BETA
MH  - HEART MITOCHONDRIAL ATPASE
MH  - Peptide Fragments
MH  - pH
MH  - Proton-Translocating ATPases
MH  - RESIDUE
MH  - TRANSFER
RP  - NOT IN FILE
NT  - UI - 85102027LA - engRN - 0 (Oxadiazoles)RN - 0 (Peptide Fragments)RN - 10199-89-0 (4-Chloro-7-nitrobenzofurazan)RN - 56-87-1 (Lysine)RN - EC 3.4.23.1 (Pepsin A)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19850225IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:2857132
SO  - FEBS Lett 1985 Jan 7 ;179(2):283-288

678
UI  - 20899
AU  - Takeda K
AU  - Miki J
AU  - Kanazawa H
AU  - Tsuchiya T
AU  - Futai M
TI  - Change of inhibitor sensitivities of Escherichia coli F1-ATPase due to a mutational substitution of Phe for Ser at residue 174 of the beta subunit
AB  - The F1-ATPase from the uncD11 mutant of E. coli (Kanazawa, H., Horiuchi, Y., Takagi, M., Ishino, Y., & Futai, M. (1980) J. Biochem. 88, 695-703), showed different enzymological properties from the wild- type enzyme. The mutant F1-ATPase had biphasic kinetics and essentially the same Km values as the wild-type enzyme, although its Vmax values were lower. The mutant enzyme showed altered sensitivities to dicyclohexylcarbodiimide (DCCD), azide and quercetin; it was less sensitive than the wild-type to quercetin and DCCD, and its Mg2+- dependent ATPase activity was slightly more resistant to azide than that of the wild-type, whereas its Ca2+-dependent activity was more sensitive. On the other hand, the mutant and wild-type F1 were inhibited equally by 4-chloro-7-nitro-2,1,3-benzoxadiazole (NBD-Cl). The fact that the Mg2+- and Ca2+-dependent F1-ATPase activities of the wild-type and mutant responded differently to quercetin and azide suggested that their mechanisms of action were different. Previous studies (Noumi, T., Mosher, M.E., Natori, S., Futai, M., & Kanazawa, H. (1984) J. Biol. Chem. 259, 10071-10075) indicated that Ser is replaced by Phe at residue 174 of the beta subunit of the mutant. Thus the Ser residue or its neighboring area(s) may constitute the binding site of DCCD, quercetin and azide
MH  - A
MH  - ATPase
MH  - Azides
MH  - BETA-SUBUNIT
MH  - BINDING
MH  - COLI F1 ATPASE
MH  - Dicyclohexylcarbodiimide
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - Kinetics
MH  - M
MH  - Macromolecular Systems
MH  - mechanism
MH  - MECHANISMS
MH  - mutant
MH  - RESIDUE
MH  - Site
MH  - SUBUNIT
MH  - SYSTEM
MH  - SYSTEMS
RP  - NOT IN FILE
NT  - UI - 85289131LA - engRN - 0 (Azides)RN - 0 (Macromolecular Systems)RN - 117-39-5 (Quercetin)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19851009IS - 0021-924XSB - IMCY - JAPAN
UR  - PM:2863263
SO  - J Biochem (Tokyo ) 1985 May ;97(5):1401-1407

679
UI  - 190
AU  - Tommasino M
AU  - Capaldi RA
TI  - Effect of dicyclohexylcarbodiimide on unisite and multisite catalytic activities of the adenosinetriphosphatase of Escherichia coli
AB  - The inhibitory effect of dicyclohexylcarbodiimide (DCCD) on the activity of the adenosine-triphosphatase of Escherichia coli (ECF1) has been examined in detail. DCCD reacted with ECF1 predominantly in beta subunits with a maximum of 2 mol of reagent per mole of ECF1 being incorporated in these subunits. Ninety-five percent inhibition of steady-state or multistate ATPase activity required incorporation of 1 mol of DCCD per mole of enzyme into beta subunits. Seventy-five percent inhibition of the initial rate of unisite catalysis was only obtained after incorporation of 2 mol of DCCD per mole of ECF1 into beta subunits. Analyses of the kinetics of unisite catalysis and nucleotide binding experiments both indicate that DCCD binds outside the substrate ATP binding site. Inhibition by this reagent appears to be due in part to an effect on the catalytic sites but mainly to the blocking of cooperativity between these sites
RP  - NOT IN FILE
NT  - UI - 86026233LA - engRN - 0 (Carbodiimides)RN - 0 (Carbon Radioisotopes)RN - 25612-73-1 (Adenylyl Imidodiphosphate)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - HL 24526/HL/NHLBIDA - 19851205IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:2864953
SO  - Biochemistry 1985 Jul 16 ;24(15):3972-3976

680
UI  - 9922
AU  - Wainio WW
TI  - An assessment of the chemiosmotic hypothesis of mitochondrial energy transduction.
AB  - It is argued that a proton concentration difference and/or a membrane potential is not the form into which the free energy of the oxidation-reduction reactions of the mitochondrial respiratory chain is first transduced. It is suggested that the search for a chemical intermediate should be continued in spite of the conclusion by some investigators that the chemical hypothesis is untenable. It is asked whether pH changes when measured in solutions containing mitochondria can be interpreted as evidence for H+ movements, also, whether there is a continuous, renewable and stable electrochemical proton concentration difference (delta mu H+) across the mitochondrial membrane, and whether in fact the delta mu H+ is a necessary intermediate in the synthesis of ATP. The four postulates of Mitchell's chemiosmotic hypothesis of energy transduction are discussed point by point. It is agreed that "The systems are plugged through a topologically closed insulating membrane," which probably is not "a nonaqueous osmotic barrier," and which probably does not have an unusually "low permeability to solutes and to H+ and OH- in particular" when compared with other membranes. There is disagreement with the statement that "Respiratory and photoredox systems are chemiosmotic membrane-located protonmotive chains" in that it is suggested by others that chemiosmosis is chemically nonexistent and that thermodynamically it would lack control. The subsequent statement, "having a characteristic----H+/2 epsilon- stoichiometry," is rendered uncertain by the experimental findings of values greater than 2H+/2 epsilon-/site and probably as large as 4H+/2 epsilon-/site. The proposal that "The synthetase is a chemiosmotic membrane-located reversible motive ATPase" requires the assumption that the ATP synthetase is the same enzyme as the ATPase, but functioning in the reverse direction. It is considered possible that there are two enzymes in the multi-subunit ATPase complex: one the hydrolase, and the other the synthetase. The further proposal, "having characteristic----H+/P stoichiometry" requires that the ratio be 2 according to Mitchell. However, values of 3, as well as larger values, have been reported by others, which introduces a large element of uncertainty. There is no disagreement with the statement that "There are proton-linked (or hydroxyl ion-linked) solute porter systems for osmotic stabilization and metabolite transport." In fact, this may be the principal reason for having proton efflux or "proton-pumping.''(ABSTRACT TRUNCATED AT 400 WORDS)
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - atp
MH  - ATPase
MH  - COMPLEX
MH  - Electron Transport
MH  - Energy Transfer
MH  - Enzymes
MH  - H(+)-Transporting ATP Synthase
MH  - H+
MH  - Hydrogen-Ion Concentration
MH  - Membranes
MH  - metabolism
MH  - Mitochondria
MH  - Models,Biological
MH  - Movement
MH  - Oxidation-Reduction
MH  - Permeability
MH  - proton
MH  - Solutions
MH  - Support,Non-U.S.Gov't
MH  - Thermodynamics
RP  - NOT IN FILE
SO  - Int Rev Cytol 1985  ;96:29-50():29-50

681
UI  - 367
AU  - Walker JE
AU  - Fearnley IM
AU  - Gay NJ
AU  - Gibson BW
AU  - Northrop FD
AU  - Powell SJ
AU  - Runswick MJ
AU  - Saraste M
AU  - Tybulewicz VL
TI  - Primary structure and subunit stoichiometry of F1-ATPase from bovine mitochondria
AB  - The enzyme complex F1-ATPase has been isolated from bovine heart mitochondria by gel filtration of the enzyme released by chloroform from sub-mitochondrial particles. The five individual subunits alpha, beta, gamma, delta and epsilon that comprise the complex have been purified from it, and their amino acid sequences determined almost entirely by direct protein sequence analysis. A single overlap in the gamma-subunit was obtained by DNA sequence analysis of a complementary DNA clone isolated from a bovine cDNA library using a mixture of 32 oligonucleotides as the hybridization probe. The alpha, beta, gamma, delta and epsilon subunits contain 509, 480, 272, 146 and 50 amino acids, respectively. Two half cystine residues are present in the alpha- subunit and one in each of the gamma- and epsilon-chains; they are absent from the beta- and delta-subunits. The stoichiometry of subunits in the complex is estimated to be alpha 3 beta 3 gamma 1 delta 1 epsilon 1 and the molecular weight of the complex is 371,135. Mild trypsinolysis of the F1-ATPase complex, which has little effect on the hydrolytic activity of the enzyme, releases peptides from the N- terminal regions of the alpha- and beta-chains only; the C-terminal regions are unaffected. Sequence analysis of the released peptides demonstrates that the N terminals of the alpha- and beta-chains are ragged. In 65% of alpha-chains, the terminus is pyrrolidone carboxylic acid; in the remainder this residue is absent and the chains commence at residue 2, i.e. lysine. In the beta-subunit a minority of chains (16%) have N-terminal glutamine, or its deamidation product, glutamic acid (6%), or the cyclized derivative, pyrrolidone carboxylic acid (5%). A further 28% commence at residue 2, alanine, and 45% at residue 3, serine. The delta-chains also are heterogeneous; in 50% of chains the N-terminal alanine residue is absent. The sequences of the alpha- and beta-chains show that they are weakly homologous, as they are in bacterial F1-ATPases. The sequence of the bovine delta-subunit of F1- ATPase shows that it is the counterpart of the bacterial epsilon- subunit. The bovine epsilon-subunit is not related to any known bacterial or chloroplast H+-ATPase subunit, nor to any other known sequence. The counterpart of the bacterial delta-subunit is bovine oligomycin sensitivity conferral protein, which helps to bind F1 to the inner mitochondrial membrane.(ABSTRACT TRUNCATED AT 400 WORDS)
RP  - NOT IN FILE
NT  - UI - 86011574LA - engRN - 0 (Amino Acids)RN - 0 (Macromolecular Systems)RN - 0 (Sulfhydryl Compounds)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19851114IS - 0022-2836SB - IMCY - ENGLANDJC - J6V
UR  - PM:2864455
SO  - J Mol Biol 1985 Aug 20 ;184(4):677-701

682
UI  - 674
AU  - Weber J
AU  - Lucken U
AU  - Schafer G
TI  - Total number and differentiation of nucleotide binding sites on mitochondrial F1-ATPase. An approach by photolabeling and equilibrium binding studies
AB  - In this study 3'-O-[3-(4-azido-2-nitrophenyl)propionyl]-ADP was used as a photoaffinity analog for nucleotide binding sites on nucleotide- depleted F1-ATPase. Catalytic and binding properties of the labeled enzyme were investigated. The analog behaves as a competitive inhibitor in the dark (Ki = 50 microM). Photoirradiation of F1 in the presence of the analog leads to inactivation depending linearly on the incorporation of label. Complete inactivation is achieved at a stoichiometry of 3 mol/mol F1. The label is distributed between alpha and beta subunits in a ratio of 30%:70%. Although three sites were blocked covalently by photolabeling, three reversible sites of much higher affinity than the labeled sites were preserved. Mild alkaline treatment of photoinactivated enzyme leads to almost complete reactivation which is due to hydrolysis of the 3'-ester bond and release of the ADP moiety from the covalently bound analog. The conclusions drawn are as follows. The total number of sites which can be simultaneously occupied by nucleotides on F1 is six. Adopting the finding [Grubmeyer, C. & Penefsky, H. S. (1981) J. Biol. Chem. 256, 3718-3727] that the high-affinity sites are the catalytic ones which can be covalently labeled by 3'-O-[5-azidonaphthoyl(1)]-ADP [Lubben, M., Lucken, U., Weber, J. & Schafer, G. (1984) Eur. J. Biochem. 143, 483-490], it appears likely that azidonitrophenylpropionyl-ADP is a specific photolabel for the lower-affinity sites on nucleotide-depleted F1. This means that both types of sites can be differentiated by specific photoaffinity analogs. The labeled low-affinity sites interact with the catalytic sites, abolishing enzyme turnover, when steadily occupied by ADP kept in place by the covalently linking residue, which by itself has no inhibitory effect on the enzyme
RP  - NOT IN FILE
NT  - UI - 85154039LA - engRN - 0 (Adenine Nucleotides)RN - 0 (Affinity Labels)RN - 58-64-0 (Adenosine Diphosphate)RN - 64655-48-7 (NAP(4)-ADP)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19850515IS - 0014-2956SB - IMCY - GERMANY, WESTJC - EMZ
UR  - PM:2858390
SO  - Eur J Biochem 1985 Apr 1 ;148(1):41-47

683
UI  - 6297
AU  - Williams RJ
TI  - Chloroplast stacking
MH  - chloroplast
MH  - Chloroplasts
MH  - grana
MH  - membrane stacking
RP  - IN FILE
AV  - membrane stacking
SO  - TIBS 1985  ;():341-341

684
UI  - 6298
AU  - Williams RJP
TI  - Further comment from Williams
MH  - grana
MH  - membrane stacking
RP  - IN FILE
AV  - membrane stacking
SO  - TIBS 1985  ;():341-341

685
UI  - 467
AU  - Wise JG
AU  - Senior AE
TI  - Catalytic properties of the Escherichia coli proton adenosinetriphosphatase: evidence that nucleotide bound at noncatalytic sites is not involved in regulation of oxidative phosphorylation
AB  - Nucleotide-depleted F1-ATPase from Escherichia coli was reconstituted with F1-depleted membranes and shown to catalyze high rates of oxidative phosphorylation of ADP and GDP. Adenine nucleotide became bound to the nonexchangeable nucleotide sites on membrane-bound F1 during ATP synthesis, but binding of guanine nucleotides to nonexchangeable sites during GTP synthesis was not detectable. It was possible to reload the nonexchangeable sites on nucleotide-depleted F1 with radioactive adenine nucleotide prior to membrane reconstitution. The radioactive adenine nucleotide did not exchange significantly during oxidative phosphorylation of ADP or GDP. The amount of nonexchangeable adenine nucleotide found in membrane-bound F1 was the same when the nonexchangeable sites were reloaded either prior to membrane reconstitution of the F1 or after membrane reconstitution with nucleotide-free F1 followed by a burst of oxidative phosphorylation of ADP. The results showed that occupation of the nonexchangeable sites on F1 by tightly bound nucleotide is not required for oxidative phosphorylation of GDP (a physiological activity of F1 in the bacterial cell). Also, the results confirm directly that the adenine-specific nonexchangeable sites on F1 are noncatalytic sites. Using this experimental approach, it was possible to look for a regulatory effect of the nonexchangeable nucleotide on oxidative phosphorylation. Nucleotide-depleted F1 was first reloaded with (i) ATP, (ii) ADP, (iii) 5'-adenylyl imidodiphosphate, or (iv) zero nucleotide, and was then reconstituted with F1-depleted membranes. The reconstituted membranes were compared in respect to rates of oxidative phosphorylation of GDP and Km values of GDP and Pi. No regulatory role for the nonexchangeable nucleotide was evident.(ABSTRACT TRUNCATED AT 250 WORDS)
RP  - NOT IN FILE
NT  - UI - 86077745LA - engRN - 0 (Guanine Nucleotides)RN - 0 (Phosphorus Radioisotopes)RN - 146-91-8 (Guanosine Diphosphate)RN - 25612-73-1 (Adenylyl Imidodiphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 86-01-1 (Guanosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSDA - 19860213IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:2866799
SO  - Biochemistry 1985 Nov 19 ;24(24):6949-6954

686
UI  - 9895
AU  - Beavis AD
AU  - Lehninger AL
TI  - The upper and lower limits of the mechanistic stoichiometry of mitochondrial oxidative phosphorylation. Stoichiometry of oxidative phosphorylation.
AB  - Determination of the intrinsic or mechanistic P/O ratio of oxidative phosphorylation is difficult because of the unknown magnitude of leak fluxes. Applying a new approach developed to overcome this problem (see our preceding paper in this journal), the relationships between the rate of O2 uptake [( Jo)3], the net rate of phosphorylation (Jp), the P/O ratio, and the respiratory control ratio (RCR) have been determined in rat liver mitochondria when the rate of phosphorylation was systematically varied by three specific means. (a) When phosphorylation is titrated with carboxyatractyloside, linear relationships are observed between Jp and (Jo)3. These data indicate that the upper limit of the mechanistic P/O ratio is 1.80 for succinate and 2.90 for 3-hydroxybutyrate oxidation. (b) Titration with malonate or antimycin yields linear relationships between Jp and (Jo)3. These data give the lower limit of the mechanistic P/O ratio of 1.63 for succinate and 2.66 for 3-hydroxybutyrate oxidation. (c) Titration with a protonophore yields linear relationships between Jp, (Jo)3, and (Jo)4 and between P/O and 1/RCR. Extrapolation of the P/O ratio to 1/RCR = 0 yields P/O ratios of 1.75 for succinate and 2.73 for 3-hydroxybutyrate oxidation which must be equal to or greater than the mechanistic stoichiometry. When published values for the H+/O and H+/ATP ejection ratios are taken into consideration, these measurements suggest that the mechanistic P/O ratio is 1.75 for succinate oxidation and 2.75 for NADH oxidation.
MH  - Adenosine Triphosphate
MH  - analogs & derivatives
MH  - Animal
MH  - Antimycin A
MH  - Atractyloside
MH  - biosynthesis
MH  - Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone
MH  - drug effects
MH  - In Vitro
MH  - Liver
MH  - metabolism
MH  - Mitochondria
MH  - Nad
MH  - Oxidation-Reduction
MH  - Oxidative Phosphorylation
MH  - Oxygen Consumption
MH  - pharmacology
MH  - Phosphorylation
MH  - Protons
MH  - Rats
MH  - succinate
MH  - Succinates
MH  - Support,U.S.Gov't,P.H.S.
MH  - Thermodynamics
RP  - NOT IN FILE
SO  - Eur J Biochem 1986 Jul 15 ;158(2):315-322

687
UI  - 876
AU  - Bogucka K
TI  - [The proton-transporting ATPase complex]
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - Amino Acids
MH  - Animal
MH  - Bacteria
MH  - Biological Transport
MH  - biosynthesis
MH  - Catalysis
MH  - Energy Metabolism
MH  - Human
MH  - metabolism
MH  - Mitochondria
MH  - Protons
RP  - NOT IN FILE
SO  - Postepy Biochem 1986  ;32(3):301-328

688
UI  - 772
AU  - Bullough DA
AU  - Yoshida M
AU  - Allison WS
TI  - Sequence of the radioactive tryptic peptide obtained after inactivating the F1-ATPase of the thermophilic bacterium PS3 with 5'-p- fluorosulfonylbenzoyl[3H]adenosine at 65 degrees C
AB  - Following a lag of about 30 min, the F1-ATPase from the thermophilic bacterium, PS3 (TF1), was inactivated slowly by 0.8 mM 5'-p- fluorosulfonylbenzoyladenosine (FSBA) at 23 degrees C and pH 7.0. When the enzyme was treated with 0.2 mM FSBA at pH 7.0 and 23 degrees C for 15 min and gel-filtered, no enzyme activity was lost. However, the lag in inactivation was abolished when the enzyme was subsequently incubated with 2.0 mM FSBA at 23 degrees C in the pH range from 6.8 to 10.0. The pH-inactivation profile obtained under these conditions revealed a pK alpha of about 9.3 which was associated with the inactivation. When pretreated TF1 was inactivated at 23 degrees C with [3H]FSBA by about 90%, greater than 20 mol of [3H]SBA was incorporated per mole of enzyme. TF1 was inactivated rapidly by 0.8 mM FSBA at pH 6.4 and 65 degrees C, and no lag was observed. Following inactivation of TF1 with 0.8 mM [3H]FSBA at 65 degrees C and pH 6.4, about 10 mol of [3H]SBA was incorporated per mole of enzyme. When a tryptic digest of the labeled enzyme was fractionated by reversed-phase high-performance liquid chromatography, a single major radioactive peptide was isolated. When subjected to automatic Edman degradation, this peptide was shown to have the amino acid sequence: A-L-A-P-E-I-V-G-E-E-H-X-Q-V-A-R, where X indicates that a phenylthiohydantoin derivative was not detected in cycle 12. However, from the DNA sequence of the gene encoding the subunit of TF1 (Y. Kagawa, M. Ishizuka, T. Saishu, and S. Nakao (1985) Abstracts International Symposium on Energy Transducing ATPases, Kobe, Japan, p. 84), this position has been shown to be occupied by tyrosine. This tyrosine is homologous with beta-Tyr-368 of the bovine mitochondrial F1-ATPase (MF1) the modification of which is responsible for the inactivation MF1 by FSBA
RP  - NOT IN FILE
NT  - UI - 86129433LA - engRN - 0 (5'-(4-fluorosulfonylbenzoyl)adenosine)RN - 0 (Affinity Labels)RN - 10028-17-8 (Tritium)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-61-7 (Adenosine)RN - EC 3.4.21.4 (Trypsin)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM-16974/GM/NIGMSDA - 19860313IS - 0003-9861SB - IMCY - UNITED STATESJC - 6SK
UR  - PM:2868693
SO  - Arch Biochem Biophys 1986 Feb 1 ;244(2):865-871

689
UI  - 20823
AU  - Cain BD
AU  - Simoni RD
TI  - Impaired proton conductivity resulting from mutations in the a subunit of F1F0 ATPase in Escherichia coli
AB  - Mutations in the uncB gene which encodes the a subunit of F1F0-ATPase in Escherichia coli were isolated and characterized. Eight mutations caused premature polypeptide chain termination. Two mutations were single amino acid substitutions resulting in the replacements of serine 206 with leucine (ser-206----leu) and histidine 245 with tyrosine (his- 245----tyr). The ser-206----leu mutation does not alter F1 binding and allows ATP driven membrane energization at a low level. Stripping of F1 from membranes containing the ser-206----leu mutation does not render the membranes permeable to protons indicating impaired proton conductivity. The his-245----tyr mutation also blocks Fo-mediated proton conduction but has normal F1 binding properties. F1 bound to membranes with both ser-206----leu and his-245----tyr mutant a subunits is sensitive to dicyclohexylcarbodiimide. Apparently, both missense mutations impair proton conduction without altering assembly of the F1F0-ATPase complex. The direct involvement of the a subunit in proton translocation is discussed
MH  - A
MH  - ACID
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATPase
MH  - BINDING
MH  - COMPLEX
MH  - Dicyclohexylcarbodiimide
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1
MH  - Histidine
MH  - membrane
MH  - Membranes
MH  - mutant
MH  - Nad
MH  - proton
MH  - Protons
MH  - SUBUNIT
MH  - translocation
RP  - NOT IN FILE
NT  - UI - 86278048LA - engRN - 0 (DNA, Bacterial)RN - 0 (Plasmids)RN - 0 (Protons)RN - 53-84-9 (NAD)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleID - GM09521/GM/NIGMSID - GM18539/GM/NIGMSDA - 19860917IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:2874137
SO  - J Biol Chem 1986 Aug 5 ;261(22):10043-10050

690
UI  - 20964
AU  - Deckers-Hebestreit G
AU  - Steffens K
AU  - Altendorf K
TI  - Conformation-specific antiserum raised against subunit c of ATP synthase (F1F0) from Escherichia coli
AB  - Subunit c of the membrane-integrated, proton-translocating F0 portion of the ATP synthase (F1F0) from Escherichia coli has been isolated under nondenaturing conditions (Schneider, E., and Altendorf, K. (1985) EMBO J. 4, 515-518) and antibodies have been raised in rabbits. The primary antisera did not recognize the antigen when present in the same buffer as used for the immunization. Surprisingly, in one of the three antisera a strong antibody binding was observed when intact F0, a.c complex or reconstituted subunit c was provided as the antigen. Incorporation of subunit c into liposomes together with subunits a and b forming an active, H+-translocating complex was not required for the recognition by the antiserum. Subunit c prepared by chloroform/methanol extraction or by chromatography in the presence of sodium dodecyl sulfate was not recognized by the anti-c antiserum when incorporated into liposomes
MH  - A
MH  - ACTIVE
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BINDING
MH  - buffer
MH  - COMPLEX
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - Liposomes
MH  - Macromolecular Systems
MH  - Sodium
MH  - SUBUNIT
MH  - SYNTHASE
MH  - SYSTEM
MH  - SYSTEMS
RP  - NOT IN FILE
NT  - UI - 87033712LA - engRN - 0 (Antigen-Antibody Complex)RN - 0 (Immune Sera)RN - 0 (Macromolecular Systems)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19861215IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:2876991
SO  - J Biol Chem 1986 Nov 15 ;261(32):14878-14881

691
UI  - 20963
AU  - Deckers-Hebestreit G
AU  - Altendorf K
TI  - Accessibility of F0 subunits from Escherichia coli ATP synthase. A study with subunit specific antisera
AB  - Antisera have been raised against denatured and non-denatured subunits a, b and c of the F0 complex of the ATP synthase from Escherichia coli. The subunit specificity of the antibodies has been established with immunoblot analysis or enzyme-linked immunosorbent assay (ELISA). In inside-out oriented membrane vesicles the binding avidities of both sets of antisera, against denatured and non-denatured subunits of F0, were similar in the presence as well as in the absence of the F1 part. F1-depleted everted membrane vesicles always produced an efficient binding of the different antisera. In the presence of F1 no antibody recognition could be observed with the anti-a antisera, while anti-b and anti-c antisera showed strong binding. However, a higher membrane protein concentration was necessary for the same antibody binding as in F1-stripped vesicles. In membrane vesicles with right-side-out orientation the recognition of the three F0 subunits was dependent on the antisera set used. Antisera raised against denatured subunits showed no binding to the membrane vesicles, only in case of anti- (dodecylsulfate-denatured b) antiserum could a slight affinity be detected. An antigen-antibody recognition with all three F0 subunits occurred when the antisera against non-denatured subunits were incubated with membrane vesicles of right-side-out orientation. The membrane protein concentration which was necessary to produce a significant binding was 10-100-fold higher compared to that of F1- depleted everted membrane vesicles
MH  - A
MH  - analysis
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BINDING
MH  - COLI ATP SYNTHASE
MH  - COMPLEX
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - F1
MH  - membrane
MH  - membrane vesicles
MH  - protein
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 87054010LA - engRN - 0 (Antibodies)RN - 0 (Immune Sera)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19870114IS - 0014-2956SB - IMCY - GERMANY, WEST
UR  - PM:2877880
SO  - Eur J Biochem 1986 Nov 17 ;161(1):225-231

692
UI  - 466
AU  - Duncan TM
AU  - Parsonage D
AU  - Senior AE
TI  - Structure of the nucleotide-binding domain in the beta-subunit of Escherichia coli F1-ATPase
AB  - We propose a working model for the tertiary structure of the nucleotide- binding domain of the beta-subunit of E. coli F1-ATPase, derived from secondary structure prediction and from comparison of the amino acid sequence with the sequences of other nucleotide-binding proteins of known three-dimensional structure. The model is consistent with previously published results of specific chemical modification studies and of analyses of mutations in the beta-subunit and its implications for subunit interactions and catalytic mechanism in F1-ATPases are discussed
RP  - NOT IN FILE
NT  - UI - 87030933LA - engRN - 0 (Carrier Proteins)RN - 0 (adenosine cyclic-3',5'-monophosphate binding proteins)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSID - GM29805/GM/NIGMSDA - 19861211IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:2876918
SO  - FEBS Lett 1986 Nov 10 ;208(1):1-6

693
UI  - 72
AU  - Engelbrecht S
AU  - Lill H
AU  - Junge W
TI  - Reconstitution of CF1-depleted thylakoid membranes with complete and fragmented chloroplast ATPase. The role of the delta subunit for proton conduction through CF0
AB  - Chloroplast ATPase (CF1) was isolated from spinach, pea and maize thylakoids by EDTA extraction followed by anion-exchange chromatography. CF1 was purified and resolved by HPLC into integral CF1, and CF1 lacking the delta & epsilon subunits: CF1(-delta) and CF1(- epsilon). Washing Mono-Q-bound CF1 with alcohol-containing buffers followed by elution without alcohol produced the beta subunit and in separate peaks CF1(-delta) and CF1(-epsilon). Elution from Mono Q in the presence of tenside yielded a beta delta fragment, CF1(-delta) and CF1(-delta epsilon). Chloroplasts were CF1-depleted by EDTA extraction. Reconstitution of photophosphorylation in these 'EDTA vesicles' was obtained by addition of CF1 and its fragments. CF1, CF1(-delta) and CF1(-delta epsilon) were active with cross-reactivity between spinach, pea and maize. delta-containing CF1 always reconstituted higher activities than delta-deficient CF1. The beta delta fragment and dicyclohexylcarbodiimide (DCCD)-inhibited CF1 also were reconstitutively active while beta and DCCD-inhibited CF1(-delta) were not. These results support the notion that subunit delta can function as a stopcock to the CF0 proton channel as proposed by Junge, W., Hong, Y. Q., Qian, L. P. and Viale, A. [(1984) Proc. Natl Acad. Sci. USA 81, 3078-3082]
RP  - NOT IN FILE
NT  - UI - 87053986LA - engRN - 0 (Macromolecular Systems)RN - 0 (Peptide Fragments)RN - 0 (Protons)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19870115IS - 0014-2956SB - IMCY - GERMANY, WESTJC - EMZ
UR  - PM:2877879
SO  - Eur J Biochem 1986 Nov 3 ;160(3):635-643

694
UI  - 364
AU  - Fearnley IM
AU  - Walker JE
TI  - Two overlapping genes in bovine mitochondrial DNA encode membrane components of ATP synthase
AB  - Two hydrophobic proteins have been purified to homogeneity from a mixture of about 13 proteins that are extracted from bovine mitochondria with a chloroform:methanol mixture. Sequence analysis shows that the smaller is a protein of 66 amino acids and is the product of a mitochondrial gene, A6L. The larger, a protein of 226 amino acids, is ATPase-6, a membrane component of ATP synthase, also encoded in mitochondrial DNA. The protein sequences determined establish that the genes for the two proteins overlap by 40 bases and indicate that translation of the second gene, ATPase-6, is initiated within the coding region of A6L. The A6L and the ATPase-6 proteins have also been isolated from the ATP synthase complex and so appear to be bona fide components of the enzyme. The function of A6L is unknown. However, weak structural homology suggests a functional similarity to the yeast mitochondrial protein, aapI, which is required for assembly of the fungal ATP synthase complex. Homologies between ATPase-6 and subunit a of the Escherichia coli ATP synthase complex indicate that the ATPase-6 protein has a similar role in the mitochondrial complex to its bacterial counterpart, being essential for the formation of an active proton channel
RP  - NOT IN FILE
NT  - UI - 87004570LA - engRN - 0 (DNA, Mitochondrial)RN - 0 (Peptide Fragments)RN - 0 (Proteolipids)RN - 506-68-3 (Cyanogen Bromide)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19861117IS - 0261-4189SB - IMCY - ENGLANDJC - EMB
UR  - PM:2875870
SO  - EMBO J 1986 Aug ;5(8):2003-2008

695
UI  - 50
AU  - Fillingame RH
AU  - Foster DL
TI  - Purification of F1F0 H(+)-ATPase from Escherichia coli
RP  - NOT IN FILE
NT  - UI - 90173859LA - engRN - 0 (Indicators and Reagents)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM-23105/GM/NIGMSDA - 19900330IS - 0076-6879SB - IMCY - UNITED STATESJC - MVA
UR  - PM:2908464
SO  - Methods Enzymol 1986  ;126():545-557

696
UI  - 117
AU  - Fillingame RH
AU  - Mosher ME
TI  - Use of lambda-unc transducing phages in genetic analysis of H(+)-ATPase mutants of Escherichia coli
RP  - NOT IN FILE
NT  - UI - 90173860LA - engRN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM-23105/GM/NIGMSDA - 19900330IS - 0076-6879SB - IMCY - UNITED STATESJC - MVA
UR  - PM:2908465
SO  - Methods Enzymol 1986  ;126():558-568

697
UI  - 118
AU  - Fillingame RH
AU  - Porter B
AU  - Hermolin J
AU  - White LK
TI  - Synthesis of a functional F0 sector of the Escherichia coli H+-ATPase does not require synthesis of the alpha or beta subunits of F1
AB  - The uncB, E, F, and H genes of the Escherichia coli unc operon were cloned behind the lac promoter of plasmid pUC9, generating plasmid pBP101. These unc loci code, respectively, for the chi, omega, and psi subunits of the F0 sector and the delta subunit of the F1 sector of the H+-ATP synthase complex. Induction of expression of the four unc genes by the addition of isopropyl-beta-D-thiogalactoside resulted in inhibition of growth. During isopropyl-beta-D-thiogalactoside induction, the three subunits of F0 were integrated into the cytoplasmic membrane with a resultant increase in H+ permeability. A functional F0 was formed from plasmid pBP101 in a genetic background lacking all eight of the unc structural genes coding the F1F0 complex. In the unc deletion background, a reasonable correlation was observed between the amount of F0 incorporated into the membrane and the function measured, i.e., high-affinity binding of F1 and rate of F0- mediated H+ translocation. This correlation indicates that most or all of the F0 assembled in the membrane is active. Although the F0 assembled under these conditions binds F1, only partial restoration of NADH-dependent or ATP-dependent quenching of quinacrine fluorescence was observed with these membranes. Proteolysis of a fraction of the psi subunit may account for this partial deficiency. The experiments described demonstrate that a functional F0 can be assembled in vivo in E. coli strains lacking genes for the alpha, beta, gamma, and epsilon subunits of F1
RP  - NOT IN FILE
NT  - UI - 86085678LA - engRN - 0 (Bacterial Proteins)RN - 0 (Plasmids)RN - 53-84-9 (NAD)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM-23105/GM/NIGMSDA - 19860220IS - 0021-9193SB - IMCY - UNITED STATESJC - HH3
UR  - PM:2867086
SO  - J Bacteriol 1986 Jan ;165(1):244-251

698
UI  - 20896
AU  - Futai M
AU  - Kanazawa H
TI  - Use of isolated subunits of F1 from Escherichia coli for genetic and biochemical studies
MH  - ATPase
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1
MH  - indicator
MH  - Indicators and Reagents
MH  - Macromolecular Systems
MH  - SUBUNIT
MH  - SYSTEM
MH  - SYSTEMS
RP  - NOT IN FILE
NT  - UI - 90173863LA - engRN - 0 (Indicators and Reagents)RN - 0 (Macromolecular Systems)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19900330IS - 0076-6879SB - IMCY - UNITED STATES
UR  - PM:2908468
SO  - Methods Enzymol 1986  ;126():588-594

699
UI  - 1162
AU  - Graber P
AU  - Junesch U
AU  - Schmidt G
AU  - Fromme P
TI  - Proton transport-coupled ATP synthesis catalyzed by the chloroplast ATPase
MH  - atp
MH  - ATP synthesis
MH  - ATPase
MH  - chloroplast
MH  - Energy Transfer
MH  - ion
MH  - proton
MH  - synthesis
MH  - TRANSFER
T2  - Ion Interact. Energy Transfer Biomembr., [Proc. Int. Workshop], Meeting Date 1985, 147-56. Edited by: Papageorgiou, George C.; Barber, James; Papa, Sergio. Plenum: New York, N. Y
Y2  - -32676  
RP  - ON REQUEST (03/18/92)
SO  -  1986  ;():

700
UI  - 1161
AU  - Graber P
AU  - Fromme P
AU  - Junesch U
AU  - Schmidt G
AU  - Thulke G
TI  - Kinetics of proton-transport-coupled ATP synthesis catalyzed by the chloroplast ATP synthase
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - chloroplast
MH  - Kinetics
MH  - SYNTHASE
MH  - synthesis
RP  - ON REQUEST (03/18/92)
SO  - Ber Bunsen-Ges Phys Chem 1986  ;90():1034-1040

701
UI  - 7970
AU  - Grber P
AU  - Fromme P
AU  - Junesch U
AU  - Schmidt G
AU  - Thulke G
TI  - Kinetics of Proton-Transport-Coupled ATP Synthesis Catalyzed by the Chloroplast ATP Synthase
MH  - ACTIVATION
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - chloroplast
MH  - Kinetics
MH  - regulation
MH  - SYNTHASE
MH  - synthesis
RP  - IN FILE
NT  - Ju, ATP 4
SO  - BBG 1986  ;90():1034-1040

702
UI  - 21309
AU  - Grzesiek S
AU  - Dencher NA
TI  - Dependency of delta pH-relaxation across vesicular membranes on the buffering power of bulk solutions and lipids
AB  - The dependency of delta pH-relaxation kinetics across the membrane of sonicated small phospholipid vesicles on the concentration of internally entrapped buffer has been investigated by means of the pH- indicator dye pyranine. A very high contribution of lipid headgroups to the internal buffering power of the liposomes is observed, amounting to an equivalent phosphate buffer concentration of 110 mM. This localized two-dimensional proton/hydroxide ion reservoir must be considered in any determination of the H+/OH- permeability coefficient. Furthermore, it could have significance for energy-transduction across biological membranes. From the established linear relation between delta pH- relaxation rates and buffering power, net H+/OH- permeabilities of 3 X 10(-3) cm/s for soybean phospholipid (SBPL) and 1 X 10(-4) cm/s for diphytanoyl phosphatidylcholine (diphytanoyl PC) vesicles at pH 7.2 as well as buffering powers per lipid molecule of 6 X 10(-2) (pH-unit)-1 (SBPL) and 4 X 10(-2) (pH-unit)-1 (diphytanoyl PC) are calculated. In the case of diphytanoyl PC vesicles, delta pH-decay is accelerated by the presence of chloride ions
MH  - A
MH  - Arylsulfonates
MH  - buffer
MH  - Buffers
MH  - delta
MH  - DYE
MH  - Hydroxides
MH  - indicator
MH  - ion
MH  - Ions
MH  - Kinetics
MH  - Lipids
MH  - liposome
MH  - Liposomes
MH  - membrane
MH  - Membranes
MH  - Permeability
MH  - pH
MH  - phosphate
MH  - proton
MH  - Protons
MH  - relaxation
MH  - Solutions
MH  - united states
MH  - Valinomycin
MH  - vesicles
RP  - NOT IN FILE
NT  - UI - 86296979LA - engRN - 0 (Arylsulfonates)RN - 0 (Buffers)RN - 0 (Hydroxides)RN - 0 (Liposomes)RN - 0 (Protons)RN - 0 (Solutions)RN - 2001-95-8 (Valinomycin)RN - 6358-69-6 (pyranine)PT - Journal ArticleDA - 19861020IS - 0006-3495SB - IMCY - UNITED STATES
UR  - PM:3017468
SO  - Biophys J 1986 Aug ;50(2):265-276

703
UI  - 19890
AU  - Gurevich YY
AU  - Kharkats YI
TI  - Ion transfer through a phase boundary: A stochastic approach
MH  - A
MH  - ion
MH  - TRANSFER
RP  - IN FILE
SO  - J Electroanal Chem Interfacial Electrochem 1986  ;200():3

704
UI  - 21093
AU  - Hicks DB
AU  - Krulwich TA
TI  - The membrane ATPase of alkalophilic Bacillus firmus RAB is an F1-type ATPase
AB  - At the optimal pH for growth (pH 10.5), alkalophilic Bacillus firmus RAB, an obligate aerobe, exhibits normal rates of oxidative phosphorylation despite the low transmembrane proton electrochemical gradient, about -60 mV (delta psi = -180 mV and delta pH = +120 mV). This bioenergetic problem might be resolved by use of an Na+ coupled ATP synthase; otherwise an F1F0-ATPase must be able to utilize low driving forces in this organism. The ATPase activity was extracted from everted membrane vesicles by low ionic strength treatment and purified to homogeneity by hydrophobic interaction chromatography and sucrose density gradient centrifugation. The ATPase preparation had the characteristic F1-ATPase subunit structure, with Mr values of 51,500 (alpha), 48,900 (beta), 34,400 (gamma), 23,300 (delta), and 14,500 (epsilon); the identity of the alpha and beta subunits was confirmed by immunoblotting with anti-beta of Escherichia coli and anti-B. firmus RAB F1. Methanol and octyl glucoside, agents that stimulated the low basal membrane ATPase activity 10- to 12-fold, dramatically elevated the MgATPase activity of the purified F1, more than 150-fold, to 50 mumol min-1 mg protein-1. Anti-F1 inhibited membrane ATPase activity greater than or equal to 80%. The membranes exhibited no Na+-stimulated or vanadate-sensitive ATPase activity when prepared in the absence or presence of Na+ or ATP. These findings, which are consistent with previous studies, establish that in alkalophilic bacteria, ATP hydrolysis, and presumably ATP synthesis is catalyzed by an F1F0-ATPase rather than a Na+ ATPase
MH  - A
MH  - alpha
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - ATPase
MH  - Bacillus
MH  - Bacteria
MH  - BETA
MH  - BETA-SUBUNIT
MH  - delta
MH  - DELTA-PH
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - Hydrolysis
MH  - membrane
MH  - membrane vesicles
MH  - Membranes
MH  - Methanol
MH  - Oxidative Phosphorylation
MH  - pH
MH  - Phosphorylation
MH  - Potassium
MH  - proton
MH  - Proton-Translocating ATPases
MH  - PSI
MH  - Sodium
MH  - structure
MH  - SUBUNIT
MH  - Sucrose
MH  - SYNTHASE
MH  - synthesis
MH  - vesicles
RP  - NOT IN FILE
NT  - UI - 86304482LA - engRN - 0 (Metals)RN - 7440-09-7 (Potassium)RN - 7440-23-5 (Sodium)RN - EC 3.6.1.- (Ca(2+) Mg(2+)-ATPase)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleID - GM28454/GM/NIGMSDA - 19861023IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:2875076
SO  - J Biol Chem 1986 Sep 25 ;261(27):12896-12902

705
UI  - 769
AU  - Hisabori T
AU  - Yoshida M
AU  - Sakurai H
TI  - ADP binding to TF1 and its subunits induces ultraviolet spectral changes
AB  - Adenine nucleotide binding sites on the coupling factor ATPase of thermophilic bacterium PS3 (TF1) were investigated by UV spectroscopy and by equilibrium dialysis. When ADP was mixed with TF1 in the presence and in the absence of Mg2+, an UV absorbance change was induced (t1/2 approximately 1 min) with a peak at about 278 nm and a trough at about 250 nm. Similar spectral changes were induced by ADP with the isolated beta subunits in the presence and in the absence of Mg2+, and with the isolated alpha subunits in the presence of Mg2+ although the magnitudes of the changes were different. From equilibrium dialysis measurement we identified two classes of nucleotide binding sites in TF1 in the presence of Mg2+, three high-affinity sites (Kd = 61 nM) and three low-affinity sites (Kd = 87 microM). In the absence of Mg2+, TF1 has one high-affinity site (Kd less than 10 nM) and five low- affinity sites (Kd = 100 microM). Moreover, we found a single Mg2+- dependent ADP binding site on the isolated alpha subunit and a single Mg2+-independent ADP binding site on the isolated beta subunit. From the above observations, we concluded that the three Mg2+-dependent high- affinity sites for ADP are located on the alpha subunit in TF1 and that the single high-affinity site is located on one of the beta subunits in TF1 in the absence of Mg2+
RP  - NOT IN FILE
NT  - UI - 87057128LA - engRN - 58-64-0 (Adenosine Diphosphate)RN - 7439-95-4 (Magnesium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19870109IS - 0021-924XSB - IMCY - JAPANJC - HIF
UR  - PM:2877979
SO  - J Biochem (Tokyo ) 1986 Sep ;100(3):663-670

706
UI  - 844
AU  - Jackson JB
AU  - Nicholls DG
TI  - Methods for the determination of membrane potential in bioenergetic systems
RP  - NOT IN FILE
NT  - UI - 86284279LA - engRN - 0 (Carotenoids)RN - 0 (Radioisotopes)RN - 7440-09-7 (Potassium)RN - 7440-17-7 (Rubidium)PT - Journal ArticleDA - 19860916IS - 0076-6879SB - IMCY - UNITED STATESJC - MVA
UR  - PM:3526088
SO  - Methods Enzymol 1986  ;127():557-577

707
UI  - 21271
AU  - Junge W
AU  - Polle A
TI  - Theory of proton flow along appressed thylakoid membranes under both non-stationary and stationary conditions
MH  - Diffusion
MH  - Kinetics
MH  - membrane
MH  - Membranes
MH  - P
MH  - proton
MH  - stacking
MH  - theory
MH  - thylakoid
MH  - thylakoid membrane
MH  - wox
RP  - IN FILE
NT  - P 30; K
SO  - Biochim Biophys Acta 1986  ;848():265-273

708
UI  - 20895
AU  - Kanazawa H
AU  - Noumi T
AU  - Futai M
TI  - Analysis of Escherichia coli mutants of the H(+)-transporting ATPase: determination of altered site of the structural genes
MH  - analysis
MH  - ATPase
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - Macromolecular Systems
MH  - mutant
MH  - Site
MH  - SYSTEM
MH  - SYSTEMS
RP  - NOT IN FILE
NT  - UI - 90173864LA - engRN - 0 (Macromolecular Systems)RN - 0 (Plasmids)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19900330IS - 0076-6879SB - IMCY - UNITED STATES
UR  - PM:2908469
SO  - Methods Enzymol 1986  ;126():595-603

709
UI  - 21279
AU  - Kell DB
TI  - Localized protonic coupling: overview and critical evaluation of techniques.
MH  - coupling
RP  - NOT IN FILE
SO  - Methods Enzymol 1986  ;127():538-557

710
UI  - 19873
AU  - Kironde FA
AU  - Cross RL
TI  - Adenine nucleotide-binding sites on beef heart F1-ATPase. Conditions that affect occupancy of catalytic and noncatalytic sites
AB  - Beef heart mitochondrial F1 contains a total of six adenine nucleotide- binding sites including at least two different types of sites. Three "exchangeable" sites exchange rapidly during hydrolysis of MgATP, whereas three "nonexchangeable" sites do not (Cross, R. L. and Nalin, C. M. (1982) J. Biol. Chem. 257, 2874-2881). When F1 that has been stored as a suspension in (NH4)2SO4/ATP/EDTA/sucrose/Tris, pH 8.0, is pelleted, rinsed with (NH4)2SO4, dissolved, and desalted, it retains three bound adenine nucleotides. We find that two of these endogenous nucleotides are bound at nonexchangeable sites and one at an exchangeable site. The vacant nonexchangeable site is highly specific for adenine nucleotide and is rapidly filled by ADP upon addition of ADP or during ATP hydrolysis. ADP bound at this site can be removed by reprecipitating the enzyme with (NH4)2SO4. The single nucleotide retained by desalted F1 at an exchangeable site is displaced during hydrolysis of ATP, GTP, or ITP. The binding of PPi at two sites on the enzyme also promotes its dissociation. Neither procedure affects retention of nucleotide at the nonexchangeable sites. These observations, combined with the finding that PPi is much more easily removed from exchangeable sites than ADP, have led to the development of a procedure for preparing F1 with uniform and well-defined nucleotide site occupancy. This involves sequential exposure to MgATP, PPi, and high concentrations of Pi. Unbound ligand is removed between each step. The resulting enzyme, F1[3,0], has three occupied nonexchangeable sites and three vacant exchangeable sites. Evidence that nonexchangeable and exchangeable sites represent noncatalytic and catalytic sites, respectively, suggest that this form of the enzyme will prove useful in numerous applications, including transient kinetic measurements and affinity labeling of active site residues
MH  - A
MH  - ACTIVE
MH  - Adenine Nucleotides
MH  - ADENINE-NUCLEOTIDES
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - ADP
MH  - atp
MH  - ATP synthase
MH  - BINDING
MH  - Binding Sites
MH  - development
MH  - Diphosphates
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - Guanosine Triphosphate
MH  - H(+)-Transporting ATP Synthase
MH  - Hydrolysis
MH  - Magnesium
MH  - NONCATALYTIC SITES
MH  - nucleotide binding
MH  - nucleotide binding sites
MH  - NUCLEOTIDE-BINDING-SITES
MH  - Nucleotides
MH  - pH
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 86304427LA - engRN - 0 (Adenine Nucleotides)RN - 0 (Diphosphates)RN - 28141-84-6 (magnesium GTP)RN - 56-65-5 (Adenosine Triphosphate)RN - 86-01-1 (Guanosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM 23152/GM/NIGMSDA - 19861023IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:2875073
SO  - J Biol Chem 1986 Sep 25 ;261(27):12544-12549

711
UI  - 21094
AU  - Krulwich TA
AU  - Guffanti AA
TI  - Regulation of internal pH in acidophilic and alkalophilic bacteria
MH  - Bacteria
MH  - pH
MH  - regulation
MH  - Sodium
RP  - NOT IN FILE
NT  - UI - 86230075LA - engRN - 7440-23-5 (Sodium)PT - Journal ArticleDA - 19860630IS - 0076-6879SB - IMCY - UNITED STATES
UR  - PM:3713537
SO  - Methods Enzymol 1986  ;125():352-365

712
UI  - 21095
AU  - Krulwich TA
TI  - Bioenergetics of alkalophilic bacteria
AB  - The central problem for organisms which grow optimally, and in some cases obligately, at pH values of 10 to 11, is the maintenance of a relatively acidified cytoplasm. A key component of the pH homeostatic mechanism is an electrogenic Na+/H+ antiporter which--by virtue of kinetic properties and/or its concentration in the membrane--catalyzes net proton uptake while the organisms extrude protons during respiration. The antiporter is also capable of maintaining a constant pHin during profound elevations in pHout as long as Na+ entry is facilitated by the presence of solutes which are taken up with Na+. Secondary to the problem of acidifying the interior is the adverse effect of the large pH gradient, acid in, on the total pmf of alkalophile cells. For the purposes of solute uptake and motility, the organisms appear to largely bypass the problem of a low pmf by utilizing a sodium motive force for energization. However, ATP synthesis appears not to resolve the energetics problem by using Na+ or by incorporating the proton-translocating ATPase into intracellular organelles. The current data suggest that effective proton pumping carried out by the alkalophile respiratory chain at high pH may deliver at least some portion of the protons to the proton-utilizing catalysts, i.e., the F1F0-ATPase and the Na+/H+ antiporter, by some localized pathway
MH  - A
MH  - ACID
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthesis
MH  - ATPase
MH  - Bacteria
MH  - bioenergetics
MH  - Carrier Proteins
MH  - Cells
MH  - CONSTANT
MH  - Cytoplasm
MH  - electrogenic
MH  - mechanism
MH  - Organelles
MH  - pH
MH  - pmf
MH  - protein
MH  - Proteins
MH  - proton
MH  - Proton-Translocating ATPases
MH  - Protons
MH  - Respiration
MH  - secondary
MH  - Sodium
MH  - synthesis
RP  - NOT IN FILE
NT  - UI - 86200150LA - engRN - 0 (Carrier Proteins)RN - 0 (Sodium-Hydrogen Antiporter)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleID - GM28454/GM/NIGMSDA - 19860602IS - 0022-2631SB - IMCY - UNITED STATES
UR  - PM:2871195
SO  - J Membr Biol 1986  ;89(2):113-125

713
UI  - 884
AU  - Lane MD
AU  - Pedersen PL
AU  - Mildvan AS
TI  - The mitochondrion updated
MH  - Adenosine Triphosphate
MH  - Energy Metabolism
MH  - Human
MH  - metabolism
MH  - Mitochondria
RP  - NOT IN FILE
SO  - Science 1986 Oct 31 ;234(4776):526-527

714
UI  - 312
AU  - Lill H
AU  - Engelbrecht S
AU  - Schonknecht G
AU  - Junge W
TI  - The proton channel, CF0, in thylakoid membranes. Only a low proportion of CF1-lacking CF0 is active with a high unit conductance (169 fS)
AB  - We investigated the conductance of pea thylakoid membranes and their capacity for photophosphorylation as function of the extraction of chloroplast coupling factor CF1. The degree of extraction was varied via the incubation time in EDTA-containing hypo-osmolar medium and was measured by rocket electroimmunodiffusion. The conductance of thylakoid membranes was measured by flash kinetic spectrophotometry. The time course of extraction followed the time course of thylakoid swelling. Contrary to expectation increasing loss of CF1 did not primarily increase the velocity of proton efflux from each vesicle. Instead proton-tight vesicles were converted to leaky ones, which lost phosphorylating activity. Two subpopulations occurred, although both types of vesicles, leaky and proton-tight ones, were CF1-depleted to a similar degree. This implied that only a small fraction of CF1-lacking CF0 was functional as a proton channel. Tight vesicles had no functional channels while leaky ones had at least one. We determined the proportion of tight vesicles in three independent ways: via the residual phosphorylation activity, via measurements of proton efflux and via measurements of the electric relaxation across the membrane. The results obtained were identical. A statistical evaluation of the data led us to the following conclusions. EDTA treatment produced vesicles containing approximately 10(5) chlorophyll molecules, equivalent to a total of approximately 100 CF0CF1 per vesicle. Even at the highest degree of extraction (75% of total CF1 extracted) only 2.5 out of 75 exposed CF0 per vesicle were proton-conducting. The unit conductance of one open CF0 channel was 169 +/- 18 fS at pH 7.5 and room temperature. At an electrical driving force of 100 mV this was equivalent to the passage of approximately 10(5) protons/s. The most important consequence of this relatively high unit conductance was that a single open CF0 channel was capable of dissipating the protonmotive force of one vesicle, thereby deactivating the whole remaining catalytic capacity of this vesicle
RP  - NOT IN FILE
NT  - UI - 87053985LA - engRN - 0 (Ion Channels)RN - 60-00-4 (Edetic Acid)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19870115IS - 0014-2956SB - IMCY - GERMANY, WESTJC - EMZ
UR  - PM:2430802
SO  - Eur J Biochem 1986 Nov 3 ;160(3):627-634

715
UI  - 580
AU  - Matsuno-Yagi A
AU  - Hatefi Y
TI  - Kinetic modalities of ATP synthesis. Regulation by the mitochondrial respiratory chain
AB  - Two interconvertible kinetic modes are described for ATP synthesis by bovine heart submitochondrial particles. One mode is characterized by low apparent Km values for ADP (6-10 microM) and Pi (less than or equal to 0.25 mM), and a limited capacity for ATP synthesis (apparent Vmax approximately 500 nmol ATP.min-1.mg of protein-1). ATP synthesis occurs predominantly in this mode when the coupled activity of the respiratory chain relative to the number of functional ATP synthase complexes is low. The second kinetic mode is characterized by high apparent Km values for ADP (50-100 microM) and Pi (approximately 2.0 mM) and a high capacity for ATP synthesis (Vmax greater than 1800 nmol ATP.min-1.mg of protein-1). This mode of ATP synthesis predominates when the available free energy relative to the number of functional ATP synthase units is high. These results suggest that energy pressure in mitochondria might regulate ATP synthesis such that at low levels of energy the ATP synthase operates economically (low substrate Km values, low turnover capacity for ATP synthesis), while at high levels of energy these kinetic constraints are relaxed (high substrate Km values, high turnover capacity for ATP synthesis). The implications of these findings are discussed in relation to the cooperative-type kinetics of ATP synthesis and hydrolysis, the differential effects of a number of F0-F1 inhibitors on the rates of ATP synthesis and hydrolysis, and the controversy as to whether protonic energy in mitochondria is localized or delocalized
RP  - NOT IN FILE
NT  - UI - 87033581LA - engRN - 0 (Oligomycins)RN - 0 (Peptides)RN - 53-84-9 (NAD)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - 56645-91-1 (efrapeptin)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleID - AM08126/AM/NIADDKDA - 19861201IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:2945814
SO  - J Biol Chem 1986 Oct 25 ;261(30):14031-14038

716
UI  - 534
AU  - McEnery MW
AU  - Pedersen PL
TI  - Diethylstilbestrol. A novel F0-directed probe of the mitochondrial proton ATPase
AB  - At low concentrations, diethylstilbestrol (DES) is shown to be a potent F0-directed inhibitor of the F0F1-ATPase of rat liver mitochondria. In analogy to other F0-directed inhibitors, DES inhibits both the ATPase and ATP-dependent proton-translocation activities of the purified and membrane bound enzyme. When added at low concentrations with dicyclohexylcarbodiimide (DCCD), a covalent inhibitor, DES acts synergistically to inhibit ATPase activity of the complex. At higher concentrations, DES restores DCCD-inhibited ATPase activity. However, there is no restoration of ATP-dependent proton translocation. Under these conditions DCCD remains covalently bound to the F0F1-ATPase complex and F1 remains bound to Fo. Significantly, when the F0F1-ATPase is inhibited by the Fo-directed inhibitor venturicidin rather than DCCD, DES is also able to restore ATPase activity. In contrast, DES is unable to restore ATPase activity to F0F1 preparations inhibited by the Fo-directed inhibitors oligomycin or tricyclohexyltin. However, combinations of [DES + DCCD] or [DES + venturicidin] can restore ATPase activity to F0F1 preparations inhibited by either oligomycin or tricyclohexyltin. Results presented here indicate that the F0 moiety of the rat liver mitochondrial proton ATPase contains a distinct binding site for DES. In addition, they suggest that at saturating concentrations simultaneous occupancy of the DES binding site and sites for either DCCD or venturicidin promote "uncoupled" ATP hydrolysis
RP  - NOT IN FILE
NT  - UI - 86111844LA - engRN - 0 (Liposomes)RN - 0 (Oligomycins)RN - 0 (Protons)RN - 0 (Trialkyltin Compounds)RN - 0 (Venturicidins)RN - 13121-70-5 (cyhexatin)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-53-1 (Diethylstilbestrol)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - CA 10951/CA/NCIDA - 19860312IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:2868005
SO  - J Biol Chem 1986 Feb 5 ;261(4):1745-1752

717
UI  - 20892
AU  - Miki J
AU  - Takeyama M
AU  - Noumi T
AU  - Kanazawa H
AU  - Maeda M
AU  - Futai M
TI  - Escherichia coli H+-ATPase: loss of the carboxyl terminal region of the gamma subunit causes defective assembly of the F1 portion
AB  - Mutant genes for the gamma subunit of H+-translocating ATPase (H+- ATPase) were cloned from eight different strains of Escherichia coli isolated in this laboratory. Determination of their nucleotide sequences revealed that they are amber nonsense mutations: a Gln codon at position 15, 158, 227, 262, and 270, respectively, was replaced by a termination codon in these strains. As terminal Met is missing in the gamma subunit, these results indicate that these strains are capable of synthesizing fragments of gamma subunits of 13, 156, 225, 260, and 268 amino acid residues, respectively. Studies on the properties of membranes of these strains suggested the importance of the region between Gln 269 and the carboxyl terminus (residue 286) for forming a stable F1 complex with ATPase activity and the region between Gln 226 and Gln 261 for normal interaction of F1 with F0. The sequence from Gln 261 to Gln 269 also seemed to be important for stability of F1 assembly on the membranes. The high frequency of the nonsense mutations suggested that the number of essential residues is limited in this subunit. Comparison of the homologies of the amino acid sequences of the gamma subunits from four different sources confirmed this notion: 19% of amino acid residues are identically conserved in these four strains, and the conserved regions are the amino terminal and carboxyl terminal regions
MH  - A
MH  - ACID
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - Amino Acid Sequence
MH  - ATPase
MH  - COMPLEX
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - F1
MH  - H+
MH  - H+-ATPase
MH  - membrane
MH  - Membranes
MH  - mutant
MH  - proton
MH  - Protons
MH  - RESIDUE
MH  - SUBUNIT
RP  - NOT IN FILE
NT  - UI - 87098777LA - engRN - 0 (Plasmids)RN - 0 (Protons)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19870205IS - 0003-9861SB - IMCY - UNITED STATES
UR  - PM:2879511
SO  - Arch Biochem Biophys 1986 Dec ;251(2):458-464

718
UI  - 20894
AU  - Noumi T
AU  - Oka N
AU  - Kanazawa H
AU  - Futai M
TI  - Mutational replacements of conserved amino acid residues in the beta subunit resulted in defective assembly of H+-translocating ATPase (F0F1) in Escherichia coli
AB  - Mutant genes for the beta subunit of H+-translocating ATPase (F0F1) were cloned from Escherichia coli strains isolated in this laboratory. Determination of their nucleotide sequence revealed four missense mutations (strain KF39, Glu-41----Lys; strain KF16 and KF42, Glu-185---- Lys; strain KF48, Gly-223----Asp; KF26 and 4 other strains, Ser-292---- Phe). Two nonsense mutants (strain KF40, Gln-361----end; strain KF20, Gln-397----end) were also identified. Glu-41, Glu-185, and Ser-292 are conserved in the amino acid sequences of the beta subunits so far studied, and Gly-223, Gln-361, and Gln-397 are conserved in beta subunits from bacteria and mitochondria, but not in those from chloroplasts. The amounts of F1 subunits in the membranes of these strains were studied by immunochemical assay and two-dimensional gel electrophoresis. In the mutants studied, the amounts of alpha and beta subunits in the membranes were 69-21 and 46-2%, respectively, of the amounts in wild-type membranes, the amount depending on the strain. No delta and epsilon subunits were detected in membranes of a missense mutant KF16, although reduced amounts of alpha and beta subunits could be detected, suggesting that the F1 portion may not be connected to F0 through the delta and epsilon subunits. The altered residues in missense mutants or missing domains in nonsense mutants may be important for the subunit-subunit interactions or assembly of the entire complex. Genetic experiments on introduction of suppressor tRNA into strains KF40 and KF20 suggested that F1 could be active even when residue 361 or 397 was replaced by a Ser, Leu, or Tyr residue
MH  - A
MH  - ACID
MH  - ACTIVE
MH  - Amino Acid Sequence
MH  - ATPase
MH  - Bacteria
MH  - BETA-SUBUNIT
MH  - chloroplast
MH  - Chloroplasts
MH  - COMPLEX
MH  - Electrophoresis
MH  - EPSILON-SUBUNIT
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - F0F1
MH  - F1
MH  - Macromolecular Systems
MH  - membrane
MH  - Membranes
MH  - Mitochondria
MH  - mutant
MH  - RESIDUE
MH  - SUBUNIT
MH  - SYSTEM
MH  - SYSTEMS
RP  - NOT IN FILE
NT  - UI - 86196154LA - engRN - 0 (Codon)RN - 0 (Macromolecular Systems)RN - 0 (Plasmids)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19860619IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:2871027
SO  - J Biol Chem 1986 May 25 ;261(15):7070-7075

719
UI  - 20893
AU  - Noumi T
AU  - Taniai M
AU  - Kanazawa H
AU  - Futai M
TI  - Replacement of arginine 246 by histidine in the beta subunit of Escherichia coli H+-ATPase resulted in loss of multi-site ATPase activity
AB  - A mutant strain KF43 of Escherichia coli defective in the beta subunit of H+-translocating ATPase (F0F1) was examined. In this mutant, replacement of Arg246 by His was identified by DNA sequencing of the mutant gene and confirmed by tryptic peptide mapping. The mutant F1- ATPase was defective in multi-site hydrolysis of ATP but was active in uni-site hydrolysis. Studies on the kinetics of uni-site hydrolysis indicated that the k1 (rate of ATP binding) was similar to that of the wild-type, but the k-1 (rate of release of ATP) could not be measured. The mutant enzyme had a k3 (rate of release of inorganic phosphate) about 15-fold higher than that of the wild-type and showed 3 orders of magnitude lower promotion from uni- to multi-site catalysis. These results suggest that Arg246 or the region in its vicinity is important in multi-site hydrolysis of ATP and is also related to the binding of inorganic phosphate. Reconstitution experiments using isolated subunits suggested that hybrid enzymes (alpha beta gamma complexes) carrying both the mutant and wild-type beta subunits were inactive in multi-site hydrolysis of ATP, supporting the notion that three intact beta subunits are required for activity of the F1 molecule
MH  - A
MH  - ACTIVE
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATPase
MH  - BETA-SUBUNIT
MH  - BINDING
MH  - Catalysis
MH  - COMPLEX
MH  - Enzymes
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0F1
MH  - F1
MH  - F1 ATPASE
MH  - H+-ATPase
MH  - Histidine
MH  - Hydrolysis
MH  - INORGANIC-PHOSPHATE
MH  - Kinetics
MH  - mutant
MH  - Peptide Mapping
MH  - Phosphates
MH  - reconstitution
MH  - SUBUNIT
RP  - NOT IN FILE
NT  - UI - 86250861LA - engRN - 0 (DNA, Bacterial)RN - 0 (Phosphates)RN - 56-65-5 (Adenosine Triphosphate)RN - 71-00-1 (Histidine)RN - 74-79-3 (Arginine)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19860813IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:2873142
SO  - J Biol Chem 1986 Jul 15 ;261(20):9196-9201

720
UI  - 640
AU  - Penefsky HS
TI  - Rate constants and equilibrium constants for the elementary steps of ATP hydrolysis by beef heart mitochondrial ATPase
RP  - NOT IN FILE
NT  - UI - 90173866LA - engRN - 0 (Phosphorus Radioisotopes)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19900330IS - 0076-6879SB - IMCY - UNITED STATESJC - MVA
UR  - PM:2908471
SO  - Methods Enzymol 1986  ;126():608-618

721
UI  - 9914
AU  - Penin F
AU  - Deleage G
AU  - Godinot C
AU  - Gautheron DC
TI  - Efficient reconstitution of mitochondrial energy-transfer reactions from depleted membranes and F1-ATPase as a function of the amount of bound oligomycin sensitivity-conferring protein (OSCP).
AB  - Pig heart mitochondrial membranes depleted of F1 and OSCP by various treatments were analyzed for their content in alpha and beta subunits of F1 and in OSCP using monoclonal antibodies. Membrane treatments and conditions of rebinding of F1 and OSCP were optimized to reconstitute efficient NADH- and ATP-dependent proton fluxes, ATP synthesis and oligomycin-sensitive ATPase activity. F1 and OSCP can be rebound independently to depleted membranes but to avoid unspecific binding of F1 to depleted membranes (ASUA) which is not efficient for ATP synthesis, F1 must be rebound before the addition of OSCP. The rebinding of OSCP to depleted membranes reconstituted with F1 inhibits the ATPase activity of rebound F1, while it restores the ATP-driven proton flux measured by the quenching of ACMA fluorescence. The rebinding of OSCP also renders the ATPase activity of bound F1 sensitive to uncouplers. The rebinding of OSCP alone or F1 alone, does not modify the NADH-dependent proton flux, while the rebinding of both F1 and OSCP controls this flux, inducing an inhibition of the rate of NADH oxidation. Similarly, oligomycin, which seals the F0 channel even in the absence of F1 and OSCP, inhibits the rate of NADH oxidation. OSCP is required to adjust the fitting of F1 to F0 for a correct channelling of protons efficient for ATP synthesis. All reconstituted energy-transfer reactions reach their optimal value for the same amount of OSCP. This amount is consistent with a stoichiometry of two OSCP per F1 in the F0-F1 complex.
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - analysis
MH  - Animal
MH  - atp
MH  - ATP synthesis
MH  - ATPase
MH  - BETA-SUBUNIT
MH  - BINDING
MH  - Carrier Proteins
MH  - COMPLEX
MH  - Energy Transfer
MH  - F0
MH  - F0F1
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - fluorescence
MH  - H(+)-Transporting ATP Synthase
MH  - Hydrolysis
MH  - In Vitro
MH  - Membrane Proteins
MH  - Membranes
MH  - metabolism
MH  - Mitochondria
MH  - MONOCLONAL-ANTIBODIES
MH  - Nad
MH  - Oligomycins
MH  - Oxidation-Reduction
MH  - pharmacology
MH  - physiology
MH  - proton
MH  - Protons
MH  - SUBUNIT
MH  - Support,Non-U.S.Gov't
MH  - Swine
RP  - NOT IN FILE
SO  - Biochim Biophys Acta 1986 Nov 5 ;852(1):55-67

722
UI  - 9915
AU  - Perlin DS
AU  - San FM
AU  - Slayman CW
AU  - Rosen BP
TI  - H+/ATP stoichiometry of proton pumps from Neurospora crassa and Escherichia coli.
AB  - A kinetic method has been used to measure the apparent stoichiometry of H+ ions translocated per ATP split by membrane-bound [H+]-ATPases. In this method, membrane vesicles are suspended in well-buffered medium, ATP is added, and a fluorescent probe of delta pH (acridine orange) is used to detect the formation of a steady-state pH gradient. At the steady state, it is assumed that proton pumping in one direction is exactly balanced by the leak of protons in the opposite direction. The pump is then rapidly turned off by the addition of an appropriate inhibitor, and the initial rate of relaxation of delta pH is used to infer the pump rate. This rate is divided by the rate of ATP hydrolysis, measured under the same condition, to give the apparent H+/ATP stoichiometry. The method has been applied to two different [H+]-ATPases, the plasma-membrane ATPase of Neurospora (a Mr = 100,000 integral membrane protein) and the ATPase of Escherichia coli (which belongs to the F0F1 group). The Neurospora ATPase displayed an apparent stoichiometry close to 1 H+/ATP (0.82-1.23), in agreement with previous estimates from electrophysiological measurements on whole cells. In contrast, the E. coli ATPase yielded an apparent stoichiometry close to 2 H+/ATP (1.90), consistent with several published values obtained by both kinetic and thermodynamic methods for bacterial, mitochondrial, and chloroplast ATPases.
MH  - Acridine Orange
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATPase
MH  - Cell Membrane
MH  - Cells
MH  - chloroplast
MH  - Comparative Study
MH  - DELTA-PH
MH  - enzymology
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0F1
MH  - H(+)-Transporting ATP Synthase
MH  - H+
MH  - Hydrogen-Ion Concentration
MH  - Hydrolysis
MH  - Ion Channels
MH  - Ions
MH  - Kinetics
MH  - membrane vesicles
MH  - metabolism
MH  - Methods
MH  - Neurospora
MH  - Neurospora crassa
MH  - proton
MH  - Proton Pump
MH  - Protons
MH  - Spectrometry,Fluorescence
MH  - Support,Non-U.S.Gov't
MH  - Support,U.S.Gov't,Non-P.H.S.
MH  - Support,U.S.Gov't,P.H.S.
RP  - NOT IN FILE
SO  - Arch Biochem Biophys 1986 Jul ;248(1):53-61

723
UI  - 9916
AU  - Petronilli V
AU  - Pietrobon D
AU  - Zoratti M
AU  - Azzone GF
TI  - Free energy coupling between H+-generating and H+-consuming pumps. Ratio between output and input forces.
AB  - The delta Gp/delta mu H ratio has been measured in mitochondria close to state 4 in the presence of various uncoupler or K+/valinomycin concentrations in media containing either 1 mM or 50 mM Pi. Care has been taken to control the factors affecting delta Gp and delta mu H which could lead to an artefactual increase of the delta Gp/delta mu H ratio above the highest accepted value for the H+/ATP stoichiometry (n = 4, synthesis + transport). In particular, to avoid overestimation of delta Gp due to inactivation of the ATPases at low delta mu H or to the presence of adenylate kinase, the static head state was approached from the side of net ATP synthesis and delta Gp was measured in a state close to static head but still maintaining a residual rate of aerobic phosphorylation. For each concentration of uncoupler or K+, the Pi concentration and/or the adenylate energy charge (EC) as a function of time have been measured as indicators of net ATP synthesis. Only the values of delta Gp measured during a decrease in Pi concentration and/or an increase in EC have been considered to be meaningful for calculations of delta Gp/delta mu H ratios. Both uncouplers and K+ transport cause a marked depression of delta mu H and a parallel depression of the rate of ATP synthesis. However the low rate of ATP synthesis taking place under conditions of low delta mu H eventually results, especially at high Pi concentrations, in a relatively large delta Gp. The delta Gp/delta mu H ratios obtained at the lower delta mu H values exceed 4 and approach 6. Although slightly higher delta Gp/delta mu H ratios are obtained with valinomycin-treated than with uncoupler-treated mitochondria, the pattern of the rise of the force ratio as delta mu H decreases is similar in both cases. An increase of the delta Gp/delta mu H ratio above 4, the maximal accepted H+/ATP stoichiometry is thermodynamically incompatible with the delocalized protonic coupling model.
MH  - Adenosine Triphosphate
MH  - Animal
MH  - atp
MH  - ATP synthesis
MH  - ATPase
MH  - biosynthesis
MH  - Energy Metabolism
MH  - Hydrogen-Ion Concentration
MH  - In Vitro
MH  - metabolism
MH  - Mitochondria
MH  - Mitochondria,Liver
MH  - model
MH  - pharmacology
MH  - Phosphorylation
MH  - Potassium
MH  - Protons
MH  - Rats
MH  - Thermodynamics
MH  - Time
MH  - transport
MH  - Uncoupling Agents
MH  - Valinomycin
RP  - NOT IN FILE
SO  - Eur J Biochem 1986 Mar 3 ;155(2):423-431

724
UI  - 1165
AU  - Roegner M
AU  - Graber P
TI  - Spectroscopic characterization of the ATPase of the thermophilic bacterium PS3 and its isolated subunits
MH  - ATPase
MH  - Bacteria
MH  - PS3
MH  - SUBUNIT
RP  - ON REQUEST (03/18/92)
SO  - J Biochem (Tokyo) 1986  ;99():993-1003

725
UI  - 1164
AU  - Roegner M
AU  - Graber P
AU  - Luecken U
AU  - Tiedge H
AU  - Weber J
AU  - Schaefer G
TI  - Subunit-subunit interactions in TF1 as revealed by ligand binding to isolated and integrated .alpha. and .beta. subunits
MH  - BINDING
MH  - SUBUNIT
RP  - ON REQUEST (03/18/92)
SO  - Biochim Biophys Acta 1986  ;849():121-130

726
UI  - 21015
AU  - Rogner M
AU  - Graber P
TI  - Kinetics of ATP hydrolysis catalyzed by isolated TF1 and reconstituted TF0F1 ATPase
AB  - The rate of ATP hydrolysis catalyzed by isolated TF1 and reconstituted TF0F1 was measured as a function of the ATP concentration in the presence of inhibitors [ADP, Pi and 3'-O-(1-naphthoyl)ATP]. ATP hydrolysis can be described by Michaelis-Menten kinetics with Km(TF1) = 390 microM and Km (TF0F1) = 180 microM. The inhibition constants are for ADP Ki(TF1) = 20 microM and Ki(TF0F1) = 100 microM, for 3'-O-(1- naphthoyl)ATP Ki(TF1) = 150 microM and Ki(TF0F1) = 3 microM, and for Pi Ki(TF1) = 60 mM. From these results it is concluded that upon binding of TF0 to TF1 the mechanism of ATP hydrolysis catalyzed by TF1 is not changed qualitatively; however, the kinetic constants differ quantitatively
MH  - A
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - ADP
MH  - atp
MH  - ATPase
MH  - BINDING
MH  - CONSTANT
MH  - function
MH  - Hydrolysis
MH  - Kinetics
MH  - mechanism
RP  - NOT IN FILE
NT  - UI - 87004633LA - engRN - 0 (Anilino Naphthalenesulfonates)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19861030IS - 0014-2956SB - IMCY - GERMANY, WEST
UR  - PM:2875871
SO  - Eur J Biochem 1986 Sep 1 ;159(2):255-261

727
UI  - 365
AU  - Runswick MJ
AU  - Walker JE
AU  - Gibson BW
AU  - Williams DH
TI  - The frayed N-terminal of the inhibitor protein of bovine mitochondrial F1-ATPase
AB  - The N-terminal region of the bovine mitochondrial F1-ATPase inhibitor protein is frayed. In three independently isolated samples about 62% of chains start at glycine-1. A further 22% of chains start at residue 2, serine, and the remainder at residue 3, glutamic acid. No evidence can be found for alpha-N-formylglycine reported previously. The fraying may be a consequence of proteolytic processing of the precursor of the inhibitor protein during entry into the mitochondrion
RP  - NOT IN FILE
NT  - UI - 86295637LA - engRN - 0 (ATPase inhibitory protein)RN - 0 (Peptide Fragments)RN - 0 (Proteins)RN - EC 3.4.- (Endopeptidases)RN - EC 3.4.22.8 (clostripain)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19860917IS - 0264-6021SB - IMCY - ENGLANDJC - 9YO
UR  - PM:2874795
SO  - Biochem J 1986 Apr 15 ;235(2):515-519

728
UI  - 19891
AU  - Samec Z
AU  - Kharkats YI
AU  - Gurevich YY
TI  - Stochastic approach to the ion transfer kinetics across the interface between two immiscible electrolyte solutions. Comparison with the experimental data
MH  - ion
MH  - Kinetics
MH  - Solutions
MH  - TRANSFER
RP  - IN FILE
SO  - J Electroanal Chem Interfacial Electrochem 1986  ;204():257

729
UI  - 20965
AU  - Schneider E
AU  - Altendorf K
TI  - Proton-conducting portion (F0) from Escherichia coli ATP synthase: preparation, dissociation into subunits, and reconstitution of an active complex
MH  - ACTIVE
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - COLI ATP SYNTHASE
MH  - COMPLEX
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - indicator
MH  - Indicators and Reagents
MH  - Liposomes
MH  - Macromolecular Systems
MH  - Proteolipids
MH  - proteoliposome
MH  - reconstitution
MH  - SUBUNIT
MH  - SYNTHASE
MH  - SYSTEM
MH  - SYSTEMS
RP  - NOT IN FILE
NT  - UI - 90173861LA - engRN - 0 (Indicators and Reagents)RN - 0 (Liposomes)RN - 0 (Macromolecular Systems)RN - 0 (Proteolipids)RN - 0 (proteoliposomes)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19900330IS - 0076-6879SB - IMCY - UNITED STATES
UR  - PM:2908466
SO  - Methods Enzymol 1986  ;126():569-578

730
UI  - 533
AU  - Schwerzmann K
AU  - Pedersen PL
TI  - Regulation of the mitochondrial ATP synthase/ATPase complex
AB  - The mitochondrial ATP synthase/ATPase (F0F1 ATPase) is perhaps the most complex enzyme known. In animal systems it consists of a minimum of 11 different polypeptide chains, 10 (or more) of which appear to be essential for function, and 1 called the "ATPase inhibitor peptide" which is involved in regulation. Recent studies from a variety of laboratories indicate that the ATP synthase/ATPase complex is regulated by several interrelated factors including the thermodynamic poise of the proton gradient across the inner mitochondrial membrane; the ATPase inhibitor peptide; ADP (and/or ADP and Pi); divalent cations; and perhaps the redox state of SH groups on the F1 molecule. The central focus of this review is the ATPase inhibitor peptide. A model involving four distinct conformational states of F1 seems essential to account for the inhibitor's mode of action. The model depicts the ATPase inhibitor protein as acting at the asymmetric center of the F1 moiety. In addition, it accounts for the "unidirectional" role of the inhibitor peptide as a "down regulator" of ATP hydrolysis and for its binding/debinding dependence on the proton motive force and other regulatory factors. Finally, it is suggested that during any physiological process, where there is an energy demand followed by a resting phase, the F1 molecule may follow a "cyclic" path involving the four distinct conformational states of the enzyme
RP  - NOT IN FILE
NT  - UI - 87024594LA - engRN - 0 (ATPase inhibitory protein)RN - 0 (Proteins)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewID - 1-F05 TWO 2755/TW/FICID - CA 10951/CA/NCIDA - 19861114IS - 0003-9861SB - IMCY - UNITED STATESJC - 6SK
UR  - PM:2876680
SO  - Arch Biochem Biophys 1986 Oct ;250(1):1-18

731
UI  - 535
AU  - Schwerzmann K
AU  - Pedersen PL
TI  - Synthesis and use of an azido-labeled form of the ATPase inhibitor peptide of rat liver mitochondria
RP  - NOT IN FILE
NT  - UI - 90173872LA - engRN - 0 (ATPase inhibitory protein)RN - 0 (Affinity Labels)RN - 0 (Azides)RN - 0 (Carbon Radioisotopes)RN - 0 (Cross-Linking Reagents)RN - 0 (Proteins)RN - 53053-08-0 (hydroxysuccinimidyl-4-azidobenzoate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - 1-F05 TWO 2755/TW/FICID - CA 10951/CA/NCIDA - 19900330IS - 0076-6879SB - IMCY - UNITED STATESJC - MVA
UR  - PM:2908475
SO  - Methods Enzymol 1986  ;126():660-666

732
UI  - 19805
AU  - Stempel KE
AU  - Boyer PD
TI  - Refinements in oxygen-18 methodology for the study of phosphorylation mechanisms
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - mechanism
MH  - Oxygen
MH  - Oxygen Isotopes
MH  - Phosphates
MH  - Phosphorylation
RP  - NOT IN FILE
NT  - UI - 90173867LA - engRN - 0 (Oxygen Isotopes)RN - 0 (Phosphates)RN - 56-65-5 (Adenosine Triphosphate)PT - Journal ArticleID - GM11094/GM/NIGMSDA - 19900330IS - 0076-6879SB - IMCY - UNITED STATESJC - MVA
UR  - PM:3272343
SO  - Methods Enzymol 1986  ;126():618-639

733
UI  - 771
AU  - Verburg JG
AU  - Yoshida M
AU  - Allison WS
TI  - The use of dithionite reduction to identify the essential tyrosine residue in the F1-ATPase from the thermophilic bacterium, PS3, that reacts with 7-chloro-4-nitrobenzofurazan
AB  - When the F1-ATPase from the thermophilic bacterium, PS3, was inactivated by greater than 90% with 7-chloro-4-nitro[14C]benzofurazan ([14C]Nbf-Cl) at pH 7.4, 1.4 mol of [14C]Nbf were incorporated per mol of enzyme. After pepsin digestion of the labeled enzyme at pH 3.0, a single, major peak of radioactivity was resolved by reversed-phase high- performance liquid chromatography under acidic conditions were peptidyl Nbf-O-tyrosine is stable. This radioactive peak, designated RP-1, eluted with a retention time of 95 min. When the material in RP-1 was subjected to reversed-phase high-performance liquid chromatography under the same conditions after treatment with sodium dithionite, a single, major peak of radioactivity, designated RP-2, was resolved with a retention time of 52 min. Automatic Edman degradation of this material revealed that it has the amino acid sequence I-Y*-V-P-A-D-(D), where Y* presumably represents peptidyl [14C]Nbf-O-tyrosine. These results provide the basis for a facile method to purify peptides containing [14C]Nbf-O-tyrosine in which the labeled residues can be identified by amino acid sequence analysis using the Edman degradation
RP  - NOT IN FILE
NT  - UI - 86129467LA - engRN - 0 (Oxadiazoles)RN - 0 (Sulfites)RN - 10199-89-0 (4-Chloro-7-nitrobenzofurazan)RN - 14844-07-6 (Dithionite)RN - 55520-40-6 (Tyrosine)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - 2T32 7313-11/PHSID - GM-16974/GM/NIGMSDA - 19860317IS - 0003-9861SB - IMCY - UNITED STATESJC - 6SK
UR  - PM:2868697
SO  - Arch Biochem Biophys 1986 Feb 15 ;245(1):8-13

734
UI  - 828
AU  - Wagner R
AU  - Ponse G
AU  - Strotmann H
TI  - Binding of 2'(3')-O-(2,4,6-trinitrophenyl)-adenosine 5'-diphosphate opens the pathway for protons through the chloroplast ATPase complex
AB  - The effect of 2'(3')-O-(2,4,6-trinitrophenyl)-adenosine 5'-diphosphate (TNP-ADP) on photophosphorylation and on the proton conductivity of the thylakoid membrane has been investigated. The results show that TNP-ADP is a potent competitive inhibitor of photophosphorylation (Ki = 1-2 microM). Moreover, in the absence of ADP and Pi, TNP-ADP accelerates basal electron transport of chloroplasts. Addition of ADP, which promotes release of the analogue from CF1, completely reverses this effect of TNP-ADP; likewise Pi alone reverses stimulation of electron transport by TNP-ADP. Dicyclohexylcarbodiimide treatment, which is known to close CF0 to H+, completely abolishes the effect of TNP-ADP. The measurements of the alkalization of the medium and the acidification of the thylakoid lumen following single turnover flashes showed that binding of TNP-ADP to CF1 increased membrane permeability for H+. Further results suggest that binding of TNP-ADP to the catalytic site of CF1 opens the CF0-CF1 complex for H+. Since ADP, as well as Pi alone, reverses the effect, it is concluded that TNP-ADP induces a conformation of the CF0-CF1 complex similar to the one triggered by simultaneous binding of ADP plus Pi. This may be achieved by interaction of the TNP residue with the Pi binding site. Thus it seems that the status of the catalytic site(s) in CF1 can be transmitted to the CF0 part to control proton flux through the ATPase complex in an economically reasonable way
RP  - NOT IN FILE
NT  - UI - 87054007LA - engRN - 0 (Protons)RN - 58-64-0 (Adenosine Diphosphate)RN - 84430-17-1 (2',3'-(O-(2,4,6-trinitrocyclohexadienylidine))adenosine 5'- diphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19870114IS - 0014-2956SB - IMCY - GERMANY, WESTJC - EMZ
UR  - PM:3023082
SO  - Eur J Biochem 1986 Nov 17 ;161(1):205-209

735
UI  - 910
AU  - Watt AH
AU  - Routledge PA
TI  - Adenosine: an importance beyond ATP
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - Energy Metabolism
MH  - Human
MH  - metabolism
MH  - physiology
RP  - NOT IN FILE
SO  - Br Med J (Clin Res Ed ) 1986 Dec 6 ;293(6560):1455-1456

736
UI  - 536
AU  - Williams N
AU  - Pedersen PL
TI  - Purification of alpha and beta subunits and subunit pairs from rat liver mitochondrial F1-ATPase
RP  - NOT IN FILE
NT  - UI - 90173855LA - engRN - 0 (Indicators and Reagents)RN - 0 (Macromolecular Systems)RN - 0 (Peptides)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - CA 10951/CA/NCIDA - 19900330IS - 0076-6879SB - IMCY - UNITED STATESJC - MVA
UR  - PM:2908462
SO  - Methods Enzymol 1986  ;126():484-489

737
UI  - 537
AU  - Williams N
AU  - Pedersen PL
TI  - Rapid purification of F1-ATPase from rat liver mitochondria using a modified chloroform extraction procedure coupled to high-performance liquid chromatography
RP  - NOT IN FILE
NT  - UI - 90173854LA - engRN - 0 (Indicators and Reagents)RN - 67-66-3 (Chloroform)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - CA 10951/CA/NCIDA - 19900330IS - 0076-6879SB - IMCY - UNITED STATESJC - MVA
UR  - PM:2908461
SO  - Methods Enzymol 1986  ;126():477-484

738
UI  - 954
AU  - Williams RJ
TI  - The connections between particle flow and mechanical, electrical and chemical work
AB  - An analysis is made of the generation of different types of field within biological systems. The fields are interactive being electrical, entropic (concentration gradients), chemical potential or mechanical in character. It is the primary disposition of proteins both in membranes and other organized systems which create the initial pattern of fields, but subsequently the distribution of proteins and the fields are mutually dependent. The value of the patterns is discussed
MH  - Adenosine Triphosphate
MH  - Biological Transport
MH  - Calcium
MH  - Calmodulin
MH  - Diffusion
MH  - Electron Transport
MH  - Energy Metabolism
MH  - Ion Channels
MH  - Membranes
MH  - metabolism
MH  - Oxidation-Reduction
MH  - Phosphoglycerate Kinase
MH  - Protein Conformation
MH  - Proteins
MH  - Protons
MH  - Receptors,Cell Surface
MH  - Thermodynamics
MH  - Troponin
RP  - NOT IN FILE
SO  - J Theor Biol 1986 Jul 7 ;121(1):1-22

739
UI  - 9925
AU  - Wojtczak L
AU  - Zolkiewska A
AU  - Duszynski J
TI  - Energy-storage capacity of the mitochondrial proton-motive force.
AB  - Resting state respiration of rat-liver mitochondria in the presence of oligomycin was rapidly blocked with cyanide and the dissipation of the membrane potential was followed with a tetraphenylphosphonium-sensitive electrode. From the rate of this dissipation and the electric capacitance of the mitochondrial membrane the energy stored in form of the membrane potential was calculated as about 7 microJ/mg protein. In the absence of oligomycin, dissipation of the membrane potential was slower, as it was partly compensated by proton ejection by mitochondrial ATPase hydrolyzing endogenous ATP. This allowed to calculate the total energy storage capacity of the proton-motive force. It amounted to the equivalence of 3.3 nmol ATP/mg protein or about 130 microJ/mg protein. The stoichiometry of proton-pumping ATPase utilizing endogenous ATP was estimated as three protons per molecule ATP.
MH  - Adenine Nucleotides
MH  - Animal
MH  - atp
MH  - ATPase
MH  - Cyanides
MH  - drug effects
MH  - Electron Transport
MH  - Energy Metabolism
MH  - Hydrogen-Ion Concentration
MH  - Membrane Potentials
MH  - Mitochondria
MH  - Mitochondria,Liver
MH  - Oligomycins
MH  - pharmacology
MH  - physiology
MH  - proton
MH  - Proton-Motive Force
MH  - Protons
MH  - Rats
MH  - Respiration
RP  - NOT IN FILE
SO  - Biochim Biophys Acta 1986 Sep 10 ;851(2):313-321

740
UI  - 19804
AU  - Wu D
AU  - Boyer PD
TI  - Bound adenosine 5'-triphosphate formation, bound adenosine 5'- diphosphate and inorganic phosphate retention, and inorganic phosphate oxygen exchange by chloroplast adenosinetriphosphatase in the presence of Ca2+ or Mg2+
AB  - When the heat-activated chloroplast F1 ATPase hydrolyzes [3H, gamma- 32P]ATP, followed by the removal of medium ATP, ADP, and Pi, the enzyme has labeled ATP, ADP, and Pi bound to it in about equal amounts. The total of the bound [3H]ADP and [3H]ATP approaches 1 mol/mol of enzyme. Over a 30-min period, most of the bound [32P]Pi falls off, and the bound [3H]ATP is converted to bound [3H]ADP. Enzyme with such remaining tightly bound ADP will form bound ATP from relatively high concentrations of medium Pi with either Mg2+ or Ca2+ present. The tightly bound ADP is thus at a site that retains a catalytic capacity for slow single-site ATP hydrolysis (or synthesis) and is likely the site that participates in cooperative rapid net ATP hydrolysis. During hydrolysis of 50 microM [3H]ATP in the presence of either Mg2+ or Ca2+, the enzyme has a steady-state level of about one bound [3H]ADP per mole of enzyme. Because bound [3H]ATP is also present, the [3H]ADP is regarded as being present on two cooperating catalytic sites. The formation and levels of bound ATP, ADP, and Pi show that reversal of bound ATP hydrolysis can occur with either Ca2+ or Mg2+ present. They do not reveal why no phosphate oxygen exchange accompanies cleavage of low ATP concentrations with Ca2+ in contrast to Mg2+ with the heat- activated enzyme. Phosphate oxygen exchange does occur with either Mg2+ or Ca2+ present when low ATP concentrations are hydrolyzed with the octyl glucoside activated ATPase. Ligand binding properties of Ca2+ at the catalytic site rather than lack of reversible cleavage of bound ATP may underlie lack of oxygen exchange under some conditions
MH  - A
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - ADP
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BINDING
MH  - Calcium
MH  - chloroplast
MH  - Creatine Kinase
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - H(+)-Transporting ATP Synthase
MH  - Heat
MH  - Hydrolysis
MH  - INORGANIC-PHOSPHATE
MH  - Magnesium
MH  - Oxygen
MH  - Phosphates
MH  - Phosphorus
MH  - Phosphorus Radioisotopes
MH  - Pyruvate Kinase
MH  - SYNTHASE
MH  - synthesis
RP  - NOT IN FILE
NT  - UI - 86269923LA - engRN - 0 (Phosphates)RN - 0 (Phosphorus Radioisotopes)RN - 10028-17-8 (Tritium)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 7439-95-4 (Magnesium)RN - 7440-70-2 (Calcium)RN - EC 2.7.1.40 (Pyruvate Kinase)RN - EC 2.7.3.2 (Creatine Kinase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM 11094/GM/NIGMSDA - 19860916IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:2873834
SO  - Biochemistry 1986 Jun 3 ;25(11):3390-3396

741
UI  - 770
AU  - Yoshida M
AU  - Allison WS
TI  - Characterization of the catalytic and noncatalytic ADP binding sites of the F1-ATPase from the thermophilic bacterium, PS3
AB  - Two classes of ADP binding sites at 20 degrees C have been characterized in the F1-ATPase from the thermophilic bacterium, PS3 (TF1). One class is comprised of three sites which saturate with [3H]ADP in less than 10 s with a Kd of 10 microM which, once filled, exchange rapidly with medium ADP. The binding of ADP to these sites is dependent on Mg2+. [3H]ADP bound to these sites is removed by repeated gel filtrations on centrifuge columns equilibrated with ADP free medium. The other class is comprised of a single site which saturates with [3H]ADP in 30 min with a Kd of 30 microM. [3H]ADP bound to this site does not exchange with medium ADP nor does it dissociate on gel filtration through centrifuge columns equilibrated with ADP free medium. Binding of [3H]ADP to this site is weaker in the presence of Mg2+ where the Kd for ADP is about 100 microM. [3H]ADP dissociated from this site when ATP plus Mg2+ was added to the complex while it remained bound in the presence of ATP alone or in the presence of ADP, Pi, or ADP plus Pi with or without added Mg2+. Significant amounts of ADP in the 1:1 TF1.ADP complex were converted to ATP in the presence of Pi, Mg2+, and 50% dimethyl sulfoxide. Enzyme-bound ATP synthesis was abolished by chemical modification of a specific glutamic acid residue by dicyclohexylcarbodiimide, but not by modification of a specific tyrosine residue with 7-chloro-4-nitrobenzofurazan. Difference circular dichroism spectra revealed that the three Mg2+ -dependent, high affinity ADP binding sites that were not stable to gel filtration were on the alpha subunits and that the single ADP binding site that was stable to gel filtration was on one of the three beta subunits. It has also been demonstrated that enzyme-bound ATP is formed when the TF0.F1 complex containing bound ADP was incubated with Pi, Mg2+, and 50% dimethyl sulfoxide
RP  - NOT IN FILE
NT  - UI - 86195959LA - engRN - 0 (Benzofurans)RN - 10028-17-8 (Tritium)RN - 115491-60-6 (7-chloro-4-nitrobenzofuran)RN - 4213-45-0 (Quinacrine Mustard)RN - 55520-40-6 (Tyrosine)RN - 58-64-0 (Adenosine Diphosphate)RN - 7439-95-4 (Magnesium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM-16974/GM/NIGMSDA - 19860530IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:2871016
SO  - J Biol Chem 1986 May 5 ;261(13):5714-5721

742
UI  - 9926
AU  - Zoratti M
AU  - Favaron M
AU  - Pietrobon D
AU  - Azzone GF
TI  - Intrinsic uncoupling of mitochondrial proton pumps. 1. Non-ohmic conductance cannot account for the nonlinear dependence of static head respiration on delta microH.
AB  - The passive membrane conductance LH1 of rat liver mitochondria has been measured and compared with the quantity nJesh/delta microHsh (n = H+/e stoichiometry; Jesh = rate of electron transfer in static head) over a delta microH range. The two curves approach each other only in the lower part of the range, while they sharply diverge at large values of delta microH. Thus nJesh/delta microHsh cannot be considered to be a measure of LH1 in the upper delta microH region. Only a fraction of the static head electron flow is accounted for by futile proton cycling via leaks. Contaminating open membrane fragments or completely leaky mitochondria can be responsible for only a small part of the residual rate of oxygen consumption. We conclude that a large part of static head respiration must have yet another cause and propose it to be intrinsic uncoupling of the respiratory chain enzymes.
MH  - Animal
MH  - DEPENDENCE
MH  - Electric Conductivity
MH  - electron
MH  - Electron Transport
MH  - Enzymes
MH  - H(+)-Transporting ATP Synthase
MH  - Hydrogen-Ion Concentration
MH  - Kinetics
MH  - Liver
MH  - Membrane Potentials
MH  - metabolism
MH  - Mitochondria
MH  - Mitochondria,Liver
MH  - Oxidative Phosphorylation
MH  - Oxygen
MH  - Oxygen Consumption
MH  - physiology
MH  - proton
MH  - Proton Pump
MH  - Rats
MH  - Respiration
MH  - TRANSFER
RP  - NOT IN FILE
SO  - Biochemistry 1986 Feb 25 ;25(4):760-767

743
UI  - 188
AU  - Aggeler R
AU  - Zhang YZ
AU  - Capaldi RA
AD  - Institute of Molecular Biology, University of Oregon, Eugene 97403
TI  - Labeling of the ATP synthase of Escherichia coli from the head-group region of the lipid bilayer
AB  - The isolated and membrane-bound forms of the adenosinetriphosphatase of Escherichia coli (ECF1 and ECF1F0, respectively) have been reacted with two lysine-specific reagents, sodium hexadecyl 4-[3H]formylphenyl phosphate (HFPP) and sodium methyl 4-[3H]formylphenyl phosphate (MFPP), and with the photoreactive reagent 1,2-[3H]dipalmitoyl-sn-glycerol 3- [[[(4-azido-2-nitrophenyl)amino]ethyl]-phosphate] (arylazidoPE). HFPP and arylazidoPE are amphipathic molecules, inserting by their hexadecyl moieties (one and two chains, respectively) into the lipid bilayer, with the reactive groups intercalated among the phospholipid head groups. MFPP is the water-soluble analogue of HFPP. The labeling patterns of ECF1F0 obtained with HFPP and arylazidoPE were very similar; in both cases the a and b subunits of the F0 part were the most heavily labeled polypeptides of the complex. Models of subunit a, arranged in six transmembrane helices, place most of the lysines in the head-group region, available for reaction with HFPP. Subunits alpha and beta of the ECF1 part were very poorly labeled in comparison to the a and b subunits, together incorporating only 4% as much HFPP and 7.5% as much arylazidoPE as the two F0 subunits together on a protein mass basis. Trypsin cleavage studies localized any labeling of the alpha subunit by arylazidoPE to the N-terminal 15 residues of this polypeptide. When MFPP was used, the alpha and beta subunits were very much more reacted than the F0 subunits. This implies that most of the mass of the alpha and beta subunits in ECF1F0 is above the membrane and not in contact with the bilayer surface.(ABSTRACT TRUNCATED AT 250 WORDS)
RP  - NOT IN FILE
NT  - UI - 88107646LA - engRN - 0 (Azides)RN - 0 (Lipid Bilayers)RN - 0 (Organophosphorus Compounds)RN - 0 (Phosphatidic Acids)RN - 0 (Phosphatidylcholines)RN - 10028-17-8 (Tritium)RN - 110457-62-0 (1,2-dipalmitoyl-sn-glycerol 3-((((4-azido-2- nitrophenyl)amino)ethyl)phosphate))RN - 56-87-1 (Lysine)RN - 99281-05-7 (hexadecyl 4-formylphenyl phosphate)RN - 99281-07-9 (methyl 4-formylphenyl phosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - HL-22050/HL/NHLBIDA - 19880318IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:2892529
SO  - Biochemistry 1987 Nov 3 ;26(22):7107-7113

744
UI  - 9975
AU  - Arata H
AU  - Takenaka I
AU  - Nishimura M
TI  - Flash-induced proton release in Rhodopseudomonas sphaeroides spaeroplasts
MH  - cytochrome
MH  - midpoint potential
MH  - proton
MH  - proton release
MH  - redox
MH  - rhodopseudomonas
RP  - IN FILE
NT  - K
SO  - J Biochem 1987  ;101():261-265

745
UI  - 9896
AU  - Beavis AD
TI  - Upper and lower limits of the charge translocation stoichiometry of mitochondrial electron transport.
AB  - The upper and lower limits of the mechanistic stoichiometry (n) of electric charge translocation coupled to mitochondrial electron transport have been determined for the oxidation of succinate and beta-hydroxybutyrate using a recently described method (Beavis, A. D., and Lehninger, A. L. (1986) Eur. J. Biochem. 158, 307-314). This method requires no assumptions regarding the magnitude of proton leakage or pump slippage, but it takes advantage of the ability to predict the direction of change as the coupled fluxes are modulated by specific means. In this study, the rates of K+ uptake (JK) and O2 consumption (JO) were determined from simultaneous electrode measurements in the presence of various concentrations of valinomycin or inhibitors of electron flow. When valinomycin is varied, the rate of proton leakage or pump slippage should decrease as JO increases, with the result that the slope dJK/dJO will be greater than n. On the other hand, when an inhibitor of electron flow is varied, the rate of proton leakage or pump slippage should increase as JO increases, with the result that the slope dJK/dJO should be less than n. The data obtained using this approach indicate that n lies between 6.7 and 7.3 for succinate oxidation and between 10.2 and 11.7 for beta-hydroxybutyrate (or NADH) oxidation. It is concluded that the mechanistic stoichiometry of charge separation coupled to electron flow is 7 q+/O in the span from succinate to oxygen and 11 q+/O in the span from NADH to oxygen. These conclusions are fully consistent with the limits of the mechanistic ATP/O ratios previously determined for these spans (Beavis, A. D., and Lehninger, A. L. (1986) Eur. J. Biochem. 158, 315-322).
MH  - Animal
MH  - electron
MH  - Electron Transport
MH  - Hydroxybutyrates
MH  - Kinetics
MH  - Malonates
MH  - Mathematics
MH  - metabolism
MH  - Mitochondria,Liver
MH  - Nad
MH  - Oxygen
MH  - Oxygen Consumption
MH  - pharmacology
MH  - Potassium
MH  - proton
MH  - Rats
MH  - succinate
MH  - Support,U.S.Gov't,P.H.S.
MH  - transport
MH  - Valinomycin
RP  - NOT IN FILE
SO  - J Biol Chem 1987 May 5 ;262(13):6165-6173

746
UI  - 19796
AU  - Boyer PD
AD  - Molecular Biology Institute, University of California, Los Angeles 90024-1570
TI  - The unusual enzymology of ATP synthase
MH  - atp
MH  - ATP synthase
MH  - enzymology
MH  - H(+)-Transporting ATP Synthase
MH  - Macromolecular Systems
MH  - review
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 88163519LA - engRN - 0 (Macromolecular Systems)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 19880506IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:2894841
SO  - Biochemistry 1987 Dec 29 ;26(26):8503-8507

747
UI  - 21169
AU  - Brusilow WS
AD  - Department of Chemistry and Biochemistry, University of Maryland, College Park 20742
TI  - Proton leakiness caused by cloned genes for the F0 sector of the proton- translocating ATPase of Escherichia coli: requirement for F1 genes
AB  - To study expression of uncG, the gene coding for the gamma subunit of the Escherichia coli proton-translocating ATPase, deletions were made in the intergenic region between uncA, the gene coding for the alpha subunit, and uncG. Two deletions which fused uncA and uncG coded for alpha-gamma fusion polypeptides which were synthesized well both in vitro and in vivo, demonstrating that uncG expression is normally controlled by nucleotides in the intergenic region. Multicopy plasmids carrying these fusion genes and the genes for the other subunits of the ATPase had a harmful effect on the growth of E. coli. The effect was overcome by N,N'-dicyclohexylcarbodiimide, indicating that the cells probably leaked protons. The deleterious effect was eliminated by making a nonpolar deletion in the upstream F0 gene uncB, or by cloning each of the uncA-uncG fusion genes onto a separate plasmid, removed from the F0 genes, thus demonstrating that the fusion genes were not primarily responsible for the proton permeability. A plasmid which carried F0 genes and the gene for the delta subunit caused deleterious proton leakiness in unc+ cells but not in cells from which the unc operon was deleted. The proton leakiness caused by these different plasmids was therefore due to the production of a leaky F0 proton channel and required the presence of F1 genes. The results support a model for ATPase assembly in which F1 genes or polypeptides are involved in the formation or opening of the F0 proton channel
MH  - A
MH  - alpha
MH  - ALPHA-SUBUNIT
MH  - ATPase
MH  - Biochemistry
MH  - Cells
MH  - Chemistry
MH  - delta
MH  - DELTA-SUBUNIT
MH  - Enzymes
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - F1
MH  - In Vitro
MH  - model
MH  - Nucleotides
MH  - Permeability
MH  - proton
MH  - Proton-Translocating ATPases
MH  - Protons
MH  - SUBUNIT
RP  - NOT IN FILE
NT  - UI - 88032814LA - engRN - 0 (Plasmids)RN - 0 (Protons)RN - EC 3.1.21 (DNA Restriction Enzymes)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleID - AI20010/AI/NIAIDDA - 19871209IS - 0021-9193SB - IMCY - UNITED STATES
UR  - PM:2889719
SO  - J Bacteriol 1987 Nov ;169(11):4984-4990

748
UI  - 765
AU  - Bullough DA
AU  - Verburg JG
AU  - Yoshida M
AU  - Allison WS
TI  - Evidence for functional heterogeneity among the catalytic sites of the bovine heart mitochondrial F1-ATPase
AB  - The characteristics of ATP hydrolysis at a single catalytic site of the bovine heart F1-ATPase (MF1) as originally described by Grubmeyer et al. (Grubmeyer, C., Cross, R.L., and Penefsky, H.S. (1982) J. Biol. Chem. 257, 12092-12100) were compared with those of various chemically modified preparations of MF1 in which the steady state activity was severely attenuated. Although it was not necessary to age our preparations of native MF1 in the presence of 2 mM Pi to observe the same characteristics of single site catalysis, such aging did shift the equilibrium of bound substrate and bound products at the single catalytic site in favor of ATP. After loading a single catalytic site on the enzyme with substoichiometric [alpha,gamma-32P]ATP, the addition of 5-20 microM ATP or ADP was effective in promoting both the hydrolysis of bound [alpha,gamma-32P]ATP and release of radioactive products. Under these conditions, the 5-20 microM ATP added as promoter was hydrolyzed at a rate commensurate with the turnover rate of the enzyme, whereas the promoted hydrolysis of the [alpha,gamma-32P]ATP, preloaded at a single catalytic site, was considerably slower. Therefore, the high affinity, single catalytic site loaded first does not directly contribute to steady state ATP hydrolysis. That the single, high affinity catalytic site is not a "normal" catalytic site is supported by the properties of enzyme modified by 5'-p- fluorosulfonylbenzoyladenosine which exhibits only slightly altered characteristics of single site catalysis and promoted single site catalysis, despite exhibiting severely attenuated steady state turnover. Other modified forms of the enzyme in which the steady state activity was severely attenuated by derivatization with 5'-p- fluorosulfonylbenzoylinosine, 7-chloro-4-nitrobenzofurazan, or 1,5- difluoro-2,4-dinitrobenzene also bound substoichiometric ATP at a single catalytic site. However, the characteristics of single site hydrolysis by these modified forms of the enzyme differed considerably from those of native MF1
RP  - NOT IN FILE
NT  - UI - 87308141LA - engRN - 0 (5'-(4-fluorosulfonylbenzoyl)adenosine)RN - 0 (Affinity Labels)RN - 0 (Benzofurans)RN - 115491-60-6 (7-chloro-4-nitrobenzofuran)RN - 327-92-4 (1,5-difluoro-2,4-dinitrobenzene)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-61-7 (Adenosine)RN - 58-63-9 (Inosine)RN - 70-34-8 (Dinitrofluorobenzene)RN - 83133-70-4 (5'-(4-fluorosulfonylbenzoyl)inosine)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM-16974/GM/NIGMSDA - 19870930IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:2887560
SO  - J Biol Chem 1987 Aug 25 ;262(24):11675-11683

749
UI  - 465
AU  - Cox GB
AU  - Hatch L
AU  - Webb D
AU  - Fimmel AL
AU  - Lin ZH
AU  - Senior AE
AU  - Gibson F
TI  - Amino acid substitutions in the epsilon-subunit of the F1F0-ATPase of Escherichia coli
AB  - A mutant strain of Escherichia coli was isolated in which Gly-48 of the mature epsilon-subunit of the energy-transducing adenosine triphosphatase was replaced by Asp. This amino acid substitution caused inhibition of ATPase activity (about 70%), loss of ATP-dependent proton translocation and lowered oxidative phosphorylation, but did not affect proton translocation through the F0. Purified F1-ATPase from the mutant strain bound to stripped membranes with the same affinity as the normal F1-ATPase. Partial revertant strains were isolated in which Pro-47 of the epsilon-subunit was replaced by Ser or Thr. Pro-47 and Gly-48 are predicted to be residues 2 and 3 in a Type II beta-turn and the Gly-48 to Asp substitution is predicted to cause a change from a Type II to a Type I or III beta-turn. Space-filling models of the beta-turn (residues 46-49) in the normal, mutant and partial revertant epsilon- subunits indicate that the peptide oxygen between Pro-47 and Gly-48 is in a different position to the peptide oxygen between Pro-47 and Asp-48 and that the substitution of Pro-47 by either Ser or Thr restores an oxygen close to the original position. It is suggested that the peptide oxygen between Pro-47 and Gly-48 of the epsilon-subunit is involved either structurally in inter-subunit H-bonding or directly in proton movements through the F1-ATPase
RP  - NOT IN FILE
NT  - UI - 87101121LA - engRN - 0 (Amino Acids)RN - 0 (DNA, Bacterial)RN - 0 (Macromolecular Systems)RN - 0 (Succinates)RN - 110-15-6 (Succinic Acid)RN - 83-89-6 (Quinacrine)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19870318IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:2879566
SO  - Biochim Biophys Acta 1987 Feb 11 ;890(2):195-204

750
UI  - 363
AU  - Cozens AL
AU  - Walker JE
TI  - The organization and sequence of the genes for ATP synthase subunits in the cyanobacterium Synechococcus 6301. Support for an endosymbiotic origin of chloroplasts
AB  - The nucleotide sequence has been determined of two regions of DNA cloned from the cyanobacterium Synechococcus 6301. The larger, 8890 base-pairs in length, contains a cluster of seven genes for subunits of ATP synthase. The order of the genes is a:c:b':b:delta:alpha:gamma, b' being a duplicated and diverged form of b. As in the Escherichia coli unc operon, the a gene is preceded by a gene for a small hydrophobic and basic protein. The hydrophobic profile of the potential gene product suggests that its secondary structure is similar to the uncI protein. The smaller DNA fragment, 4737 base-pairs in length, is separated from the larger by at least 15 X 10(3) base-pairs of DNA. It contains a cluster of two genes encoding ATP synthase subunits beta and epsilon. Both clusters of ATP synthase genes are preceded by sequences resembling the -10 (Pribnow) box of E. coli promoters and are followed by sequences able to form stable stem-loop structures that might serve to terminate transcription. These features and the small intergenic non- coding sequences suggest that the clusters are operons, for which the names atp1 and atp2 are proposed. The order of genes within the two clusters is very similar to the gene order in the E. coli unc operon. However, it is most closely related to the arrangement of genes for ATP synthetase subunits a:c:b:alpha and beta:epsilon in two clusters in pea chloroplast DNA. This close relationship between chloroplasts and the cyanobacterium is also evident from comparisons of the sequences of ATP synthase subunits; the Synechococcus proteins are much more closely related to chloroplast homologues than to those in other bacteria or in mitochondria. It is further supported by the cyanobacterial b and b' proteins which, in common with their chloroplast counterpart, subunit I, have extra amino-terminal extensions relative to the E. coli b protein. This extension is known to be removed by post-translational processing in the chloroplast, but its function is obscure. It also seems likely that the cyanobacterial and chloroplast ATP synthases have important similarities in subunit composition. For example, the presence of two related genes, b and b', in the cyanobacterium suggests that its ATP synthase is a complex of nine polypeptides, and that it may have single copies of related b and b' proteins rather than two copies of identical b subunits as found in the E. coli enzyme.4+off
RP  - NOT IN FILE
NT  - UI - 87311713LA - engRN - 0 (DNA, Bacterial)RN - 0 (Macromolecular Systems)RN - 0 (Multienzyme Complexes)RN - EC 2.7 (Phosphotransferases)RN - EC 2.7.4.- (ATP synthetase complex)PT - Journal ArticleDA - 19871009IS - 0022-2836SB - IMCY - ENGLANDJC - J6V
UR  - PM:3041005
SO  - J Mol Biol 1987 Apr 5 ;194(3):359-383

751
UI  - 19799
AU  - Cross RL
AU  - Cunningham D
AU  - Miller CG
AU  - Xue ZX
AU  - Zhou JM
AU  - Boyer PD
TI  - Adenine nucleotide binding sites on beef heart F1 ATPase: photoaffinity labeling of beta-subunit Tyr-368 at a noncatalytic site and beta Tyr- 345 at a catalytic site
AB  - 2-Azidoadenine [32P]nucleotide was bound specifically at catalytic or noncatalytic nucleotide binding sites on beef heart mitochondrial F1 ATPase. In both cases, photolysis resulted in nearly exclusive labeling of the beta subunit. The modified enzyme was digested with trypsin, and labeled peptides were purified by reversed-phase high-pressure liquid chromatography. Amino acid sequence analysis of the major 32P-labeled tryptic fragments showed beta-subunit Tyr-368 to be present at noncatalytic sites and beta Tyr-345 to be present at catalytic sites. From the relationship between the degree of inhibition and extent of modification, it is estimated that one-third of the catalytic sites or two-thirds of the noncatalytic sites must be modified to give near- complete inhibition of catalytic activity
MH  - A
MH  - Adenine Nucleotides
MH  - ADENINE-NUCLEOTIDES
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Amino Acid Sequence
MH  - analysis
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BETA-SUBUNIT
MH  - BINDING
MH  - Binding Sites
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - H(+)-Transporting ATP Synthase
MH  - NONCATALYTIC SITES
MH  - nucleotide binding
MH  - nucleotide binding sites
MH  - NUCLEOTIDE-BINDING-SITES
MH  - Nucleotides
MH  - SUBUNIT
MH  - SYNTHASE
MH  - Trypsin
RP  - NOT IN FILE
NT  - UI - 87289675LA - engRN - 0 (Adenine Nucleotides)RN - 0 (Affinity Labels)RN - 55520-40-6 (Tyrosine)RN - 58-64-0 (Adenosine Diphosphate)RN - 64020-53-7 (2-azidoadenosine 3',5'-diphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM07165/GM/NIGMSID - GM11094/GM/NIGMSID - GM23152/GM/NIGMSDA - 19870918IS - 0027-8424SB - IMCY - UNITED STATESJC - PV3
UR  - PM:2886991
SO  - Proc Natl Acad Sci U S A 1987 Aug ;84(16):5715-5719

752
UI  - 20960
AU  - Deckers-Hebestreit G
AU  - Schmid R
AU  - Kiltz HH
AU  - Altendorf K
AD  - Fachbereich Biologie/Chemie, Mikrobiologie, Universitat Osnabruck, FRG
TI  - F0 portion of Escherichia coli ATP synthase: orientation of subunit c in the membrane
AB  - Incubation of right-side-out oriented membrane vesicles of Escherichia coli with tetranitromethane resulted in the nitration of tyrosine residues (Tyr-10 and Tyr-73) of subunit c from the ATP synthase. Cleavage of the protein with cyanogen bromide and separation of the resulting fragments, especially of the tyrosine-containing peptides, clearly demonstrated that the distribution of the nitro groups is similar at any time and at any pH value chosen for the analysis. Furthermore, the percentage of 3-nitrotyrosine present in the two peptide fragments was in good agreement with that obtained for the intact polypeptide chain. While the modification of the tyrosine residues in subunit c with the lipophilic tetranitromethane is independent of the orientation of the membrane vesicles, the subsequent partial conversion of the 3-nitrotyrosine to the amino form only occurred when membrane vesicles with right-side-out orientation were treated with the ionic, water-soluble sodium dithionite, which at certain concentrations cannot penetrate biological membranes. Cleavage of subunit c isolated from nitrated and subsequently reduced membrane vesicles and separation of the resulting fragments by high-pressure liquid chromatography showed that the 3-nitrotyrosine in the Tyr-73- containing peptides has been completely reduced, while the nitro group in peptides containing Tyr-10 remained nearly unaffected
MH  - analysis
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - COLI ATP SYNTHASE
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - LIQUID
MH  - Macromolecular Systems
MH  - membrane
MH  - membrane vesicles
MH  - Membranes
MH  - Peptide Fragments
MH  - pH
MH  - protein
MH  - RESIDUE
MH  - Sodium
MH  - SUBUNIT
MH  - SYNTHASE
MH  - SYSTEM
MH  - SYSTEMS
MH  - Time
RP  - NOT IN FILE
NT  - UI - 88050833LA - engRN - 0 (Macromolecular Systems)RN - 0 (Peptide Fragments)RN - 506-68-3 (Cyanogen Bromide)RN - 509-14-8 (Tetranitromethane)RN - 55520-40-6 (Tyrosine)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19871228IS - 0006-2960SB - IMCY - UNITED STATES
UR  - PM:2890375
SO  - Biochemistry 1987 Aug 25 ;26(17):5486-5492

753
UI  - 20889
AU  - Futai M
AU  - Shimomura S
AU  - Maeda M
TI  - Binding of Mg2+ to the beta subunit or F1 of H+-ATPase from Escherichia coli
AB  - The bindings of Mg2+ to the F1 portion of Escherichia coli H+-ATPase and its isolated alpha and beta subunits were studied with 8- anilinonaphthalene-1-sulfonate (ANS). The fluorescence of ANS increased upon addition of F1 or its alpha subunit or beta subunit, as reported previously (M. Hirano, K. Takeda, H. Kanazawa, and M. Futai (1984) Biochemistry 23, 1652-1656). The fluorescence of ANS bound to F1 or its beta subunit increased significantly with further addition of Mg2+, whereas that of the alpha subunit increased only slightly. Ca2+ and Mn2+ had similar effects on the fluorescence of ANS with F1 and its beta subunit. The Mg2+-induced fluorescence enhancement (delta F) was high at an alkaline pH and was lowered by addition of ethylenediaminetetraacetic acid. Dicyclohexylcarbodiimide and azide had no effect on the delta F. Binding analysis showed that the concentration dependence of Mg2+ on the fluorescence enhancement of the beta subunit is similar to that of F1. These results suggest that both the beta subunit and F1 have binding sites for Mg2+ and that the delta F observed with F1 may be due to the binding of Mg2+ to the beta subunit
MH  - ACID
MH  - ALPHA-SUBUNIT
MH  - analysis
MH  - ANS
MH  - ATPase
MH  - BETA-SUBUNIT
MH  - BINDING
MH  - Binding Sites
MH  - Biochemistry
MH  - DEPENDENCE
MH  - Dicyclohexylcarbodiimide
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1
MH  - fluorescence
MH  - H+-ATPase
MH  - M
MH  - Magnesium
MH  - pH
MH  - Site
MH  - SUBUNIT
RP  - NOT IN FILE
NT  - UI - 87212004LA - engRN - 0 (Anilino Naphthalenesulfonates)RN - 7439-95-4 (Magnesium)RN - 82-76-8 (1-anilino-8-naphthalenesulfonate)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19870529IS - 0003-9861SB - IMCY - UNITED STATES
UR  - PM:2883932
SO  - Arch Biochem Biophys 1987 Apr ;254(1):313-318

754
UI  - 20891
AU  - Futai M
AU  - Noumi T
AU  - Maeda M
AD  - Department of Organic Chemistry and Biochemistry, Osaka University, Japan
TI  - Molecular biological studies on structure and mechanism of proton translocating ATPase (H+-ATPase, F0F1)
AB  - Recent results on ATPase, mainly from E. coli, obtained by biochemical and molecular biological approaches are reviewed, with special emphasis on results obtained in this laboratory. The advantages of using E. coli in studies of this important enzyme in oxidative phosphorylation are indicated: variant enzymes with specific amino acid replacements can be obtained and their functions and structures can be compared with those of the wild-type enzyme. Structural aspects of this complex enzyme are discussed, including the primary amino acid sequences and molecular assembly of subunits, and mechanistic aspects of the catalytic mechanism and proton translocation
MH  - ACID
MH  - Amino Acid Sequence
MH  - ATPase
MH  - Biochemistry
MH  - Chemistry
MH  - COMPLEX
MH  - Enzymes
MH  - F0F1
MH  - function
MH  - H+-ATPase
MH  - mechanism
MH  - Oxidative Phosphorylation
MH  - Phosphorylation
MH  - proton
MH  - review
MH  - SUBUNIT
MH  - translocation
RP  - NOT IN FILE
NT  - UI - 88147515LA - engRN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticlePT - ReviewPT - Review, AcademicDA - 19880328IS - 0065-227XSB - IMCY - IRELAND
UR  - PM:2894114
SO  - Adv Biophys 1987  ;23():1-37

755
UI  - 189
AU  - Gogol EP
AU  - Lucken U
AU  - Capaldi RA
TI  - The stalk connecting the F1 and F0 domains of ATP synthase visualized by electron microscopy of unstained specimens
AB  - E. coli F1F0 ATP synthase has been reconstituted into membranes and visualized by electron microscopy of unstained samples preserved in thin layers of amorphous ice. Unlike previous observations in negative stain, these specimens are not exposed to potentially denaturing or perturbing conditions, having been rapidly frozen from well-defined conditions in which the enzyme is fully active. The structures visualized in views normal to the lipid bilayer clearly show the presence of a narrow stalk approx. 45 A long, connecting the F1 to the membrane-embedded F0
RP  - NOT IN FILE
NT  - UI - 87276499LA - engRN - 0 (Lipid Bilayers)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - HL24526/HL/NHLBIDA - 19870903IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:2886365
SO  - FEBS Lett 1987 Jul 27 ;219(2):274-278

756
UI  - 1145
AU  - Graber P
TI  - Primary charge separation and energy transduction in photosynthesis
MH  - Photosynthesis
RP  - ON REQUEST (03/18/92)
SO  - Ettore Majorana Int Sci Ser : Phys Sci 1987  ;32():379-429

757
UI  - 1144
AU  - Graber P
TI  - Regulation of proton-transport coupled ATP synthesis by the chloroplast ATPase
MH  - atp
MH  - ATP synthesis
MH  - ATPase
MH  - chloroplast
MH  - regulation
MH  - synthesis
T2  - Perspect. Biol. Energy Transduction, [Pap. Yamada Conf. Energy Transduction ATPases], 11th, Meeting Date 1985, 415-22. Edited by: Mukohata, Yasuo; Morales, Manuel F.; Fleischer, Sidney. Academic: Tokyo, Japan
Y2  - -32676  
RP  - ON REQUEST (03/18/92)
SO  -  1987  ;():

758
UI  - 1154
AU  - Graber P
AU  - Junesch U
AU  - Thulke G
TI  - The chloroplast ATP-synthase: the rate of the catalytic reaction
MH  - ATP synthase
MH  - chloroplast
T2  - Prog. Photosynth. Res., Proc. Int. Congr. Photosynth., 7th, Meeting Date 1986, Volume 3, 177-84. Edited by: Biggins, John. Nijhoff: Dordrecht, Neth
Y2  - -32676  
RP  - ON REQUEST (03/18/92)
SO  -  1987  ;():

759
UI  - 974
AU  - Grant WD
TI  - The enigma of the alkaliphile
AB  - Alkaliphilic bacteria provide a model system in which to test the basic principles of the chemiosmotic theory. The proton-motive force (delta mu H +) in these bacteria under alkaline conditions appears to be too low to power ATP synthesis assuming normal stoichiometry and a normal H+/F1F0 ATPase. Current data suggest that these bacteria might obviate the problems by harnessing a sodium-motive force (delta mu Na+)
MH  - Adenosine Triphosphate
MH  - Alkalies
MH  - Bacteria
MH  - biosynthesis
MH  - Energy Metabolism
MH  - Hydrogen-Ion Concentration
MH  - Intracellular Fluid
MH  - isolation &amp
MH  - metabolism
MH  - purification
MH  - Sodium
RP  - NOT IN FILE
NT  - Department of Microbiology, University of Leicester, UK
SO  - Microbiol Sci 1987 Aug ;4(8):251-255

760
UI  - 21092
AU  - Hicks DB
AU  - Krulwich TA
AD  - Department of Biochemistry, Mount Sinai School of Medicine, City University of New York, New York 10029
TI  - Purification and characterization of the F1 ATPase from Bacillus subtilis and its uncoupler-resistant mutant derivatives
AB  - The F1 ATPase of Bacillus subtilis BD99 was extracted from everted membrane vesicles by low-ionic-strength treatment and purified by DEAE- cellulose chromatography, hydrophobic interaction chromatography, and anion-exchange high-performance liquid chromatography. The subunit structure of the enzyme was examined by sodium dodecyl sulfate- polyacrylamide gel electrophoresis in the absence and presence of urea. In the absence of urea, the alpha and beta subunits comigrated and the ATPase was resolved into four bands. The mobility of the beta subunit, identified by immunoblotting with anti-beta from Escherichia coli F1, was altered dramatically by the presence of urea, causing it to migrate more slowly than the alpha subunit. The catalytic activity of the ATPase was strongly metal dependent; in the absence of effectors, the Ca2+-ATPase activity was 15- to 20-fold higher than the Mg2+ -ATPase activity. On the other hand, sulfite anion, methanol, and optimally, octylglucoside stimulated the Mg2+ -ATPase activity up to twice the level of Ca2+ -ATPase activity (specific activity, about 80 mumol of Pi per min per mg of protein). The F1 ATPase was also isolated from mutants of B. subtilis that had been isolated and characterized in this laboratory by their ability to grow in the presence of protonophores. The specific activities of the ATPase preparations from the mutant and the wild type were very similar for both Mg2+- and Ca2+ -dependent activities. Kinetic parameters (Vmax and Km for Mg-ATP) for octylglucoside-stimulated Mg2+ -ATPase activity were similar in both preparations.(ABSTRACT TRUNCATED AT 250 WORDS)
MH  - alpha
MH  - ALPHA-SUBUNIT
MH  - ATPase
MH  - Bacillus
MH  - BETA
MH  - BETA-SUBUNIT
MH  - Biochemistry
MH  - Carbonyl Cyanide m-Chlorophenyl Hydrazone
MH  - Electrophoresis
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - LIQUID
MH  - membrane
MH  - membrane vesicles
MH  - Methanol
MH  - mutant
MH  - protein
MH  - Proton-Translocating ATPases
MH  - protonophore
MH  - purification
MH  - Sodium
MH  - structure
MH  - SUBUNIT
MH  - vesicles
RP  - NOT IN FILE
NT  - UI - 88007423LA - engRN - 555-60-2 (Carbonyl Cyanide m-Chlorophenyl Hydrazone)RN - 57-13-6 (Urea)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleID - GM-28454/GM/NIGMSDA - 19871105IS - 0021-9193SB - IMCY - UNITED STATES
UR  - PM:2888751
SO  - J Bacteriol 1987 Oct ;169(10):4743-4749

761
UI  - 20887
AU  - Hsu SY
AU  - Noumi T
AU  - Takeyama M
AU  - Maeda M
AU  - Ishibashi S
AU  - Futai M
TI  - Beta-subunit of Escherichia coli F1-ATPase. An amino acid replacement within a conserved sequence (G-X-X-X-X-G-K-T/S) of nucleotide-binding proteins
AB  - A mutant strain KF87 of E. coli with a defective beta-subunit (Ala-151-- --Val) of F1-ATPase was isolated. The mutation is within the conserved sequence (G-X-X-X-X-G-K-T/S) of nucleotide-binding proteins. The mutant F1-ATPase had a much higher rate of uni-site hydrolysis of ATP than the wild type, and about 6% of the wild-type multi-site activity. The mutant enzyme showed defective transmission of conformational change(s) between the ligand- and aurovertin-binding sites
MH  - A
MH  - ACID
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATPase
MH  - Aurovertins
MH  - BETA-SUBUNIT
MH  - BINDING
MH  - Carrier Proteins
MH  - COLI F1 ATPASE
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - Hydrolysis
MH  - mutant
MH  - nucleotide binding
MH  - Phosphates
MH  - protein
MH  - Proteins
MH  - Site
RP  - NOT IN FILE
NT  - UI - 87247274LA - engRN - 0 (Aurovertins)RN - 0 (Carrier Proteins)RN - 0 (DNA, Recombinant)RN - 0 (Phosphates)RN - 0 (adenosine cyclic-3',5'-monophosphate binding proteins)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19870810IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:2885226
SO  - FEBS Lett 1987 Jun 29 ;218(2):222-226

762
UI  - 1153
AU  - Junesch U
AU  - Graber P
TI  - Influence of the redox state and the activation of the chloroplast ATP synthase on proton-transport-coupled ATP synthesis/hydrolysis
MH  - ACTIVATION
MH  - atp
MH  - ATP synthase
MH  - chloroplast
MH  - redox
MH  - SYNTHASE
RP  - ON REQUEST (03/18/92)
SO  - Biochim Biophys Acta 1987  ;893():275-288

763
UI  - 1155
AU  - Junesch U
AU  - Graber P
TI  - The activation of the reduced chloroplast ATP-synthase by .DELTA.pH
MH  - ACTIVATION
MH  - ATP synthase
MH  - chloroplast
T2  - Prog. Photosynth. Res., Proc. Int. Congr. Photosynth., 7th, Meeting Date 1986, Volume 3, 173-6. Edited by: Biggins, John. Nijhoff: Dordrecht, Neth
Y2  - -32676  
RP  - ON REQUEST (03/18/92)
SO  -  1987  ;():

764
UI  - 20829
AU  - Junge W
AU  - McLaughlin S
TI  - The role of fixed and mobile buffers in the kinetics of proton movement
MH  - buffer
MH  - Buffers
MH  - Diffusion
MH  - Kinetics
MH  - Movement
MH  - proton
MH  - SURFACE
MH  - theory
RP  - NOT IN FILE
SO  - Biochim Biophys Acta 1987  ;890():1-5

765
UI  - 8269
AU  - Junge W
TI  - Complete tracking of transient proton flow through active chloroplast ATP synthase
MH  - ACTIVE
MH  - atp
MH  - ATP synthase
MH  - chloroplast
MH  - proton
MH  - SYNTHASE
RP  - IN FILE
NT  - G (Protonen)
SO  - Proc Natl Acad Sci USA 1987  ;84():7084-7088

766
UI  - 19803
AU  - Kandpal RP
AU  - Boyer PD
TI  - Escherichia coli F1 ATPase is reversibly inhibited by intra- and intersubunit crosslinking: an approach to assess rotational catalysis
AB  - Reaction of the multisubunit F1 ATPase from Escherichia coli (EF1) with a bifunctional cleavable crosslinker, 3,3'- dithiobis(succinimidylpropionate) (DSP), has been used to explore the possibility that during catalysis a rotational movement of catalytic subunits relative to noncatalytic subunits occurs. The premise is that such rotational catalysis is tenable if intersubunit crosslinking of a major subunit with one of the minor subunits inhibits the enzyme activity and if upon cleavage of the crosslinks, the enzyme regains activity. The results presented in this paper show that crosslinking of about 5-6 reactive groups on EF1 with DSP is accompanied by a loss of 2/3 of the enzyme activity. Both intra- and intersubunit crosslinks are formed. The most prominent intersubunit crosslinks are those of gamma and delta subunits with the alpha subunit. Nearly complete recovery of activity can be attained by cleaving the disulfide bond in the crosslinker with dithiothreitol. Because the chemical modification of enzyme groups remains after the crosslinker is cleaved, the loss in activity before cleavage can be ascribed to conformational restraints. The results show that catalysis by the EF1 ATPase is highly sensitive to the restrictions of crosslinking, and are consistent with the view that catalysis is accompanied by appreciable movements of the major subunits with respect to the minor subunits, as suggested for rotational catalysis
MH  - A
MH  - ALPHA-SUBUNIT
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Catalysis
MH  - COLI F1 ATPASE
MH  - CROSS-LINKING
MH  - DELTA-SUBUNIT
MH  - Dithiothreitol
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - H(+)-Transporting ATP Synthase
MH  - Macromolecular Systems
MH  - Movement
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 87101118LA - engRN - 0 (Cross-Linking Reagents)RN - 0 (Macromolecular Systems)RN - 0 (Succinimides)RN - 3483-12-3 (Dithiothreitol)RN - 57757-57-0 (dithiobis(succinimidylpropionate))RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM 11094/GM/NIGMSDA - 19870304IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:2879565
SO  - Biochim Biophys Acta 1987 Jan 16 ;890(1):97-105

767
UI  - 19801
AU  - Kandpal RP
AU  - Stempel KE
AU  - Boyer PD
TI  - Characteristics of the formation of enzyme-bound ATP from medium inorganic phosphate by mitochondrial F1 adenosinetriphosphatase in the presence of dimethyl sulfoxide
AB  - Addition of dimethyl sulfoxide promotes the formation of enzyme-bound ATP from medium Pi by mitochondrial F1 adenosinetriphosphatase that has tightly bound ADP present. Measurements are reported of medium Pi in equilibrium H18OH exchange and of the dependence of formation of enzyme- bound ATP on Pi concentration. Attainment of an apparent equilibrium between medium Pi and bound ATP requires longer than 30 min, even though the rates of Pi binding and release after apparent equilibrium is reached would suffice for a faster approach to equilibrium. Slow protein conformational changes or other unknown modulating factors may be responsible for the slow rate of bound ATP formation. After apparent equilibrium is reached, each Pi that binds to the enzyme reversibly forms ATP about 50 times before being released to the medium. The rate of interconversion of bound ATP to bound ADP and Pi is much slower than that in the absence of dimethyl sulfoxide as measured with sufficiently low ATP concentrations so that single-site catalysis is favored. Although the interconversion rate is slowed, the equilibrium constant for bound ATP formation from bound ADP and Pi is not far from unity. Dimethyl sulfoxide favors the formation of enzyme-bound ATP by promoting the competent binding of Pi to enzyme with ADP bound at a catalytic site rather than by promoting formation of bound ATP from bound ADP and Pi
MH  - A
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - ADP
MH  - atp
MH  - ATP FORMATION
MH  - ATP synthase
MH  - BINDING
MH  - Catalysis
MH  - conformational change
MH  - DEPENDENCE
MH  - F1
MH  - H(+)-Transporting ATP Synthase
MH  - INORGANIC-PHOSPHATE
MH  - Oxygen
MH  - Oxygen Isotopes
MH  - Phosphates
MH  - SYNTHASE
MH  - Time
RP  - NOT IN FILE
NT  - UI - 87242322LA - engRN - 0 (Oxygen Isotopes)RN - 0 (Phosphates)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 67-68-5 (Dimethyl Sulfoxide)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM 11094/GM/NIGMSDA - 19870724IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:2885026
SO  - Biochemistry 1987 Mar 24 ;26(6):1512-1517

768
UI  - 20959
AU  - Kauffer S
AU  - Schmid R
AU  - Steffens K
AU  - Deckers-Hebestreit G
AU  - Altendorf K
AD  - Arbeitsgruppe Mikrobiologie, Universitat Osnabruck, Federal Republic of Germany
TI  - Evolutionary relationship between Enterobacteriaceae: comparison of the ATP synthases (F1F0) of Escherichia coli and Klebsiella pneumoniae
AB  - The ATP synthase complex of Klebsiella pneumoniae (KF1F0) has been purified and characterized. SDS-gel electrophoresis of the purified F1F0 complexes revealed an identical subunit pattern for E. coli (EF1F0) and K. pneumoniae. Antibodies raised against EF1 complex and purified EF0 subunits recognized the corresponding polypeptides of EF1F0 and KF1F0 in immunoblot analysis. Protease digestion of the individual subunits generated an identical cleavage pattern for subunits alpha, beta, gamma, epsilon, a, and c of both enzymes. Only for subunit delta different cleavage products were obtained. The isolated subunit c of both organisms showed only a slight deviation in the amino acid composition. These data suggest that extensive homologies exist in primary and secondary structure of both ATP synthase complexes reflecting a close phylogenetic relationship between the two enterobacteric tribes
MH  - A
MH  - ACID
MH  - analysis
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - COMPLEX
MH  - Electrophoresis
MH  - Enterobacteriaceae
MH  - Enzymes
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - Multienzyme Complexes
MH  - secondary
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 88048773LA - engRN - 0 (Multienzyme Complexes)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19871214IS - 0302-8933SB - IMCY - GERMANY, WEST
UR  - PM:2890332
SO  - Arch Microbiol 1987 Sep ;148(3):187-192

769
UI  - 19872
AU  - Kironde FA
AU  - Cross RL
TI  - Adenine nucleotide binding sites on beef heart F1-ATPase. Asymmetry and subunit location
AB  - Previously we have shown that beef heart mitochondrial F1 contains a total of six adenine nucleotide binding sites. Three "catalytic" sites exchange bound ligand rapidly during hydrolysis of MgATP, whereas three "noncatalytic" sites do not. The noncatalytic sites behave asymmetrically in that a single site releases bound ligand upon precipitation of F1 with ammonium sulfate. In the present study, we find this same site to be the only noncatalytic site that undergoes rapid exchange of bound ligand when F1 is incubated in the presence of EDTA at pH 8.0. Following 1000 catalytic turnovers/F1, the site retains the unique capacity for EDTA-induced exchange, indicating that the asymmetric determinants are permanent and that the three noncatalytic sites on soluble F1 do not pass through equivalent states during catalysis. Measurements of the rate of ligand binding at the unique noncatalytic site show that uncomplexed nucleotide binds preferentially. At pH 7.5, in the presence of Mg2+, the rate constant for ADP binding is 9 X 10(3) M-1 s-1 and for dissociation is 4 X 10(-4) s-1 to give a Kd = 50 nM. The rate of dissociation is 10 times faster in the presence of EDTA or during MgATP hydrolysis, and it increases rapidly at pH below 7. EDTA-induced exchange is inhibited by Mg2+, Mn2+, Co2+, and Zn2+ but not by Ca2+ and is unaffected by dicyclohexylcarbodiimide modification. The unique noncatalytic site binds 2-azido-ADP. Photolysis results in the labeling of the beta subunit. Photolabeling of a single high-affinity catalytic site under conditions for uni-site catalysis also results in the labeling of beta, but a different pattern of labeled peptides is obtained in proteolytic digests. The results demonstrate the presence of two different nucleotide binding domains on the beta subunit of mitochondrial F1
MH  - A
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - ADP
MH  - ADP BINDING
MH  - atp
MH  - ATP synthase
MH  - BETA-SUBUNIT
MH  - BINDING
MH  - Binding Sites
MH  - Catalysis
MH  - Dicyclohexylcarbodiimide
MH  - Edetic Acid
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - H(+)-Transporting ATP Synthase
MH  - Hydrolysis
MH  - Macromolecular Systems
MH  - Magnesium
MH  - NONCATALYTIC SITES
MH  - nucleotide binding
MH  - nucleotide binding sites
MH  - NUCLEOTIDE-BINDING-SITES
MH  - pH
MH  - SUBUNIT
MH  - SYNTHASE
MH  - Time
MH  - uni-site catalysis
MH  - UNISITE CATALYSIS
RP  - NOT IN FILE
NT  - UI - 87137637LA - engRN - 0 (Macromolecular Systems)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 60-00-4 (Edetic Acid)RN - 7439-95-4 (Magnesium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM 23152/GM/NIGMSDA - 19870420IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:2880850
SO  - J Biol Chem 1987 Mar 15 ;262(8):3488-3495

770
UI  - 761
AU  - Konishi J
AU  - Wakagi T
AU  - Oshima T
AU  - Yoshida M
AD  - Department of Life Science, Tokyo Institute of Technology, Kanagawa
TI  - Purification and properties of the ATPase solubilized from membranes of an acidothermophilic archaebacterium, Sulfolobus acidocaldarius
AB  - A novel ATPase was solubilized from membranes of an acidothermophilic archaebacterium, Sulfolobus acidocaldarius, with low ionic strength buffer containing EDTA. The enzyme was purified to homogeneity by hydrophobic chromatography and gel filtration. The molecular weight of the purified enzyme was estimated to be 360,000. Polyacrylamide gel electrophoresis of the purified enzyme in the presence of sodium dodecyl sulfate revealed that it consisted of three kinds of subunits, alpha, beta, and gamma, whose molecular weights were approximately 69,000, 54,000, and 28,000, respectively, and the most probable subunit stoichiometry was alpha 3 beta 3 gamma 1. The purified ATPase hydrolyzed ATP, GTP, ITP, and CTP but not UTP, ADP, AMP, or p- nitrophenylphosphate. The enzyme was highly heat stable and showed an optimal temperature of 85 degrees C. It showed an optimal pH of around 5, very little activity at neutral pH, and another small activity peak at pH 8.5. The ATPase activity was significantly stimulated by bisulfite and bicarbonate ions, the optimal pH remaining unchanged. The Lineweaver-Burk plot was linear, and the Km for ATP and the Vmax were estimated to be 1.6 mM and 13 mumol Pi.mg.-1.min-1, respectively, at pH 5.2 at 60 degrees C in the presence of bisulfite. The chemical modification reagent, 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole, caused inactivation of the ATPase activity although the enzyme was not inhibited by N,N'-dicyclohexylcarbodiimide, N-ethyl-maleimide, azide or vanadate. These results suggest that the ATPase purified from membranes of S. acidocaldarius resembles other archaebacterial ATPases, although a counterpart of the gamma subunit has not been found in the latter. The relationship of the S. acidocaldarius ATPase to other ion- transporting ATPases, such as F0F1 type or E1E2 type ATPases, was discussed
RP  - NOT IN FILE
NT  - UI - 88198056LA - engRN - 0 (Bicarbonates)RN - 0 (Buffers)RN - 0 (Ions)RN - 0 (Nucleotides)RN - 0 (Sulfites)RN - 0 (bisulfites)RN - 10199-89-0 (4-Chloro-7-nitrobenzofurazan)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19880602IS - 0021-924XSB - IMCY - JAPANJC - HIF
UR  - PM:2966145
SO  - J Biochem (Tokyo ) 1987 Dec ;102(6):1379-1387

771
UI  - 766
AU  - Konishi J
AU  - Yohda M
AU  - Hashimoto T
AU  - Yoshida M
AD  - Department of Life Science, Tokyo Institute of Technology, Kanagawa
TI  - Single site catalysis of the F1-ATPase from Saccharomyces cerevisiae and the effect of inorganic phosphate on it
AB  - The kinetical characteristics of ATP hydrolysis by mitochondrial F1- ATPase from Saccharomyces cerevisiae (yeast) have been studied under conditions where only a single catalytic site per enzyme molecule bound ATP. Four major features were observed, that is, fast ATP binding to the enzyme, slow product release from the enzyme, an equilibrium close to unity between ATP and products on the enzyme, and promotion of ATP hydrolysis on the second addition of a large excess of ATP (cold chase). These are essentially the same as the kinetical characteristics observed for beef heart mitochondrial F1-ATPase, which were called as unisite catalysis by Grubmeyer et al. (Grubmeyer, C. et al. (1982) J. Biol. Chem. 257, 12092-12100), although the release of a hydrolysis product, Pi, from the yeast enzyme appeared to occur significantly faster than that from the beef enzyme, which resulted in a decreased extent of cold chase promotion of ATP hydrolysis of the yeast enzyme. The yeast F1-ATPase showed unisite catalysis even in the absence of Pi in the reaction mixtures, while it was reported for the beef F1-ATPase that the presence of Pi in the reaction mixture was essential for unisite catalysis (Penefsky, H.S. & Grubmeyer, C. (1984) in H+-ATPase (ATP Synthase) (Papa, S. et al., eds.) pp. 195-204, The ICSU Press). Another difference in the Pi effect on the kinetics was that ATP hydrolysis was initiated without a lag time in the absence of Pi in the case of the yeast enzyme when a 1,000-fold molar excess of ATP per enzyme molecular was mixed with the enzyme.(ABSTRACT TRUNCATED AT 250 WORDS)
RP  - NOT IN FILE
NT  - UI - 88032945LA - engRN - 0 (Phosphates)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19871217IS - 0021-924XSB - IMCY - JAPANJC - HIF
UR  - PM:2889726
SO  - J Biochem (Tokyo ) 1987 Aug ;102(2):273-279

772
UI  - 99
AU  - Laubinger W
AU  - Dimroth P
AD  - Institut fur Physiologische Chemie, Technischen Universitat Muchen, Federal Republic of Germany
TI  - Characterization of the Na+-stimulated ATPase of Propionigenium modestum as an enzyme of the F1F0 type
AB  - The ATP-hydrolyzing activity of Propionigenium modestum was extracted from the membranes with Triton X-100 or by incubation with EDTA at low ionic strength. The ATPase in the Triton extract was highly sensitive to dicyclohexylcarbodiimide but not to vanadate. These properties are characteristic for enzymes of the F1 F0 type. The ATPase was specifically activated by Na+ ions yielding a 15-fold increase in catalytic activity at 5 mM Na+ concentration. The additional presence of 1% Triton X-100 caused a further 1.5-fold activation. In the absence of Na+ Triton stimulated the ATPase about 13-fold. The Triton- stimulated ATPase was further activated about 1.5-2-fold by Na+ addition. The ATPase extracted by the low-ionic-strength treatment was purified to homogeneity by fractionation with poly(ethylene glycol) and gel chromatography. The enzyme had the characteristic F1-ATPase subunit structure with Mr values of 58,000 (alpha), 56,000 (beta), 37,600 (gamma), 22,700 (delta), and 14,000 (epsilon). The F1-ATPase was not stimulated by Na+ ions. The membrane-bound ATPase was reconstituted from the purified F1 part and F1-depleted membranes, thus further indicating an F1 F0 structure for the ATPase of P. modestum. Upon reconstitution the ATPase recovered its stimulation by Na+ ions, suggesting that the binding site for Na+ is localized on the membrane- bound F0 part of the enzyme complex
RP  - NOT IN FILE
NT  - UI - 88029463LA - engRN - 0 (Detergents)RN - 7440-23-5 (Sodium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19871208IS - 0014-2956SB - IMCY - GERMANY, WESTJC - EMZ
UR  - PM:2889596
SO  - Eur J Biochem 1987 Oct 15 ;168(2):475-480

773
UI  - 19763
AU  - Lundin M
AU  - Pereira dS
AU  - Baltscheffsky H
TI  - Energy-dependent formation of free ATP in yeast submitochondrial particles, and its stimulation by oligomycin
AB  - Yeast submitochondrial particles, in a Pi- and NADH-dependent reaction, produced low concentrations of free ATP in the absence of added ADP. This formation of free ATP, as measured by the luciferin-luciferase method, was strongly stimulated by oligomycin. For maximal stimulation, oligomycin was to be added not earlier than 5-10 min after the addition of NADH. Upon addition of antimycin or FCCP the system was completely inhibited. The amount of free ATP formed corresponded to one-third of the amount of bound ATP in submitochondrial particles. The stimulatory effect of oligomycin disappeared if the submitochondrial particles were spun down after oligomycin stimulation and then resuspended in the reaction medium, whereas submitochondrial particles with no oligomycin added initially were stimulated by oligomycin after the same procedure. A different picture emerged with addition of ADP. If the submitochondrial particles were preenergized with NADH in the presence of oligomycin before the addition of ADP the formation of free ATP upon subsequent addition of ADP was inhibited by oligomycin. In the presence of oligomycin, but lacking preenergization with NADH, a stimulation of free ATP formation was achieved with added ADP. A possible explanation for the stimulating effect of oligomycin on ATP formation in the absence of added ADP is that it enhances the release of bound ATP in an energy-requiring process. The release of only about one-third of the bound ATP could indicate that one of three nucleotide-binding subunits involved in the mechanism of ATP formation by ATP synthase is in a state suitable for such an energy-dependent release of ATP
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - ADP
MH  - Antimycin A
MH  - atp
MH  - ATP synthase
MH  - Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone
MH  - mechanism
MH  - method
MH  - Nad
MH  - nucleotide binding
MH  - Oligomycins
MH  - Submitochondrial Particles
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 87128937LA - engRN - 0 (Oligomycins)RN - 370-86-5 (Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone)RN - 53-84-9 (NAD)RN - 56-65-5 (Adenosine Triphosphate)RN - 642-15-9 (Antimycin A)PT - Journal ArticleDA - 19870410IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:3545293
SO  - Biochim Biophys Acta 1987 Mar 4 ;890(3):279-285

774
UI  - 9911
AU  - Luvisetto S
AU  - Pietrobon D
AU  - Azzone GF
TI  - Uncoupling of oxidative phosphorylation. 1. Protonophoric effects account only partially for uncoupling.
AB  - The mechanism of uncoupling of oxidative phosphorylation by carbonyl cyanide p-trifluoromethoxy)phenylhydrazone (FCCP), a typical weak acid protonophore, oleic acid, a fatty acid, and chloroform, a general anesthetic, has been investigated by measuring in mitochondria their effect on (i) the transmembrane proton electrochemical potential gradient (delta mu H) and the rates of electron transfer and adenosine 5'-triphosphate (ATP) hydrolysis in static head, (ii) delta mu H and the rates of electron transfer and ATP synthesis in state 3, and (iii) the membrane proton conductance. Both FCCP and oleic acid increase the membrane proton conductance, and accordingly, they cause a depression of delta mu H [generated by either the redox proton pumps or the adenosinetriphosphatase (ATPase) proton pumps]. Although their effects on ATP synthesis/hydrolysis, respiration, and delta mu H are qualitatively consistent with a pure protonophoric uncoupling mechanism and an additional inhibitory action of oleic acid on both the ATPases and the electron-transfer enzymes, a quantitative comparison between the dissipative proton influx and the rate of either electron transfer or ATP hydrolysis (multiplied by either the H+/e- or the H+/ATP stoichiometry, respectively) at the same delta mu H shows that the increase in membrane conductance induced by FCCP and oleic acid accounts for the stimulation of the rate of ATP hydrolysis but not for that of the rate of electron transfer. Chloroform (at concentrations that fully inhibit ATP synthesis) only very slightly increases the proton conductance of the mitochondrial membrane and causes only a little depression of delta mu H.(ABSTRACT TRUNCATED AT 250 WORDS)
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - Animal
MH  - atp
MH  - ATP synthesis
MH  - ATPase
MH  - Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone
MH  - Chloroform
MH  - drug effects
MH  - electron
MH  - Enzymes
MH  - Hydrolysis
MH  - Intracellular Membranes
MH  - Kinetics
MH  - mechanism
MH  - metabolism
MH  - Mitochondria
MH  - Mitochondria,Liver
MH  - Nitriles
MH  - Oleic Acids
MH  - Oxidative Phosphorylation
MH  - pharmacology
MH  - Phosphorylation
MH  - physiology
MH  - proton
MH  - Proton Pump
MH  - Protons
MH  - Rats
MH  - Respiration
MH  - Support,Non-U.S.Gov't
MH  - TRANSFER
MH  - Uncoupling Agents
RP  - NOT IN FILE
NT  - CNR Unit for the Study of the Physiology of Mitochondria, University of Padova, ItalyPMID- 0002827753
SO  - Biochemistry 1987 Nov 17 ;26(23):7332-7338

775
UI  - 462
AU  - Maggio MB
AU  - Pagan J
AU  - Parsonage D
AU  - Hatch L
AU  - Senior AE
TI  - The defective proton-ATPase of uncA mutants of Escherichia coli. Identification by DNA sequencing of residues in the alpha-subunit which are essential for catalysis or normal assembly
AB  - A group of mutant uncA alleles, affecting essential residues of the alpha-subunit of Escherichia coli proton-ATPase, have been identified by intragenic complementation mapping, cloning, and DNA sequencing. One of the mutations, uncA450, abolishes normal assembly of F1-ATPase. The amino acid substitution found was Glu-299----Lys, which is predicted to lie in an alpha-helix in alpha-subunit. The reversal of the charge at residue 299 is a likely cause of defective assembly. The uncA462 allele causes impairment of catalysis while allowing normal assembly of membrane-bound F1-ATPase. The amino acid substitution found was Ser-347- ---Phe. Three mutations which impair catalysis but do not cause structural perturbation of either membrane-bound or solubilized F1ATPase were characterized as follows: uncA401, Ser-373----Phe; uncA447, Gly-351----Asp; uncA453, Ser-375----Phe. We predict here that the nucleotide-binding domain of alpha-subunit is formed by the amino acids in the sequence from residue 160 to approximately residue 340. The mutations which cause impairment of catalysis lie in a short segment between residues 347-375 of alpha-subunit, at the C-terminal end of the predicted nucleotide-binding domain. This segment is suggested to be important for beta-alpha-beta intersubunit conformational interaction involved in positive catalytic cooperativity in F1-ATPase
RP  - NOT IN FILE
NT  - UI - 87250533LA - engRN - 0 (DNA, Bacterial)RN - 0 (Macromolecular Systems)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSDA - 19870814IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:2885325
SO  - J Biol Chem 1987 Jul 5 ;262(19):8981-8984

776
UI  - 1000
AU  - Mitchell P
AD  - Glynn Research Institute, Bodmin, England
TI  - A new redox loop formality involving metal-catalysed hydroxide-ion translocation. A hypothetical Cu loop mechanism for cytochrome oxidase
AB  - A new hypothetical type of redox loop is described, which translocates hydroxide instead of protons. Conventional protonmotive redox loops use carriers of protons with electrons (e.g. QH2/Q systems) to couple electron transfer to the translocation of protons. The putative hydroxidemotive redox loop uses carriers of hydroxide ions against electrons (e.g. transition-metal centres) to couple electron transfer to the translocation of hydroxide ions. This simple idea leads to the proposal of a hydroxidemotive Cu loop mechanism that may possibly be applicable to the CuA or CuB centre of cytochrome oxidase, and might thus account for the coupling of electron transfer to net proton translocation in that osmoenzyme
MH  - Cytochrome-c Oxidase
MH  - Electrons
MH  - Ions
MH  - Protons
RP  - NOT IN FILE
NT  - UI - 88005161LA - engRN - 0 (Hydroxides)RN - 7440-50-8 (Copper)RN - EC 1.9.3.1 (Cytochrome-c Oxidase)PT - Journal ArticlePT - ReviewPT - Review, AcademicDA - 19871118IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:2820802
SO  - FEBS Lett 1987 Oct 5 ;222(2):235-245

777
UI  - 1011
AU  - Mitchell P
TI  - A new redox loop formality involving metal-catalysed hydroxide-ion translocation. A hypothetical Cu loop mechanism for cytochrome oxidase
AB  - A new hypothetical type of redox loop is described, which translocates hydroxide instead of protons. Conventional protonmotive redox loops use carriers of protons with electrons (e.g. QH2/Q systems) to couple electron transfer to the translocation of protons. The putative hydroxidemotive redox loop uses carriers of hydroxide ions against electrons (e.g. transition-metal centres) to couple electron transfer to the translocation of hydroxide ions. This simple idea leads to the proposal of a hydroxidemotive Cu loop mechanism that may possibly be applicable to the CuA or CuB centre of cytochrome oxidase, and might thus account for the coupling of electron transfer to net proton translocation in that osmoenzyme
MH  - Copper
MH  - Cytochrome-c Oxidase
MH  - Electron Transport
MH  - Electrons
MH  - Hydroxides
MH  - Ions
MH  - metabolism
MH  - Oxidation-Reduction
MH  - pharmacology
MH  - Protons
MH  - Support,Non-U.S.Gov't
RP  - NOT IN FILE
NT  - Glynn Research Institute, Bodmin, England
SO  - FEBS Lett 1987 Oct 5 ;222(2):235-245

778
UI  - 762
AU  - Mukohata Y
AU  - Ihara K
AU  - Yoshida M
AU  - Konishi J
AU  - Sugiyama Y
AU  - Yoshida M
AD  - Department of Biology, Faculty of Science, Osaka University, Toyomaka, Japan
TI  - The halobacterial H+-translocating ATP synthase relates to the eukaryotic anion-sensitive H+-ATPase
AB  - The H+-translocating ATP synthase of Halobacterium halobium (Y. Mukohata and M. Yoshida (1987) J. Biochem. 102, 797-802) includes a catalytic moiety of 320 kDa which is isolated as an azide-insensitive ATPase (T. Nanba and Y. Mukohata (1987) J. Biochem. 102, 591-598). The polyclonal antibody against this archaebacterial ATPase cross-reacts with the anion-sensitive H+-ATPase of red beet, Beta vulgaris, tonoplast as well as with another archaebacterial ATPase from Sulfolobus acidocaldarius. The affinity is much higher than to F1- ATPase from spinach chloroplasts or to Ca2+-ATPase from sarcoplasmic reticulum of rabbit skeletal muscle
RP  - NOT IN FILE
NT  - UI - 88105465LA - engRN - 0 (Macromolecular Systems)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19880208IS - 0003-9861SB - IMCY - UNITED STATESJC - 6SK
UR  - PM:2892466
SO  - Arch Biochem Biophys 1987 Dec ;259(2):650-653

779
UI  - 768
AU  - Mukohata Y
AU  - Yoshida M
TI  - Activation and inhibition of ATP synthesis in cell envelope vesicles of Halobacterium halobium
AB  - The characteristics of ATP synthesis in cell envelope vesicles of Halobacterium halobium were further studied. The results confirmed the previous conclusion (Mukohata et al. (1986) J. Biochem. 99, 1-8) that the ATP synthase in this extremely halophilic archaebacterium can not be an ordinary type of F0F1-ATPase, which has been thought to be ubiquitous among all the aerobic organisms on our biosphere. The ATP synthesis was activated most in 1 M NaCl and/or KCl, and at 40 degrees C, and at 80 mM MgCl2 where F0F1-ATPase loses its activity completely. The synthesis was negligible at 10 degrees C, and at 5 mM MgCl2. The Km for ADP was about 0.3 mM in the presence of 20 mM Pi, 1 M NaCl, 80 mM MgCl2, and 10 mM PIPES at pH 6.8 and 20 degrees C. The ATP synthesis was not inhibited by NaN3 and quercetin (specific inhibitors for F0F1- ATPase) or vanadate (for E1E2-ATPase) or ouabain (for Na+,K+-ATPase) or P1,P5-di(adenosine-5')pentaphosphate (AP5A, for adenylate kinase). The ATP synthesis was not inhibited by modification (pretreatment) with NaN3 or 5'-p-fluorosulfonylbenzoyladenosine (FSBA). On the contrary, the ATP synthesis was rather non-specifically inhibited by N- ethylmaleimide (NEM), trinitrobenzenesulfonate (TNBS), phenylglyoxal, and pyridoxal phosphate. 7-Chloro-4-nitrobenz-2-oxa-1,3-diazole (NBD- Cl) as well as N,N'-dicyclohexylcarbodiimide (DCCD) was found to be a specific inhibitor at least partly, because the NBD-Cl inhibition was partly prevented by ADP added to the modification mixture
RP  - NOT IN FILE
NT  - UI - 87222246LA - engRN - 0 (Salts)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 7439-95-4 (Magnesium)PT - Journal ArticleDA - 19870706IS - 0021-924XSB - IMCY - JAPANJC - HIF
UR  - PM:3584088
SO  - J Biochem (Tokyo ) 1987 Feb ;101(2):311-318

780
UI  - 764
AU  - Mukohata Y
AU  - Yoshida M
AD  - Department of Biology, Faculty of Science, Osaka University
TI  - The H+-translocating ATP synthase in Halobacterium halobium differs from F0F1-ATPase/synthase
AB  - Cell envelope vesicles of Halobacterium halobium synthesize ATP by utilizing base-acid transition (an outside acidic pH jump) under optimal conditions (1 M NaCl, 80 mM MgCl2, pH 6.8) even in the presence of azide (a specific inhibitor of F0F1-ATPase) (Mukohata & Yoshida (1987) J. Biochem. 101, 311-318). An azide-insensitive ATPase was isolated from the inner face of the vesicle membrane, and shown to hydrolyze ATP under very specific conditions (1.5 M Na2SO4, 10 mM MnCl2, pH 5.8) (Nanba & Mukohata (1987) J. Biochem. 102, 591-598). This ATPase activity could also be detected when the vesicle components were solubilized by detergent. The relationship between ATP synthesis and the membrane-bound ATPase was investigated by modification of the vesicles with 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole (NBD-Cl) or N- ethylmaleimide (NEM). The inhibition pattern of ATP synthesis in the modified vesicles and that of ATP hydrolysis of the solubilized modified vesicles were compared under the individual optimum conditions. The inhibition patterns were almost identical, suggesting that the ATP synthesis and hydrolysis are catalyzed by a single enzyme complex. The ATP synthase includes the above ATPase (300-320 kDa), which is composed of two pairs of 86 and 64 kDa subunits. This is a novel H+-translocating ATP synthase functioning in the extremely halophilic archaebacterium. This "archae-ATP-synthase" differs from F0F1-ATPase/synthase, which had been thought to be ubiquitous among all respiring organisms on our biosphere
RP  - NOT IN FILE
NT  - UI - 88139231LA - engRN - 0 (Multienzyme Complexes)RN - 10199-89-0 (4-Chloro-7-nitrobenzofurazan)RN - 128-53-0 (Ethylmaleimide)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 2.7 (Phosphotransferases)RN - EC 2.7.4.- (ATP synthetase complex)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19880325IS - 0021-924XSB - IMCY - JAPANJC - HIF
UR  - PM:2893789
SO  - J Biochem (Tokyo ) 1987 Oct ;102(4):797-802

781
UI  - 20886
AU  - Noumi T
AU  - Azuma M
AU  - Shimomura S
AU  - Maeda M
AU  - Futai M
AD  - Department of Organic Chemistry and Biochemistry, Osaka University, Japan
TI  - Escherichia coli H+-ATPase. Glutamic acid 185 in beta subunit is essential for its structure and assembly
AB  - The uncD gene for the beta subunit of Escherichia coli H+-ATPase was cloned downstream of the lac promoter and mutagenized (Glu-185----Gln or Lys) by an oligonucleotide-directed procedure. The recombinant plasmid was introduced into a strain in which the unc operon for subunits of H+-ATPase was deleted. The wild-type or mutant beta subunit synthesized amounted to about 10% total cell protein and was mainly found in the cytoplasmic fraction. These subunits could be purified to almost homogeneity by conventional procedures. The wild-type and two mutant beta subunits had essentially the same Kd values for 8- anilinonaphthalene-1-sulfonate, aurovertin, and ATP, although the fluorescence intensities of 8-anilinonaphthalene-1-sulfonate and aurovertin were significantly less when bound to the two mutant beta subunits than when bound to the wild-type subunit. The three beta subunits showed essentially the same circular dichroism spectra, indicating alpha-helical contents of about 16-18%. Thus, the mutations did not cause marked change of the secondary structure of the subunit. However, measurements of theta 208 during linear increase in temperature suggested that replacement of Glu-185 by Gln or Lys slightly changed the stability of the secondary structure. Only trace amounts of alpha beta gamma complexes could be reconstituted using the two mutant beta subunits. These results suggest that Glu-185 or the region in its vicinity may be essential for subunit assembly. The methods developed in this study should be useful for further studies on the beta subunit
MH  - A
MH  - ACID
MH  - atp
MH  - ATPase
MH  - BETA-SUBUNIT
MH  - Biochemistry
MH  - Chemistry
MH  - COMPLEX
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - fluorescence
MH  - glutamic acid
MH  - H+-ATPase
MH  - Macromolecular Systems
MH  - method
MH  - Methods
MH  - mutant
MH  - protein
MH  - secondary
MH  - spectra
MH  - SUBUNIT
MH  - SYSTEM
MH  - SYSTEMS
MH  - Temperature
RP  - NOT IN FILE
NT  - UI - 88033072LA - engRN - 0 (Glutamates)RN - 0 (Macromolecular Systems)RN - 0 (Plasmids)RN - 56-86-0 (Glutamic Acid)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19871210IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:2889734
SO  - J Biol Chem 1987 Nov 5 ;262(31):14978-14982

782
UI  - 20890
AU  - Noumi T
AU  - Maeda M
AU  - Futai M
TI  - Mode of inhibition of sodium azide on H+-ATPase of Escherichia coli
AB  - Sodium azide inhibited multi-site (steady-state) ATPase activity of E. coli F1 more than 90%, but did not affect uni-site (single-site) ATPase activity. Thus azide inhibited multi-site ATPase activity by lowering catalytic cooperativity. Consistent with this observation, azide changed the ligand-induced fluorescence response of aurovertin bound to F1
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - ATPase
MH  - Aurovertins
MH  - Azides
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1
MH  - fluorescence
MH  - H+-ATPase
MH  - Magnesium
MH  - Sodium
RP  - NOT IN FILE
NT  - UI - 87162509LA - engRN - 0 (Aurovertins)RN - 0 (Azides)RN - 26628-22-8 (Sodium Azide)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 7439-95-4 (Magnesium)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19870504IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:2881810
SO  - FEBS Lett 1987 Mar 23 ;213(2):381-384

783
UI  - 20888
AU  - Noumi T
AU  - Tagaya M
AU  - Miki-Takeda K
AU  - Maeda M
AU  - Fukui T
AU  - Futai M
TI  - Loss of unisite and multisite catalyses by Escherichia coli F1 through modification with adenosine tri- or tetraphosphopyridoxal
AB  - Pyridoxal phosphate (PLP) and adenosine diphospho (AP2-PL)-, triphospho (AP3-PL)-, and tetraphospho (AP4-PL)-pyridoxals (Tagaya, M., and Fukui, T. (1986) Biochemistry 25, 2958-2964) were tested as potential affinity probes for F1 ATPase of Escherichia coli. Both AP3-PL and AP4-PL bound and inhibited F1 ATPase, whereas PLP and AP2-PL were weak inhibitors. The concentrations of AP3-PL and AP4-PL for half-maximal inactivations of the multisite (steady state) ATPase activity were both 18 microM. The binding of these reagents to a reactive lysyl residue(s) was confirmed from the difference absorption spectra, and the stoichiometry of binding of [3H]AP3-PL to F1 at the saturating level was about 1 mol/mol F1. The analogue bound to both the alpha subunit (about two- thirds of the radioactivity) and the beta subunit (about one-third of the radioactivity). No inactivation of multisite ATPase activity or binding of AP3-PL was observed in the presence of ATP. F1 modified with about one mol of AP3-PL had essentially no uni- and multisite hydrolysis of ATP. The rate of binding of ATP decreased to 10(-2) of that of unmodified F1, and the rate of release of ATP was about two times faster. The equilibrium F1 X ATP in equilibrium F1 X ADP X Pi was shifted toward F1 X ATP, and no promotion of ATP hydrolysis at unisite was observed with excess ATP. These results suggest that the AP3-PL or AP4-PL bound to an active site, and catalysis by the two remaining sites was completely abolished
MH  - A
MH  - absorption
MH  - ACTIVE
MH  - Adenine Nucleotides
MH  - ADENINE-NUCLEOTIDES
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - ADP
MH  - ALPHA-SUBUNIT
MH  - atp
MH  - ATPase
MH  - BETA-SUBUNIT
MH  - BINDING
MH  - Biochemistry
MH  - Catalysis
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - Hydrolysis
MH  - M
MH  - Macromolecular Systems
MH  - Nucleotides
MH  - Site
MH  - spectra
MH  - SUBUNIT
MH  - SYSTEM
MH  - SYSTEMS
MH  - Time
RP  - NOT IN FILE
NT  - UI - 87222393LA - engRN - 0 (Adenine Nucleotides)RN - 0 (Macromolecular Systems)RN - 101418-63-7 (adenosine triphosphopyridoxal)RN - 101418-64-8 (adenosine tetraphosphopyridoxal)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19870702IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:2884220
SO  - J Biol Chem 1987 Jun 5 ;262(16):7686-7692

784
UI  - 767
AU  - Ohtsubo M
AU  - Yoshida M
AU  - Ohta S
AU  - Kagawa Y
AU  - Yohda M
AU  - Date T
TI  - In vitro mutated beta subunits from the F1-ATPase of the thermophilic bacterium, PS3, containing glutamine in place of glutamic acid in positions 190 or 201 assembles with the alpha and gamma subunits to produce inactive complexes
AB  - Using site-directed mutagenesis, Glu-190 or Glu-201 of the beta subunit of the F1-ATPase from the thermophilic bacterium PS3 were replaced with glutamine. It was possible to reconstitute complexes of the mutated beta subunits with alpha and gamma subunits, but the complexes did not have ATPase activity. It is concluded that carboxylic acid side chains of Glu-190 and Glu-201 of the beta subunit are essential for catalytic activity of F1-ATPase
RP  - NOT IN FILE
NT  - UI - 87298498LA - engRN - 0 (Glutamates)RN - 0 (Macromolecular Systems)RN - 56-85-9 (Glutamine)RN - 56-86-0 (Glutamic Acid)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19870916IS - 0006-291XSB - IMCY - UNITED STATESJC - 9Y8
UR  - PM:2887165
SO  - Biochem Biophys Res Commun 1987 Jul 31 ;146(2):705-710

785
UI  - 463
AU  - Parsonage D
AU  - Wilke-Mounts S
AU  - Senior AE
TI  - Directed mutagenesis of the beta-subunit of F1-ATPase from Escherichia coli
AB  - Oligonucleotide-directed mutagenesis was used to generate six mutant strains of Escherichia coli which had the following specific amino acid substitutions in the beta-subunit of F1-ATPase: (i) Lys-155----Gln; (ii) Lys-155----Glu; (iii) Gly-149----Ile; (iv) Gly-154----Ile; (v) Tyr- 297----Phe;(vi) Tyr-354----Phe. The effects of each mutation on growth of cells on succinate plates or limiting (3 mM) glucose and on cell membrane ATPase activity and ATP-driven pH gradient formation were studied. The results showed Lys-155 to be essential for catalysis, as has been predicted previously from sequence homology and structural considerations; however, the results appear to contradict the hypothesis that Lys-155 interacts with one of the substrate phosphate groups because the Lys-155----Glu mutation was less detrimental than Lys-155----Gln. Gly-149 and Gly-154 have been predicted to be involved in essential conformational changes in F1-ATPase by virtue of their position in a putative glycine-rich flexible loop structure. The mutation of Gly-154----Ile caused strong impairment of catalysis, but the Gly-149----Ile mutation produced only moderate impairment. The two tyrosine residues chosen for mutation were residues which have previously received much attention due to their being the sites of reaction of the inactivating chemical modification reagents 4-chloro-7- nitrobenzofurazan (Tyr-297) and p-fluorosulfonylbenzoyl-5'-adenosine (Tyr-354). We found that mutation of Tyr-297----Phe caused only minor impairment of catalysis, and mutation of Tyr-354----Phe produced no impairment. Therefore, a direct role for either of these tyrosine residues in catalysis is unlikely
RP  - NOT IN FILE
NT  - UI - 87250385LA - engRN - 0 (Plasmids)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSDA - 19870730IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:2885316
SO  - J Biol Chem 1987 Jun 15 ;262(17):8022-8026

786
UI  - 530
AU  - Pedersen PL
AU  - Williams N
AU  - Hullihen J
AD  - Laboratory for Molecular and Cellular Bioenergetics, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205
TI  - Mitochondrial ATP synthase: dramatic Mg2+-induced alterations in the structure and function of the F1-ATPase moiety
AB  - The ATPase activity of the F1 moiety of rat liver ATP synthase is inactivated when incubated prior to assay at 25 degrees C in the presence of MgCl2. The concentration of MgCl2 (130 microM) required to induce half-maximal inactivation is over 30 times higher than the apparent Km (MgCl2) during catalysis. Moreover, the relative efficacy of divalent cations in inducing inactivation during prior incubation follows an order significantly different from that promoting catalysis. Inactivation of F1-ATPase activity by Mg2+ is accompanied by the dramatic dissociation from the F1 complex of alpha subunits and part of the gamma-subunit population. The latter form a precipitate while the beta, delta, and epsilon subunits, and the remaining part of the gamma- subunit population, remain soluble. Dissociation is not a sudden "all or none" event but parallels loss of ATPase activity until alpha subunits have almost completely dissociated together with about 50% of the gamma-subunit population. Mg2+-induced loss of F1-ATPase activity cannot be prevented by including either the hydrolytic substrates ATP, GTP, or ITP in the incubation medium or the product ADP. Ethylenediaminetetraacetic acid, mercaptoethanol, and dithiothreitol are also ineffective in preventing loss of ATPase activity. Significantly, KPi at high concentration (greater than or equal to 200 mM) is effective in partially protecting F1 against inactivation. However, the most effective means of preventing Mg2+-induced inactivation of F1-ATPase activity is to rebind F1 to its F0 moiety in F1-depleted particles. When bound to F0, F1 is protected completely against divalent cation induced inactivation.(ABSTRACT TRUNCATED AT 250 WORDS)
RP  - NOT IN FILE
NT  - UI - 88163539LA - engRN - 0 (Cations, Divalent)RN - 0 (Macromolecular Systems)RN - 7439-95-4 (Magnesium)RN - 7786-30-3 (Magnesium Chloride)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - CA 10951/CA/NCIDA - 19880506IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:2894844
SO  - Biochemistry 1987 Dec 29 ;26(26):8631-8637

787
UI  - 639
AU  - Penefsky HS
TI  - Molecular mechanism of ATP synthesis in oxidative phosphorylation
AB  - The experiments described in this paper may perhaps point the way towards a reaction mechanism for oxidative phosphorylation. However, we are not yet in a position to write a detailed chemical equation, supported by experimental evidence, for the mechanism of ATP synthesis. Continued pursuit of some of the implications of these experiments will be very much dependent on information presently unavailable. For example, it would be of great value to have three-dimensional X-ray crystal structures for F0 as well as F1. It will also be important to know the pathway of proton translocation through the ATPase complex. We shall surely require entirely new experimental tools to probe many of these questions
RP  - NOT IN FILE
NT  - UI - 87162839LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19870513IS - 0300-5127SB - IMCY - ENGLANDJC - E48
UR  - PM:2881816
SO  - Biochem Soc Trans 1987 Feb ;15(1):97-99

788
UI  - 531
AU  - Petrone G
AU  - Garboczi DN
AU  - Pedersen PL
AD  - Department of Biological Chemistry, Johns Hopkins University School of Medicine, Baltimore 21205
TI  - Mitochondrial ATP synthase complex: interaction of its F1 adenosinetriphosphatase moiety with the heavy atom iodine
AB  - Studies were carried out to determine whether a simple electron-dense "heavy atom" like iodine could be introduced selectively into one or more of the subunits of the mitochondrial ATP synthase complex of rat liver. Surprisingly, very low amounts of iodine are incorporated into the isolated F1 moiety of this complex under conditions which result in a marked loss of catalytic activity. ATPase activity is inactivated in a concentration-dependent manner at pH 7.5 with half-maximal inactivation occurring at about 40 microM iodine. A maximum of only 10 atoms of iodine are incorporated per F1 molecule under conditions where inhibition of ATPase activity is linearly related to iodine incorporation. The molecular size of F1 after iodination is unchanged, indicating that inactivation is due to modification of essential amino acid residues rather than subunit dissociation. Treatment of F1, with 20-50 microM [125I]iodine followed sequentially by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography showed that the beta subunit is preferentially labeled. Significantly, about two atoms of iodine per beta subunit are incorporated. Some iodine amounting to less than 23% of the total radioactivity placed on the gels is recovered in the alpha and gamma subunits whereas no radioactivity is detected in the delta and epsilon subunits. Iodination of F1 appears to modify essential residues other than those involved in substrate or product binding per se. Thus, nucleotide binding to F1 is unaltered by iodine, and neither phosphate, MgADP, nor MgATP protects F1 against inhibition by this agent.(ABSTRACT TRUNCATED AT 250 WORDS)
RP  - NOT IN FILE
NT  - UI - 88000623LA - engRN - 0 (Iodine Radioisotopes)RN - 0 (Nucleotides)RN - 7553-56-2 (Iodine)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - CA 10951/CA/NCIDA - 19871116IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:2888483
SO  - Biochemistry 1987 Jun 30 ;26(13):4016-4021

789
UI  - 78
AU  - Rao R
AU  - Perlin DS
AU  - Senior AE
TI  - The defective proton-ATPase of uncA mutants of Escherichia coli: ATP- binding and ATP-induced conformational change in mutant alpha-subunits
AB  - Mutations in the uncA gene of Escherichia coli cause loss of both oxidative phosphorylation and ATP-driven generation of the transmembrane proton gradient. The uncA gene encodes the alpha-subunit of the F1-sector of the E. coli membrane proton-ATPase. F1-alpha- subunit from normal (unc+) E. coli binds ATP tightly (KD = 0.1 microM) and undergoes a large ATP-induced conformational change, but the functional role of the ATP-binding site is currently unknown. There is disagreement in the literature as to whether the ATP-binding site is present or lacking in F1-alpha-subunit from uncA mutant strains. One obstacle in studying this question is the difficulty of purifying mutant alpha-subunits in native form. In order to circumvent this difficulty we have studied ATP binding and ATP-induced conformational changes in mixtures of F1 subunits obtained by dissociating uncA mutant F1. Anti-alpha antibody was used in conjunction with immunoblotting to identify the alpha-subunits in the mixtures. Retention of native conformation by the alpha-subunits was demonstrated by the fact that the dissociated alpha-subunits were fully competent to repolymerize with other F1 subunits to yield intact F1 aggregate. The results show that, contrary to previous reports, alpha-subunits from three catalytically defective uncA mutants do indeed bind ATP and do undergo an ATP-induced conformational change. The binding affinity of alpha- subunit for ATP was lower than normal in each of the three mutants, but this is not likely to be a significant factor under physiological conditions
RP  - NOT IN FILE
NT  - UI - 87240212LA - engRN - 0 (Macromolecular Systems)RN - 0 (Peptide Fragments)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.4.21.4 (Trypsin)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM-25349/GM/NIGMSDA - 19870710IS - 0003-9861SB - IMCY - UNITED STATESJC - 6SK
UR  - PM:2884928
SO  - Arch Biochem Biophys 1987 Jun ;255(2):309-315

790
UI  - 461
AU  - Rao R
AU  - Senior AE
AD  - Department of Biochemistry, University of Rochester Medical Center, New York 14642
TI  - The properties of hybrid F1-ATPase enzymes suggest that a cyclical catalytic mechanism involving three catalytic sites occurs
AB  - Maximal rates of ATP hydrolysis catalyzed by F1-ATPase enzymes are known to involve strong positive catalytic site cooperativity. There are three potential catalytic nucleotide-binding sites on F1. Two important and unanswered questions are (i) whether all three potential catalytic sites must interact cooperatively to yield maximal rates of ATP hydrolysis and (ii) whether a cyclical three-site mechanism operates as suggested by several authors. We have studied these two questions here by measuring the ATPase activities of hybrid enzymes containing normal beta-, gamma-, delta-, and epsilon-subunits together with different combinations of mutant and normal alpha-subunits. The mutant alpha-subunits were derived from uncA401, uncA447, and uncA453 mutant E. coli F1-ATPase, in which positive cooperativity between catalytic sites is strongly attenuated by defined mis-sense mutations. Our data show that three normal catalytic sites are required to interact in order to achieve maximal ATPase rates and suggest that a cyclical mechanism does operate. Hybrid enzyme containing one-third mutant alpha-subunit and two-thirds normal alpha-subunits had substantial but submaximal activity, showing that cooperativity between three sites in a noncyclical fashion, or between pairs of sites, can achieve effective catalysis
RP  - NOT IN FILE
NT  - UI - 88087026LA - engRN - 0 (Isoenzymes)RN - 0 (Macromolecular Systems)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSID - GM29805/GM/NIGMSDA - 19880129IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:2891693
SO  - J Biol Chem 1987 Dec 25 ;262(36):17450-17454

791
UI  - 1152
AU  - Roegner M
AU  - Graber P
TI  - Subunit-subunit-interactions in F0F1 as revealed by ligand binding and intrinsic fluorescence
MH  - BINDING
MH  - F0F1
MH  - fluorescence
T2  - Prog. Photosynth. Res., Proc. Int. Congr. Photosynth., 7th, Meeting Date 1986, Volume 3, 25-8. Edited by: Biggins, John. Nijhoff: Dordrecht, Neth
Y2  - -32676  
RP  - ON REQUEST (03/18/92)
SO  -  1987  ;():

792
UI  - 1156
AU  - Schmidt G
AU  - Graber P
TI  - The rate of ATP synthesis and ATP hydrolysis catalyzed by reconstituted CF0F1 liposomes
MH  - atp
MH  - ATP synthesis
MH  - CF0F1
MH  - Hydrolysis
MH  - liposome
MH  - Liposomes
MH  - RECONSTITUTED CF0F1
MH  - synthesis
T2  - Prog. Photosynth. Res., Proc. Int. Congr. Photosynth., 7th, Meeting Date 1986, Volume 3, 91-4. Edited by: Biggins, John. Nijhoff: Dordrecht, Neth
Y2  - -32676  
RP  - ON REQUEST (03/18/92)
SO  -  1987  ;():

793
UI  - 8687
AU  - Schmidt G
AU  - Grber P
TI  - The rate of ATP hydrolysis catalyzed by reconstituted CF0F1- liposomes
MH  - atp
MH  - CF0F1
MH  - Hydrolysis
MH  - liposome
MH  - Liposomes
MH  - RECONSTITUTED CF0F1
RP  - ON REQUEST (05/29/92)
SO  - Z Naturforsch 1987  ;42():231-236

794
UI  - 1160
AU  - Schmidt G
AU  - Graber P
TI  - The rate of ATP synthesis catalyzed by reconstituted CF0F1- liposomes: dependence on .DELTA.pH and .DELTA..psi
MH  - atp
MH  - ATP synthesis
MH  - CF0F1
MH  - DEPENDENCE
MH  - liposome
MH  - Liposomes
MH  - RECONSTITUTED CF0F1
MH  - synthesis
RP  - ON REQUEST (03/18/92)
SO  - Biochim Biophys Acta 1987  ;890():392-394

795
UI  - 1158
AU  - Schmidt G
AU  - Graber P
TI  - The rate of ATP hydrolysis catalyzed by reconstituted CF0F1- liposomes
MH  - atp
MH  - CF0F1
MH  - Hydrolysis
MH  - liposome
MH  - Liposomes
MH  - RECONSTITUTED CF0F1
RP  - ON REQUEST (03/18/92)
SO  - Z Naturforsch ,C: Biosci 1987  ;42():231-236

796
UI  - 20958
AU  - Schneider E
AU  - Altendorf K
TI  - Bacterial adenosine 5'-triphosphate synthase (F1F0): purification and reconstitution of F0 complexes and biochemical and functional characterization of their subunits
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - ATPase
MH  - Bacterial Proteins
MH  - Carrier Proteins
MH  - COMPLEX
MH  - Dicyclohexylcarbodiimide
MH  - F0
MH  - protein
MH  - Proteins
MH  - Proteolipids
MH  - purification
MH  - reconstitution
MH  - review
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 88142699LA - engRN - 0 (Bacterial Proteins)RN - 0 (Carrier Proteins)RN - 0 (Proteolipids)RN - 0 (dicyclohexylcarbodiimide-binding protein)RN - 0 (dicyclohexylcarbodiimide-binding proteolipid)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticlePT - ReviewPT - Review, AcademicDA - 19880330IS - 0146-0749SB - IMCY - UNITED STATES
UR  - PM:2893973
SO  - Microbiol Rev 1987 Dec ;51(4):477-497

797
UI  - 20962
AU  - Steffens K
AU  - Schneider E
AU  - Deckers-Hebestreit G
AU  - Altendorf K
TI  - Fo portion of Escherichia coli ATP synthase. Further resolution of trypsin-generated fragments from subunit b
AB  - F1-stripped everted membrane vesicles of the ATP synthase-overproducing Escherichia coli strain KY 7485 were treated with trypsin for different lengths of time. Subsequently, the Fo complex was isolated and analyzed by sodium dodecyl sulfate-gel electrophoresis, as well as immunoblotting using antibodies raised against subunit b. By these techniques 3 degradation products with apparent molecular masses of about 16 kDa could be detected in accordance with previous findings (Perlin, D.S., and Senior, A.E. (1985) Arch. Biochem. Biophys. 236, 603- 611). Labeling of isolated trypsin-treated Fo fractions with the thiol- specific reagent N-(7-dimethylamino-4-methylcoumarinyl)-maleimide, which has been demonstrated recently to specifically modify subunit b (Schneider, E., and Altendorf, K. (1985) Eur. J. Biochem. 153, 105-109) revealed that the 16-kDa digestion products were degraded into two stable fragments of 12 and 8.3 kDa. These polypeptides do not react with the anti-b antibodies. Treatment of purified liposome-integrated Fo with trypsin resulted in a similar cleavage pattern. In both cases protease digestion inhibited F1 binding while proton-translocating activity remained unaffected. However, liposomes reconstituted with Fo isolated from trypsin-treated membranes were impaired in both binding of F1 and proton translocation. These activities could be restored when reconstitution was carried out in the presence of native subunit b. From this we conclude that the C-terminal region of subunit b is necessary for proper reconstitution of Fo into liposomes
MH  - A
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BINDING
MH  - COLI ATP SYNTHASE
MH  - COMPLEX
MH  - Electrophoresis
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1
MH  - Liposomes
MH  - Macromolecular Systems
MH  - membrane
MH  - membrane vesicles
MH  - Membranes
MH  - Proteolipids
MH  - proteoliposome
MH  - proton
MH  - reconstitution
MH  - resolution
MH  - Sodium
MH  - SUBUNIT
MH  - SYNTHASE
MH  - SYSTEM
MH  - SYSTEMS
MH  - Time
MH  - translocation
MH  - Trypsin
RP  - NOT IN FILE
NT  - UI - 87194787LA - engRN - 0 (Liposomes)RN - 0 (Macromolecular Systems)RN - 0 (Proteolipids)RN - 0 (proteoliposomes)RN - EC 3.4.21.4 (Trypsin)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19870605IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:2883181
SO  - J Biol Chem 1987 Apr 25 ;262(12):5866-5869

798
UI  - 20961
AU  - Steffens K
AU  - Di Gioia A
AU  - Deckers-Hebestreit G
AU  - Altendorf K
TI  - Structural and functional relationship of ATP synthases (F1F0) from Escherichia coli and the thermophilic bacterium PS3
AB  - Functional compatibility between the F1 and F0 parts of ATP synthases from Escherichia coli (EF1F0) and the thermophilic bacterium PS3 (TF1F0) was analyzed. F1-stripped everted membrane vesicles from both organisms bound the homologous or heterologous F1 part to the same extent. Titration of the reconstituted membrane vesicles with dicyclohexylcarbodiimide revealed a similar sensitivity of the homologous and hybrid F1F0 complexes towards the inhibitor. Furthermore, the heterologous enzymes exhibited ATP-dependent H+ translocation comparable to that of homologous F1F0. Antisera raised against EF1 or subunits a, b, and c of EF0 were analyzed for cross- reactivity with TF1 and TF0. Common antigenic sites have been detected with immunoblot analysis for subunit beta and subunit c of EF1F0 and the corresponding subunits from TF1F0. A weak binding of the anti-a and anti-b antisera with the TF0 part has been observed in an enzyme-linked immunosorbent assay. Based on these findings the structural and functional relationship between the mesophilic and thermophilic ATP synthase complexes is discussed
MH  - A
MH  - analysis
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Bacteria
MH  - BINDING
MH  - COMPLEX
MH  - Dicyclohexylcarbodiimide
MH  - Enzymes
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - F1
MH  - H+
MH  - membrane
MH  - membrane vesicles
MH  - PS3
MH  - Site
MH  - SUBUNIT
MH  - SYNTHASE
MH  - translocation
RP  - NOT IN FILE
NT  - UI - 87194857LA - engRN - 0 (Epitopes)RN - 0 (Immune Sera)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19870605IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:2437118
SO  - J Biol Chem 1987 May 5 ;262(13):6334-6338

799
UI  - 19802
AU  - Stroop SD
AU  - Boyer PD
TI  - Catalytic and regulatory effects of light intensity on chloroplast ATP synthase
AB  - The incorporation of water oxygens into ATP made by photophosphorylation is known to be increased markedly when either Pi or ADP concentration is lowered. The present studies show a similar increase in oxygen exchange when light intensity is lowered even with ample ADP and Pi present. The number of reversals of bound ATP formation prior to release increases about 1 to about 27 in the presence of dithiothreitol and to 5 in its absence. The equilibrium of the bound reactants still favors ATP at low light intensity, as shown by measurement of the amount of bound ATP rapidly labeled from [32P]Pi during steady-state photophosphorylation. Changes observed in the interconversion rate in the absence of added thiol are likely involved in the regulation of the dark ATPase activity in the chloroplast. The interconversion rate of bound ATP to bound ADP and Pi in the presence of thiol is about the same at low and high light intensities. This rate of bound ATP formation is not sufficient, however, to account for the maximum rate of photophosphorylation. Thus, when adequate protonmotive force is present, the rate of conversion of bound ADP and Pi to bound ATP, and possibly that of bound ATP to bound ADP and Pi, must be increased, with proton translocation being completed only when bound ATP is present to be released. These observations are consistent with the predictions of the binding change mechanism with sequential participation of catalytic sites and are accommodated by a simplified general scheme for the binding change mechanism that is presented here.(ABSTRACT TRUNCATED AT 250 WORDS)
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - ADP
MH  - atp
MH  - ATP FORMATION
MH  - ATP synthase
MH  - ATPase
MH  - BINDING
MH  - CHANGE MECHANISM
MH  - chloroplast
MH  - Dithiothreitol
MH  - H(+)-Transporting ATP Synthase
MH  - Light
MH  - mechanism
MH  - Oxygen
MH  - Photophosphorylation
MH  - proton
MH  - regulation
MH  - SYNTHASE
MH  - translocation
MH  - Water
RP  - NOT IN FILE
NT  - UI - 87185454LA - engRN - 0 (Disulfides)RN - 3483-12-3 (Dithiothreitol)RN - 56-65-5 (Adenosine Triphosphate)RN - 7782-44-7 (Oxygen)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM 119034/GM/NIGMSID - GM07185/GM/NIGMSDA - 19870605IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:2882778
SO  - Biochemistry 1987 Mar 10 ;26(5):1479-1484

800
UI  - 77
AU  - Walker JE
AU  - Runswick MJ
AU  - Poulter L
AD  - Medical Research Council, Laboratory of Molecular Biology, Cambridge, UK
TI  - ATP synthase from bovine mitochondria. The characterization and sequence analysis of two membrane-associated sub-units and of the corresponding cDNAs
AB  - ATP synthase from bovine mitochondria is a complex of 13 different polypeptides, whereas the Escherichia coli enzyme is simpler and contains eight subunits only. Two of the bovine subunits, b and d, which had not been characterized, have been isolated from the purified enzyme. Subunits with sizes corresponding to bovine subunits b and d are evident in preparations of the enzyme from mitochondria of other species. Partial protein sequences have been determined by direct methods. On the basis of some of this information, two oligonucleotide mixtures, 17 and 18 bases in length, have been synthesized and used as hybridization probes in the isolation of clones of the cognate cDNAs. The sequences of the two proteins have been deduced from their DNA sequences. Subunit b is 214 amino acid residues in length and has a free N terminus. Subunit d is 160 amino acid residues long. Its N- terminal alanine is blocked by an N-acetyl group, as demonstrated by fast atom bombardment mass spectrometry of N-terminal peptides. The sequence near the N terminus of the b subunit is made predominantly of hydrophobic residues, whereas the remainder of the protein is mainly hydrophilic. This N-terminal hydrophobic region may be folded into an alpha-helical structure spanning the lipid bilayer. In its distribution of hydrophobic residues, this protein resembles the b subunits of ATP synthase complexes in bacteria and chloroplasts. The b subunit in E. coli forms an important structural link between the extramembrane sector of the enzyme F1, and the intrinsic membrane domain, FO. It is proposed that the bovine mitochondrial subunit b serves a similar function. If this is so, the mitochondrial enzyme, as the chloroplast ATP synthase, contains equivalent subunits to all eight of those that constitute the E. coli enzyme. Subunit d has no extensive hydrophobic sequences, and is not apparently related to any subunit described in the simpler ATP synthases in bacteria and chloroplasts
RP  - NOT IN FILE
NT  - UI - 88062745LA - engRN - 0 (Amino Acids)RN - 0 (Macromolecular Systems)RN - 9007-49-2 (DNA)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19880107IS - 0022-2836SB - IMCY - ENGLANDJC - J6V
UR  - PM:2890767
SO  - J Mol Biol 1987 Sep 5 ;197(1):89-100

801
UI  - 82
AU  - Walker JE
AU  - Cozens AL
AU  - Dyer MR
AU  - Fearnley IM
AU  - Powell SJ
AU  - Runswick MJ
TI  - Structure and genes of ATP synthase
RP  - NOT IN FILE
NT  - UI - 87162796LA - engRN - 0 (Macromolecular Systems)RN - 0 (Multienzyme Complexes)RN - EC 2.7 (Phosphotransferases)RN - EC 2.7.4.- (ATP synthetase complex)PT - Journal ArticleDA - 19870513IS - 0300-5127SB - IMCY - ENGLANDJC - E48
UR  - PM:3030835
SO  - Biochem Soc Trans 1987 Feb ;15(1):104-106

802
UI  - 76
AU  - Walker JE
AU  - Gay NJ
AU  - Powell SJ
AU  - Kostina M
AU  - Dyer MR
AD  - MRC Laboratory of Molecular Biology, Cambridge, UK
TI  - ATP synthase from bovine mitochondria: sequences of imported precursors of oligomycin sensitivity conferral protein, factor 6, and adenosinetriphosphatase inhibitor protein
AB  - Oligomycin sensitivity conferral protein (OSCP), factor 6 (F6), and ATPase inhibitor protein are all components of the ATP synthase complex of bovine mitochondria. They are encoded in nuclear DNA. Complementary DNA clones encoding the precursors of these proteins have been isolated from a bovine library by using mixtures of synthetic oligonucleotides as hybridization probes, and their DNA sequences have been determined. The deduced protein sequences show that the OSCP, F6, and inhibitor proteins have N-terminal presequences of 23, 32, and 25 amino acids, respectively. These presequences are not present in the mature proteins. It is assumed that they serve to direct the proteins into the mitochondrial matrix. The cDNA clones have also been employed as hybridization probes to investigate the genetic complexity of the three proteins in cows and humans. These experiments indicate that the bovine and human inhibitor and bovine F6 proteins are encoded by single genes but suggest the possibility of the presence in both species of more than one gene (or pseudogenes) for the OSCP
RP  - NOT IN FILE
NT  - UI - 88163536LA - engRN - 0 (ATPase inhibitory protein)RN - 0 (Carrier Proteins)RN - 0 (Membrane Proteins)RN - 0 (Oligomycins)RN - 0 (Proteins)RN - 0 (oligomycin sensitivity-conferring protein)RN - 9007-49-2 (DNA)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19880506IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:2894843
SO  - Biochemistry 1987 Dec 29 ;26(26):8613-8619

803
UI  - 673
AU  - Weber J
AU  - Rogner M
AU  - Schafer G
TI  - Novel approaches towards characterization of the high-affinity nucleotide binding sites on mitochondrial F1-ATPase by the fluorescence probes 3'-O-(1-naphthoyl)adenosine di- and triphosphate
AB  - The fluorescence properties of 3'-O-(1-naphthoyl)adenosine di- and triphosphates (termed N-ADP and N-ATP, respectively) were investigated in detail. Of special importance for the use of these analogues as environmental probes is their high quantum yield (0.58 in water) and the polarity dependence of shape and wavelength position of the emission spectrum. Upon binding of N-ADP and N-ATP to mitochondrial F1- ATPase, the fluorescence intensity is markedly decreased, due to polarity changes and 'ground-state' quenching. Using this signal for equilibrium binding studies, three (at least a priori) equivalent nucleotide-binding sites were detected on the enzyme. The perspective intrinsic dissociation constants are as follows: N-ADP/Mg2+ 120 nM; N- ATP/Mg2+ 160 nM; N-ADP/EDTA 560 nM; N-ATP/EDTA 3500 nM. For bound ligand the environment was found to be rather unipolar; the rotational mobility of the fluorophore is restricted, its accessibility for iodide anions (as a quencher) is hindered. These facts show a location of the binding sites quite deeply embedded in the protein. The conformation of the binding domains is strongly dependent on the absence or presence of Mg2+, as can be seen from the relative efficiencies of the singlet- singlet energy transfer from tyrosine residues in the protein to bound naphthoyl moieties. Investigation of the binding kinetics revealed this process as biphasic (in presence of Mg2+). After the first fast step (kon greater than 1 X 10(6) M-1 X s-1), in which the analogue is bound to the enzyme, a slow local conformational rearrangement occurs
RP  - NOT IN FILE
NT  - UI - 87214188LA - engRN - 0 (Fluorescent Dyes)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 71160-02-6 (3'-O-(naphthoyl-1)adenosine diphosphate)RN - 76152-01-7 (naphthoyl-ATP)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19870722IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:2883993
SO  - Biochim Biophys Acta 1987 Jun 9 ;892(1):30-41

804
UI  - 532
AU  - Williams N
AU  - Hullihen J
AU  - Pedersen PL
TI  - Ligand binding studies of the F1 moiety of rat liver ATP synthase: implications about the enzyme's structure and mechanism
AB  - F1-ATPase of rat liver was examined for its capacity to interact with both metal ions and nucleotides and for the effect of covalent ATPase inhibitors on these interactions. As isolated, rat liver F1 contains about 2 mol of Mg2+/mol of F1, 1 mol of which can be removed or exchanged. The remaining mole of Mg2+ per mole of F1 remains very tightly associated with F1 and is recovered in the alpha gamma fraction after cold denaturation. Rat liver F1 also contains as isolated a nearly equivalent amount of nucleotide (approximately 1.7 mol/mol of F1) which is readily removed by incubation at room temperature followed by column centrifugation. The "2 Mg2+ enzyme" binds almost 3 mol of 5'- adenylyl imidodiphosphate (AMP-PNP)/mol of F1 in the presence or absence of added divalent cation. When divalent cation is present as Co2+, an equivalent activator to Mg2+ in the ATPase reaction, 1 mol of F1 binds 3 mol of both AMP-PNP and Co2+. under these conditions, the very tight Mg2+ site remains loaded, the exchangeable Mg2+ site is replaced with AMP-PNPCo, and two additional AMP-PNPCo sites are filled. At this point, ADP can be loaded onto the enzyme as a fourth nucleotide at a site separate and distinct from the AMP-PNP sites. Significantly, rat liver F1 contains only a single readily detectable ADP binding site in the presence or absence of divalent cation.(ABSTRACT TRUNCATED AT 250 WORDS)
RP  - NOT IN FILE
NT  - UI - 87157609LA - engRN - 0 (Ligands)RN - 0 (Macromolecular Systems)RN - 0 (Ribonucleotides)RN - 25612-73-1 (Adenylyl Imidodiphosphate)RN - 56-65-5 (Adenosine Triphosphate)RN - 7440-48-4 (Cobalt)RN - 7646-79-9 (cobaltous chloride)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - CA 10951/CA/NCIDA - 19870520IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:2881576
SO  - Biochemistry 1987 Jan 13 ;26(1):162-169

805
UI  - 19797
AU  - Wise JG
AU  - Hicke BJ
AU  - Boyer PD
AD  - Department of Chemistry and Biochemistry, University of California, Los Angeles 90024-1570
TI  - Catalytic and noncatalytic nucleotide binding sites of the Escherichia coli F1 ATPase. Amino acid sequences of beta-subunit tryptic peptides labeled with 2-azido-ATP
AB  - Under appropriate conditions tight, noncovalent binding of 2-azido- adenine nucleotides to either catalytic or noncatalytic binding sites on the E. coli F1-ATPase occurs. After removal of unbound ligands, UV- irradiation results primarily in the covalent incorporation of nucleotide moieties into the beta-subunit in both catalytic and noncatalytic site labeling experiments. Minor labeling of the alpha- subunit was also observed. After trypsin digestion and purification of the labeled peptides, microsequencing studies identified two adjacent beta-subunit tryptic peptides labeled by 2-azido-ADP or -ATP. These beta-subunit peptides were labeled on tyrosine-331 (catalytic sites) and tyrosine-354 (noncatalytic sites) in homology with the labeling patterns of the mitochondrial and chloroplast enzymes
MH  - 2-AZIDO-ATP
MH  - Adenine Nucleotides
MH  - ADENINE-NUCLEOTIDES
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - Amino Acid Sequence
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Azides
MH  - BETA-SUBUNIT
MH  - BINDING
MH  - Binding Sites
MH  - Biochemistry
MH  - Chemistry
MH  - chloroplast
MH  - COLI F1 ATPASE
MH  - Enzymes
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - H(+)-Transporting ATP Synthase
MH  - Ligands
MH  - NONCATALYTIC SITES
MH  - nucleotide binding
MH  - nucleotide binding sites
MH  - NUCLEOTIDE-BINDING-SITES
MH  - Nucleotides
MH  - Peptide Fragments
MH  - purification
MH  - SUBUNIT
MH  - SYNTHASE
MH  - Trypsin
RP  - NOT IN FILE
NT  - UI - 88030084LA - engRN - 0 (Affinity Labels)RN - 0 (Azides)RN - 0 (Peptide Fragments)RN - 56-65-5 (Adenosine Triphosphate)RN - 72884-75-4 (2-azidoadenosine 5'-triphosphate)RN - EC 3.4.21.4 (Trypsin)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM11094/GM/NIGMSDA - 19871209IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:2889623
SO  - FEBS Lett 1987 Nov 2 ;223(2):395-401

806
UI  - 464
AU  - Wood JM
AU  - Wise JG
AU  - Senior AE
AU  - Futai M
AU  - Boyer PD
TI  - Catalytic properties of the F1-adenosine triphosphatase from Escherichia coli K-12 and its genetic variants as revealed by 18O exchanges
AB  - We have examined intermediate Pi-water oxygen exchange during [gamma- 18O]ATP hydrolysis by the F1 adenosine triphosphatase from Escherichia coli K-12. Water oxygen incorporation into each Pi released was increased as ATP concentration was lowered as observed previously for the same reaction catalyzed by the enzyme from eukaryotic sources. Heterogeneous distributions of 18O in product Pi were produced by coexisting epsilon subunit-replete and epsilon subunit-depleted enzyme molecules. The epsilon-replete enzyme showed a much higher probability for oxygen exchange. These data imply that the epsilon subunit inhibits net ATP hydrolysis by imposing conformational constraints which reduce the cooperative conformational interactions that promote ADP and Pi release. Four enzyme variants altered in alpha or beta subunit structure with reduced net hydrolytic activity showed sharply increased oxygen exchange during ATP hydrolysis. Heterogeneity was apparent in the 18O distribution of the product Pi, however. That behavior could reflect hindered conformational interactions and/or increased affinity of the alpha 3 beta 3 gamma delta complex for the epsilon subunit. In contrast, enzyme from mutant uncA401 showed very little oxygen exchange accompanying hydrolysis of 20 microM ATP. This is the only enzyme so far reported with this unusual property. Its rate limitation appears to be in the hydrolytic rather than the product release step of the catalytic sequence
RP  - NOT IN FILE
NT  - UI - 87137441LA - engRN - 0 (Isoenzymes)RN - 0 (Macromolecular Systems)RN - 0 (Oxygen Isotopes)RN - 7782-44-7 (Oxygen)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM-25349/GM/NIGMSDA - 19870330IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:2880843
SO  - J Biol Chem 1987 Feb 15 ;262(5):2180-2186

807
UI  - 19800
AU  - Xue ZX
AU  - Zhou JM
AU  - Melese T
AU  - Cross RL
AU  - Boyer PD
AD  - Department of Chemistry and Biochemistry, University of California, Los Angeles 90024
TI  - Chloroplast F1 ATPase has more than three nucleotide binding sites, and 2-azido-ADP or 2-azido-ATP at both catalytic and noncatalytic sites labels the beta subunit
AB  - The photolabeling of chloroplast F1 ATPase, following exposure to Mg2+ and 2-azido-ATP and separation from medium nucleotides, results in derivatization of two separate peptide regions of the beta subunit. Up to 3 mol of the analogue can be incorporated per mole of CF1, with covalent binding of one moiety or two moieties per beta subunit that can be either AMP, ADP, or ATP derivatives. These results, the demonstration of noncovalent tight binding of at least four [3H]adenine nucleotides to the enzyme and the presence of three beta subunits per enzyme, point to six potential adenine nucleotide binding sites per molecule. The tightly bound 2-azido nucleotides on CF1, found after exposure of the heat-activated and EDTA-treated enzyme to Mg2+ and 2- azido-ATP, differ in their ease of replacement during subsequent hydrolysis of ATP. Some of the bound nucleotides are not readily replaced during catalytic turnover and covalently label one peptide region of the beta subunit. They are on noncatalytic sites. Other tightly bound nucleotides are readily replaced during catalytic turnover and label another peptide region of the beta subunit. They are at catalytic sites. No alpha-subunit labeling is detected upon photolysis of the bound 2-azido nucleotides. However, one or both of the sites could be at an alpha-beta-subunit interface with the 2-azido region close to the beta subunit, or both binding sites may be largely or entirely on the beta subunit
MH  - 2-AZIDO-ATP
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - ADP
MH  - ALPHA-SUBUNIT
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Azides
MH  - BETA-SUBUNIT
MH  - BINDING
MH  - Binding Sites
MH  - Biochemistry
MH  - BOUND NUCLEOTIDES
MH  - Chemistry
MH  - chloroplast
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - H(+)-Transporting ATP Synthase
MH  - Hydrolysis
MH  - NONCATALYTIC SITES
MH  - nucleotide binding
MH  - nucleotide binding sites
MH  - NUCLEOTIDE-BINDING-SITES
MH  - Nucleotides
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 88000583LA - engRN - 0 (Azides)RN - 0 (Nucleotides)RN - 10028-17-8 (Tritium)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 64020-53-7 (2-azidoadenosine 3',5'-diphosphate)RN - 72884-75-4 (2-azidoadenosine 5'-triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM 11094/GM/NIGMSID - GM 23152/GM/NIGMSDA - 19871116IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:2888481
SO  - Biochemistry 1987 Jun 30 ;26(13):3749-3753

808
UI  - 19798
AU  - Xue ZX
AU  - Miller CG
AU  - Zhou JM
AU  - Boyer PD
AD  - Department of Chemistry and Biochemistry, University of California, Los Angeles 90024-1570
TI  - Catalytic and noncatalytic nucleotide binding sites of chloroplast F1 ATPase. Photoaffinity labeling and peptide sequencing
AB  - Exposure of chloroplast F1 ATPase to 2-azido-ATP results in the noncovalent tight binding of 2-azido-ATP or 2-azido-ADP to noncatalytic or to catalytic sites. Subsequent photolysis results in covalent labeling of adjacent tryptic peptides of the beta-subunit. Binding at noncatalytic sites results in labeling of tyrosine 385 by an ATP or an ADP moiety. Binding at catalytic sites results in labeling of tyrosine 362 by only an ADP moiety. Similar labeling patterns are observed for the heat-activated or the membrane-bound enzymes
MH  - 2-AZIDO-ATP
MH  - Adenine Nucleotides
MH  - ADENINE-NUCLEOTIDES
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - ADP
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BETA-SUBUNIT
MH  - BINDING
MH  - Binding Sites
MH  - Biochemistry
MH  - Chemistry
MH  - chloroplast
MH  - Enzymes
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - H(+)-Transporting ATP Synthase
MH  - NONCATALYTIC SITES
MH  - nucleotide binding
MH  - nucleotide binding sites
MH  - NUCLEOTIDE-BINDING-SITES
MH  - Nucleotides
MH  - Peptide Fragments
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 88030083LA - engRN - 0 (Adenine Nucleotides)RN - 0 (Affinity Labels)RN - 0 (Peptide Fragments)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM11094/GM/NIGMSDA - 19871209IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:2889622
SO  - FEBS Lett 1987 Nov 2 ;223(2):391-394

809
UI  - 579
AU  - Yagi T
AU  - Hatefi Y
TI  - Thiols in oxidative phosphorylation: thiols in the F0 of ATP synthase essential for ATPase activity
AB  - It was shown previously that the ATP synthase complex of bovine heart mitochondria contains an essential set of thiols or dithiols in its membrane sector (F0), whose modification by various reagents results in uncoupling [Yagi, T., and Hatefi, Y. (1984) Biochemistry 23, 2449- 2455]. The sensitivity to modifiers was increased by membrane energization, and the uncoupling was reversed by membrane-permeable thiol compounds when modifiers other than alkylating agents were used to uncouple. The present paper demonstrates that there exists in the F0 of bovine ATP synthase another set of essential thiols, whose modification results in reversible inhibition of ATPase activity. These thiols are most susceptible to modification by mercurials (p- chloromercuribenzoate greater than p-chloromercuribenzene sulfonate) and do not appear to be modified by N-ethylmaleimide. The reversible modification of these thiols by mercurials protects the ATP synthase against irreversible inhibition in F0 by N,N-dicyclohexylcarbodiimide. The possible location of these two sets of thiols in the F0 of bovine ATP synthase is discussed
RP  - NOT IN FILE
NT  - UI - 87211980LA - engRN - 0 (Chloromercuribenzoates)RN - 0 (Oligomycins)RN - 0 (Sulfhydryl Compounds)RN - 0 (Sulfhydryl Reagents)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 554-77-8 (4-Chloromercuribenzenesulfonate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - AM 08126/AM/NIADDKID - GM 33712/GM/NIGMSDA - 19870529IS - 0003-9861SB - IMCY - UNITED STATESJC - 6SK
UR  - PM:2883930
SO  - Arch Biochem Biophys 1987 Apr ;254(1):102-109

810
UI  - 763
AU  - Yohda M
AU  - Yoshida M
AD  - Department of Chemical Engineering, University of Tokyo
TI  - Single-site catalysis of F1-ATPase from thermophilic bacterium PS3 and its dominance in steady-state catalysis at low ATP concentration
AB  - Single-site catalysis by F1-ATPase from a thermophilic bacterium PS3 (TF1) was examined by incubating the enzyme with a submolar amount of radioactive ATP. The profile of single-site catalysis by TF1 at 23 degrees C was different from that of beef heart mitochondrial F1-ATPase (MF1). ATP hydrolysis on the enzyme and release of the products was rapid, and subsequent addition of non-radioactive ATP (cold chase) did not promote the hydrolysis of radioactive ATP, indicating that the rate- limiting step was not the step of product release but the step of ATP binding to the enzyme. Thus, the characteristic features of so-called uni-site catalysis were not observed. At 60 degrees C, whether in the presence or absence of phosphate ion, a small amount of bound [alpha, gamma-32P]ATP and cold chase promotion were observed. However, since bound 32P1 was not detected by centrifugal gel filtration, it is not yet certain whether TF1 has typical uni-site characteristics. Based on the hydrolytic turnover rate for single-site catalysis and analysis of the kinetics of steady-state catalysis, it is proposed that single-site catalysis is dominant even in steady-state catalysis at ATP concentrations of less than about 20 microM
RP  - NOT IN FILE
NT  - UI - 88139241LA - engRN - 0 (Adenine Nucleotides)RN - 0 (Phosphates)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19880325IS - 0021-924XSB - IMCY - JAPANJC - HIF
UR  - PM:2893790
SO  - J Biochem (Tokyo ) 1987 Oct ;102(4):875-883

811
UI  - 452
AU  - al Shawi MK
AU  - Parsonage D
AU  - Senior AE
AD  - Department of Biochemistry, University of Rochester Medical Center, New York 14642
TI  - Directed mutagenesis of the strongly conserved aspartate 242 in the beta-subunit of Escherichia coli proton-ATPase
AB  - Oligonucleotide-directed mutagenesis was used to substitute Asn or Val for residue Asp-242 in the beta-subunit of Escherichia coli F1-ATPase. Asp-242 is strongly conserved in beta-subunits of F1-ATPase enzymes, in a region of sequence which shows homology with numerous nucleotide- binding proteins. By analogy with adenylate kinase (Fry, D.C., Kuby, S.A., and Mildvan, A.S. (1986) Proc. Natl. Acad. Sci. U.S.A. 83, 907- 911), beta-Asp-242 of F1-ATPase might participate in catalysis through electrostatic effects on the substrate Mg2+ or through hydrogen bonding to the substrate(s); an acid-base catalytic role is also plausible. The substitutions Asn and Val were chosen to affect the charge, hydrogen- bonding ability, and hydrophobicity of residue beta-Asp-242. Both mutations significantly impaired oxidative phosphorylation rates in vivo and membrane ATPase and ATP-driven proton-pumping activities in vitro. Asn-242 was more detrimental than Val-242. Purified soluble mutant F1-ATPases had normal molecular size and subunit composition, and displayed 7% (beta-Asn-242) and 17% (beta-Val-242) of normal specific Mg-ATPase activity. The relative MgATPase activities of both mutant enzymes showed similar pH dependence to normal. Relative MgATPase and CaATPase activities of normal and mutant enzymes were compared at widely varied pMg and pCa. The mutations had little effect on KM MgATP, but KM CaATP was reduced. The data showed that the carboxyl side-chain of beta-Asp-242 is not involved in catalysis either as a general acid-base catalyst or through direct involvement in any protonation/deprotonation-linked mechanism, nor is it likely to be directly involved in liganding to substrate Mg2+ during the reaction. Specificity constants (kcat/KM) for MgATP and CaATP were reduced in both mutant enzymes, showing that the mutations destabilized interactions between the catalytic nucleotide-binding domain and the transition state
RP  - NOT IN FILE
NT  - UI - 89066792LA - engRN - 0 (Macromolecular Systems)RN - 56-84-8 (Aspartic Acid)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSID - GM29805/GM/NIGMSDA - 19890124IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:2904440
SO  - J Biol Chem 1988 Dec 25 ;263(36):19633-19639

812
UI  - 451
AU  - al Shawi MK
AU  - Senior AE
AD  - Department of Biochemistry, University of Rochester Medical Center, New York 14642
TI  - Complete kinetic and thermodynamic characterization of the unisite catalytic pathway of Escherichia coli F1-ATPase. Comparison with mitochondrial F1-ATPase and application to the study of mutant enzymes
AB  - A complete analysis is presented of the component rate constants of the "unisite" reaction pathway in normal Escherichia coli F1-ATPase. Gibbs free energy profiles of the unisite reaction pathway were constructed for both normal E. coli F1 and bovine-heart mitochondrial F1, and comparison indicated that E. coli F1 is an ancestral form of the mitochondrial enzyme. Similar kinetic and thermodynamic analyses of the unisite reaction pathway were done for mutant beta-Asn-242 and beta-Val- 242 E. coli F1-ATPases. Both mutations affected unisite binding and hydrolysis of MgATP but had little effect on release of products or binding of MgADP. It was apparent that a primary effect of the mutations was on the interaction between the catalytic nucleotide- binding domain and the substrate MgATP. The catalytic transition state [F1-ATP]++ was the most destabilized step in the reaction sequence. Measurements of delta delta G[F1.ATP]++ and linear free energy plots for the catalytic step were consistent with the view that, in normal enzyme, residue beta-Asp-242 accepts an H-bond from the transition- state substrate in order to facilitate catalysis. Both mutations impaired positive catalytic cooperativity. This was caused by energetic destabilization of the catalytic transition state and was an indirect effect, not a direct effect on signal transmission per se between catalytic nucleotide-binding domains on beta-subunits. Therefore, impairment of unisite catalysis and of positive catalytic cooperativity appeared to be linked. This may provide a unifying explanation as to why a series of other, widely separated mis-sense mutations within the catalytic nucleotide-binding domain on F1-beta-subunit, which have been reported to affect unisite catalysis, also impair positive catalytic cooperativity. Linear free energy plots for the ATP-binding step of unisite catalysis demonstrated that beta-Asn-242 and beta-Val-242 mutant enzymes did not suffer any gross disruptive change in structure of the catalytic nucleotide-binding domain, reinforcing the view that impairment of catalysis was due to a localized effect. Such analyses confirmed that six other F1-beta-subunit mutants, previously generated and characterized in this laboratory and thought to have inhibitory side-chain substitutions in the catalytic nucleotide-binding domain, are also devoid of gross structural disruption
RP  - NOT IN FILE
NT  - UI - 89066793LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSID - GM29805/GM/NIGMSDA - 19890124IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:2904441
SO  - J Biol Chem 1988 Dec 25 ;263(36):19640-19648

813
UI  - 21013
AU  - Boekema E
AU  - van Heel M
AU  - Graber P
AD  - Fritz-Haber-Institut der Max-Planck Gesellschaft, Berlin
TI  - Structure of the ATP-synthase from chloroplasts studied by electron microscopy and image processing
MH  - ATP synthase
MH  - ATPase
MH  - chloroplast
MH  - Chloroplasts
MH  - electron
MH  - electron microscopy
MH  - ELECTRON-MICROSCOPY
MH  - image processing
MH  - Microscopy
RP  - NOT IN FILE
NT  - UI - 88336211LA - engRN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19881027IS - 0361-7742SB - IMCY - UNITED STATES
UR  - PM:2901753
SO  - Prog Clin Biol Res 1988  ;273():75-80

814
UI  - 21012
AU  - BOEKEMA EJ
AU  - Schmidt G
AU  - Graber P
AU  - Berden JA
AD  - Fritz-Haber-Institut der Max-Planck-Gesellschaft, Berlin, Bundesrepublik Deutschland
TI  - Structure of the ATP-synthase from chloroplasts and mitochondria studied by electron microscopy
AB  - The structure of the ATP-synthase, F0F1, from spinach chloroplasts and beef heart mitochondria has been investigated by electron microscopy with negatively stained specimens. The detergent-solubilized ATP- synthase forms string-like structures in which the F0 parts are aggregated. In most cases, the F1 parts are arranged at alternating sides along the string. The F0 part has an approximate cylindrical shape with heights of 8.3 and 8.9 nm and diameters of 6.2 and 6.4 nm for the chloroplast and mitochondrial enzyme, respectively. The F1 parts are disk-like structures with a diameter of about 11.5 nm and a height of about 8.5 nm. The F1 parts are attached to the strings, composed of F0 parts, in most cases, with their smallest dimension parallel to the strings. The stalk connecting F0 and F1 has a length of 3.7 nm and 4.3 nm and a diameter of 2.7 nm and 4.3 nm for the chloroplast and mitochondrial enzyme, respectively
MH  - A
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - chloroplast
MH  - Chloroplasts
MH  - electron
MH  - electron microscopy
MH  - ELECTRON-MICROSCOPY
MH  - F0
MH  - F0F1
MH  - F1
MH  - Lipid Bilayers
MH  - Microscopy
MH  - Mitochondria
MH  - SPECIMENS
MH  - Spinach
MH  - SPINACH-CHLOROPLASTS
MH  - stalk
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 88266186LA - engRN - 0 (Lipid Bilayers)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19880811IS - 0341-0382SB - IMCY - GERMANY, WEST
UR  - PM:2898840
SO  - Z Naturforsch [C] 1988 Mar ;43(3-4):219-225

815
UI  - 19048
AU  - Bowman EJ
AU  - Siebers A
AU  - Altendorf K
TI  - Bafilomycins: a class of inhibitors of membrane ATPases from microorganisms, animal cells, and plant cells
MH  - A
MH  - A1
MH  - Adenosinetriphosphatase
MH  - Animal
MH  - antagonists & inhibitors
MH  - ANTIBIOTIC
MH  - Antibiotics
MH  - ATPase
MH  - Bacteria
MH  - biology
MH  - Brain
MH  - cell
MH  - Cells
MH  - Chromaffin Granules
MH  - enzymology
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - inhibitor
MH  - inhibitors
MH  - lactones
MH  - membrane
MH  - Membranes
MH  - Mitochondria
MH  - Neurospora
MH  - pharmacology
MH  - plant
MH  - Support,Non-U.S.Gov't
MH  - Support,U.S.Gov't,P.H.S.
MH  - universities
MH  - vacuoles
RP  - NOT IN FILE
NT  - Various membrane ATPases have been tested for their sensitivity to bafilomycin A1, a macrolide antibiotic. F1F0 ATPases from bacteria and mitochondria are not affected by this antibiotic. In contrast, E1E2 ATPases--e.g., the K+-dependent (Kdp) ATPase from Escherichia coli, the Na+,K+-ATPase from ox brain, and the Ca2+- ATPase from sarcoplasmic reticulum--are moderately sensitive to this inhibitor. Finally, membrane ATPases from Neurospora vacuoles, chromaffin granules, and plant vacuoles are extremely sensitive. From this we conclude that bafilomycin A1 is a valuable tool for distinguishing among the three different types of ATPases and represents the first relatively specific potent inhibitor of vacuolar ATPases Department of Biology, University of California, Santa Cruz 95064PMID- 0002973058
SO  - Proc Natl Acad Sci U S A 1988  ;85(21):7972-7976

816
UI  - 19795
AU  - Boyer PD
TI  - Bioenergetic coupling to protonmotive force: should we be considering hydronium ion coordination and not group protonation?
MH  - England
MH  - ion
MH  - proton
MH  - Protons
MH  - review
RP  - NOT IN FILE
NT  - UI - 89223002LA - engRN - 0 (Protons)PT - Journal ArticlePT - ReviewPT - Review, TutorialID - GM11940/GM/NIGMSDA - 19890608IS - 0968-0004SB - IMCY - ENGLANDJC - WEF
UR  - PM:2854307
SO  - Trends Biochem Sci 1988 Jan ;13(1):5-7

817
UI  - 392
AU  - Bulygin VV
AU  - Vinogradov AD
AD  - Department of Biochemistry, School of Biology, Moscow State University, USSR
TI  - Three adenine nucleotide binding sites in F1-F0 mitochondrial ATPase as revealed by presteady-state and steady-state kinetics of ATP hydrolysis. Evidence for two inhibitory ADP-specific noncatalytic sites
AB  - Preincubation of submitochondrial particles with ADP in the presence of Mg2+ results in the complete inhibition of ATPase which is slowly reactivated in the assay mixture containing ATP and the ATP regenerating system. Significantly, the rate of activation increases as the concentration of ADP in the preincubation mixture rises from 1 microM to 20 microM and reaches a constant value at higher ADP concentrations. The first-order rate constant for the activation process in the assay mixture is ATP-dependent at any level of inhibitory ADP. The data obtained strongly suggest that two ADP- specific inhibitory sites and one ATP-specific hydrolytic site are present in F1-F0 ATPase. Taking into account the (3 alpha.3 beta).gamma.delta.epsilon structure of F1, it is concluded that the synchronous discharge of ADP from two inhibitory sites during the activation occurs after ATP binds to the ATPase catalytic site
RP  - NOT IN FILE
NT  - UI - 88313025LA - engRN - 0 (Adenine Nucleotides)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19881006IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:2900778
SO  - FEBS Lett 1988 Aug 29 ;236(2):497-500

818
UI  - 20821
AU  - Cain BD
AU  - Simoni RD
AD  - Department of Biological Sciences, Stanford University, California 94305-5020
TI  - Interaction between Glu-219 and His-245 within the a subunit of F1F0- ATPase in Escherichia coli
AB  - Oligonucleotide-directed mutagenesis was used to generate mutations in the a subunit gene (uncB) altering the glutamic acid 219 and the histidine 245 codons. Substitutions of aspartic acid, glutamine, histidine, and leucine for glutamic acid at position 219 neither alter the hydrolytic activity of membrane-bound F1 nor the association of F1 with F0. However, the efficiency of F0-mediated proton translocation was reduced to varying degrees. Replacement of glutamic acid 219 with leucine reduced the ATP-driven proton pumping activity of intact F1F0 to undetectable levels. Roughly 5% of normal activity was observed when glutamine occupied position 219. Surprisingly higher activity, approaching 20% of wild type levels, is seen when histidine replaced glutamic acid 219. The aspartic acid substitution resulted in little loss of enzyme function. Substitution of glutamic acid for histidine 245 results in a reduction to about 45% of normal proton translocation. Construction of the double mutant with substitution of histidine at position 219 and glutamic acid at position 245 yields a complex with better proton translocation than with either mutant separately. The possibility that a functionally important interaction between histidine 245 and glutamic acid 219 of the a subunit may be directly involved in the proton translocation mechanism of F1F0-ATP synthase is discussed
MH  - A
MH  - ACID
MH  - ATPase
MH  - COMPLEX
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - F1
MH  - F1F0-ATP SYNTHASE
MH  - function
MH  - glutamic acid
MH  - Glutamine
MH  - Histidine
MH  - Macromolecular Systems
MH  - mechanism
MH  - mutagenesis
MH  - mutant
MH  - proton
MH  - SUBUNIT
MH  - SYNTHASE
MH  - SYSTEM
MH  - SYSTEMS
MH  - translocation
RP  - NOT IN FILE
NT  - UI - 88198222LA - engRN - 0 (Glutamates)RN - 0 (Macromolecular Systems)RN - 0 (Plasmids)RN - 56-86-0 (Glutamic Acid)RN - 71-00-1 (Histidine)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleID - GM09521/GM/NIGMSID - GM18539/GM/NIGMSDA - 19880609IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:2896197
SO  - J Biol Chem 1988 May 15 ;263(14):6606-6612

819
UI  - 19871
AU  - Cross RL
AD  - Department of Biochemistry and Molecular Biology, State University of New York, Health Science Center, Syracuse 13210
TI  - The number of functional catalytic sites on F1-ATPases and the effects of quaternary structural asymmetry on their properties
AB  - Recent structural and kinetic studies of F1 and F0F1 are reviewed with regard to their implications for the binding change mechanism for ATP synthesis by oxidative phosphorylation and photophosphorylation. It is concluded that at least two and probably all three of the catalytic sites on F1 are functionally equivalent despite permanent structural asymmetry in the soluble enzyme. A rotary mechanism in which all three catalytic subunits experience all possible interactions with the single- copy subunits during turnover is thought not to apply to soluble F1 but remains an attractive model for the membrane bound enzyme
MH  - A
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - BINDING
MH  - Biochemistry
MH  - CHANGE MECHANISM
MH  - F0F1
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - H(+)-Transporting ATP Synthase
MH  - mechanism
MH  - membrane
MH  - model
MH  - Oxidative Phosphorylation
MH  - Phosphorylation
MH  - Photophosphorylation
MH  - review
MH  - SUBUNIT
MH  - SYNTHASE
MH  - synthesis
RP  - NOT IN FILE
NT  - UI - 89123234LA - engRN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialID - GM 23152/GM/NIGMSDA - 19890316IS - 0145-479XSB - IMCY - UNITED STATESJC - HIO
UR  - PM:2906058
SO  - J Bioenerg Biomembr 1988 Aug ;20(4):395-405

820
UI  - 19870
AU  - Cunningham D
AU  - Cross RL
AD  - Department of Biochemistry and Molecular Biology, State University of New York Health Science Center, Syracuse 13210
TI  - Catalytic site occupancy during ATP hydrolysis by MF1-ATPase. Evidence for alternating high affinity sites during steady-state turnover
AB  - The mechanism of ATP hydrolysis by the solubilized mitochondrial ATPase (MF1) has been studied under conditions where catalytic turnover occurs at one site, uni-site catalysis (obtained when enzyme is in excess of substrate), or at two sites, bi-site catalysis (obtained when substrate is in excess of enzyme). Pulse-chase experiments support the conclusion that the sites which participate in bi-site catalysis are the same as those which participate in uni-site catalysis. Upon addition of ATP in molar excess to MF1, label that was bound under uni-site conditions dissociates at a rate equal to the rate of bi-site catalysis. Similarly, when medium ATP is removed, label that was bound under bi- site conditions dissociates at a rate equal to the rate of uni-site catalysis. Evidence that a high affinity catalytic site equivalent to the one observed under uni-site conditions participates as an intermediate in bi-site catalysis includes the demonstration of full occupancy of a catalytically competent site during steady-state turnover at nanomolar concentrations of ATP. Improved measurements of the interaction of ADP at a high affinity catalytic site have lead to the revision of several of the rate constants that define uni-site catalysis. The rate constant for unpromoted dissociation of ADP is equal to that for Pi (4 X 10(-3) s-1). The rate of binding ADP at a high affinity chaseable site (Kd = 1 nM) is equal to the rate of binding ATP (4 X 10(6) M-1 s-1). The rate of catalysis obtained when substrate binding at one site promotes product release from an adjacent site (bi-site catalysis) is up to 100,000-fold faster than unpromoted product release (uni-site catalysis)
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - ADP
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BINDING
MH  - Biochemistry
MH  - Catalysis
MH  - H(+)-Transporting ATP Synthase
MH  - Hydrolysis
MH  - mechanism
MH  - SYNTHASE
MH  - uni-site catalysis
MH  - UNISITE CATALYSIS
RP  - NOT IN FILE
NT  - UI - 89066677LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM 23152/GM/NIGMSDA - 19890120IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:2904435
SO  - J Biol Chem 1988 Dec 15 ;263(35):18850-18856

821
UI  - 21289
AU  - Decker ER
AU  - Levitt DG
AD  - Department of Physiology, University of Minnesota, Minneapolis 55455
TI  - Use of weak acids to determine the bulk diffusion limitation of H+ ion conductance through the gramicidin channel
AB  - The addition of 2 M formic acid at pH 3.75 increased the single channel H+ ion conductance of gramicidin channels 12-fold at 200 mV. Other weak acids (acetic, lactic, oxalic) produce a similar, but smaller increase. Formic acid (and other weak acids) also blocks the K+ conductance at pH 3.75, but not at pH 6.0 when the anion form predominates. This increased H+ conductance and K+ block can be explained by formic acid (HF) binding to the mouth of the gramicidin channel (Km = 1 M) and providing a source of H+ ions. A kinetic model is derived, based on the equilibrium binding of formic acid to the channel mouth, that quantitatively predicts the conductance for different mixtures of H+, K+, and formic acid. The binding of the neutral formic acid to the mouth of the gramicidin channel is directly supported by the observation that a neutral molecule with a similar structure, formamide (and malonamide and acrylamide), blocks the K+ conductance at pH 6.0. The H+ conductance in the presence of formic acid provides a lower bound for the intrinsic conductance of the gramicidin channel when there is no diffusion limitation at the channel mouth. The 12-fold increase in conductance produced by formic acid suggests that greater than 90% of the total resistance to H+ results from diffusion limitation in the bulk solution
MH  - A
MH  - ACID
MH  - BINDING
MH  - conductance
MH  - Diffusion
MH  - H+
MH  - ion
MH  - Ion Channels
MH  - Ions
MH  - M
MH  - model
MH  - pH
MH  - physiology
MH  - Potassium
MH  - proton
MH  - Protons
MH  - structure
RP  - NOT IN FILE
NT  - UI - 88135044LA - engRN - 0 (Amides)RN - 0 (Carboxylic Acids)RN - 0 (Formic Acids)RN - 0 (Ion Channels)RN - 0 (Protons)RN - 1405-97-6 (Gramicidin)RN - 64-18-6 (formic acid)RN - 7440-09-7 (Potassium)PT - Journal ArticleDA - 19880325IS - 0006-3495SB - IMCY - UNITED STATES
UR  - PM:2449253
SO  - Biophys J 1988 Jan ;53(1):25-32

822
UI  - 758
AU  - Denda K
AU  - Konishi J
AU  - Oshima T
AU  - Date T
AU  - Yoshida M
AD  - Department of Life Science, Tokyo Institute of Technology, Yokohama, Japan
TI  - Molecular cloning of the beta-subunit of a possible non-F0F1 type ATP synthase from the acidothermophilic archaebacterium, Sulfolobus acidocaldarius
AB  - The gene which encodes the beta subunit of the novel membrane- associated ATPase has been identified and characterized. The beta subunit, which is most likely the soluble part of the non-F0F1 type H+- ATPase, was obtained from the archaebacterium, Sulfolobus acidocaldarius. In terms of its location, it follows just after the gene for its alpha subunit. It is comprised of 1398 nucleotides, corresponding to a protein of 465 amino acids, and the consensus sequence in the nucleotide binding proteins is poorly conserved. Together with previously described results, the distant homology of the S. acidocaldarius ATPase alpha and beta subunits when compared to those of F0F1-ATPases indicates that this archaebacterial ATPase belongs to an ion-translocating ATPase family uniquely different than F0F1-ATPases even if S. acidocaldarius ATPase and F0F1-ATPases have been derived from a common ancestral ATPase
RP  - NOT IN FILE
NT  - UI - 89034240LA - engRN - 0 (DNA, Bacterial)RN - 0 (Macromolecular Systems)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19881220IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:2903160
SO  - J Biol Chem 1988 Nov 25 ;263(33):17251-17254

823
UI  - 759
AU  - Denda K
AU  - Konishi J
AU  - Oshima T
AU  - Date T
AU  - Yoshida M
AD  - Department of Life Science, Tokyo Institute of Technology, Yokohama, Japan
TI  - The membrane-associated ATPase from Sulfolobus acidocaldarius is distantly related to F1-ATPase as assessed from the primary structure of its alpha-subunit
AB  - Isolation of novel membrane-associated ATPases, presumably soluble parts of the H+-ATPases, from archaebacteria has been recently reported, and their properties were found to be significantly different from the usual F1-ATPase. In order to assess the relationship of the archaebacterial ATPases to the F1-ATPases and other known ATPases, the amino acid sequence of the alpha subunit of the ATPase from Sulfolobus acidocaldarius, an acidothermophilic archaebacterium, was compared with the sequences of other ATPases. The gene encoding its alpha subunit was cloned from the genomic library of S. acidocaldarius, and the nucleotide sequence was determined. The 591-amino acid sequence deduced from the nucleotide sequence contains a small number of short stretches that shows sequence similarity to the alpha and beta subunits of F1- ATPase. However, the overall similarity is too weak to consider it to be a typical member of the F1-ATPase family when the highly conserved sequences of the F1-ATPase subunits among various organisms are taken into account. Moreover, most of these stretches overlap the consensus sequences that are commonly found in some nucleotide-binding proteins. There is no significant sequence similarity to the ion-translocating ATPases, which form phosphorylated intermediates, such as animal Na+,K+- ATPases. Thus, the S. acidocaldarius ATPase and probably other archaebacterial ATPases also appear to belong to a new group of ion- translocating ATPases that has only a distant relationship to F1-ATPase
RP  - NOT IN FILE
NT  - UI - 88198130LA - engRN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19880606IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:2896191
SO  - J Biol Chem 1988 May 5 ;263(13):6012-6015

824
UI  - 311
AU  - Engelbrecht S
AU  - Junge W
AD  - Biophysik, Fachbereich Biologie/Chemie, Universitat Osnabruck, Federal Republic of Germany
TI  - Purified subunit delta of chloroplast coupling factor CF1 reconstitutes photophosphorylation in partially CF1-depleted membranes
AB  - The ATP synthase of chloroplasts consists of the proton channel, CF0, and the catalytic part, CF1, which carries nucleotide-binding sites on subunits alpha and beta. The still poorly understood interaction between CF0 and the catalytic sites on CF1 is mediated by the smaller subunits gamma, delta and epsilon of CF1. We investigated the ability of purified delta to block proton leakage through CF0 channels after their exposure by removal of the CF1 counterpart. Thylakoids were partially depleted of CF1 by EDTA treatment. This increased their proton permeability and thereby reduced the rate of photophosphorylation. Subunit delta was isolated and purified by FPLC [Engelbrecht, S. and Junge, W. (1987) FEBS Lett. 219, 321-325]. Addition of delta to EDTA-treated thylakoids reconstituted high rates of phenazine-methosulfate-mediated photophosphorylation. Since delta does not interact with nucleotides by itself, the reconstitution was due to a reduction of the proton leakage through open CF0 channels. The molar ratio of purified delta over exposed CF0, which started to elicit this effect, was 3:1. However, if delta was added together with purified CF1 lacking delta, in a 1:1 molar ratio, the relative amount over exposed CF0 was as low as 0.06. This corroborated our previous conclusion [Lill, H., Engelbrecht, S., Schonknecht, G. and Junge, W. (1986) Eur. J. Biochem. 160, 627-634] that only a very small fraction of exposed CF0 was actually proton-conducting but with a very high unit conductance. CF1 including delta was apparently rebound preferentially to open CF0 channels. Although the ability of delta to control proton conduction through CF0 was evident, it remains to be established whether delta acts as a gated proton valve or as a conformational transducer in the integral CF0CF1 ATPase
RP  - NOT IN FILE
NT  - UI - 88151990LA - engRN - 0 (Macromolecular Systems)RN - 0 (Membrane Proteins)RN - 0 (Protons)RN - 0 (thylakoid membrane phosphoprotein)RN - 56-65-5 (Adenosine Triphosphate)RN - 60-00-4 (Edetic Acid)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19880419IS - 0014-2956SB - IMCY - GERMANY, WESTJC - EMZ
UR  - PM:2894310
SO  - Eur J Biochem 1988 Feb 15 ;172(1):213-218

825
UI  - 20879
AU  - Eya S
AU  - Noumi T
AU  - Maeda M
AU  - Futai M
AD  - Department of Organic Chemistry and Biochemistry, Osaka University, Japan
TI  - Intrinsic membrane sector (Fo) of H+-ATPase (FoF1) from Escherichia coli. Mutations in the alpha subunit give Fo with impaired proton translocation and F1 binding
AB  - Mutant alleles for the alpha subunit of H+-translocating ATPase (FoF1) were cloned from Escherichia coli strains isolated in this laboratory. Determination of their DNA sequence revealed four nonsense mutations (KF3 and KF9, Gln-20----end; KF24, Trp-111----end; KF2, Trp-231----end; KF70, Gln-252----end) and one missense mutation (KF45, Pro-143----Ser). The membranes of all the mutants except strain KF9 (KF3) had 50-70% of ATPase activities of the wild-type. Unlike the F1-ATPase of the wild- type, those of the mutants were insensitive to dicyclohexylcarbodiimide and were easier to solubilize from membranes. As membranes of strain KF24 had F1-ATPase activity, these results suggest that at least a part of the F1-binding sites could be formed without a region between residues 111 and the carboxyl terminus of the alpha subunit. However, normal interactions between Fo and F1 require regions between residues 252 and 271 (carboxyl terminus) and in the vicinity of Pro-143. Membranes of strain KF45 were capable of forming a low ATP-driven H+ gradient, whereas other membranes were not. The possibility that the region between residues 252 and 271 is involved in H+ translocation is discussed
MH  - A
MH  - ALPHA-SUBUNIT
MH  - ATPase
MH  - BINDING
MH  - Biochemistry
MH  - Chemistry
MH  - Dicyclohexylcarbodiimide
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - H+
MH  - H+-ATPase
MH  - membrane
MH  - Membranes
MH  - mutant
MH  - proton
MH  - RESIDUE
MH  - Site
MH  - SUBUNIT
MH  - translocation
RP  - NOT IN FILE
NT  - UI - 88273090LA - engRN - 0 (Codon)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19880819IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:2899073
SO  - J Biol Chem 1988 Jul 25 ;263(21):10056-10062

826
UI  - 362
AU  - Falk G
AU  - Walker JE
AD  - Department of Biochemistry, Arrhenius Laboratory, University of Stockholm, Sweden
TI  - DNA sequence of a gene cluster coding for subunits of the F0 membrane sector of ATP synthase in Rhodospirillum rubrum. Support for modular evolution of the F1 and F0 sectors
AB  - A region was cloned from the genome of the purple non-sulphur photobacterium Rhodospirillum rubrum that contains genes coding for the membrane protein subunits of the F0 sector of ATP synthase. The clone was identified by hybridization with a synthetic oligonucleotide designed on the basis of the known protein sequence of the dicyclohexylcarbodi-imide-reactive proteolipid, or subunit c. The complete nucleotide sequence of 4240 bp of this region was determined. It is separate from an operon described previously that encodes the five subunits of the extrinsic membrane sector of the enzyme, F1- ATPase. It contains a cluster of structural genes encoding homologues of all three membrane subunits a, b and c of the Escherichia coli ATP synthase. The order of the genes in Rsp. rubrum is a-c-b'-b where b and b' are homologues. A similar gene arrangement for F0 subunits has been found in two cyanobacteria, Synechococcus 6301 and Synechococcus 6716. This suggests that the ATP synthase complexes of all these photosynthetic bacteria contain nine different polypeptides rather than eight found in the E. coli enzyme; the chloroplast ATP synthase complex is probably similar to the photosynthetic bacterial enzymes in this respect. The Rsp. rubrum b subunit is modified after translation. As shown by N-terminal sequencing of the protein, the first seven amino acid residues are removed before or during assembly of the ATP synthase complex. The subunit-a gene is preceded by a gene coding for a small hydrophobic protein, as has been observed previously in the atp operons in E. coli, bacterium PS3 and cyanobacteria. A number of features suggest that the Rsp. rubrum cluster of F0 genes is an operon. On its 5' side are found sequences resembling the -10 (Pribnow) and -35 boxes of E. coli promoters, and the gene cluster is followed by a sequence potentially able to form a stable stem-loop structure, suggesting that it acts as a rho-independent transcription terminator. These features and the small intergenic non-coding sequences suggest that the genes are cotranscribed, and so the name atp2 is proposed for this second operon coding for ATP synthase subunits in Rsp. rubrum. The finding that genes for the F0 and F1 sectors of the enzyme are in separate clusters supports the view that these represent evolutionary modules
RP  - NOT IN FILE
NT  - UI - 89025651LA - engRN - 0 (DNA, Bacterial)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19881122IS - 0264-6021SB - IMCY - ENGLANDJC - 9YO
UR  - PM:2902844
SO  - Biochem J 1988 Aug 15 ;254(1):109-122

827
UI  - 9131
AU  - Feierabend B
AU  - Schumann J
TI  - Kinetics of nucleotide exchange and of ATP hydrolysis by isolated chloroplast coupling factor CF1 in the presence of inhibitors
MH  - atp
MH  - chloroplast
MH  - coupling
MH  - COUPLING FACTOR
MH  - Hydrolysis
MH  - inhibitor
MH  - inhibitors
MH  - Kinetics
RP  - IN FILE
SO  - Biochim Biophys Acta 1988  ;933():351-357

828
UI  - 20883
AU  - Futai M
AU  - Noumi T
AU  - Maeda M
AD  - Department of Organic Chemistry and Biochemistry, Osaka University, Japan
TI  - Molecular genetics of F1-ATPase from Escherichia coli
AB  - We have reviewed recent molecular biological studies on F1-ATPase of Escherichia coli and emphasized the advantages of using the bacterium in studies on this important enzyme. All subunits had homologies of varied degrees with those from other organisms. Mutations of F1 subunits caused defects in catalysis and assembly. Defects of the mutant enzymes were studied extensively together with the determination of the amino acid substitutions. Extensive molecular biological studies may help greatly in understanding the normal mechanism and assembly of the complex
MH  - ACID
MH  - ATPase
MH  - Bacteria
MH  - Biochemistry
MH  - Catalysis
MH  - Chemistry
MH  - COMPLEX
MH  - Enzymes
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - genetics
MH  - mechanism
MH  - mutant
MH  - review
MH  - SUBUNIT
RP  - NOT IN FILE
NT  - UI - 88153634LA - engRN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticlePT - ReviewPT - Review, AcademicDA - 19880412IS - 0145-479XSB - IMCY - UNITED STATES
UR  - PM:2894372
SO  - J Bioenerg Biomembr 1988 Feb ;20(1):41-58

829
UI  - 20878
AU  - Futai M
AU  - Noumi T
AU  - Maeda M
AD  - Department of Organic Chemistry and Biochemistry, Osaka University, Japan
TI  - Mechanism of F1-ATPase studied by the genetic approach
AB  - E. coli F1-ATPase has been studied mainly by the genetic approach. Mutations in either the alpha or beta subunit modified the kinetics of multisite and uni-site hydrolysis of ATP. The mechanism of F1-ATPase and the essential amino acid residues of beta subunits are discussed
MH  - ACID
MH  - atp
MH  - ATPase
MH  - BETA-SUBUNIT
MH  - Biochemistry
MH  - Chemistry
MH  - COLI F1 ATPASE
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - Hydrolysis
MH  - Kinetics
MH  - Macromolecular Systems
MH  - mechanism
MH  - RESIDUE
MH  - review
MH  - SUBUNIT
MH  - SYSTEM
MH  - SYSTEMS
RP  - NOT IN FILE
NT  - UI - 89123238LA - engRN - 0 (Macromolecular Systems)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 19890316IS - 0145-479XSB - IMCY - UNITED STATES
UR  - PM:2906061
SO  - J Bioenerg Biomembr 1988 Aug ;20(4):469-480

830
UI  - 527
AU  - Garboczi DN
AU  - Fox AH
AU  - Gerring SL
AU  - Pedersen PL
AD  - Department of Biological Chemistry, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205
TI  - Beta subunit of rat liver mitochondrial ATP synthase: cDNA cloning, amino acid sequence, expression in Escherichia coli, and structural relationship to adenylate kinase
AB  - The amino acid sequence of all but a few N-terminal residues of the beta subunit of rat liver ATP synthase has been determined from cDNA clones. Rat liver F1-beta is shown to contain 17 amino acid differences from that reported for F1-beta of bovine heart, 2 differences of which involve differences in charge. This may account in part for the observation that bovine heart F1 binds nucleotides with much greater affinity than the rat liver enzyme. Rat liver F1-beta also contains homologous regions with another nucleotide binding protein, adenylate kinase, for which high-resolution structural studies are available. Adjacent to one of these homologous regions is an eight amino acid stretch which bears striking homology to the phosphorylation region of the (Na+,K+)-ATPase. The combination of these two homology regions may constitute at least part of a nucleotide binding domain in F1-beta. Significantly, both rat liver and bovine heart beta contain these regions of homology, whereas the 17 amino acid differences between the two enzymes lie outside this region. The possibility of a second nucleotide binding domain which differs between the two enzymes is discussed. A cDNA clone containing all the regions of homology as well as 11 of the 17 amino acid differences between the bovine heart and rat liver beta subunits has been ligated into the bacterial expression vector pKK223-3. After transformation of a protease-deficient strain of Escherichia coli, this cDNA clone is expressed as a 36-kilodalton protein.(ABSTRACT TRUNCATED AT 250 WORDS)
RP  - NOT IN FILE
NT  - UI - 88163632LA - engRN - 0 (Macromolecular Systems)RN - 9007-49-2 (DNA)RN - EC 2.7 (Phosphotransferases)RN - EC 2.7.4.3 (Adenylate Kinase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - CA 10951/CA/NCIDA - 19880505IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:2894849
SO  - Biochemistry 1988 Jan 26 ;27(2):553-560

831
UI  - 525
AU  - Garboczi DN
AU  - Hullihen JH
AU  - Pedersen PL
AD  - Department of Biological Chemistry, Johns Hopkins School of Medicine, Baltimore, Maryland 21205
TI  - Mitochondrial ATP synthase. Overexpression in Escherichia coli of a rat liver beta subunit peptide and its interaction with adenine nucleotides
AB  - The C-terminal two-thirds of the rat liver ATP synthase beta subunit has been overexpressed and exported to the Escherichia coli periplasm under the direction of the alkaline phosphatase (phoA) promoter and leader peptide. The processed soluble protein contains the 358 amino acids from glutamate 122 to the rat liver beta C-terminal serine 479, including all the regions that have been predicted by chemical and genetic modification studies to be involved in nucleotide, Pi, and Mg2+ binding. Through a simple sequence of Tris/EDTA/lysozyme treatment, osmotic lysis, and alkaline pH washes, the processed beta subunit fragment can be prepared in greater than 95% purity and at a yield of greater than 20 mg/liter of culture. It interacts with 2'(3')-O-(2,4,6- trinitrophenyl) adenosine 5'-triphosphate (TNP-ATP) which exhibits a strong enhancement of fluorescence upon binding. A similar enhancement is observed upon interaction with TNP-ADP. Enhancement observed with both TNP-nucleotides is markedly reduced by prior addition of either ATP or ADP and almost completely prevented by the ATP synthase inhibitor 7-chloro-4-nitrobenz-2-oxa-1,3-diazole. Both TNP-ATP and TNP- ADP bind at a stoichiometry of approximately 1 mol of nucleotide/mol of beta subunit fragment. Under the same conditions, TNP-AMP does not exhibit a fluorescence enhancement. This work demonstrates that, in the absence of interaction with other ATP synthase subunits, the rat liver beta subunit sequence from glutamate 122 to the C terminus exhibits no more than one readily detectable nucleotide binding domain. This success in producing a "functional" beta subunit fragment has significance for the pursuit of genetic and physical studies focused on the structure and function of the rat liver ATP synthase beta subunit
RP  - NOT IN FILE
NT  - UI - 89008480LA - engRN - 0 (Adenine Nucleotides)RN - 0 (Macromolecular Systems)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - CA 10951/CA/NCIDA - 19881121IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:2902092
SO  - J Biol Chem 1988 Oct 25 ;263(30):15694-15698

832
UI  - 528
AU  - Garboczi DN
AU  - Shenbagamurthi P
AU  - Kirk W
AU  - Hullihen J
AU  - Pedersen PL
AD  - Department of Biological Chemistry, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205
TI  - Mitochondrial ATP synthase. Interaction of a synthetic 50-amino acid, beta-subunit peptide with ATP
AB  - A 50-amino acid peptide predicted by chemical modification studies of F1 and by comparison with adenylate kinase to comprise part of an ATP- binding domain within the beta-subunit of mitochondrial ATP synthase has been synthesized and purified. In the numbering system used for bovine heart beta, the peptide consists of amino acid residues from aspartate 141 at the N-terminal end to threonine 190 at the carboxyl end. In Tris-Cl buffer, pH 7.4, the peptide undergoes a dramatic reaction with ATP resulting in precipitate formation. Analysis of the precipitate shows it to contain both peptide and ATP. Similar to the ATPase activity of F1 and the binding of nucleotide to the enzyme, the capacity of ATP to induce precipitation of the peptide is decreased markedly by lowering pH. Interaction of the peptide with the fluorescent ATP analog, TNP-ATP (2'(3')-O-(2,4-6-trinitrophenyl)- adenosine 5'-triphosphate), can be demonstrated in solution at low concentrations. A 7-fold enhancement in fluorescence is observed when 2.5 microM TNP-ATP interacts with 2.5 microM peptide. Divalent cation is neither required for ATP-induced precipitation of the peptide nor for demonstrating interaction between TNP-ATP and peptide, just as Mg2+ is not required for nucleotide binding to F1. These results indicate that the beta-subunit peptide studied here comprises at least part of a nucleotide-binding domain within the mitochondrial ATP synthase complex
RP  - NOT IN FILE
NT  - UI - 88087205LA - engRN - 0 (Peptide Fragments)RN - 56-65-5 (Adenosine Triphosphate)RN - 61368-63-6 (2',3'-O-(2,4,6-trinitro-cyclohexadienylidine)adenosine 5'- triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - CA 10951/CA/NCIDA - 19880223IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:2891704
SO  - J Biol Chem 1988 Jan 15 ;263(2):812-816

833
UI  - 187
AU  - Gavilanes-Ruiz M
AU  - Tommasino M
AU  - Capaldi RA
AD  - Institute of Molecular Biology, University of Oregon, Eugene 97403
TI  - Structure-function relationships of the Escherichia coli ATP synthase probed by trypsin digestion
AB  - Trypsin cleavage has been used to probe structure-function relationships of the Escherichia coli ATP synthase (ECF1F0). Trypsin cleaved all five subunits, alpha, beta, gamma, delta, and epsilon, in isolated ECF1. Cleavage of the alpha subunit involved the removal of the N-terminal 15 residues, the beta subunit was cleaved near the C- terminus, the gamma subunit was cleaved near Ser202, and the delta and epsilon subunits appeared to be cleaved at several sites to yield small peptide fragments. Trypsin cleavage of ECF1 enhanced the ATPase activity between 6- and 8-fold in different preparations, in a time course that followed the cleavage of the epsilon subunit. This removal of the epsilon subunit increased multisite ATPase activity but not unisite ATPase activity, showing that the inhibitory role of the epsilon subunit is due to an effect on cooperativity. The detergent lauryldimethylamine oxide was found to increase multisite catalysis and also increase unisite catalysis more than 2-fold. Prolonged trypsin cleavage left a highly active ATPase containing only the alpha and beta subunits along with two fragments of the gamma subunit. All of the subunits of ECF1 were cleaved by trypsin in preparations of ECF1F0 at the same sites as in isolated ECF1. Two subunits, the beta and epsilon subunits, were cleaved at the same rate in ECF1F0 as in ECF1 alone. The alpha, gamma, and delta subunits were cleaved significantly more slowly in ECF1F0.(ABSTRACT TRUNCATED AT 250 WORDS)
RP  - NOT IN FILE
NT  - UI - 88163640LA - engRN - 0 (Macromolecular Systems)RN - 0 (Peptide Fragments)RN - EC 3.4.21.4 (Trypsin)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - FO5TW03633/TW/FICID - HL24526/HL/NHLBIDA - 19880505IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:2894850
SO  - Biochemistry 1988 Jan 26 ;27(2):603-609

834
UI  - 21014
AU  - Graber P
AU  - Fromme P
AU  - Schmidt G
AU  - Boekema E
AD  - Max-Volmer-Institut, Technische Universitat, Berlin, FRG
TI  - Structure of the ATP-synthase from chloroplasts as revealed from biochemical studies and electron microscopy
MH  - ATP synthase
MH  - ATPase
MH  - Carrier Proteins
MH  - chloroplast
MH  - Chloroplasts
MH  - electron
MH  - electron microscopy
MH  - ELECTRON-MICROSCOPY
MH  - Microscopy
MH  - protein
MH  - Proteins
RP  - NOT IN FILE
NT  - UI - 88336209LA - engRN - 0 (Carrier Proteins)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19881027IS - 0361-7742SB - IMCY - UNITED STATES
UR  - PM:2901752
SO  - Prog Clin Biol Res 1988  ;273():67-74

835
UI  - 19792
AU  - Guerrero KJ
AU  - Boyer PD
AD  - Department of Chemistry and Biochemistry, University of California, Los Angeles 90024-1570
TI  - Tightly bound 2-azido-adenine nucleotides at catalytic and noncatalytic sites of the rat liver F1 ATPase label adjacent tryptic peptides of the beta subunit
AB  - UV irradiation of rat liver F1 ATPase, previously exposed to Mg2+ and [beta, gamma-32P]-2-azido-ATP and separated from medium nucleotides, covalently modifies two tyrosine residues in adjacent tryptic peptides of the beta subunit. This results from the occupancy by 2-azido-ATP or 2-azido-ADP of two distinct types of nucleotide binding sites, the catalytic and noncatalytic sites. The two modified peptides are identical to the ones modified by 2-azido-adenine nucleotides in the beef heart F1 ATPase. Both catalytic and noncatalytic sites are labeled when the ATPase is exposed to [beta-32P]-2-azido-ADP in the presence or the absence of 5'-adenylyimidodiphosphate (AMP-PNP), showing that two distinct types of ADP binding sites are present on the liver enzyme. Similar incorporation of 2-azido-adenine nucleotides is obtained when membrane-bound rat liver F1 ATPase is incubated with Mg2+ and [beta, gamma-32P]-2-azido-ATP
MH  - 2-AZIDO-ATP
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - ADP
MH  - ADP BINDING
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Azides
MH  - BETA-SUBUNIT
MH  - BINDING
MH  - Binding Sites
MH  - Biochemistry
MH  - Chemistry
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - H(+)-Transporting ATP Synthase
MH  - Liver
MH  - Macromolecular Systems
MH  - NONCATALYTIC SITES
MH  - nucleotide binding
MH  - nucleotide binding sites
MH  - NUCLEOTIDE-BINDING-SITES
MH  - Nucleotides
MH  - Peptide Fragments
MH  - SUBUNIT
MH  - SYNTHASE
MH  - Trypsin
RP  - NOT IN FILE
NT  - UI - 88309128LA - engRN - 0 (Affinity Labels)RN - 0 (Azides)RN - 0 (Macromolecular Systems)RN - 0 (Peptide Fragments)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 64020-53-7 (2-azidoadenosine 3',5'-diphosphate)RN - 72884-75-4 (2-azidoadenosine 5'-triphosphate)RN - EC 3.4.21.4 (Trypsin)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM-11094/GM/NIGMSDA - 19880920IS - 0006-291XSB - IMCY - UNITED STATESJC - 9Y8
UR  - PM:2900637
SO  - Biochem Biophys Res Commun 1988 Aug 15 ;154(3):854-860

836
UI  - 21090
AU  - Guffanti AA
AU  - Krulwich TA
AD  - Department of Biochemistry, Mount Sinai School of Medicine, City University of New York, New York 10029
TI  - ATP synthesis is driven by an imposed delta pH or delta mu H+ but not by an imposed delta pNa+ or delta mu Na+ in alkalophilic Bacillus firmus OF4 at high pH
AB  - Starved whole cells of alkalophilic Bacillus firmus OF4 that are equilibrated at either pH 10.2, 9.5, or 8.5 synthesize ATP in response to a pH gradient that is imposed by rapid dilution of the cyanide- treated cells into buffer at pH 7.5. If a valinomycin-mediated potassium diffusion potential (positive out) is generated simultaneously with the pH gradient, then the rate of ATP synthesis and the level of synthesis achieved is much higher than upon imposition of a pH gradient alone. By contrast, imposition of a large chemical gradient of Na+, either in the presence or absence of a concomitant diffusion potential, fails to result in ATP synthesis. We conclude that this organism does not possess a sodium-motive ATPase that can be made to synthesize detectable levels of ATP by imposition of a suitably large chemical or electrochemical gradient of Na+. On the other hand, a proton-translocating ATPase is in evidence when protons are provided at very high pH, corroborating our earlier work on extremely alkalophilic bacilli. Oxidative phosphorylation must, then, be catalyzed in these organisms by a proton-translocating ATPase even though the putative bulk driving forces for such a catalyst are low under optimal growth conditions. Stable, imposed pH gradients of 1 unit, comparable to the magnitude of the total electrochemical proton gradient of growing cells, result in much lower ATP concentrations than observed in such cells. We hypothesize that ATP synthesis in growing cells utilizes protons that are made available by some localized pathway between proton pumps and the ATP synthase
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - ATPase
MH  - Bacillus
MH  - Biochemistry
MH  - buffer
MH  - Cells
MH  - delta
MH  - DELTA-PH
MH  - Diffusion
MH  - diffusion potential
MH  - H+
MH  - Oxidative Phosphorylation
MH  - pH
MH  - Phosphorylation
MH  - Potassium
MH  - proton
MH  - Proton Pump
MH  - Proton-Translocating ATPases
MH  - Protons
MH  - Sodium
MH  - SYNTHASE
MH  - synthesis
RP  - NOT IN FILE
NT  - UI - 89008343LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - 7440-23-5 (Sodium)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleID - GM28454/GM/NIGMSDA - 19881110IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:2902088
SO  - J Biol Chem 1988 Oct 15 ;263(29):14748-14752

837
UI  - 576
AU  - Hekman C
AU  - Matsuno-Yagi A
AU  - Hatefi Y
AD  - Department of Basic and Clinical Research, Research Institute of Scripps Clinic, La Jolla, California 92037
TI  - Energy-induced modulation of the kinetics of oxidative phosphorylation and reverse electron transfer
AB  - The kinetics of ATP synthesis by bovine heart submitochondrial particles (SMP) are modulated by the rate of energy production by the respiratory chain between two fixed limits characterized by apparent KmADP = 2-4 microM and Vmax approximately 200 nmol of ATP min-1 (mg of SMP protein)-1 at low energy levels and apparent KmADP = 120-160 microM and Vmax = 11,000 nmol of ATP min-1 (mg of SMP protein)-1 at high energy levels. These data indicate that KmADP and Vmax increase approximately 50-fold each; therefore, there is essentially no change in the catalytic efficiency of the ATP synthase complex in going from one extreme to the other. At intermediate rates of energy production, the kinetic data required introduction of a third, intermediate KmADP. A KmADP of 10-15 microM fitted all the data reported here and previously [Matsuno-Yagi, A., & Hatefi, Y. (1986) J. Biol. Chem. 261, 14031-14038]. However, this is not meant to suggest that there is a fixed intermediate KmADP, as the transition from one fixed limit to the other may be fluid or involve more than one intermediate state. In addition, it has been shown that kinetic plots of SMP-catalyzed and ATP- driven reverse electron transfer from succinate to NAD are curvilinear and resolvable into a minimum of two apparent KmNAD values of about 20- 30 and 200-300 microM. These results have been discussed in relation to the three potentially active catalytic sites of F1-ATPase and the structure of the NADH:ubiquinone oxidoreductase complex, the curvilinear kinetics of ATP hydrolysis, and changes in KmADP and KmPi in photophosphorylation as affected by the duration and intensity of light
RP  - NOT IN FILE
NT  - UI - 89088122LA - engRN - 0 (Succinates)RN - 110-15-6 (Succinic Acid)RN - 53-84-9 (NAD)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - DK08126/DK/NIDDKDA - 19890221IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:2905168
SO  - Biochemistry 1988 Sep 20 ;27(19):7559-7565

838
UI  - 21264
AU  - Jackson JB
TI  - Bacterial photosynthesis
MH  - Photosynthesis
T2  - Bacterial energy trunsduction
A2  - Anthony C
Y2  - -32676  
PB  - London: Academic Press
RP  - NOT IN FILE
SO  -  1988  ;():317-376

839
UI  - 21041
AU  - Junge W
TI  - Complete tracking of transient proton flow through active chloroplast ATP synthase [Erratum to document cited in CA108(5): 34961c]
MH  - ACTIVE
MH  - atp
MH  - ATP synthase
MH  - chloroplast
MH  - conductance
MH  - proton
MH  - SYNTHASE
RP  - NOT IN FILE
SO  - Proc Natl Acad Sci U S A 1988  ;85():7240-7240

840
UI  - 19759
AU  - Keen NT
AU  - Tamaki S
AU  - Kobayashi D
AU  - Trollinger D
AD  - Department of Plant Pathology, University of California, Riverside 92521
TI  - Improved broad-host-range plasmids for DNA cloning in gram-negative bacteria
AB  - Improved broad-host-range plasmid vectors were constructed based on existing plasmids RSF1010 and RK404. The new plasmids pDSK509, pDSK519, and pRK415, have several additional cloning sites and improved antibiotic-resistance genes which facilitate subcloning and mobilization into various Gram-negative bacteria. Several new polylinker sites were added to the Escherichia coli plasmids pUC118 and pUC119, resulting in the new plasmids, pUC128 and pUC129. These plasmids facilitate the transfer of cloned DNA fragments to the broad- host-range vectors. Finally, the broad-host-range cosmid cloning vector pLAFR3 was improved by the addition of a double cos casette to generate the new plasmid, pLAFR5. This latter cosmid simplifies vector preparation and has permitted the rapid cloning of genomic DNA fragments generated with Sau3A. The resulting clones may be introduced into other Gram-negative bacteria by conjugation
MH  - A
MH  - Bacteria
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - plant
MH  - TRANSFER
RP  - NOT IN FILE
NT  - UI - 89196911LA - engRN - 0 (Cosmids)RN - 0 (DNA Transposable Elements)RN - 0 (DNA, Recombinant)RN - 0 (Plasmids)PT - Journal ArticleDA - 19890519IS - 0378-1119SB - IMCY - NETHERLANDSJC - FOP
UR  - PM:2853689
SO  - Gene 1988 Oct 15 ;70(1):191-197

841
UI  - 21278
AU  - Kell DB
TI  - Protonmotive energy-transducing systems: some physical principles and experimental approaches.
MH  - SYSTEMS
MH  - SYSTEM
T2  - Bacterial Energy Transduction
A2  - Anthony CJ
PB  - London: Academic Press
RP  - NOT IN FILE
SO  -  1988  ;():429-490

842
UI  - 21091
AU  - Krulwich TA
AU  - Hicks DB
AU  - Seto-Young D
AU  - Guffanti AA
AD  - Department of Biochemistry, Mount Sinai School of Medicine, New York, New York
TI  - The bioenergetics of alkalophilic bacilli
AB  - A summary, cum speculation, of the major bioenergetic characteristics of alkalophilic bacilli is presented in Figure 5. Further progress will depend heavily on the purification and characterization of the relevant proteins that catalyze the ion fluxes and on the development of much more potent genetic approaches to the outstanding issues of this interesting group of bacteria
MH  - A
MH  - Bacteria
MH  - Biochemistry
MH  - bioenergetics
MH  - development
MH  - ion
MH  - protein
MH  - Proteins
MH  - purification
MH  - review
RP  - NOT IN FILE
NT  - UI - 89029925LA - engPT - Journal ArticlePT - ReviewPT - Review, AcademicID - GM28454/GM/NIGMSDA - 19881201IS - 1040-841XSB - IMCY - UNITED STATES
UR  - PM:3053049
SO  - Crit Rev Microbiol 1988  ;16(1):15-36

843
UI  - 20876
AU  - Kuki M
AU  - Noumi T
AU  - Maeda M
AU  - Amemura A
AU  - Futai M
AD  - Department of Organic Chemistry and Biochemistry, Osaka University, Japan
TI  - Functional domains of epsilon subunit of Escherichia coli H+-ATPase (F0F1)
AB  - Mutants of the uncC gene for the epsilon subunit (138 amino acid residues) of Escherichia coli H+-ATPase were isolated: strain KF53 (Gln- 72----end) and KF148(SD-) (two base substitutions in the Shine-Dalgarno sequence, GGAGG----AAAGG). These strains did not have F1 bound to membranes and were unable to grow by oxidative phosphorylation. A series of plasmids carrying truncated uncC genes were constructed and introduced into strain KF148(SD-). Analyses of KF148(SD-) cells with different plasmids indicated that the amino-terminal fragment of the epsilon subunit of 78-80 amino acid residues was capable of forming active membrane-bound F1-ATPase, whereas that of 73 residues was not, indicating that the carboxyl-terminal half of the epsilon subunit is not necessary for the active enzyme. Furthermore, results indicated that residues between 73 and 78-80 may have a critical role(s) in binding F1 to F0. Truncated epsilon subunits of 80 and 93 residues were identified in purified F1 from cells carrying the respective uncC genes, and only the latter subunit had intrinsic activity to inhibit ATPase of F1, suggesting that residues between 80 and 93 are essential for the inhibitory activity
MH  - A
MH  - ACID
MH  - ACTIVE
MH  - ATPase
MH  - BASE
MH  - BINDING
MH  - Biochemistry
MH  - Cells
MH  - Chemistry
MH  - EPSILON-SUBUNIT
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - F0F1
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - H+-ATPase
MH  - Macromolecular Systems
MH  - membrane
MH  - Membranes
MH  - mutant
MH  - Oxidative Phosphorylation
MH  - Phosphorylation
MH  - RESIDUE
MH  - SUBUNIT
MH  - SYSTEM
MH  - SYSTEMS
RP  - NOT IN FILE
NT  - UI - 89034268LA - engRN - 0 (Macromolecular Systems)RN - 0 (Plasmids)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19881220IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:2903163
SO  - J Biol Chem 1988 Nov 25 ;263(33):17437-17442

844
UI  - 96
AU  - Laubinger W
AU  - Dimroth P
AD  - Institut fur Physiologische Chemie der Technischen Universitat Munchen, Federal Republic of Germany
TI  - Characterization of the ATP synthase of Propionigenium modestum as a primary sodium pump
AB  - The ATP synthase (F1F0) of Propionigenium modestum has been purified to a specific ATPase activity of 5.5 units/mg of protein, which is about 6 times higher than that of the bacterial membranes. Analysis by SDS gel electrophoresis indicated that in addition to the five subunits of the F1 ATPase, subunits of Mr 26,000 (a), 23,000 (b), and 7500 (c) have been purified. The ATPase activity of F1F0 was specifically activated about 10-fold by Na+ions. The enzyme was strongly inhibited by dicyclohexylcarbodiimide, venturicidin, tributyltin chloride, and azide. After incubation with [14C]dicyclohexylcarbodiimide, about 3-4 mol of the inhibitor was bound per 500,000 g of the enzyme. The radioactive label was specifically bound to submit c. These subunits form stable aggregates which resist dissociation by SDS at 100 degrees C. The monomer is formed upon heating with SDS to 121 degrees C or by extraction of the membranes with chloroform/methanol. The ATP synthase was incorporated into liposomes by a freeze-thaw-sonication procedure. The reconstituted proteoliposomes catalyzed the transport of Na+ions upon ATP hydrolysis. The transport was completely abolished by dicyclohexylcarbodiimide. Whereas monensin prevented the accumulation of Na+ions, the uptake rate was stimulated 4-5-fold in the presence of valinomycin or carbonyl cyanide m=chlorophenylhydrazone. These results indicate an electrogenic Na+ transport and also that it is a primary event and not accomplished by a H+-translocating ATP synthase in combination with a Na+/H+ antiporter
RP  - NOT IN FILE
NT  - UI - 89088118LA - engRN - 0 (Amino Acids)RN - 0 (Liposomes)RN - 0 (Macromolecular Systems)RN - 0 (Sodium Channels)RN - 151-21-3 (Sodium Dodecyl Sulfate)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - 7440-23-5 (Sodium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19890221IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:2905167
SO  - Biochemistry 1988 Sep 20 ;27(19):7531-7537

845
UI  - 21288
AU  - Levitt DG
AU  - Decker ER
AD  - Department of Physiology, University of Minnesota, Minneapolis 55455
TI  - Electrostatic radius of the gramicidin channel determined from voltage dependence of H+ ion conductance
AB  - The results of Decker and Levitt (1987) suggest that the conductance of H+ ion through the gramicidin channel is limited primarily by diffusion in the bulk solution at the channel mouth. It is assumed in this paper that the H+ conductance is 100% diffusion limited. This means that all the factors that influence the H+ flux are external to the channel and are presumed to be known. In particular, the diffusion coefficient of H+ in this region is assumed to be equal to the bulk solution value and the only force acting on the ion is that due to the applied voltage. A model of the H+ flux is derived, based on the Nernst-Planck equation. It has three adjustable parameters: the electrostatic radius, the capture distance, and the radius of the H+ ion. The acceptable range of the parameters was determined by comparing the predictions of the model with the experimental measurements of the H+ conductance at pH 3.75. The best fit was obtained for an electrostatic radius in the range 2.3- 2.7 A. This is in good agreement with earlier predictions (2.5 A) based on the assumption that the dielectric constant of the channel water is equal to that of bulk water. The addition of 1 M choline Cl- (an impermeant) increases the H+ current at low voltage and decreases it at high voltage. The increase can be explained by the small surface charge that results from the separation of charge produced by exclusion of the large choline cation (relative to Cl-) from the membrane surface. The decrease at high voltages can be accounted for by the change in the profile of the applied potential produced by the increase in ionic strength
MH  - A
MH  - Choline
MH  - conductance
MH  - CONSTANT
MH  - DEPENDENCE
MH  - Diffusion
MH  - H+
MH  - ion
MH  - Ion Channels
MH  - M
MH  - membrane
MH  - model
MH  - pH
MH  - physiology
MH  - SURFACE
MH  - voltage
MH  - Water
RP  - NOT IN FILE
NT  - UI - 88135045LA - engRN - 0 (Ion Channels)RN - 1405-97-6 (Gramicidin)PT - Journal ArticleID - 5RO1 GM25938/GM/NIGMSDA - 19880325IS - 0006-3495SB - IMCY - UNITED STATES
UR  - PM:2449254
SO  - Biophys J 1988 Jan ;53(1):33-38

846
UI  - 310
AU  - Lill H
AU  - Engelbrecht S
AU  - Junge W
AD  - Biophysik, Fachbereich Biologie/Chemie, Universitat Osnabruck, West Germany
TI  - Delta subunit of chloroplast coupling factor 1 inhibits proton leakage through coupling factor O
AB  - The ATP synthase of chloroplasts consists of a proton-conducting portion, CF0, and a catalytic portion, CF1. The smaller subunits of CF1, in particular delta, may play a key role in the coupling of proton transport to ATP synthesis. Purified subunit delta, when added to partially CF1-depleted thylakoid membranes, can restore photophosphorylation (Engelbrecht, S., and Junge, W. (1987) Eur. J. Biochem. 172, 213-218). We report here that it does so by blocking proton conduction through CF0. Thylakoids were CF1-depleted by incubation in hypoosmolar NaCl/EDTA solutions. Variation of the NaCl concentrations and of the incubation times not only changed the overall degree of CF1 depletion but also the subunit composition of solubilized CF1, namely CF1 containing delta and CF1(-delta). This was quantified by immunoelectrophoresis and by fast protein liquid chromatography. Proton conduction was measured by flash spectrophotometry by using standard electrochromic and pH-indicating absorption changes. The removal of integral CF1 was correlated with high electric conductance of thylakoid membranes, an increased extent of rapid proton leakage, and loss of ATP synthesis activity, which exceeded the percentual loss of CF1. The removal of predominantly CF1(-delta) resulted in comparatively lesser effects on the loss of ATP synthesis and on the extent and velocity of proton leakage. On the same line, addition of integral CF1 and of purified delta diminished the electric leak in CF1- depleted thylakoids. Both approaches, the controlled removal of CF1 and CF1(-delta), respectively, and addition of delta and CF1 showed that delta can act as a "stopcock" to the exposed proton channel CF0
RP  - NOT IN FILE
NT  - UI - 89008305LA - engRN - 0 (Macromolecular Systems)RN - 0 (Protons)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19881107IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:2902083
SO  - J Biol Chem 1988 Oct 5 ;263(28):14518-14522

847
UI  - 458
AU  - Maggio MB
AU  - Parsonage D
AU  - Senior AE
AD  - Department of Biochemistry, University of Rochester Medical Center, New York 14642
TI  - A mutation in the alpha-subunit of F1-ATPase from Escherichia coli affects the binding of F1 to the membrane
AB  - The mutation Gly-29----Asp in the alpha-subunit of the F1-ATPase from Escherichia coli was characterized and shown to cause the following effects. 1) Oxidative phosphorylation was markedly impaired in vivo 2) Membrane ATPase and ATP-driven proton-pumping activities were decreased markedly. 3) Membranes were proton-permeable, and membrane-bound ATPase was dicyclohexylcarbodiimide-insensitive. Therefore, it appeared that integration between F1 and F0 was abnormal. This was confirmed directly by the demonstration that the mutant F1 bound poorly to stripped membranes from a normal strain. Purified, soluble mutant F1 had normal ATPase activity. These results suggest that residue Gly-29, which is strongly conserved in alpha-subunits of F1-ATPases, lies in a region of the alpha-subunit important for membrane binding. Thus, three regions of the F1-alpha-subunit have now been recognized, specialized for membrane binding, nucleotide binding, and alpha/beta intersubunit signal transmission, respectively. The approximate locations of the three regions are described
RP  - NOT IN FILE
NT  - UI - 88169571LA - engRN - 0 (DNA, Single-Stranded)RN - 0 (Macromolecular Systems)RN - 0 (Plasmids)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSID - GM29805/GM/NIGMSDA - 19880502IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:2895104
SO  - J Biol Chem 1988 Apr 5 ;263(10):4619-4623

848
UI  - 577
AU  - Matsuno-Yagi A
AU  - Hatefi Y
AD  - Department of Basic and Clinical Research, Research Institute of Scripps Clinic, La Jolla, California 92037
TI  - Role of energy in oxidative phosphorylation
AB  - This article reviews the current status of information regarding the role of energy in the process of oxidative phosphorylation by mitochondria. The available data suggest that in submitochondrial particles (SMP) energy is utilized for the binding of ADP and Pi and for the release of ATP bound at the catalytic sites of F1-ATPase. The process of ATP synthesis on the surface of F1 from F1-bound ADP and Pi appears to be associated with negligible free energy change. The rate of energy production by the respiratory chain modulates the kinetics of ATP synthesis between a low Km (for ADP and Pi)-low Vmax mode and a high Km-high Vmax mode. The Km extremes for ADP are 2-3 microM and 120- 150 microM, and Vmax for ATP synthesis at high rates of energy production by bovine-heart SMP is about 440 S-1 (mole F1)-1 at 30 degrees C, which corresponds to 11 mumol ATP (min.mg of protein)-1. The interaction of dicyclohexylcarbodiimide (DCCD) or oligomycin at the proteolipid (subunit c) of the membrane sector (F0) of the ATP synthase complex alters the mode of ATP binding at the catalytic sites of F1, probably to one of lower affinity. It has been suggested that protonic energy might be conveyed to the catalytic sites of F1 in an analogous manner, i.e., via conformation changes in the ATP synthase complex initiated by proton-induced alterations in the structure of the DCCD- binding proteolipid. Finally, the relationship between the steady-state membrane potential (delta psi) and the rates of electron transfer and ATP synthesis has been discussed. It has been shown, in agreement with the delocalized chemiosmotic mechanism, that under appropriate conditions delta psi is exquisitely sensitive to changes in the rates of energy production and consumption
RP  - NOT IN FILE
NT  - UI - 89123239LA - engRN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialID - DK 08126/DK/NIDDKDA - 19890316IS - 0145-479XSB - IMCY - UNITED STATESJC - HIO
UR  - PM:2906062
SO  - J Bioenerg Biomembr 1988 Aug ;20(4):481-502

849
UI  - 578
AU  - Matsuno-Yagi A
AU  - Hatefi Y
AD  - Department of Basic and Clinical Research, Research Institute of Scripps Clinic, La Jolla, California 92037
TI  - Estimation of the turnover number of bovine heart F0F1 complexes for ATP synthesis
AB  - In mitochondria and submitochondrial particles (SMP), the rate of ATP synthesis is restricted by the rate of energy production by the respiratory chain. Fractional inactivation of the ATP synthase complexes (F0F1) of bovine heart SMP by covalent modifiers increased the rate of ATP synthesis per mole of active F0F1. Thus, by use of SMP containing fractionally inactivated F0F1 complexes, a synthetic rate of 420 mol of ATP (mol of F0F1.s)-1 was measured, which extrapolated to a Vmax of 440 s-1. At this extrapolated point, the turnover rate of F0F1 complexes was independent of the rate of energy production by the respiratory chain. These results have been discussed in relation to the effect of fractional inactivation of the F0F1 complexes of SMP on the steady-state free energy of the system. The above rate of ATP synthesis is comparable to the rate of ATP hydrolysis by SMP (400-520 s-1) in the absence of energy coupling constraints and control by the ATPase inhibitor protein. More interestingly, this rate is also comparable to the rate of ATP synthesis by chloroplast F0F1 under high light intensity (approximately 420 s-1). Under the conditions specified, bovine heart SMP and chloroplasts show similar apparent Km values for ADP. Thus, it appears that the mammalian and chloroplast ATP synthase complexes are similar not only in structure but also in catalytic efficiency for ATP synthesis
RP  - NOT IN FILE
NT  - UI - 88163590LA - engRN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - DK08126/DK/NIDDKDA - 19880428IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:2894847
SO  - Biochemistry 1988 Jan 12 ;27(1):335-340

850
UI  - 19794
AU  - Melese T
AU  - Xue ZX
AU  - Stempel KE
AU  - Boyer PD
AD  - Molecular Biology Institute, University of California, Los Angeles 90024-1570
TI  - Catalytic properties of chloroplast F1-ATPase modified at catalytic or noncatalytic sites by 2-azido adenine nucleotides
AB  - When heat-activated F1-ATPase from chloroplasts was repeatedly exposed to Mg2+ and 2-azido-ATP, followed by separation from medium nucleotides and photolysis, a total of two sites per enzyme, both catalytic and noncatalytic, were labeled. In a coupled assay with pyruvate kinase about half the activity was lost when one site per enzyme was modified. However, increased modification resulted in no further loss of activity. In contrast, methanol-sulfite activation of the enzyme showed a loss of most of the catalytic capacity when one site per enzyme was modified. Predominant labeling of either one catalytic or one noncatalytic site caused a loss of most of the activity in either assay. An indication that the enzyme modified at one site retained some catalytic activity was verified by measurement of the [18O]Pi species formed when [gamma-18O]ATP was hydrolyzed by partially derivatized enzyme. With either catalytic or noncatalytic site modification, the distributions of [18O]Pi species formed showed that the modified enzyme had different catalytic characteristics. An interpretation is that with modification by azido nucleotides at either catalytic or noncatalytic sites, capacity for rapid catalysis is largely lost but the remaining sites retain weak modified catalytic properties
MH  - 2-AZIDO-ATP
MH  - A
MH  - ACTIVATION
MH  - Adenine Nucleotides
MH  - ADENINE-NUCLEOTIDES
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthase
MH  - Azides
MH  - Catalysis
MH  - chloroplast
MH  - Chloroplasts
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - H(+)-Transporting ATP Synthase
MH  - Magnesium
MH  - NONCATALYTIC SITES
MH  - Nucleotides
MH  - Phosphates
MH  - Pyruvate Kinase
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 88186905LA - engRN - 0 (Affinity Labels)RN - 0 (Azides)RN - 0 (Phosphates)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 64020-53-7 (2-azidoadenosine 3',5'-diphosphate)RN - 72884-75-4 (2-azidoadenosine 5'-triphosphate)RN - 7439-95-4 (Magnesium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM11904/GM/NIGMSDA - 19880518IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:2895774
SO  - J Biol Chem 1988 Apr 25 ;263(12):5833-5840

851
UI  - 20882
AU  - Miki J
AU  - Maeda M
AU  - Mukohata Y
AU  - Futai M
AD  - Department of Organic Chemistry and Biochemistry, Osaka University, Japan
TI  - The gamma-subunit of ATP synthase from spinach chloroplasts. Primary structure deduced from the cloned cDNA sequence
AB  - cDNA clones encoding the gamma-subunit of chloroplast ATP synthase were isolated from a spinach library using synthetic oligonucleotide probes. The predicted amino acid sequence indicated that the mature chloroplast gamma-subunit consists of 323 amino acid residues and is highly homologous (55% identical residues) with the sequence of the cyanobacterial subunit. The positions of the four cysteine residues were identified. The carboxyl-terminal region of the chloroplast gamma- subunit is highly homologous with those of the gamma-subunits from six other sources (bacteria and mitochondria) sequenced thus far
MH  - A
MH  - ACID
MH  - Amino Acid Sequence
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Bacteria
MH  - Biochemistry
MH  - Chemistry
MH  - chloroplast
MH  - Chloroplasts
MH  - Cysteine
MH  - GAMMA- SUBUNIT
MH  - Mitochondria
MH  - Peptide Fragments
MH  - RESIDUE
MH  - Spinach
MH  - SPINACH-CHLOROPLASTS
MH  - SUBUNIT
MH  - SYNTHASE
MH  - Trypsin
RP  - NOT IN FILE
NT  - UI - 88211863LA - engRN - 0 (DNA, Recombinant)RN - 0 (Peptide Fragments)RN - 0 (Plasmids)RN - 9007-49-2 (DNA)RN - EC 3.4.21.4 (Trypsin)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19880616IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:2896606
SO  - FEBS Lett 1988 May 9 ;232(1):221-226

852
UI  - 20885
AU  - Miki J
AU  - Maeda M
AU  - Futai M
AD  - Department of Organic Chemistry and Biochemistry, Osaka University, Japan
TI  - Temperature-sensitive Escherichia coli mutant with an altered initiation codon of the uncG gene for the H+-ATPase gamma subunit
AB  - A mutant of Escherichia coli showing temperature-sensitive growth on succinate was isolated, and its mutation in the initiation codon (ATG to ATA) of the uncG gene (coding for the gamma subunit of H+-ATPase F0F1) was identified. This strain could grow on succinate as the sole carbon source at 25 and 30 degrees C, but not at 37 or 42 degrees C. When this strain was grown at 25 degrees C on succinate or glycerol, its membranes had about 15% of the ATPase activity of wild-type membranes, whereas when it was grown at 42 degrees C, its membranes had about 2% of the wild-type ATPase activity. Membranes of the mutant grown at 25 or 42 degrees C could bind F1 functionally, resulting in about 40% of the specific activity of wild-type membranes. The gamma subunit was identified in an EDTA extract of membranes of the mutant grown at 25 degrees C, but was barely detectable in the same amount of extract from the mutant grown at 42 degrees C. These results indicate that initiation of protein synthesis from the AUA codon is temperature sensitive and that the gamma subunit is essential for assembly of F1 in vivo as shown by in vitro reconstitution experiments (S. D. Dunn and M. Futai, J. Biol. Chem. 255:113-118, 1980)
MH  - A
MH  - ATPase
MH  - Biochemistry
MH  - Chemistry
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0F1
MH  - F1
MH  - H+-ATPase
MH  - In Vitro
MH  - M
MH  - membrane
MH  - Membranes
MH  - mutant
MH  - protein
MH  - reconstitution
MH  - SUBUNIT
MH  - succinate
MH  - Succinates
MH  - synthesis
MH  - Temperature
RP  - NOT IN FILE
NT  - UI - 88086865LA - engRN - 0 (Codon)RN - 0 (DNA, Bacterial)RN - 0 (RNA, Messenger)RN - 0 (Succinates)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19880210IS - 0021-9193SB - IMCY - UNITED STATES
UR  - PM:2891679
SO  - J Bacteriol 1988 Jan ;170(1):179-183

853
UI  - 1008
AU  - Mitchell P
TI  - Strategy of research on the chemiosmotic mechanism of cytochrome oxidase
AB  - In recent papers on protonmotive redox mechanisms in cytochrome oxidase in [(1987) FEBS Lett. 222, 235-245] and [Glynn Biological Research Reports (1987) 3, 1-7], I have suggested that a copper centre may enable the H2O/OH or H2O/O couple to act as the hydrogen-carrying arm of a redox loop by means of a (CuOH2)+/(CuOH)+ or (CuOH2)+/(CuO)+ system at the centre. I here explain that critical comments by Malmstrom [(1988) FEBS Lett. 231, 268-269] on the first of these papers, which might also be levelled at the second, depend on a misunderstanding. I also respond to Malmstrom's comment about testing conformationally coupled proton-pump mechanisms
MH  - Cytochrome-c Oxidase
MH  - metabolism
MH  - Oxidation-Reduction
MH  - Proton Pump
MH  - Protons
MH  - Research Design
RP  - NOT IN FILE
NT  - Glynn Research Institute, Bodmin, Cornwall, England
SO  - FEBS Lett 1988 Apr 11 ;231(1):270-271

854
UI  - 1010
AU  - Mitchell P
TI  - Possible protonmotive osmochemistry in cytochrome oxidase
MH  - Cytochrome-c Oxidase
MH  - Electron Transport
MH  - Kinetics
MH  - Ligands
MH  - metabolism
MH  - Models,Theoretical
MH  - Osmolar Concentration
MH  - Oxidation-Reduction
MH  - Protons
MH  - Support,Non-U.S.Gov't
RP  - NOT IN FILE
NT  - Glynn Research Institute, Bodmin, Cornwall, England
SO  - Ann N Y Acad Sci 1988  ;550:185-98.():185-198

855
UI  - 20880
AU  - Noumi T
AU  - Maeda M
AU  - Futai M
AD  - Department of Organic and Biochemistry, Osaka University, Japan
TI  - A homologous sequence between H+-ATPase (F0F1) and cation-transporting ATPases. Thr-285----Asp replacement in the beta subunit of Escherichia coli F1 changes its catalytic properties
AB  - A sequence of 10 amino acids (I-C-S-D-K-T-G-T-L-T) of ion motive ATPases such as Na+/K+-ATPase is similar to the sequence of the beta subunit of H+-ATPases, including that of Escherichia coli (I-T-S-T-K-T- G-S-I-T) (residues 282-291). The Asp (D) residue phosphorylated in ion motive ATPase corresponds to Thr (T) of the beta subunit. This substitution may be reasonable because there is no phosphoenzyme intermediate in the catalytic cycle of F1-ATPase. We replaced Thr-285 of the beta subunit by an Asp residue by in vitro mutagenesis and reconstituted the alpha beta gamma complex from the mutant (or wild- type) beta and wild-type alpha and gamma subunits. The uni- and multisite ATPase activities of the alpha beta gamma complex with mutant beta subunits were about 20 and 30% of those with the wild-type subunit. The rate of ATP binding (k1) of the mutant complex under uni- site conditions was about 10-fold less than that of the wild-type complex. These results suggest that Thr-285, or the region in its vicinity, is essential for normal catalysis of the H+-ATPase. The mutant complex could not form a phosphoenzyme under the conditions where the H+/K+-ATPase is phosphorylated, suggesting that another residue(s) may also be involved in formation of the intermediate in ion motive ATPase. The wild-type alpha beta gamma complex had slightly different kinetic properties from the wild-type F1, possibly because it did not contain the epsilon subunit
MH  - A
MH  - ACID
MH  - Adenosinetriphosphatase
MH  - Amino Acids
MH  - atp
MH  - ATPase
MH  - BETA-SUBUNIT
MH  - BINDING
MH  - Biochemistry
MH  - Catalysis
MH  - COMPLEX
MH  - EPSILON-SUBUNIT
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0F1
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - H+-ATPase
MH  - In Vitro
MH  - intermediate
MH  - ion
MH  - Macromolecular Systems
MH  - mutagenesis
MH  - mutant
MH  - RESIDUE
MH  - Site
MH  - SUBUNIT
MH  - SYSTEM
MH  - SYSTEMS
RP  - NOT IN FILE
NT  - UI - 88243734LA - engRN - 0 (Macromolecular Systems)RN - 56-84-8 (Aspartic Acid)RN - 72-19-5 (Threonine)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19880727IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:2897962
SO  - J Biol Chem 1988 Jun 25 ;263(18):8765-8770

856
UI  - 20884
AU  - Noumi T
AU  - Takeyama M
AU  - Kuki M
AU  - Maeda M
AU  - Futai M
AD  - Department of Organic Chemistry and Biochemistry, Institute of Scientific and Industrial Research, Osaka, Japan
TI  - Studies on essential amino acid residues and functional regions of H+- ATPase (F0F1) from Escherichia coli by gene manipulation
AB  - We discussed application of in vitro mutagenesis on H+-ATPase (F0F1) of Escherichia coli. The oligonucleotide-directed site specific mutagenesis and construction of a set of truncated subunits were useful for identifying essential residues of beta subunit and a functional region of epsilon subunit, respectively, of this complicated membrane enzyme
MH  - A
MH  - ACID
MH  - ATPase
MH  - BETA-SUBUNIT
MH  - Biochemistry
MH  - Chemistry
MH  - England
MH  - EPSILON-SUBUNIT
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0F1
MH  - H+
MH  - H+-ATPase
MH  - In Vitro
MH  - membrane
MH  - mutagenesis
MH  - RESIDUE
MH  - Site
MH  - SUBUNIT
RP  - NOT IN FILE
NT  - UI - 89144946LA - engRN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19890329IS - 0261-3166SB - IMCY - ENGLAND
UR  - PM:2906427
SO  - Nucleic Acids Symp Ser 1988  ;(19):175-178

857
UI  - 869
AU  - Ort DR
AU  - Good NE
TI  - Textbooks ignore photosystem II-dependent ATP formation: is the Z scheme to blame?
MH  - Adenosine Triphosphate
MH  - biosynthesis
MH  - Energy Metabolism
MH  - Photophosphorylation
MH  - Photosynthesis
RP  - NOT IN FILE
SO  - Trends Biochem Sci 1988 Dec ;13(12):467-469

858
UI  - 459
AU  - Parsonage D
AU  - Wilke-Mounts S
AU  - Senior AE
AD  - Department of Biochemistry, University of Rochester Medical Center, New York 14642
TI  - Directed mutagenesis of the dicyclohexylcarbodiimide-reactive carboxyl residues in beta-subunit of F1-ATPase of Escherichia coli
AB  - Previous studies in which dicyclohexylcarbodiimide (DCCD) was used to inactivate F1-ATPase enzymes have suggested that two glutamate residues in the beta-subunit are essential for catalysis. In the Escherichia coli F1-ATPase, these are residues beta-Glu-181 and beta-Glu-192. Oligonucleotide-directed mutagenesis was used to change these residues to beta-Gln-181 and beta-Gln-192. The beta-Gln-181 mutation produced strong impairment of oxidative phosphorylation in vivo and also of ATPase and ATP-driven proton-pumping activities in membranes assayed in vitro. A low level of each activity was detected and an F1-ATPase appeared to be assembled normally on the membranes. Therefore, it is suggested that the carboxyl side chain at residue beta-181 is important, although not absolutely required, for catalysis in both directions on E. coli F1-ATPase. The beta-Gln-192 mutation produced partial inhibition of oxidative phosphorylation in vivo and membrane ATPase activity was reduced by 78%. These results contrast with the complete or near-complete inactivation seen when E. coli F1-ATPase is reacted with DCCD and imply that DCCD-inactivation is attributable more to the attachment of the bulky DCCD molecule than to the derivatization of the carboxyl side chain of residue beta-Glu-192. M. Ohtsubo and colleagues (Biochem. Biophys. Res. Commun. (1987) 146, 705-710) described mutagenesis of the F1-beta-subunit of thermophilic bacterium PS3. Mutations (Glu----Gln) of the residues homologous to Glu-181 and Glu-192 of E. coli F1-beta-subunit both caused total inhibition of ATPase activity. Therefore, there was a marked difference in results obtained when the same residues were modified in the PS3 and E. coli F1- beta-subunits
RP  - NOT IN FILE
NT  - UI - 88132919LA - engRN - 0 (Carbodiimides)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSID - GM29805/GM/NIGMSDA - 19880323IS - 0003-9861SB - IMCY - UNITED STATESJC - 6SK
UR  - PM:2893590
SO  - Arch Biochem Biophys 1988 Feb 15 ;261(1):222-225

859
UI  - 457
AU  - Parsonage D
AU  - al Shawi MK
AU  - Senior AE
AD  - Department of Biochemistry, University of Rochester Medical Center, New York 14642
TI  - Directed mutations of the strongly conserved lysine 155 in the catalytic nucleotide-binding domain of beta-subunit of F1-ATPase from Escherichia coli
AB  - The amino acid sequence -Gly-X-X-X-X-Gly-Lys- occurs in many, diverse, nucleotide-binding proteins, and there is evidence that it forms a flexible loop which interacts with one or other of the phosphate groups of bound nucleotide. This sequence occurs as -Gly-Gly-Ala-Gly-Val-Gly- Lys- in the beta-subunit of the enzyme F1-ATPase, where it is thought to form part of the catalytic nucleotide-binding domain. Mutants of Escherichia coli were generated in which residue beta-lysine 155, at the end of the above sequence, was replaced by glutamine or glutamate. Properties of the soluble purified F1-ATPase from each mutant were studied. The results showed: 1) replacement of lysine 155 by Gln or Glu decreased the steady-state rate of ATP hydrolysis by 80 and 66%, respectively. 2) Characteristics of ATP hydrolysis at a single site were not markedly changed in the mutant enzymes, implying that lysine 155 is not directly involved in bond cleavage during ATP hydrolysis or bond formation during ATP synthesis. 3) The binding affinity for MgATP was weakened considerably in the mutants (Lys much much greater than Gln greater than Glu), whereas the binding affinity for MgADP was affected only mildly (Lys = Gln greater than Glu), suggesting that lysine 155 interacts with the gamma-phosphate of ATP bound at a single high affinity catalytic site. 4) The major determinant of inhibition of steady-state ATPase turnover rate in the mutant enzymes was an attenuation of positive catalytic cooperativity. 5) The data are consistent with the idea that during multisite catalysis residue 155 of beta-subunit undergoes conformational movement which changes substrate and product binding affinities
RP  - NOT IN FILE
NT  - UI - 88169589LA - engRN - 0 (Macromolecular Systems)RN - 56-65-5 (Adenosine Triphosphate)RN - 56-85-9 (Glutamine)RN - 56-87-1 (Lysine)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM-25349/GM/NIGMSID - GM-29805/GM/NIGMSDA - 19880502IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:2895106
SO  - J Biol Chem 1988 Apr 5 ;263(10):4740-4744

860
UI  - 454
AU  - Parsonage D
AU  - Wilke-Mounts S
AU  - Senior AE
AD  - Dept of Biochemistry, University of Rochester Medical Center, NY 14642
TI  - E. coli F1-ATPase: site-directed mutagenesis of the beta-subunit
AB  - Residues beta Glu-181 and beta Glu-192 of E. coli F1-ATPase (the DCCD- reactive residues) were mutated to Gln. Purified beta Gln-181 F1 showed 7-fold impairment of 'unisite' Pi formation from ATP and a large decrease in affinity for ATP. Thus the beta-181 carboxyl group in normal F1 significantly contributes to catalytic site properties. Also, positive catalytic site cooperativity was attenuated from 5 X 10(4)- to 548-fold in beta Gln-181 F1. In contrast, purified beta Gln-192 F1 showed only 6-fold reduction in 'multisite' ATPase activity. Residues beta Gly-149 and beta Gly-154 were mutated to Ile singly and in combination. These mutations, affecting residues which are strongly conserved in nucleotide-binding proteins, were chosen to hinder conformational motion in a putative 'flexible loop' in beta-subunit. Impairment of purified F1-ATPase ranged from 5 to 61%, with the double mutant F1 less impaired than either single mutant. F1 preparations containing beta Ile-154 showed 2-fold activation after release from membranes, suggesting association with F0 restrained turnover on F1 in these mutants
RP  - NOT IN FILE
NT  - UI - 88211839LA - engRN - 0 (Phosphates)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19880616IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:2896602
SO  - FEBS Lett 1988 May 9 ;232(1):111-114

861
UI  - 996
AU  - Paternostre MT
AU  - Roux M
AU  - Rigaud JL
TI  - Mechanisms of membrane protein insertion into liposomes during reconstitution procedures involving the use of detergents. 1. Solubilization of large unilamellar liposomes (prepared by reverse-phase evaporation) by triton X-100, octyl glucoside, and sodium cholate
AB  - The mechanisms governing the solubilization by Triton X-100, octyl glucoside, and sodium cholate of large unilamellar liposomes prepared by reverse-phase evaporation were investigated. The solubilization process is described by the three-stage model previously proposed for these detergents [Lichtenberg, D., Robson, R.J., & Dennis, E.A.(1983) Biochim. Biophys. Acta 737, 285-304]. In stage I, detergent monomers are incorporated into the phospholipid bilayers until they saturate the liposomes. At that point, i.e., stage II, mixed phospholipid-detergent micelles begin to form. By stage III, the lamellar to micellar transition is complete and all the phospholipids are present as mixed micelles. The turbidity of liposome preparations was systematically measured as a function of the amount of detergent added for a wide range of phospholipid concentrations (from 0.25 to 20 mM phospholipid). The results allowed a quantitative determination of RSat, the effective detergent to lipid molar ratios in the saturated liposomes, which were 0.64, 1.3, and 0.30 for Triton X-100, octyl glucoside, and sodium cholate, respectively. The corresponding ratios in the mixed micelles, RSol, were 2.5, 3.8, and 0.9 mol of detergent/mol of phospholipid. The monomer concentrations of the three detergents in the aqueous phase were also determined at the lamellar to micellar transitions (0.18, 17, and 2.8 mM, respectively). These transitions were also investigated by 31P NMR spectroscopy, and complete agreement was found with turbidity measurements. Freeze-fracture electron microscopy and permeability studies in the sublytic range of detergent concentrations indicated that during stage I of solubilization detergent partitioning between the aqueous phase and the lipid bilayer greatly affects the basic permeability of the liposomes without significantly changing the morphology of the preparations. A rough approximation of the partition coefficients was derived from the turbidity and permeability data (K = 3.5, 0.09, and 0.11 mM-1 for Triton X-100, octyl glucoside, and sodium cholate, respectively). It is concluded that when performed systematically, turbidity measurements constitute a very convenient and powerful technique for the quantitative study of the liposome solubilization process by detergents
MH  - Bacteriorhodopsin
MH  - Cholic Acid
MH  - Cholic Acids
MH  - Detergents
MH  - Fluoresceins
MH  - Freeze Fracturing
MH  - Glucosides
MH  - Glycosides
MH  - Liposomes
MH  - Magnetic Resonance Spectroscopy
MH  - Membrane Proteins
MH  - metabolism
MH  - Micelles
MH  - Microscopy
MH  - Microscopy,Electron
MH  - Models,Biological
MH  - Nephelometry and Turbidimetry
MH  - Octoxynol
MH  - Phospholipids
MH  - Polyethylene Glycols
MH  - Protons
MH  - Sodium
MH  - Support,Non-U.S.Gov't
MH  - Surface-Active Agents
RP  - NOT IN FILE
NT  - Departement de Biologie, CEN Saclay, Gif-sur-Yvette, France
SO  - Biochemistry 1988 Apr ;%19;27(8):2668-2677

862
UI  - 638
AU  - Penefsky HS
AD  - Department of Biochemistry, Public Health Research Institute, New York, NY 10016
TI  - Mechanism of action of the mitochondrial proton pumping ATPase in ATP synthesis and hydrolysis
RP  - NOT IN FILE
NT  - UI - 88336177LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19881027IS - 0361-7742SB - IMCY - UNITED STATESJC - PZ5
UR  - PM:2901746
SO  - Prog Clin Biol Res 1988  ;273():261-268

863
UI  - 637
AU  - Penefsky HS
AD  - Department of Biochemistry, Public Health Research Institute, New York, New York 10016
TI  - Rate of chase-promoted hydrolysis of ATP in the high affinity catalytic site of beef heart mitochondrial ATPase
AB  - Incubation of [gamma-32P]ATP with a molar excess of the soluble, homogeneous ATPase from beef heart mitochondria (F1) results in binding of substrate primarily in a single, very high affinity (KA = 10(12) M- 1) catalytic site and in a slow rate of hydrolysis characteristic of single site catalysis. Subsequent addition of millimolar concentrations of nonradioactive ATP as a cold chase, sufficient to fill catalytic sites on the enzyme, results in an acceleration of hydrolysis of bound radioactive ATP of as much as 10(6)-fold, that is, to Vmax rates (Cross, R.L., Grubmeyer, C., and Penefsky, H.S. (1982) J. Biol. Chem. 257, 12101-12105). For this reason, it was proposed that the high affinity catalytic site is a normal catalytic site on the molecule. Recently, Bullough et al. (Bullough, D.A., Verburg, J.G., Yoshida, M., and Allison, W.A. (1987) J. Biol. Chem. 262, 11675-11683) reported that when 5 to 20 microM concentrations of nonradioactive ATP were added as a cold chase to an enzyme-substrate complex consisting of F1 and ATP bound to the high affinity catalytic site, hydrolysis of the chase was commensurate with the turnover rate of the enzyme, whereas the hydrolysis of bound ATP was considerably slower. These authors suggested that the high affinity catalytic site on F1 is not a normal catalytic site. This paper shows, in experiments with a rapid mixing- chemical quench apparatus, that hydrolysis of ATP bound in the high affinity catalytic site is accelerated to Vmax rates following addition of 5 microM ATP as a cold chase. Hydrolysis of bound ATP appears to precede that of the chase. The weight of the available evidence continues to support the original suggestion that the high affinity catalytic site of beef heart F1 is a normal catalytic site
RP  - NOT IN FILE
NT  - UI - 88198132LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM21737/GM/NIGMSDA - 19880606IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:2896192
SO  - J Biol Chem 1988 May 5 ;263(13):6020-6022

864
UI  - 456
AU  - Rao R
AU  - al Shawi MK
AU  - Senior AE
AD  - Department of Biochemistry, University of Rochester Medical Center, New York 14642
TI  - Trinitrophenyl-ATP and -ADP bind to a single nucleotide site on isolated beta-subunit of Escherichia coli F1-ATPase. In vitro assembly of F1-subunits requires occupancy of the nucleotide-binding site on beta-subunit by nucleoside triphosphate
AB  - The stoichiometry of nucleotide binding to the isolated alpha- and beta- subunits of Escherichia coli F1-ATPase was investigated using two experimental techniques: (a) titration with fluorescent trinitrophenyl (TNP) derivatives of AMP, ADP, and ATP and (b) the centrifuge column procedure using the particular conditions of Khananshvili and Gromet- Elhanan (Khananshvili, D., and Gromet-Elhanan, Z. (1985) FEBS Lett. 178, 10-14). Both procedures showed that alpha-subunit contains one nucleotide-binding site, confirming previous work. TNP-ADP and TNP-ATP bound to a maximal level of 1 mol/mol beta-subunit, consistent with previous equilibrium dialysis studies which showed isolated beta- subunit bound 1 mol of ADP or ATP per mol (Issartel, J. P., and Vignais, P. V. (1984) Biochemistry 23, 6591-6595). However, binding of only approximately 0.1 mol of ATP or ADP per mol of beta-subunit was detected using centrifuge columns. Our results are consistent with the conclusion that each of the alpha- and beta-subunits contains one nucleotide-binding domain. Because the subunit stoichiometry is alpha 3 beta 3 gamma delta epsilon, this can account for the location of the six known nucleotide-binding sites in E. coli F1-ATPase. Studies of in vitro assembly of isolated alpha-, beta-, and gamma- subunits into an active ATPase showed that ATP, GTP, and ITP all supported assembly, with half-maximal reconstitution of ATPase occurring at concentrations of 100-200 microM, whereas ADP, GDP, and IDP did not. Also TNP-ATP supported assembly and TNP-ADP did not. The results demonstrate that (a) the nucleotide-binding site on beta-subunit has to be filled for enzyme assembly to proceed, whereas occupancy of the alpha-subunit nucleotide-binding site is not required, and (b) that enzyme assembly requires nucleoside triphosphate
RP  - NOT IN FILE
NT  - UI - 88186866LA - engRN - 0 (2',3'-O-(2,4,6-trinitrophenyl)adenosine 5'-triphosphate)RN - 0 (Macromolecular Systems)RN - 132-06-9 (Inosine Triphosphate)RN - 25612-73-1 (Adenylyl Imidodiphosphate)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 77450-67-0 (2',3' O-(2,4,6-trinitrophenyl)adenosine 5'-diphosphate)RN - 86-01-1 (Guanosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSID - GM29805/GM/NIGMSDA - 19880518IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:2895769
SO  - J Biol Chem 1988 Apr 25 ;263(12):5569-5573

865
UI  - 453
AU  - Rao R
AU  - Pagan J
AU  - Senior AE
AD  - Department of Biochemistry, University of Rochester Medical Center, New York 14642
TI  - Directed mutagenesis of the strongly conserved lysine 175 in the proposed nucleotide-binding domain of alpha-subunit from Escherichia coli F1-ATPase
AB  - The alpha-subunit of Escherichia coli F1-ATPase contains an adenine- specific noncatalytic nucleotide-binding domain. A recent proposal (Maggio, M. B., Pagan, J., Parsonage, D., Hatch, L., and Senior, A. E. (1987) J. Biol. Chem. 262, 8981-8984) suggested that this domain is formed by residues 160-340, approximately, in alpha-subunit. Within this proposed domain is a sequence Gly-X-X-X-X-Gly-Lys which is conserved in a large and diverse group of nucleotide-binding proteins and is thought to interact with phosphate groups of bound nucleotide. In this work, residue alpha Lys-175, the terminal residue of the above conserved sequence in F1-alpha-subunit, was mutagenized to Ile or Glu. The specific activity of purified mutant F1-ATPase was reduced by 2.5- fold (Ile) or 3-fold (Glu). Apparent binding of ATP to alpha-subunit, as measured by the centrifuge column procedure, was strongly impaired and ATP-induced conformational change in alpha-subunit, as measured by protection against trypsin proteolysis, was nearly abolished in both mutants. The results suggest that residue alpha Lys-175 is located within the nucleotide-binding domain of alpha-subunit, and that this residue is functionally involved in nucleotide binding. The results support previous suggestions that the alpha-subunit nucleotide-binding site is not involved, directly or indirectly, in catalysis
RP  - NOT IN FILE
NT  - UI - 89034048LA - engRN - 0 (Macromolecular Systems)RN - 0 (Nucleotides)RN - 0 (Plasmids)RN - 56-87-1 (Lysine)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSID - GM29805/GM/NIGMSDA - 19881202IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:2903146
SO  - J Biol Chem 1988 Nov 5 ;263(31):15957-15963

866
UI  - 455
AU  - Rao R
AU  - Cunningham D
AU  - Cross RL
AU  - Senior AE
AD  - Department of Biochemistry and Molecular Biology, State University of New York Health Science Center, Syracuse 13210
TI  - Pyridoxal 5'-diphospho-5'-adenosine binds at a single site on isolated alpha-subunit from Escherichia coli F1-ATPase and specifically reacts with lysine 201
AB  - Pyridoxal 5'-diphospho-5'-adenosine (PLP-AMP), an adenine nucleotide affinity analog, was found to bind in a saturable fashion to isolated alpha-subunit from Escherichia coli F1-ATPase with a stoichiometry of one mol/mol and a Kd approximately 150 microM. The binding was shown to be specific by the following criteria: 1) ATP reduced the binding of PLP-AMP by 80%, and 2) PLP-AMP, like ATP, induced a conformational change which increased the mobility of alpha-subunit in nondenaturing polyacrylamide gel electrophoresis and rendered alpha-subunit resistant to mild trypsin proteolysis. A stable adduct was formed between isolated alpha-subunit and [3H] PLP-AMP after reduction with NaBH4. alpha-Subunit labeled to the extent of 0.4-0.7 mol/mol was digested with trypsin and subjected to high pressure liquid chromatography purification, yielding a single labeled peptide. Automated amino acid sequencing showed that residue alpha-Lys-201 was specifically labeled. The results suggest that Lys-201 occupies a position proximate to the phosphate groups of bound ATP in the alpha.ATP complex. PLP-AMP did not support repolymerization of isolated alpha-, beta-, and gamma-subunits, consistent with previous reports that subunit repolymerization in vitro is dependent upon the presence of nucleoside triphosphate. Further, PLP- AMP-labeled alpha-subunit could not be reconstituted with isolated beta- and gamma-subunits in the presence of ATP, showing that occupation of the alpha-subunit nucleotide site by PLP-AMP impairs normal subunit- subunit interaction
RP  - NOT IN FILE
NT  - UI - 88186877LA - engRN - 0 (Affinity Labels)RN - 0 (Borohydrides)RN - 0 (Macromolecular Systems)RN - 16940-66-2 (sodium borohydride)RN - 4500-99-6 (pyridoxal 5'-diphospho-5'-adenosine)RN - 54-47-7 (Pyridoxal Phosphate)RN - 56-65-5 (Adenosine Triphosphate)RN - 56-87-1 (Lysine)RN - EC 3.4.21.4 (Trypsin)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM23152/GM/NIGMSID - GM25349/GM/NIGMSID - GM29805/GM/NIGMSDA - 19880518IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:2895772
SO  - J Biol Chem 1988 Apr 25 ;263(12):5640-5645

867
UI  - 995
AU  - Rigaud JL
AU  - Paternostre MT
AU  - Bluzat A
TI  - Mechanisms of membrane protein insertion into liposomes during reconstitution procedures involving the use of detergents. 2. Incorporation of the light-driven proton pump bacteriorhodopsin
AB  - A method has been developed for identifying the step in a detergent-mediated reconstitution procedure at which an integral membrane protein can be associated with phospholipids to give functional proteoliposomes. Large liposomes prepared by reverse-phase evaporation were treated with various amounts of the detergents Triton X-100, octyl glucoside, or sodium cholate as described in the preceding paper [Paternostre, M.-T., Roux, M., & Rigaud, J. L. (1988) Biochemistry (preceding paper in this issue)]. At each step of the solubilization process, we added bacteriorhodopsin, the light-driven proton pump from Halobacterium halobium. The protein-phospholipid detergent mixtures were then subjected to SM2 Bio-Beads treatments to remove the detergent, and the resulting vesicles were analyzed with respect to protein insertion and orientation in the membrane by freeze-fracture electron microscopy, sucrose density gradients, and proton pumping measurements. The nature of the detergent used for reconstitution proved to be important for determining the mechanism of protein insertion. With sodium cholate, proteoliposomes were formed only from ternary phospholipid-protein-detergent micelles. With octyl glucoside, besides proteoliposome formation from ternary mixed micelles, direct incorporation of bacteriorhodopsin into preformed liposomes destabilized by saturating levels of this detergent was observed and gave proteoliposomes with optimal proton pumping activity. With Triton X-100, protein insertion into destabilized liposomes was also observed but involved a transfer of the protein initially present in phospholipid-Triton X-100-protein micelles into Triton X-100 saturated liposomes. Our results further demonstrated that protein orientation in the resulting proteoliposomes was critically dependent upon the mechanism by which the protein was incorporated
MH  - Bacteriorhodopsin
MH  - Biochemistry
MH  - Carbon Radioisotopes
MH  - Detergents
MH  - Halobacterium
MH  - Liposomes
MH  - Membrane Proteins
MH  - metabolism
MH  - Micelles
MH  - Microscopy
MH  - Models,Biological
MH  - Octoxynol
MH  - Phosphatidylcholines
MH  - Phospholipids
MH  - Polyethylene Glycols
MH  - Proteolipids
MH  - Proton Pump
MH  - Sodium
MH  - Support,Non-U.S.Gov't
MH  - Surface-Active Agents
RP  - NOT IN FILE
NT  - Departement de Biologie, CEN Saclay, Gif-sur-Yvette, France
SO  - Biochemistry 1988 Apr ;%19;27(8):2677-2688

868
UI  - 672
AU  - Schafer G
AU  - Weber J
AU  - Tiedge H
AU  - Lubben M
AD  - Institute of Biochemistry, Medical University of Lubeck, FRG
TI  - Proton-ATPases: universal catalysts in biological energy conservation
RP  - NOT IN FILE
NT  - UI - 88336208LA - engRN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19881027IS - 0361-7742SB - IMCY - UNITED STATESJC - PZ5
UR  - PM:2901751
SO  - Prog Clin Biol Res 1988  ;273():57-66

869
UI  - 460
AU  - Senior AE
AD  - Department of Biochemistry, University of Rochester Medical Center, New York
TI  - ATP synthesis by oxidative phosphorylation
RP  - NOT IN FILE
NT  - UI - 88097736LA - engRN - 0 (Protons)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, AcademicID - GM-25349/GM/NIGMSID - GM-29805/GM/NIGMSDA - 19880217IS - 0031-9333SB - IMCY - UNITED STATESJC - P7M
UR  - PM:2892214
SO  - Physiol Rev 1988 Jan ;68(1):177-231

870
UI  - 20957
AU  - Steffens K
AU  - Hoppe J
AU  - Altendorf K
AD  - Universitat Osnabruck, Fachbereich Biologie/Chemie, Mikrobiologie, Federal Republic of Germany
TI  - F0 part of the ATP synthase from Escherichia coli. Influence of subunits a, and b, on the structure of subunit c
AB  - Four different sets of proteoliposomes were prepared from F0, subunit c, a complex of subunits a and c (ac complex) and an ac complex supplemented with subunit b. Only liposomes containing intact F0 or all subunits of F0 were active in proton translocation and F1 binding [Schneider, E. and Altendorf, K. (1985) EMBO J. 4, 515-518]. The conformation of subunit c in the different preparations was analyzed by labelling the proteoliposomes with the hydrophobic photoactivatable reagent 3-(trifluoromethyl)-3-(m-[125I]iodophenyl)diazirine ([125I]TID). Subsequent isolation and Edman degradation of this polypeptide revealed distinct radioactive labelling patterns over the entire amino acid sequence. In the F0 complex and in the ac complex subunit c retains a labelling pattern which is related to that found in TID-labelled membrane vesicles of Escherichia coli [Hoppe et al. (1984) Biochemistry 23, 5610-5616]. In the absence of subunit a, considerably more and different amino acid residues of subunit c are modified. The labelling data are discussed in relation to structural aspects of F0 and functional properties of proteoliposomes reconstituted with F0 or individual subunits
MH  - A
MH  - ACID
MH  - ACTIVE
MH  - Amino Acid Sequence
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BINDING
MH  - Biochemistry
MH  - COMPLEX
MH  - conformation
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - F1
MH  - Liposomes
MH  - Macromolecular Systems
MH  - membrane
MH  - membrane vesicles
MH  - Proteolipids
MH  - proteoliposome
MH  - proton
MH  - RESIDUE
MH  - SUBUNIT
MH  - SYNTHASE
MH  - SYSTEM
MH  - SYSTEMS
MH  - translocation
RP  - NOT IN FILE
NT  - UI - 88111649LA - engRN - 0 (Azirines)RN - 0 (Liposomes)RN - 0 (Macromolecular Systems)RN - 0 (Proteolipids)RN - 0 (proteoliposomes)RN - 81340-56-9 (3-(trifluoromethyl)-3-(3-iodophenyl)diazirine)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19880303IS - 0014-2956SB - IMCY - GERMANY, WEST
UR  - PM:2892677
SO  - Eur J Biochem 1988 Jan 4 ;170(3):627-630

871
UI  - 905
AU  - Stokes GB
TI  - Estimating the energy content of nutrients
MH  - Adenosine Triphosphate
MH  - biosynthesis
MH  - Calorimetry
MH  - Energy Metabolism
MH  - metabolism
MH  - Nutrition
MH  - Oxygen
MH  - Thermodynamics
RP  - NOT IN FILE
SO  - Trends Biochem Sci 1988 Nov ;13(11):422-424

872
UI  - 760
AU  - Suzuki K
AU  - Mizuno Y
AU  - Yoshida M
AD  - Department of Neurology, Jichi Medical School, Tochigi-ken, Japan
TI  - Inhibition of mitochondrial NADH-ubiquinone oxidoreductase activity and ATP synthesis by tetrahydroisoquinoline
AB  - Effects of tetrahydroisoquinoline (TIQ) on mitochondrial respiration, NADH-ubiquinone oxidoreductase (complex I) activity and on adenosine triphosphate (ATP) synthesis were studied using mitochondria prepared from mouse brains. Tetrahydroisoquinoline significantly inhibited mitochondrial respiration supported by glutamate + malate, pyruvate + malate or alpha-ketoglutarate. Activity of complex I and synthesis of ATP were also significantly inhibited by TIQ. Mitochondrial respiration supported by succinate and subsequent ATP synthesis were not inhibited at all by 5 mM of TIQ. Our study has revealed a novel action of TIQ, which has been proposed as a candidate for an endogenous substance that may induce Parkinson's disease
RP  - NOT IN FILE
NT  - UI - 88202595LA - engRN - 0 (Isoquinolines)RN - 56-65-5 (Adenosine Triphosphate)RN - 91-21-4 (1,2,3,4-tetrahydroisoquinoline)RN - EC 1.6.99. (Quinone Reductases)RN - EC 1.6.99.2 (NAD(P)H Dehydrogenase (Quinone))PT - Journal ArticleDA - 19880602IS - 0304-3940SB - IMCY - NETHERLANDSJC - N7N
UR  - PM:3129681
SO  - Neurosci Lett 1988 Mar 21 ;86(1):105-108

873
UI  - 20881
AU  - Tagaya M
AU  - Noumi T
AU  - Nakano K
AU  - Futai M
AU  - Fukui T
AD  - Institute of Scientific and Industrial Research, Osaka University, Japan
TI  - Identification of alpha-subunit Lys201 and beta-subunit Lys155 at the ATP-binding sites in Escherichia coli F1-ATPase
AB  - Binding of about 1 mol of adenosine triphosphopyridoxal to Escherichia coli F1-ATPase resulted in the nearly complete inactivation of the enzyme [(1987) J. Biol. Chem. 262, 7686-7692]. About two thirds of the label was bound to the alpha-subunit, and the rest to the beta-subunit. The present study revealed that Lys201 in the alpha-subunit and Lys155 in the glycine-rich region of the beta-subunit are the major sites labeled with this reagent. Thus, these two residues might be located close to the gamma-phosphate of the bound ATP
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - ALPHA-SUBUNIT
MH  - atp
MH  - ATPase
MH  - BETA-SUBUNIT
MH  - BINDING
MH  - COLI F1 ATPASE
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - Macromolecular Systems
MH  - Peptide Fragments
MH  - RESIDUE
MH  - Site
MH  - SYSTEM
MH  - SYSTEMS
RP  - NOT IN FILE
NT  - UI - 88255328LA - engRN - 0 (Macromolecular Systems)RN - 0 (Peptide Fragments)RN - 56-65-5 (Adenosine Triphosphate)RN - 56-87-1 (Lysine)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19880808IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:2898387
SO  - FEBS Lett 1988 Jun 20 ;233(2):347-351

874
UI  - 20877
AU  - Takeyama M
AU  - Noumi T
AU  - Maeda M
AU  - Futai M
AD  - Department of Organic Chemistry and Biochemistry, Osaka University, Japan
TI  - Fo portion of Escherichia coli H+-ATPase. Carboxyl-terminal region of the b subunit is essential for assembly of functional Fo
AB  - Six chromosomal uncF mutants of Escherichia coli defective in the b subunit of H+-ATPase (156 amino acid residues) were identified (KF92, Met-1----Val; KF164, Gln-64----end; KF61 and KF144, Gln-104----end; KF138, Gln-106----end; and KF79, Gln-123----end). The membranes of all these mutants had low ATPase activities (less than 5% of that of the wild type), and no functional H+ pathway, although the truncated b subunits were integrated into these membranes. These findings suggest that about 30 carboxyl-terminal amino acid residues of the b subunit are essential for formation of the F1-binding site and H+ pathway. For examination of the role(s) of the carboxyl-terminal region(s) or residue(s) of the b subunit, recombinant plasmids carrying truncated uncF genes of various lengths were constructed by in vitro muta-genesis and introduced into a recA1 derivative of strain KF92 (Met-1----Val). Analyses of the membranes from the resulting strains demonstrated that almost the entire carboxyl-terminal region of the b subunit is necessary for formation of functional Fo, since loss of the carboxyl- terminal residue resulted in significant reduction of both F1 binding and H+ translocation, and loss of two or more residues abolished both activities completely
MH  - A
MH  - ACID
MH  - ATPase
MH  - BINDING
MH  - Biochemistry
MH  - Chemistry
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1
MH  - H+
MH  - H+-ATPase
MH  - In Vitro
MH  - Macromolecular Systems
MH  - membrane
MH  - Membranes
MH  - mutagenesis
MH  - mutant
MH  - RESIDUE
MH  - Site
MH  - SUBUNIT
MH  - SYSTEM
MH  - SYSTEMS
MH  - translocation
RP  - NOT IN FILE
NT  - UI - 89034070LA - engRN - 0 (Macromolecular Systems)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19881202IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:2903150
SO  - J Biol Chem 1988 Nov 5 ;263(31):16106-16112

875
UI  - 20822
AU  - Vik SB
AU  - Cain BD
AU  - Chun KT
AU  - Simoni RD
AD  - Department of Biological Sciences, Stanford University, California 94305
TI  - Mutagenesis of the alpha subunit of the F1Fo-ATPase from Escherichia coli. Mutations at Glu-196, Pro-190, and Ser-199
AB  - In an attempt to identify amino acid residues involved in proton translocation by the Fo sector of the Escherichia coli F1Fo-ATPase, 16 mutations at the carboxyl-terminal third of the a subunit have been isolated, and their phenotypes have been partially characterized. Thirteen mutations were constructed by "cassette" mutagenesis at two highly conserved residues, aglu196 and apro190. Two mutations were products of oligonucleotide-directed mutagenesis of a portion of of oligonucleotide-directed mutagenesis of a portion of the uncB gene cloned into an M13 vector. One mutation was isolated after in vitro mutagenesis of the entire uncB gene in a plasmid vector with hydroxylamine. Amino acid substitutions for aglu196 (Asp, Gln, His, Asn, Lys, Ala, Ser, Pro) affect ATP-driven proton translocation and passive proton permeability by Fo to varying extents, but do not prevent growth on minimal succinate media. Amino acid substitutions of glutamine or arginine for apro190 affect F1Fo-ATPase assembly and eliminate ATP-driven proton translocation, while the substitution of asparagine at this position does not significantly affect either assembly or proton translocation. The substitution of amino acids threonine or alanine for aser199 causes no detectable phenotypic change from wild type. These and other mutations are discussed in terms of the assembly, structure, and function of the a subunit. It is concluded that aglu196 and apro190 are not obligate components of the proton channel, but that they affect proton translocation indirectly
MH  - A
MH  - ACID
MH  - ALPHA-SUBUNIT
MH  - Amino Acids
MH  - ATPase
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - function
MH  - glutamic acid
MH  - Glutamine
MH  - In Vitro
MH  - Macromolecular Systems
MH  - mutagenesis
MH  - Permeability
MH  - proton
MH  - RESIDUE
MH  - SUBUNIT
MH  - succinate
MH  - SYSTEM
MH  - SYSTEMS
MH  - translocation
RP  - NOT IN FILE
NT  - UI - 88198221LA - engRN - 0 (Glutamates)RN - 0 (Macromolecular Systems)RN - 0 (Plasmids)RN - 147-85-3 (Proline)RN - 56-45-1 (Serine)RN - 56-86-0 (Glutamic Acid)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleID - GM07276/GM/NIGMSID - GM09521/GM/NIGMSID - GM18539/GM/NIGMSDA - 19880609IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:2896196
SO  - J Biol Chem 1988 May 15 ;263(14):6599-6605

876
UI  - 9923
AU  - Wang MY
AU  - Chien LF
AU  - Pan RL
TI  - Radiation inactivation analysis of chloroplast CF0-CF1 ATPase.
AB  - Radiation inactivation technique was employed to measure the functional size of adenosine triphosphatase of spinach chloroplasts. The functional size for acid-base-induced ATP synthesis was 450 +/- 24 kilodaltons; for phenazine methosulfate-mediated ATP synthesis, 613 +/- 33 kilodaltons; and for methanol-activated ATP hydrolysis, 280 +/- 14 kilodaltons. The difference (170 +/- 57 kilodaltons) between 450 +/- 24 and 280 +/- 14 kilodaltons is explained to be the molecular mass of proton channel (coupling factor 0) across the thylakoid membrane. Our data suggest that the stoichiometry of subunits I, II, and III of coupling factor 0 is 1:2:15. Ca2+- and Mg2+-ATPase activated by methanol, heat, and trypsin digestion have a similar functional size. However, anions such as SO3(2-) and CO3(2-) increased the molecular mass for both ATPase's (except trypsin-activated Mg2+-ATPase) by 12-30%. Soluble coupling factor 1 has a larger target size than that of membrane-bound. This is interpreted as the cold effect during irradiation.
MH  - Adenosine
MH  - ADENOSINE-TRIPHOSPHATASE
MH  - Adenosinetriphosphatase
MH  - analysis
MH  - Anions
MH  - antagonists & inhibitors
MH  - atp
MH  - ATP synthesis
MH  - ATPase
MH  - Ca(2+) Mg(2+)-Atpase
MH  - Ca(2+)-Transporting ATPase
MH  - chloroplast
MH  - Chloroplasts
MH  - COUPLING FACTOR
MH  - Dose-Response Relationship,Radiation
MH  - Enzyme Activation
MH  - enzymology
MH  - Heat
MH  - Hydrolysis
MH  - Intracellular Membranes
MH  - Kinetics
MH  - metabolism
MH  - Methanol
MH  - Plants
MH  - proton
MH  - radiation effects
MH  - Spinach
MH  - SPINACH-CHLOROPLASTS
MH  - SUBUNIT
MH  - Support,Non-U.S.Gov't
MH  - Trypsin
RP  - NOT IN FILE
NT  - Institute of Radiation Biology, College of Nuclear Sciences, Taiwan, Republic of ChinaPMID- 0002967817
SO  - J Biol Chem 1988 Jun 25 ;263(18):8838-8843

877
UI  - 1009
AU  - West IC
AU  - Mitchell P
AU  - Rich PR
TI  - Electron conduction between b cytochromes of the mitochondrial respiratory chain in the presence of antimycin plus myxothiazol
AB  - The b haems of the bc1 complex of bovine heart mitochondria were poised with succinate and fumarate so that only the high-potential haem (b-562) was reduced, and then isolated from further redox exchange with the ubiquinone pool by adding antimycin and myxothiazol. A transmembrane electric potential difference was then developed, either by electron flow from [Ru(NH3)6]Cl2 to oxygen or by ATP hydrolysis. The small difference spectrum, caused by the electric field, indicated 32-55% oxidation of b-562 with concomitant reduction of b-566. No lag greater than 0.1 s was detectable between the initiation of respiration and the development of the difference spectrum, thus providing a direct demonstration of (fairly) rapid electron transfer between the b haems
MH  - analogs & derivatives
MH  - Animal
MH  - antagonists & inhibitors
MH  - Antimycin A
MH  - Cattle
MH  - Cytochrome b
MH  - development
MH  - drug effects
MH  - Electron Transport
MH  - Fumarates
MH  - Hydrolysis
MH  - Membrane Potentials
MH  - metabolism
MH  - Mitochondria
MH  - Mitochondria,Heart
MH  - Oxidation-Reduction
MH  - Oxygen
MH  - pharmacology
MH  - Respiration
MH  - Spectrophotometry
MH  - Succinates
MH  - Succinic Acid
MH  - Support,Non-U.S.Gov't
MH  - Thiazoles
RP  - NOT IN FILE
NT  - Glynn Research Institute, Bodmin, UK
SO  - Biochim Biophys Acta 1988 Mar 30 ;933(1):35-41

878
UI  - 529
AU  - Williams N
AU  - Hullihen J
AU  - Pedersen PL
AD  - Johns Hopkins University School of Medicine, Baltimore, MD 21205
TI  - Mitochondrial ATP synthase: role of metal binding in structure and function
RP  - NOT IN FILE
NT  - UI - 88336213LA - engRN - 0 (Adenine Nucleotides)RN - 0 (Carrier Proteins)RN - 0 (Cations, Divalent)RN - 0 (Metals)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19881027IS - 0361-7742SB - IMCY - UNITED STATESJC - PZ5
UR  - PM:2901755
SO  - Prog Clin Biol Res 1988  ;273():87-92

879
UI  - 8084
AU  - Williams RJP
TI  - Proton circuits in biological energy conversion
MH  - proton
RP  - NOT IN FILE
SO  - Annu Rev Biophys Biophys Chem 1988  ;17():71-97

880
UI  - 19791
AU  - Xue ZX
AU  - Melese T
AU  - Stempel KE
AU  - Reedy TJ
AU  - Boyer PD
AD  - Molecular Biology Institute, University of California, Los Angeles 90024-1570
TI  - Properties of chloroplast F1-ATPase partially modified by 2-azido adenine nucleotides, including demonstration of three catalytic pathways
AB  - Previous investigations on the distribution of [18O]Pi isotopomers formed by hydrolysis of [gamma-18O]ATP by the chloroplast F1-ATPase (CF1) showed that a single reaction pathway is used by all participating sites and that the pathway is modulated by ATP concentration as expected for cooperative interactions between catalytic sites. Such oxygen exchange measurements have been applied to CF1 modified at a single catalytic or noncatalytic site by 2-azido adenine nucleotides. When less than one catalytic or one noncatalytic site per enzyme is modified, hydrolysis occurs in part by the pathway of the unmodified enzyme plus at least one additional pathway at 200 microM and two additional pathways at 4 microM [gamma-18O]ATP. Thus, three sites are potentially catalytically active. The two new pathways shown by the derivatized enzyme logically can arise from nonidentical interactions of the remaining two underivatized beta subunits with the derivatized beta subunit. Reversals of bound ATP cleavage before Pi is released are increased, and the amount of product formed by the new pathways is changed when the ATP concentration is lowered. These modulations must result from the behavior of two remaining active catalytic sites rather than of one catalytic and one regulatory site. When the CF1 is derivatized more extensively, the original catalytic pathway is lost, and two catalytic pathways that do not show modulation by ATP concentration are found. The remaining beta subunits now have weak but independent catalytic capacity. In addition, the enzyme is no longer activated by Ca2+, loses MgGTPase activity, and is much less sensitive to azide
MH  - A
MH  - ACTIVE
MH  - Adenine Nucleotides
MH  - ADENINE-NUCLEOTIDES
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthase
MH  - Azides
MH  - BETA-SUBUNIT
MH  - chloroplast
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - Guanosine Triphosphate
MH  - H(+)-Transporting ATP Synthase
MH  - Hydrolysis
MH  - Magnesium
MH  - Nucleotides
MH  - Oxygen
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 89034183LA - engRN - 0 (Azides)RN - 28141-84-6 (magnesium GTP)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 64020-53-7 (2-azidoadenosine 3',5'-diphosphate)RN - 72884-75-4 (2-azidoadenosine 5'-triphosphate)RN - 86-01-1 (Guanosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM11094/GM/NIGMSDA - 19881214IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:2903156
SO  - J Biol Chem 1988 Nov 15 ;263(32):16880-16885

881
UI  - 526
AU  - Ysern X
AU  - Amzel LM
AU  - Pedersen PL
AD  - Department of Biophysics, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205
TI  - ATP synthases--structure of the F1-moiety and its relationship to function and mechanism
AB  - A great deal of progress has been made in understanding both the structure and the mechanism of F1-ATPase. The primary structure is now fully known for at least five species. Sequence comparison between chloroplast, photobacteria, aerobic bacteria, and mitochondrial representatives allow us to infer more general functional relationships and evolutionary trends. Although the F1 moiety is the most studied segment of the H+-ATPase complex, there is not a full understanding of the mechanism and regulation of its hydrolytic activity. The beta subunit is now known to contain one and probably two nucleotide binding domains, one of which is believed to be a catalytic site. Recently, two similar models have been proposed to attempt to describe the "active" part of the beta subunits. These models are mainly an attempt to use the structure of adenylate kinase to represent a more general working model for nucleotide binding phosphotransferases. Labelling experiments seem to indicate that several critical residues outside the region described by the "adenylate kinase" part of this model are also actively involved in the ATPase activity. New models will have to be introduced to include these regions. Finally, it seems that a consensus has been reached with regard to a broad acceptance of the asymmetric structure of the F1-moiety. In addition, recent experimental evidence points toward the presence of nonequivalent subunits to describe the functional activity of the F1-ATPase. A summary diagram of the conformational and binding states of the enzyme including the nonequivalent beta subunit is presented. Additional research is essential to establish the role of the minor subunits--and of the asymmetry they introduce in F1--on the physiological function of the enzyme
RP  - NOT IN FILE
NT  - UI - 89123236LA - engRN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialID - CA 10951/CA/NCIID - GM 25432/GM/NIGMSDA - 19890316IS - 0145-479XSB - IMCY - UNITED STATESJC - HIO
UR  - PM:2906060
SO  - J Bioenerg Biomembr 1988 Aug ;20(4):423-450

882
UI  - 19793
AU  - Zhou JM
AU  - Xue ZX
AU  - Du ZY
AU  - Melese T
AU  - Boyer PD
AD  - Department of Chemistry and Biochemistry, University of California, Los Angeles 90024-1570
TI  - Relationship of tightly bound ADP and ATP to control and catalysis by chloroplast ATP synthase
AB  - Whether the tightly bound ADP that can cause a pronounced inhibition of ATP hydrolysis by the chloroplast ATP synthase and F1 ATPase (CF1) is bound at catalytic sites or at noncatalytic regulatory sites or both has been uncertain. We have used photolabeling by 2-azido-ATP and 2- azido-ADP to ascertain the location, with Mg2+ activation, of tightly bound ADP (a) that inhibits the hydrolysis of ATP by chloroplast ATP synthase, (b) that can result in an inhibited form of CF1 that slowly regains activity during ATP hydrolysis, and (c) that arises when low concentrations of ADP markedly inhibit the hydrolysis of GTP by CF1. The data show that in all instances the inhibition is associated with ADP binding without inorganic phosphate (Pi) at catalytic sites. After photophosphorylation of ADP or 2-azido-ADP with [32P]Pi, similar amounts of the corresponding triphosphates are present on washed thylakoid membranes. Trials with appropriately labeled substrates show that a small portion of the tightly bound 2-azido-ATP gives rise to covalent labeling with an ATP moiety at noncatalytic sites but that most of the bound 2-azido-ATP gives rise to covalent labeling by an ADP moiety at a catalytic site. We also report the occurrence of a 1-2-min delay in the onset of the Mg2+-induced inhibition after addition of CF1 to solutions containing Mg2+ and ATP, and that this delay is not associated with the filling of noncatalytic sites. A rapid burst of Pi formation is followed by a much lower, constant steady-state rate.(ABSTRACT TRUNCATED AT 250 WORDS)
MH  - 2-AZIDO-ATP
MH  - A
MH  - ACTIVATION
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - ADP
MH  - ADP BINDING
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BINDING
MH  - Biochemistry
MH  - Catalysis
MH  - Chemistry
MH  - chloroplast
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - Guanosine Triphosphate
MH  - H(+)-Transporting ATP Synthase
MH  - Hydrolysis
MH  - INORGANIC-PHOSPHATE
MH  - Magnesium
MH  - membrane
MH  - Membranes
MH  - NONCATALYTIC SITES
MH  - Photophosphorylation
MH  - Solutions
MH  - SYNTHASE
MH  - thylakoid
MH  - thylakoid membrane
RP  - NOT IN FILE
NT  - UI - 89000727LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 7439-95-4 (Magnesium)RN - 86-01-1 (Guanosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM-11094/GM/NIGMSDA - 19881123IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:2901855
SO  - Biochemistry 1988 Jul 12 ;27(14):5129-5135

883
UI  - 448
AU  - al Shawi MK
AU  - Parsonage D
AU  - Senior AE
AD  - Department of Biochemistry, University of Rochester Medical Center, New York 14642
TI  - Kinetic characterization of the unisite catalytic pathway of seven beta- subunit mutant F1-ATPases from Escherichia coli
AB  - We have studied the kinetics of "unisite" ATP hydrolysis and synthesis in seven mutant Escherichia coli F1-ATPase enzymes. The seven mutations are distributed over a 105-residue segment of the catalytic nucleotide- binding domain in beta-subunit and are: G142S, K155Q, K155E, E181Q, E192Q, M209I, and R246C. We report forward and reverse rate constants and equilibrium constants in all seven mutant enzymes for the four steps of unisite kinetics, namely (i) ATP binding/release, (ii) ATP hydrolysis/synthesis, (iii) Pi release/binding, and (iv) ADP release/binding. The seven mutant enzymes displayed a wide range of deviations from normal in both rate and equilibrium constants, with no discernible common pattern. Notably, steep reductions in Kd ATP were seen in some cases, the value of Kd Pi was high, and K2 (ATP hydrolysis/synthesis) was relatively unaffected. Significantly, when the data from the seven mutations were combined with previous data from two other E. coli F1-beta-subunit mutations (D242N, D242V), normal E. coli F1, soluble and membranous mitochondrial F1, it was found that linear free energy relationships obtained for both ATP binding/release (log k+1 versus log K1) and ADP binding/release (log k-4 versus log K- 4). Two conclusions follow. 1) The seven mutations studied here cause subtle changes in interactions between the catalytic nucleotide-binding domain and substrate ATP or product ADP. 2) The mitochondrial, normal E. coli, and nine total beta-subunit mutant enzymes represent a continuum in which subtle structural differences in the catalytic site resulted in changes in binding energy; therefore insights into the nature of energy coupling during ATP hydrolysis and synthesis by F1- ATPase may be ascertained by detailed studies of this group of enzymes
RP  - NOT IN FILE
NT  - UI - 89359370LA - engRN - 0 (Macromolecular Systems)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSDA - 19891012IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:2527851
SO  - J Biol Chem 1989 Sep 15 ;264(26):15376-15383

884
UI  - 19789
AU  - Boyer PD
AD  - Department of Chemistry and Biochemistry, University of California, Los Angeles 90024
TI  - A perspective of the binding change mechanism for ATP synthesis
AB  - An overview of research in the field of bioenergetics that led to the development of the binding change mechanism for ATP synthesis is presented, with emphasis on research from the author's laboratory. The text follows closely the Rose Award Lecture given at the 1989 meeting of the American Society for Biochemistry and Molecular Biology. Remarkable advances have revealed that the ubiquitous membrane-bound ATP synthase has unusual composition and properties. The enzyme complex has 1, 2, 3, or 9-12 copies of eight or more protein subunits. The catalytic sites are located on three copies of an approximately 55-kDa subunit. It has the strongest positive catalytic cooperativity known for any enzyme. Examples are given of selected experimental results that have provided insights into its mechanism. These include demonstration of the characteristics, location, and function of catalytic and noncatalytic adenine nucleotide binding sites and the incisive information provided by measurement of phosphate oxygen exchanges and distribution of 18(O) in ATP or Pi formed by catalysis. Research from various laboratories gives support to the binding change mechanism in which energy from proton translocation serves principally to promote release of tightly bound ATP, with sequential participation of three catalytic sites. Some speculative suggestions about a rotational catalysis and about the different forms assumed by the ATPase are included
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - ATPase
MH  - BINDING
MH  - Binding Sites
MH  - Biochemistry
MH  - bioenergetics
MH  - Catalysis
MH  - CHANGE MECHANISM
MH  - Chemistry
MH  - COMPLEX
MH  - development
MH  - FIELD
MH  - H(+)-Transporting ATP Synthase
MH  - mechanism
MH  - nucleotide binding
MH  - nucleotide binding sites
MH  - NUCLEOTIDE-BINDING-SITES
MH  - Oxygen
MH  - proton
MH  - review
MH  - SUBUNIT
MH  - SYNTHASE
MH  - synthesis
MH  - translocation
RP  - NOT IN FILE
NT  - UI - 89325964LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - 7782-44-7 (Oxygen)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialID - GM 11094/GM/NIGMSDA - 19890901IS - 0892-6638SB - IMCY - UNITED STATESJC - FAS
UR  - PM:2526771
SO  - FASEB J 1989 Aug ;3(10):2164-2178

885
UI  - 390
AU  - Bulygin VV
AU  - Vinogradov AD
TI  - [Kinetic evidence of the interaction of three nucleotide-binding centers of mitochondrial ATP-synthetase]
AB  - It is shown that methanol significantly decreases the rate of ATP- dependent activation of submitochondrial particle ATPase blocked by low (approximately 1 microM) ADP concentrations, having an insignificant effect on the initial rate of ATP hydrolysis. The dissociation rate constant for the F1.ADP complex (Kd = approximately 2.10(-8) M) decreases thereby from 0.28 to 0.12 min-1. Within a narrow range of ADP concentrations (2-40 microM) used to inhibit ATPase, the activation rate constant measured in the presence of methanol changes from the minimum (0.12 min-1) to the maximum (0.48 min-1) value. The rate of dissociation of the enzyme-inhibitor complexes formed in the presence of low (approximately 1 microM) or high (greater than or equal to 40 microM) ADP concentrations depends on the concentration of ATP in a similar way. In the presence of EDTA, the enzyme-inhibitor complex (ADP.F1.ADP) is activated within 1-3 minutes, whereas the dissociation of the F1.ADP complex proceeds on an hour scale. The results obtained are interpreted as the interaction of at least three nucleotide-binding sites in the membrane-bound F1
RP  - NOT IN FILE
NT  - UI - 90057507LA - rusRN - 0 (Nucleotides)RN - 0 (Sulfites)RN - 60-00-4 (Edetic Acid)RN - 67-56-1 (Methanol)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19891226IS - 0320-9725SB - IMCY - USSRJC - A28
UR  - PM:2510833
SO  - Biokhimiia 1989 Aug ;54(8):1359-1367

886
UI  - 20820
AU  - Cain BD
AU  - Simoni RD
AD  - Department of Biological Sciences, Stanford University, California 94305-5020
TI  - Proton translocation by the F1F0ATPase of Escherichia coli. Mutagenic analysis of the a subunit
AB  - Cassette site-directed mutagenesis was employed to generate mutations in the a subunit (uncB (a) gene) of F1F0ATP synthase. Using sequence homology with similar subunits of other F1F0ATP synthases as a guide, 20 mutations were targeted to a region of the a subunit thought to constitute part of the proton translocation mechanism. ATP-driven proton pumping activity is lost with the substitution of lys, ile, val, or glu for arginine 210. Substitution of val, leu, gln, or glu for asparagine 214 does not completely block proton conduction, however, replacement of asparagine 214 with histidine does reduce enzyme activity below that necessary for significant function. Two or three mutations were constructed in each of four nonpolar amino acids, leucine 207, leucine 211, alanine 217, and glycine 218. Certain specific mutations in these positions result in partial loss of F1F0ATP synthase activity, but only the substitution of arginine for alanine 217 reduces ATP-driven proton pumping activity to undetectable levels. It is concluded that of the six amino acids studied, only arginine 210 is an essential component of the proton translocation mechanism. Fractionation of cell-free extracts of a subunit mutation strains generally reveals normal amounts of F1 specifically bound to the particulate fraction. One possible exception is the arginine 210 to isoleucine mutation which results in somewhat elevated levels of free F1 detectable in the soluble fraction. For nearly all a subunit mutations, F1F0-mediated ATP hydrolysis activity remains sensitive to inhibition by dicyclohexylcarbodiimide in spite of the fact that the mutations block proton translocation
MH  - A
MH  - ACID
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - Amino Acids
MH  - analysis
MH  - atp
MH  - ATPase
MH  - Dicyclohexylcarbodiimide
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1
MH  - function
MH  - Histidine
MH  - Hydrolysis
MH  - mechanism
MH  - mutagenesis
MH  - proton
MH  - Protons
MH  - site-directed
MH  - SUBUNIT
MH  - SYNTHASE
MH  - translocation
RP  - NOT IN FILE
NT  - UI - 89123453LA - engRN - 0 (DNA, Recombinant)RN - 0 (Plasmids)RN - 0 (Protons)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleID - GM18539/GM/NIGMSDA - 19890321IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:2536742
SO  - J Biol Chem 1989 Feb 25 ;264(6):3292-3300

887
UI  - 21200
AU  - Drachev LA
AU  - Drachev AL
AU  - Chekulaeva LN
AU  - Evstigneeva RP
AU  - Kaulen AD
AU  - Khitrina LV
AU  - Khodonov AA
AU  - Lazarova ZR
AU  - Mitsner BI
AD  - AN Belozersky Laboratory of Molecular Biology and Bioorganic Chemistry, Moscow State University, USSR
TI  - An investigation of the electrochemical cycle of bacteriorhodopsin analogs with the modified ring
AB  - 5,6-Epoxy-, 4-methoxy-, 4-hydroxy-, and 3,4-dehydrobacteriorhodopsins can generate delta psi coupled to a photochemical cycle with intermediate M. The kinetics of delta psi comprises three main electrogenic phases: the fast small negative, the microsecond, and the millisecond positive phases. The photocycle efficiency is lower in all the analogs. The photocycle is modified insignificantly only in 3,4- dehydrobacteriorhodopsin. In the other pigments the decay of the flash- induced bleaching in the chromophore main absorption band is slower than the decay of M or long-wave intermediates, especially in the 4- hydroxy analog. In the latter analog, such distinctions, according to delta pH measurements, are partly due to deceleration of the decay of the novel intermediate (P). In 5,6-epoxybacteriorhodopsin, at all wavelengths, the decay of the intermediates takes seconds upon M formation. According to our and literature data, no bacteriorhodopsin analogs are known to have a cycle which preserves the M-intermediate and does not transport a proton
MH  - A
MH  - absorption
MH  - ANALOGS
MH  - Bacteriorhodopsin
MH  - Chemistry
MH  - delta
MH  - DELTA-PH
MH  - electrogenic
MH  - flash
MH  - intermediate
MH  - Kinetics
MH  - M
MH  - M-intermediate
MH  - microsecond
MH  - P
MH  - pH
MH  - pigments
MH  - proton
MH  - Protons
MH  - PSI
MH  - Retinaldehyde
MH  - transport
RP  - NOT IN FILE
NT  - UI - 89192368LA - engRN - 0 (Protons)RN - 0 (Retinoids)RN - 116-31-4 (Retinaldehyde)RN - 472-87-7 (dehydroretinal)RN - 53026-44-1 (Bacteriorhodopsins)PT - Journal ArticleDA - 19890510IS - 0003-9861SB - IMCY - UNITED STATES
UR  - PM:2539044
SO  - Arch Biochem Biophys 1989 Apr ;270(1):184-197

888
UI  - 359
AU  - Dyer MR
AU  - Gay NJ
AU  - Powell SJ
AU  - Walker JE
AD  - MRC Laboratory of Molecular Biology, Cambridge, UK
TI  - ATP synthase from bovine mitochondria: complementary DNA sequence of the mitochondrial import precursor of the gamma-subunit and the genomic sequence of the mature protein
AB  - The gamma-subunit of mitochondrial ATP synthase is part of the extrinsic membrane sector of the enzyme F1-ATPase. It is a nuclear gene product. Complementary DNA clones encoding a precursor of the protein have been isolated from a bovine library. The initial partial clone was identified with a mixture of 32 synthetic oligonucleotides designed from the known protein sequence (Walker et al., 1985), and this isolate was then used to screen the library again in order to find a complete cDNA. The DNA sequence of a clone that encodes the entire mature protein has been established, and the deduced protein sequence agrees exactly with that determined by direct sequence analysis of protein isolated from bovine hearts (Walker et al., 1985). At the 3' ends of two independently isolated clones, alternative polyadenylation sites have been observed; otherwise, the DNA sequences of the clones are concordant. In common with many other mitochondrial proteins encoded in nuclear genes, the deduced protein sequence has an N-terminal extension that is absent from the mature protein. These presequences direct the protein to its appropriate mitochondrial compartment and are removed during the import process. The cDNA clone has been employed to isolate bovine genomic clones containing the gene for the gamma-subunit. From them, the DNA sequence has been established of a region encoding the mature protein and six amino acids in the presequence, but not the remainder of the proposed import sequence. This sequence extends over almost 10 kb and is divided into eight exons. Intron B between exons I and II contains a sequence that is related to long interspersed repetitive elements (LINEs) that have been described in other mammals. Human LINEs are usually flanked by directly repeated sequences with a poly(A) tract at their 3' ends, and these features are present in the bovine LINE which is truncated. This sequence contains an open reading frame encoding part of a protein that is closely related to a protein encoded in mouse LINEs, to reverse transcriptase, and to DNA binding proteins. We have also made a preliminary investigation by DNA hybridization of the number of sequences related to the bovine gene in both the bovine and human genomes. Under the experimental conditions employed, one fragment hybridized in digests of bovine DNA, and two to four bands were detected in digests of human DNA; these latter fragments have originated from either expressed genes or pseudogenes.(ABSTRACT TRUNCATED AT 400 WORDS)
RP  - NOT IN FILE
NT  - UI - 89323066LA - engRN - 0 (Macromolecular Systems)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19890829IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:2526651
SO  - Biochemistry 1989 May 2 ;28(9):3670-3680

889
UI  - 358
AU  - Dyer MR
AU  - Gay NJ
AU  - Walker JE
AD  - Medical Research Council Laboratory of Molecular Biology, Cambridge, UK
TI  - DNA sequences of a bovine gene and of two related pseudogenes for the proteolipid subunit of mitochondrial ATP synthase
AB  - The dicyclohexylcarbodi-imide-reactive proteolipid is a membrane subunit of mitochondrial ATP synthase. In cows it is encoded by two different nuclear genes known as P1 and P2. These genes are expressed in a tissue-specific fashion which reflects the embryonic origin of the tissues. The proteins that they encode are synthesized in the cytosol, and are precursors of the proteolipid that have different mitochondrial import sequences of 61 and 68 amino acids respectively. By use of gene- specific probes derived from the bovine P2 cDNA, regions containing corresponding parts of the bovine P2 gene have been isolated from a bovine genomic library, and their DNA sequences and those of flanking and intervening regions have been determined. The sequence contains four exons, which represent the cDNA sequence, spread over 3.8 kb of the bovine genome. Two of the introns are in the DNA sequence coding for the mitochondrial import sequence, and a third intron is in a sequence encoding an extramembranous structure between the two putative transmembrane alpha-helical domains of the mature proteolipid. An Alu- type repetitive element was detected at the extreme 5' end of the sequence. The bovine P1 and P2 genes for the dicyclohexylcarbodimide- reactive proteolipid of ATP synthase are members of a multiple gene family that also contains many pseudogenes. The bovine P1 gene has not been isolated, but two distinct P1 pseudogenes have been cloned and their DNA sequences have been determined. Both of them contain 'in- phase' stop codons and frame-shift mutations, and one of them bears the hallmarks of retroposition; it has no introns, it contains a poly(A) tract at its 3' end and it is flanked by direct DNA sequence repeats. The second P1 pseudogene is very unusual. It appears to be derived from a partially processed transcript and contains an intervening DNA sequence of 861 bp that corresponds in position with an intron in the human P1 gene. This pseudogene also could have been introduced by retroposition since its sequence is flanked by short direct repeats. However, it does not contain a poly(A) tract at its 3' end. An alternative, but less likely, explanation is that rather than being a retroposon, this sequence arose by duplication of an expressed gene at a time when it had only one intron
RP  - NOT IN FILE
NT  - UI - 89374049LA - engRN - 0 (Multienzyme Complexes)RN - 0 (Proteolipids)RN - 9007-49-2 (DNA)RN - EC 2.7 (Phosphotransferases)RN - EC 2.7.4.- (ATP synthetase complex)PT - Journal ArticleDA - 19890922IS - 0264-6021SB - IMCY - ENGLANDJC - 9YO
UR  - PM:2549952
SO  - Biochem J 1989 May 15 ;260(1):249-258

890
UI  - 306
AU  - Engelbrecht S
AU  - Deckers-Hebestreit G
AU  - Altendorf K
AU  - Junge W
AD  - Department of Biophysics, University of Osnabruck, Federal Republic of Germany
TI  - Cross-reconstitution of the F0F1-ATP synthases of chloroplasts and Escherichia coli with special emphasis on subunit delta
AB  - F0F1-ATP synthases catalyse ATP formation from ADP and Pi by using the free energy supplied by the transmembrane electrochemical potential of the proton. The delta subunit of F1 plays an important role at the interface between the channel portion F0 and the catalytic portion F1. In chloroplasts it can plug the protonic conductance of CF0 and in Escherichia coli it is required for binding of EF1 to EF0. We wanted to know whether or not delta of one species was effective between F0 and F1 of the other species and vice versa. To this end the respective coupling membrane (thylakoids, everted vesicles from E. coli) was (partially) depleted of F1 and purified F1, F1(-delta), and delta were added in various combinations to the F1-depleted membranes. The efficiency or reconstitution was measured in thylakoids via the rate of phenazinemethosulfate-mediated cyclic photophosphorylation and in E. coli everted vesicles via the degree of 9-amino-6-chloro-2- methoxyacridine fluorescence quenching. Addition of CF1 to partially CF1-depleted thylakoid vesicles restored photophosphorylation to the highest extent. CF1(-delta)+chloroplast delta, EF1, EF1(-delta)+E. coli delta were also effective but to lesser extent. CF1(-delta)+E. coli delta and EF1(-delta)+chloroplast delta restored photophosphorylation to a small but still significant extent. With F1-depleted everted vesicles prepared by repeated EDTA treatment of E. coli membranes, addition of CF1, CF1 (-delta)+chloroplast delta and CF1(-delta)+E. coli delta gave approximately half the extent of 9-amino-6-chloro-2- methoxyacridine fluorescence quenching as compared to EF1 or EF1(- delta)+E. coli delta by energization of the vesicles with NADH, while Ef1(-delta)+chloroplast delta was ineffective. All 'mixed' combinations were probably reconstitutively active only by plugging the protonic leak through the exposed F0 (structural reconstitution) rather than by catalytic activity. Nevertheless, the cross-reconstitution is stunning in view of the weak sequence similarity between chloroplast delta and E. coli delta. It favors a role of delta as a conformational transducer rather than as a proton conductor between F0 and F1
RP  - NOT IN FILE
NT  - UI - 89231735LA - engRN - 0 (Macromolecular Systems)RN - 60-00-4 (Edetic Acid)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19890622IS - 0014-2956SB - IMCY - GERMANY, WESTJC - EMZ
UR  - PM:2523802
SO  - Eur J Biochem 1989 May 1 ;181(2):485-491

891
UI  - 308
AU  - Engelbrecht S
AU  - Schurmann K
AU  - Junge W
AD  - Biophysik, Fachbereich Biologie/Chemie, Universitat Osnabruck, Federal Republic of Germany
TI  - Chloroplast ATP synthase contains one single copy of subunit delta that is indispensable for photophosphorylation
AB  - F0F1 ATP synthases synthesize ATP in their F1 portion at the expense of free energy supplied by proton flow which enters the enzyme through their channel portion F0. The smaller subunits of F1, especially subunit delta, may act as energy transducers between these rather distant functional units. We have previously shown that chloroplast delta, when added to thylakoids partially depleted of the coupling factor CF1, can reconstitute photophosphorylation by inhibiting proton leakage through exposed coupling factor CF0. In view of controversies in the literature, we reinvestigated two further aspects related to subunit delta, namely (a) its stoichiometry in CF0CF1 and (b) whether or not delta is required for photophosphorylation. By rocket immunoelectrophoresis of thylakoid membranes and calibration against purified delta, we confirmed a stoichiometry of one delta per CF0CF1. In CF1-depleted thylakoids photophosphorylation could be reconstituted not only by adding CF1 and subunit delta but, surprisingly, also by CF1 (-delta). We found that the latter was attributable to a contamination of CF1 (-delta) preparations with integral CF1. To lesser extent CF1 (- delta) acted by complementary rebinding to CF0 channels that were closed because they contained delta [CF0(+delta)]. This added catalytic capacity to proton-tight thylakoid vesicles. The ability of subunit delta to control proton flow through CF0 and the absolute requirement for delta in restoration of photophosphorylation suggest an essential role of this small subunit at the interface between the large portions of ATP synthase: delta may be part of the coupling site between electrochemical, conformational and chemical events in this enzyme
RP  - NOT IN FILE
NT  - UI - 89137068LA - engRN - 0 (Amino Acids)RN - 0 (Plant Proteins)RN - 299-11-6 (Methylphenazonium Methosulfate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19890404IS - 0014-2956SB - IMCY - GERMANY, WESTJC - EMZ
UR  - PM:2521825
SO  - Eur J Biochem 1989 Jan 15 ;179(1):117-122

892
UI  - 20872
AU  - Eya S
AU  - Maeda M
AU  - Tomochika K
AU  - Kanemasa Y
AU  - Futai M
AD  - Department of Organic Chemistry and Biochemistry, Osaka University, Japan
TI  - Overproduction of truncated subunit a of H+-ATPase causes growth inhibition of Escherichia coli
AB  - Genes (uncB) for wild-type and mutant a subunits of Escherichia coli H+- ATPase (F0F1) were cloned into recombinant plasmids. The subunits were expressed under the control of a weak promoter of the unc operon at 30 degrees C and strong promoters of lambda phage at 42 degrees C. At 30 degrees C, the wild type and a truncated (Glu-269----end) a subunit complemented the defect of the a subunit mutant KF24A (Trp-111----end), whereas the other mutant subunits (Trp-111----end, Trp-231----end, Gln- 252----end, and a subunit with a deletion of residues 21 to 227) did not. Three mutant subunits (Trp-231----end, Gln-252----end, and Glu-269- ---end) and the wild-type a subunit caused growth inhibition associated with cell elongation, an uneven distribution of membrane proteins, and an altered septum structure when they were expressed at 42 degrees C. These phenomena were not observed with the other mutant subunits, suggesting that overproduction of the middle region (between residues 111 and 230) of the a subunit causes growth inhibition
MH  - A
MH  - ATPase
MH  - Biochemistry
MH  - Chemistry
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0F1
MH  - H+
MH  - H+-ATPase
MH  - Macromolecular Systems
MH  - membrane
MH  - Membrane Proteins
MH  - mutant
MH  - protein
MH  - Proteins
MH  - RESIDUE
MH  - SUBUNIT
MH  - SYSTEM
MH  - SYSTEMS
RP  - NOT IN FILE
NT  - UI - 90078138LA - engRN - 0 (Macromolecular Systems)RN - 0 (Membrane Proteins)RN - 0 (Plasmids)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19900125IS - 0021-9193SB - IMCY - UNITED STATES
UR  - PM:2531735
SO  - J Bacteriol 1989 Dec ;171(12):6853-6858

893
UI  - 46
AU  - Fraga D
AU  - Fillingame RH
AD  - Department of Physiological Chemistry, University of Wisconsin Medical School, Madison 53706
TI  - Conserved polar loop region of Escherichia coli subunit c of the F1F0 H+-ATPase. Glutamine 42 is not absolutely essential, but substitutions alter binding and coupling of F1 to F0
AB  - The uncE114 mutation (Gln42----Glu) in subunit c of the Escherichia coli H+ ATP synthetase causes uncoupling of proton translocation from ATP hydrolysis (Mosher, M. E., White, L. K., Hermolin, J., and Fillingame, R. H. (1985) J. Biol. Chem. 260, 4807-4814). In the background of strain ER, the mutation led to dissociation of F1 from the membrane. Ten revertants to the uncE114 mutation were isolated, and the uncE gene was cloned and sequenced. Six of the revertants were intragenic and had substitutions of glycine, alanine, or valine for the mutant glutamate residue at position 42. The intragenic, revertant uncE genes were incorporated into an otherwise wild type chromosome of strain ER. Membrane vesicles prepared from each of the revertants showed a restoration of F1 binding to F0. The Val42 revertant differed from the other two revertants in that the ATPase activity of F1 was inhibited when membrane bound. This was shown by the stimulation of ATPase activity when F1 was released from the membrane. The Gly42 and Ala42 revertants demonstrated membrane ATPase activity that was resistant to dicyclohexylcarbodiimide treatment. Resistance was shown to be due to the increased dissociation of F1 from the membrane under ATPase assay conditions. The Ala42 revertant showed a significant reduction in ATP-dependent quenching of quinacrine fluorescence that was attributed to less efficient coupling of ATP hydrolysis to H+ translocation, whereas the other revertants showed responses very near to that of wild type. Minor changes in the F1-F0 interaction in all three revertants were indicated by an increase in H+ leakiness, as judged by reduced NADH-dependent quenching of quinacrine fluorescence. The minor defects in the revertants support the idea that residue 42 is involved in the binding and coupling of F1 to F0 but also show that the conserved glutamine (or asparagine) is not absolutely necessary in this function
RP  - NOT IN FILE
NT  - UI - 89214089LA - engRN - 0 (Bacterial Proteins)RN - 0 (Macromolecular Systems)RN - 0 (Membrane Proteins)RN - 56-65-5 (Adenosine Triphosphate)RN - 56-85-9 (Glutamine)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - 5-T32-GM07133/GM/NIGMSID - GM23105/GM/NIGMSDA - 19890602IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:2523384
SO  - J Biol Chem 1989 Apr 25 ;264(12):6797-6803

894
UI  - 1139
AU  - Fromme P
AU  - Graber P
TI  - Heterogeneity of ATP-hydrolyzing sites on reconstituted CF0F1
MH  - CF0F1
MH  - RECONSTITUTED CF0F1
MH  - Site
RP  - ON REQUEST (03/18/92)
SO  - FEBS Lett 1989  ;259():33-36

895
UI  - 20875
AU  - Futai M
AU  - Noumi T
AU  - Maeda M
AD  - Department of Organic Chemistry and Biochemistry, Osaka University, Japan
TI  - ATP synthase (H+-ATPase): results by combined biochemical and molecular biological approaches
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Biochemistry
MH  - Chemistry
MH  - H+-ATPase
MH  - review
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 89372792LA - engRN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticlePT - ReviewPT - Review, AcademicDA - 19891011IS - 0066-4154SB - IMCY - UNITED STATES
UR  - PM:2528322
SO  - Annu Rev Biochem 1989  ;58():111-136

896
UI  - 45
AU  - Girvin ME
AU  - Hermolin J
AU  - Pottorf R
AU  - Fillingame RH
AD  - Department of Physiological Chemistry, University of Wisconsin, Madison 53706
TI  - Organization of the F0 sector of Escherichia coli H+-ATPase: the polar loop region of subunit c extends from the cytoplasmic face of the membrane
AB  - The membrane-spanning F0 sector of the Escherichia coli H+-transporting ATP synthase (EC 3.6.1.34) contains multiple copies of subunit c, a 79 amino acid residue protein that is thought to insert in the membrane like a hairpin with two membrane traversing alpha-helices. The center of the protein is much more polar than the putative transmembrane alpha- helices and has been postulated to play a crucial role in coupling H+ translocation through F0 to ATP synthesis in the membrane extrinsic, F1 sector of the complex. However, the direction of insertion of subunit c in the membrane has not been established. We show here that the "polar loop" lies on the F1 binding side of the membrane. A peptide corresponding to Lys34----Ile46 of the polar loop was synthesized. Antisera were generated to the Lys34----Ile46 cognate peptide, and the polyclonal antipeptide IgG was shown to bind to a crude F0 fraction by using enzyme-linked immunosorbent assays. The antipeptide serum did not bind tightly enough to F0 to disrupt function. However, a polyclonal antiserum made to purified, whole subunit c was shown to block the binding of F1 to the F0 exposed in F1-stripped membranes. Incubation of the antisubunit c serum with the peptide reduced the inhibitory effect of the antiserum on the binding of F1 to F0. The reversal of inhibition by the peptide was specific to the antisubunit c serum in that the peptide had no effect on inhibition of F1 binding to F0 by antiserum to subunit a of F0.(ABSTRACT TRUNCATED AT 250 WORDS)
RP  - NOT IN FILE
NT  - UI - 89352487LA - engRN - 0 (Epitopes)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - F32-GM11096/GM/NIGMSID - GM-23105/GM/NIGMSDA - 19890928IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:2475164
SO  - Biochemistry 1989 May 16 ;28(10):4340-4343

897
UI  - 186
AU  - Gogol EP
AU  - Lucken U
AU  - Bork T
AU  - Capaldi RA
AD  - Institute of Molecular Biology, University of Oregon, Eugene 97403
TI  - Molecular architecture of Escherichia coli F1 adenosinetriphosphatase
AB  - The structure of the E. coli F1 ATPase (ECF1) has been studied by a novel combination of two specimen preparation and image analysis techniques. The molecular outline of the ECF1 was determined by three- dimensional reconstruction of images of negatively stained two- dimensional crystals of ECF1. Internal features were revealed by analysis of single particles of ECF1, preserved in their native state in a thin layer of amorphous ice, and examined by cryoelectron microscopy. Various projections of the unstained ECF1 were interpreted consistently with the three-dimensional structure in negative stain, yielding a more informative description of the enzyme than otherwise possible. Results show that the ECF1 is a roughly spherical complex approximately 90-100 A in diameter. Six elongated protein densities (the alpha and beta subunits, each approximately 90 A X approximately 30 A in size) comprise its hexagonally modulated periphery. At the center of the ECF1 is an aqueous cavity which extends nearly or entirely through the length of the complex. A compact protein density, located at one end of the hexagonal barrel and closely associated with one of the peripheral subunits, partially obstructs the central cavity
RP  - NOT IN FILE
NT  - UI - 89352542LA - engRN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM 39806/GM/NIGMSID - HL 24526/HL/NHLBIID - RR 02756-01/RR/NCRRDA - 19890929IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:2475169
SO  - Biochemistry 1989 May 30 ;28(11):4709-4716

898
UI  - 185
AU  - Gogol EP
AU  - Aggeler R
AU  - Sagermann M
AU  - Capaldi RA
AD  - Institute of Molecular Biology, University of Oregon, Eugene 97403
TI  - Cryoelectron microscopy of Escherichia coli F1 adenosinetriphosphatase decorated with monoclonal antibodies to individual subunits of the complex
AB  - Monoclonal antibodies directed against epitopes on each of the five subunits (alpha, beta, gamma, delta, and epsilon) of the Escherichia coli F1 ATPase (ECF1) have been prepared and used to localize the subunits in the enzyme complex. Fab' fragments, prepared by pepsin digestion of the antibodies, were bound to ECF1 and visualized by cryoelectron microscopy of the unstained, frozen hydrated ECF1-Fab' complexes. Besides aiding in the identification of the ECF1 subunits, addition of Fab's to the specimen fortuitously offers additional advantages in this technique. ECF1 labeled with anti-alpha Fab' is uniformly oriented in the amorphous ice layer, in contrast to unlabeled ECF1, which exhibits a multitude of projection views when examined in ice. Almost all complexes display a triangular projection, which image averaging reveals to be a hexagonal view of ECF1 with Fab' fragments labeling every other peripheral subunit, confirming the alternating arrangement of alpha and beta subunits in the enzyme. A density in the interior of the structure is positioned asymmetrically, adjacent to an unlabeled peripheral mass, indicating that its primary linkage is to a beta rather than an alpha subunit. The composition of the asymmetric density was explored by examining the trypsin-treated ECF1, taking advantage of the unique orientation induced by the binding of anti- alpha Fab'. Trypsin treatment releases the delta and epsilon subunits and cleaves the gamma subunit; the internal density is reduced but not eliminated, showing the contribution of the gamma subunit to the residual structure, and suggesting that the loss of the delta and epsilon subunits, or a structural rearrangement of the gamma subunit, is responsible for its smaller size.(ABSTRACT TRUNCATED AT 250 WORDS)
RP  - NOT IN FILE
NT  - UI - 89352543LA - engRN - 0 (Antibodies, Monoclonal)RN - 0 (Antigen-Antibody Complex)RN - 0 (Epitopes)RN - 0 (Immunoglobulins, Fab)RN - EC 3.4.21.4 (Trypsin)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM 39806/GM/NIGMSID - HL 24526/HL/NHLBIID - RR 02756/RR/NCRRDA - 19890929IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:2475170
SO  - Biochemistry 1989 May 30 ;28(11):4717-4724

899
UI  - 21011
AU  - Grotjohann I
AU  - Graber P
AD  - Max-Volmer-Institut, Technische Universitat Berlin, FRG
TI  - The ion channel of the ATP-synthase from chloroplasts
MH  - ATP synthase
MH  - ATPase
MH  - chloroplast
MH  - Chloroplasts
MH  - ion
MH  - Ion Channels
MH  - proton
MH  - Protons
RP  - NOT IN FILE
NT  - UI - 90000311LA - engRN - 0 (Ion Channels)RN - 0 (Protons)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19891109IS - 0277-8033SB - IMCY - UNITED STATES
UR  - PM:2477016
SO  - J Protein Chem 1989 Jun ;8(3):379-380

900
UI  - 20873
AU  - Hanada H
AU  - Noumi T
AU  - Maeda M
AU  - Futai M
AD  - Department of Organic Chemistry and Biochemistry, Osaka University, Japan
TI  - Uni-site catalysis by Escherichia coli F1-ATPase with different numbers of bound nucleotides
AB  - We prepared two types of E. coli F1 by slightly different gel filtration procedures of the purified F1: F1(II) contained about 2 mol, and F1(V) about 5 mol of bound adenine nucleotides per mol of the enzyme. Thus F1(II) had more than 2, possibly 3, vacant catalytic sites, while F1(V) had less than one vacant catalytic site. The rate of ATP hydrolysis in uni-site catalysis (in the presence of inorganic phosphate) was about 3-fold higher with F1(II) than with F1(V), suggesting that ADP and inorganic phosphate bound at the catalytic sites of F1(V) changed the kinetics of uni-site catalysis significantly
MH  - Adenine Nucleotides
MH  - ADENINE-NUCLEOTIDES
MH  - ADP
MH  - atp
MH  - ATPase
MH  - Biochemistry
MH  - BOUND NUCLEOTIDES
MH  - Catalysis
MH  - Chemistry
MH  - COLI F1 ATPASE
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - Hydrolysis
MH  - INORGANIC-PHOSPHATE
MH  - Kinetics
MH  - Nucleotides
MH  - Site
MH  - uni-site catalysis
MH  - UNISITE CATALYSIS
RP  - NOT IN FILE
NT  - UI - 90060381LA - engRN - 0 (Adenine Nucleotides)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19900108IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:2531096
SO  - FEBS Lett 1989 Nov 6 ;257(2):465-467

901
UI  - 524
AU  - Hanley-Trawick S
AU  - Carpen ME
AU  - Dunaway-Mariano D
AU  - Pedersen PL
AU  - Hullihen J
AD  - Department of Chemistry and Biochemistry, University of Maryland, College Park 20742
TI  - Investigation of the substrate structure and metal cofactor requirements of the rat liver mitochondrial ATP synthase/ATPase complex
AB  - The F1 moiety of the rat liver mitochondrial ATP synthase/ATPase complex contains as isolated 2 mol Mg2+/mol F1, 1 mol of which is nonexchangeable and the other which is exchangeable (N. Williams, J. Hullihen, and P.L. Pedersen, (1987) Biochemistry 26, 162-169). In addition, the enzyme binds 1 mol ADP/mol F1 and 3 mol AMP.PNP, the latter of which can bind in complex formation with divalent cation and displace the Mg2+ at the exchangeable site. Thus, in terms of ligand binding sites the fully loaded rat liver F1 complex contains 3 mol MgAMP.PNP, 1 mol ADP, and 1 mol Mg2+. In this study we have used several metal ATP complexes or analogs thereof to gain further insight into the ligand binding domains of rat liver F1 and the mechanism by which it catalyzes ATP hydrolysis in soluble and membrane bound form. Studies with LaATP confirmed that MgATP is the most likely substrate for rat liver F1, and provided evidence that the enzyme may contain additional Mg2+ binding sites, undetected in previous studies of F1- ATPases, that are required for catalytic activity. Thus, F1 containing the thermodynamically stable LaATP complex in place of MgATP requires added Mg2+ to induce ATP hydrolysis. As Mg2+ cannot readily displace La2+ under these conditions there appears to be a catalytically important class of Mg2+ binding sites on rat liver F1, distinct from the nonexchangeable Mg2+ site and the sites involved in binding MgATP. Additional studies carried out with exchange inert metal-nucleotide complexes involving rhodium and the Mg2+ and Cd2+ complexes of ATP beta S and ATP alpha S imply that the rate-limiting step in the ATPase reaction pathway occurs subsequent to the P gamma-O-P beta bond cleavage steps, perhaps at the level of Mg(ADP)(Pi) hydrolysis or MgADP release. Evidence is presented that Mg2+ remains coordinated to the leaving group of the reaction, i.e., the beta phosphoryl group. Finally, in contrast to soluble F1, F1 bound to F0 in the inner mitochondrial membrane failed to discriminate between the Mg2+ complexes of the ATP beta S isomers. This indicates that a fundamental difference may exist between the catalytic or kinetic mechanism of F1 and the more physiologically intact F0F1 complex
RP  - NOT IN FILE
NT  - UI - 89104409LA - engRN - 0 (Cations, Divalent)RN - 0 (Metals, Rare Earth)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - CA 10951/CA/NCIID - ES-00111/ES/NIEHSID - GM28688/GM/NIGMSDA - 19890215IS - 0003-9861SB - IMCY - UNITED STATESJC - 6SK
UR  - PM:2521440
SO  - Arch Biochem Biophys 1989 Jan ;268(1):116-123

902
UI  - 47
AU  - Hermolin J
AU  - Fillingame RH
AD  - Department of Physiological Chemistry, University of Wisconsin Medical School, Madison 53706
TI  - H+-ATPase activity of Escherichia coli F1F0 is blocked after reaction of dicyclohexylcarbodiimide with a single proteolipid (subunit c) of the F0 complex
AB  - Dicyclohexylcarbodiimide (DCCD) specifically inhibits the F1F0-H+-ATP synthase complex of Escherichia coli by covalently modifying a proteolipid subunit that is embedded in the membrane. Multiple copies of the DCCD-reactive protein, also known as subunit c, are found in the F1F0 complex. In order to determine the minimum stoichiometry of reaction, we have treated E. coli membranes with DCCD, at varying concentrations and for varying times, and correlated inhibition of ATPase activity with the degree of modification of subunit c. Subunit c was purified from the membrane, and the degree of modification was determined by two methods. In the "specific radioactivity" method, the moles of [14C]DCCD per total mole of subunit c was calculated from the radioactivity incorporated per mg of protein, and conversion of mg of protein to mol of protein based upon amino acid analysis. In the "high performance liquid chromatography (HPLC) peak area" method, the DCCD- modified subunit c was separated from unmodified subunit c on an anion exchange AX300 HPLC column, and the areas of the peaks from the chromatogram quantitated. The shape of the modification versus inhibition curve indicated that modification of a single subunit c per F0 was sufficient to abolish ATPase activity. The titration data were fit by nonlinear regression analysis to a single hit mathematical model, A = Un(1 - r) + r, where A is the relative activity, U is the ratio of unmodified/total subunit c, n is the number of subunit c per F0, and r is a residual fraction of ATPase activity that was resistant to inhibition by DCCD. The two methods gave values for n equal to 10 by the specific radioactivity method and 14 by the HPLC peak area method, and values for r of 0.28 and 0.30, respectively. Most of the r value was accounted for by the observed dissociation of 15-20% of the F1- ATPase from the membrane under ATPase assay conditions. When the minimal, experimentally justified value of r = 0.15 was used in the equation above, the calculated values of n were reduced to 8 and 11, respectively. The value of n determined here, with a probable range of uncertainty of 8-14, is consistent with, and provides an independent type of experimental support for, the suggested stoichiometry of 10 +/- 1 subunit c per F1F0, which was determined by a more precise radiolabeling method (Foster, D. L., and Fillingame, R. H. (1982) J. Biol. Chem. 257, 2009-2015)
RP  - NOT IN FILE
NT  - UI - 89139450LA - engRN - 0 (Carbodiimides)RN - 0 (Macromolecular Systems)RN - 0 (Proteolipids)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM-23105/GM/NIGMSDA - 19890403IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:2521856
SO  - J Biol Chem 1989 Mar 5 ;264(7):3896-3903

903
UI  - 21199
AU  - Holz M
AU  - Drachev LA
AU  - Mogi T
AU  - Otto H
AU  - Kaulen AD
AU  - Heyn MP
AU  - Skulachev VP
AU  - Khorana HG
AD  - Biophysics Group, Freie Universitat Berlin, Federal Republic of Germany
TI  - Replacement of aspartic acid-96 by asparagine in bacteriorhodopsin slows both the decay of the M intermediate and the associated proton movement
AB  - The photocycle, electrical charge translocation, and release and uptake of protons from the aqueous phase and release and uptake of protons from the aqueous phase were investigated for bacteriorhodopsin mutants with aspartic acid-96 replaced by asparagine or glutamic acid. At neutral pH the main effect of the Asp-96----Asn mutation is to slow by 2 orders of magnitude the decay of the M intermediate and the concomitant charge displacement associated with the reprotonation of the Schiff base from the cytoplasmic side of the membrane. The proton uptake measured with the indicator dye pyranine is likewise slowed without affecting the stoichiometry of proton pumping. The corresponding results for the Asp-96----Glu mutant, on the other hand, are very close to those for the wild-type protein. These results provide a kinetic explanation for the fact that at pH 7 and saturating light intensities the steady-state proton pumping is almost abolished in the Asp-96----Asn mutant but is close to normal in the Asp-96----Glu mutant. Thus, the pump is simply turning over much more slowly in the Asp-96----Asn mutant. The time constants of the decay of M and the associated charge translocation increase strongly with increasing pH for the Asp-96----Asn mutant but are virtually pH-independent for the Asp-96----Glu mutant and wild-type bacteriorhodopsin. At pH 5 the M decay of the Asp-96----Asn mutant is as fast as for wild type. These results suggest that Asp-96 serves as an internal proton donor in the proton-uptake pathway from the cytoplasm to the Schiff base
MH  - A
MH  - ACID
MH  - Bacteriorhodopsin
MH  - BASE
MH  - Biophysics
MH  - CONSTANT
MH  - Cytoplasm
MH  - DYE
MH  - glutamic acid
MH  - indicator
MH  - intermediate
MH  - Light
MH  - M
MH  - M-intermediate
MH  - membrane
MH  - Movement
MH  - mutant
MH  - pH
MH  - protein
MH  - proton
MH  - Protons
MH  - Schiff base
MH  - Schiff-base
MH  - Time
MH  - translocation
RP  - NOT IN FILE
NT  - UI - 89184589LA - engRN - 53026-44-1 (Bacteriorhodopsins)RN - 56-84-8 (Aspartic Acid)RN - 7006-34-0 (Asparagine)PT - Journal ArticleID - R01 GM28289-07/GM/NIGMSDA - 19890505IS - 0027-8424SB - IMCY - UNITED STATES
UR  - PM:2648392
SO  - Proc Natl Acad Sci U S A 1989 Apr ;86(7):2167-2171

904
UI  - 20956
AU  - Humbert R
AU  - Altendorf K
AD  - Department of Biological Sciences, Stanford University, California 94305-5020
TI  - Defective gamma subunit of ATP synthase (F1F0) from Escherichia coli leads to resistance to aminoglycoside antibiotics
AB  - A strain of Escherichia coli which was derived from a gentamicin- resistant clinical isolate was found to be cross-resistant to neomycin and streptomycin. The molecular nature of the genetic defect was found to be an insertion of two GC base pairs in the uncG gene of the mutant. The insertion led to the production of a truncated gamma subunit of 247 amino acids in length instead of the 286 amino acids that are present in the normal gamma subunit. A plasmid which carried the ATP synthase genes from the mutant produced resistance to aminoglycoside antibiotics when it was introduced into a strain with a chromosomal deletion of the ATP synthase genes. Removal of the genes coding for the beta and epsilon subunits abolished antibiotic resistance coded by the mutant plasmid. The relationship between antibiotic resistance and the gamma subunit was investigated by testing the antibiotic resistance of plasmids carrying various combinations of unc genes. The presence of genes for the F0 portion of the ATP synthase in the presence or absence of genes for the gamma subunit was not sufficient to cause antibiotic resistance. alpha, beta, and truncated gamma subunits were detected on washed membranes of the mutant by immunoblotting. The first 247 amino acid residues of the gamma subunit may be sufficient to allow its association with other F1 subunits in such a way that the proton gate of F0 is held open by the mutant F1
MH  - A
MH  - ACID
MH  - Amino Acids
MH  - Antibiotics
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BASE
MH  - EPSILON-SUBUNIT
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - F1
MH  - Macromolecular Systems
MH  - membrane
MH  - Membranes
MH  - mutant
MH  - proton
MH  - RESIDUE
MH  - SUBUNIT
MH  - SYNTHASE
MH  - SYSTEM
MH  - SYSTEMS
RP  - NOT IN FILE
NT  - UI - 89155445LA - engRN - 0 (Antibiotics, Aminoglycoside)RN - 0 (Macromolecular Systems)RN - 0 (Plasmids)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleID - GM 18539/GM/NIGMSDA - 19890414IS - 0021-9193SB - IMCY - UNITED STATES
UR  - PM:2522090
SO  - J Bacteriol 1989 Mar ;171(3):1435-1444

905
UI  - 20874
AU  - Inatomi K
AU  - Maeda M
AU  - Futai M
AD  - Institute of Scientific and Industrial Research, Osaka University, Japan
TI  - Dicyclohexylcarbodiimide-binding protein is a subunit of the Methanosarcina barkeri ATPase complex
AB  - Membrane ATPase of Methanosarcina barkeri was inhibited by N, N'- dicyclohexylcarbodiimide (DCCD), whereas the extrinsic alpha beta complex of the same enzyme was not. Consistent with this finding, a 6,000 dalton (6 kDa) membrane protein was preferentially labeled with radioactive DCCD. The DCCD-sensitive ATPase was solubilized from the membranes with octylglucoside and purified in the presence of this detergent. The purified ATPase contained the alpha and beta subunits and also at least four additional proteins (40, 27, 23 and 6 kDa). The 6 kDa protein in the purified enzyme reacted with DCCD, indicating that it is a subunit of an integral part of the M. barkeri ATPase complex
MH  - A
MH  - ATPase
MH  - BETA-SUBUNIT
MH  - COMPLEX
MH  - Dicyclohexylcarbodiimide
MH  - M
MH  - Macromolecular Systems
MH  - membrane
MH  - Membrane Proteins
MH  - Membranes
MH  - protein
MH  - Proteins
MH  - SUBUNIT
MH  - SYSTEM
MH  - SYSTEMS
RP  - NOT IN FILE
NT  - UI - 89350987LA - engRN - 0 (Carbodiimides)RN - 0 (Macromolecular Systems)RN - 0 (Membrane Proteins)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19890921IS - 0006-291XSB - IMCY - UNITED STATES
UR  - PM:2527501
SO  - Biochem Biophys Res Commun 1989 Aug 15 ;162(3):1585-1590

906
UI  - 9974
AU  - Jones MR
AU  - Jackson JB
AD  - School of Biochemistry, University of Birmingham, UK
TI  - Proton release by the quinol oxidase site of the cytochrome b/c1 complex following single turnover flash exitation of intact cells of Rhodobacter capsulatus.
MH  - capsulatus
MH  - Cells
MH  - COMPLEX
MH  - proton
MH  - proton release
MH  - Rhodobacter
MH  - Diffusion
RP  - NOT IN FILE
SO  - Biochim Biophys Acta 1989  ;975():34-43

907
UI  - 309
AU  - Junge W
AD  - Biophysik, FB Biologie/Chemie, Universitat Osnabruck, Osnabruck, FRG
TI  - Protons, the thylakoid membrane, and the chloroplast ATP synthase
AB  - According to the chemiosmotic theory, proton pumps and ATP synthases are coupled by lateral proton flow through aqueous phases. Three long- standing challenges to this concept, all of which have been loosely subsumed under 'localized coupling' in the literature, were examined in the light of experiments carried out with thylakoids: (1) Nearest neighbor interaction between pumps and ATP synthases. Considering the large distances between photosystem II and CFoCF1, in stacked thylakoids this is a priori absent. (2) Enhanced proton diffusion along the surface of the membrane. This could not be substantiated for the outer side of the thylakoid membrane. Even for the interface between pure lipid and water, two laboratories have reported the absence of enhanced diffusion. (3) Localized proton ducts in the membrane. Intramembrane domains that can transiently trap protons do exist in thylakoid membranes, but because of their limited storage capacity for protons, they probably do not matter for photophosphorylation under continuous light. Seemingly in favor of localized proton ducts is the failure of a supposedly permeant buffer to enhance the onset lag of photophosphorylation. However, it was found that failure of some buffers and the ability of others in this respect were correlated with their failure/ability to quench pH transients in the thylakoid lumen, as predicted by the chemiosmotic theory. It was shown that the chemiosmotic concept is a fair approximation, even for narrow aqueous phases, as in stacked thylakoids. These are approximately isopotential, and protons are taken in by the ATP synthase straight from the lumen. The molecular mechanism by which F0F1 ATPases couple proton flow to ATP synthesis is still unknown. The threefold structural symmetry of the headpiece that, probably, finds a corollary in the channel portion of these enzymes appeals to the common wisdom that structural symmetry causes functional symmetry. "Rotation catalysis" has been proposed. It is of heuristic value to visualize CFoCF1 as a mechanical coupling device. Its maximum turnover number ranges up to 400 s-1 for ATP and 1200 s-1 for protons. At about 200 mV electric driving force this implied a conductance of about 1 fS. Its channel portion (CFo), however, has revealed a very large protonic conductance of 1 pS (three orders of magnitude greater than the protonic conductance of gramicidin around neutral pH). (6) The sight and smell of food increased LH serotonin release; this effect was detectable when local fluoxetine was used to block serotonin reuptake.(ABSTRACT TRUNCATED AT 400 WORDS)
RP  - NOT IN FILE
NT  - UI - 90233653LA - engRN - 0 (Ion Channels)RN - 0 (Protons)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19900529IS - 0077-8923SB - IMCY - UNITED STATESJC - 5NM
UR  - PM:2483874
SO  - Ann N Y Acad Sci 1989  ;574():268-286

908
UI  - 757
AU  - Kasho VN
AU  - Yoshida M
AU  - Boyer PD
AD  - Molecular Biology Institute, University of California, Los Angeles 90024-1570
TI  - F1 ATPase from the thermophilic bacterium PS3 (TF1) shows ATP modulation of oxygen exchange
AB  - The ATPase from the ATP synthase of the thermophilic bacterium PS3 (TF1), unlike F1 ATPase from other sources, does not retain bound ATP, ADP, and Pi at a catalytic site under conditions for single-site catalysis [Yohda, M., & Yoshida, M. (1987) J. Biochem. 102, 875-883]. This raised a question as to whether catalysis by TF1 involved alternating participation of catalytic sites. The possibility remained, however, that there might be transient but catalytically significant retention of bound reactants at catalytic sites when the medium ATP concentration was relatively low. To test for this, the extent of water oxygen incorporation into Pi formed by ATP hydrolysis was measured at various ATP concentrations. During ATP hydrolysis at both 45 and 60 degrees C, the extent of water oxygen incorporation into the Pi formed increased markedly as the ATP concentration was lowered to the micromolar range, with greater modulation observed at 60 degrees C. Most of the product Pi formed arose by a single catalytic pathway, but measurable amounts of Pi were formed by a pathway with high oxygen exchange. This may result from the presence of some poorly active enzyme. The results are consistent with sequential participation of three catalytic sites on the TF1 as predicted by the binding change mechanism
RP  - NOT IN FILE
NT  - UI - 90057375LA - engRN - 0 (Oxygen Isotopes)RN - 0 (Phosphates)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM11904/GM/NIGMSDA - 19900105IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:2531004
SO  - Biochemistry 1989 Aug 22 ;28(17):6949-6954

909
UI  - 19788
AU  - Kasho VN
AU  - Boyer PD
AD  - Department of Chemistry and Biochemistry, University of California, Los Angeles 90024-1570
TI  - Vacuolar ATPases, like F1,F0-ATPases, show a strong dependence of the reaction velocity on the binding of more than one ATP per enzyme
AB  - Recent studies with vacuolar ATPases have shown that multiple copies catalytic subunits are present and that these have definite sequence homology with catalytic subunits of the F1,F0-ATPases. Experiments are reported that assess whether the vacuolar ATPases may have the unusual catalytic cooperativity with sequential catalytic site participation as in the binding change mechanism for the F1,F0-ATPases. The extent of reversal of bound ATP hydrolysis to bound ADP and Pi as medium ATP concentration was lowered was determined by 18O-exchange measurements for yeast and neurospora vacuolar ATPases. The results show a pronounced increase in the extent of water oxygen incorporation into the Pi formed as ATP concentration is decreased to the micromolar range. The F1,F0-ATPase from neurospora mitochondria showed an even more pronounced modulation, similar to that of other F1-type ATPases. The vacuolar ATPases thus appear to have a catalytic mechanism quite analogous to that of the F1,F0-ATPases
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - ADP
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BINDING
MH  - Biochemistry
MH  - CHANGE MECHANISM
MH  - Chemistry
MH  - DEPENDENCE
MH  - H(+)-Transporting ATP Synthase
MH  - Hydrolysis
MH  - Macromolecular Systems
MH  - mechanism
MH  - Mitochondria
MH  - Neurospora
MH  - Oxygen
MH  - Oxygen Isotopes
MH  - SUBUNIT
MH  - SYNTHASE
MH  - Water
RP  - NOT IN FILE
NT  - UI - 90046862LA - engRN - 0 (Macromolecular Systems)RN - 0 (Oxygen Isotopes)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM-11094/GM/NIGMSDA - 19891221IS - 0027-8424SB - IMCY - UNITED STATESJC - PV3
UR  - PM:2530585
SO  - Proc Natl Acad Sci U S A 1989 Nov ;86(22):8708-8711

910
UI  - 450
AU  - Kironde FA
AU  - Parsonage D
AU  - Senior AE
AD  - Department of Biochemistry, University of Rochester Medical Center, New York 14642
TI  - Random mutagenesis of the gene for the beta-subunit of F1-ATPase from Escherichia coli
AB  - ATP synthesis by oxidative phosphorylation in Escherichia coli occurs in catalytic sites on the beta-subunits of F1-ATPase. Random mutagenesis of the beta-subunit combined with phenotypic screening is potentially important for studies of the catalytic mechanism. However, when applied to haploid strains, this approach is hampered by a preponderance of mutants in which assembly of F1-ATPase in vivo is defective, precluding enzyme purification. Here we mutagenized plasmids carrying the uncD (beta-subunit) gene with hydroxylamine or N-methyl-N'- nitro-N-nitrosoguanidine and isolated, by phenotypic screening and complementation tests, six plasmids carrying mutant uncD alleles. When the mutant plasmids were used to transform a suitable uncD- strain, assembly of F1-ATPase in vivo occurred in each case. Moreover, in one case (beta Gly-223----Asp) F1-ATPase assembly proceeded although it had previously been reported that this mutation, when present on the chromosome of a haploid strain, prevented assembly of the enzyme in vivo. Therefore, this work demonstrates an improved approach for random mutagenesis of the F1-beta-subunit. Six new mutant uncD alleles were identified: beta Cys-137----Tyr; beta Gly-142----Asp; beta Gly-146---- Ser; beta Gly-207----Asp; beta-Gly-223----Asp; and a double mutant beta Pro-403----Ser,Gly-415----Asp which we could not separate. The first five of these lie within or very close to the predicted catalytic nucleotide-binding domain of the beta-subunit. The double mutant lies outside this domain; we speculate that the region around residues beta 403-415 is part of an alpha-beta intersubunit contact surface. Membrane ATPase and ATP-driven proton pumping activities were impaired by all six mutations. Purified F1-ATPase was obtained from each mutant and shown to have impaired specific ATPase activity
RP  - NOT IN FILE
NT  - UI - 89246346LA - engRN - 0 (Hydroxylamines)RN - 0 (Plasmids)RN - 70-25-7 (Methylnitronitrosoguanidine)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSID - GM29805/GM/NIGMSDA - 19890620IS - 0264-6021SB - IMCY - ENGLANDJC - 9YO
UR  - PM:2524189
SO  - Biochem J 1989 Apr 15 ;259(2):421-426

911
UI  - 21089
AU  - Krulwich TA
AU  - Guffanti AA
AD  - Department of Biochemistry, Mount Sinai School of Medicine, City University of New York, New York 10029
TI  - Alkalophilic bacteria
MH  - Alkalies
MH  - Bacteria
MH  - Biochemistry
MH  - review
RP  - NOT IN FILE
NT  - UI - 90024960LA - engRN - 0 (Alkalies)PT - Journal ArticlePT - ReviewPT - Review, AcademicDA - 19891120IS - 0066-4227SB - IMCY - UNITED STATES
UR  - PM:2679360
SO  - Annu Rev Microbiol 1989  ;43():435-463

912
UI  - 91
AU  - Laubinger W
AU  - Dimroth P
AD  - Institut fur Physiologische Chemie der Technischen Universitat Munchen, Federal Republic of Germany
TI  - The sodium ion translocating adenosinetriphosphatase of Propionigenium modestum pumps protons at low sodium ion concentrations
AB  - The purified ATPase (F1F0) of Propionigenium modestum has its pH optimum at pH 7.0 or at pH 6.0 in the presence or absence of 5 mM NaCl, respectively. The activation by 5 mM NaCl was 12-fold at pH 7.0, 3.5- fold at pH 6.0, and 1.5-fold at pH 5.0. In addition to its function as a primary Na+ pump, the ATPase was capable of pumping protons. This activity was demonstrated with reconstituted proteoliposomes by the ATP- dependent quenching of the fluorescence of 9-amino-6-chloro-2- methoxyacridine. No delta pH was formed in the presence of the uncoupler carbonyl cyanide m-chlorophenylhydrazone or by blocking the ATPase with dicyclohexylcarbodiimide. In the presence of valinomycin and K+, the delta pH increased, in accord with the operation of an electrogenic proton pump. The proton pump was only operative at low Na+ concentrations (less than 1 mM), and its activity increased as the Na+ concentration decreased. Parallel to the decrease of H+ pumping, the velocity of the Na+ transport increased about 6-fold from 0.1 to 4 mM NaCl, indicating a switch from H+ to Na+ pumping, as the Na+ concentration increases. Due to proton leaks in the proteoliposomal membranes, fluorescence quenching was released after blocking the ATPase with dicyclohexylcarbodiimide, by trapping residual ATP with glucose and hexokinase, or by the Na+-induced conversion of the proton pump onto a Na+ pump. Amiloride, an inhibitor of various Na+-coupled transport systems, was without effect on the kinetics of Na+ transport by the P. modestum ATPase
RP  - NOT IN FILE
NT  - UI - 90057410LA - engRN - 0 (Liposomes)RN - 0 (Protons)RN - 2001-95-8 (Valinomycin)RN - 2609-46-3 (Amiloride)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 7440-23-5 (Sodium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19900111IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:2554965
SO  - Biochemistry 1989 Sep 5 ;28(18):7194-7198

913
UI  - 449
AU  - Lee RS
AU  - Pagan J
AU  - Satre M
AU  - Vignais PV
AU  - Senior AE
AD  - Department of Biochemistry, University of Rochester Medical Center, NY 14642
TI  - Identification of a mutation in Escherichia coli F1-ATPase beta-subunit conferring resistance to aurovertin
AB  - A mutation conferring aurovertin resistance on Escherichia coli F1- ATPase was identified as R398----H in the F1 beta-subunit. Beta-subunit from the mutant does not bind aurovertin; therefore our results suggest the region of sequence around residue beta-398 is involved in aurovertin binding. Since nucleotide and aurovertin binding to isolated beta-subunit are not mutually exclusive, the data further suggest that the beta-subunit catalytic nucleotide-binding domain does not include residue 398. The mutation prevented aurovertin inhibition of ATPase at pH 6 and 8.5, implying charge on the arginine side-chain is not a major determinant of aurovertin binding or that the pK of R398 is shifted due to a peculiar environment. The equivalent residue is usually arginine in F1 beta-subunits of different species; notably in the aurovertin- insensitive thermophilic bacterium PS3 F1-ATPase, this residue is phenylalanine
RP  - NOT IN FILE
NT  - UI - 89338748LA - engRN - 0 (Aurovertins)RN - 0 (Pyrans)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSDA - 19890912IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:2527166
SO  - FEBS Lett 1989 Aug 14 ;253(1-2):269-272

914
UI  - 307
AU  - Lill H
AU  - Junge W
AD  - Biophysik, Fachbereich Biologie/Chemie, Universitat Osnabrauck, Federal Republic of Germany
TI  - CF0, the proton channel of chloroplast ATP synthase. After removal of CF1 it appears in two forms with highly different proton conductance
AB  - The discharge of the flash-induced transmembrane voltage through the exposed proton channel, CF0, of the chloroplast ATP synthase, CF0CF1 was investigated. EDTA treatment of thylakoid membranes exposed approximately 50% of total CF0 by removal of the CF1 counterparts. This greatly accelerated the decay of the transmembrane voltage, as was apparent from electrochromic-absorption changes of intrinsic pigments and by pH-indicating-absorption changes of added dyes. Two decay processes were discernible, one rapid with a typical half-decay time of 2 ms, and a slower one with a half-decay time variable between 20-100 ms. Both were sensitive to CF0 inhibitors, but only the rapid decay process was also inhibited by added CF1. CF1 was effective in surprisingly small amounts, which were significantly lower than those previously removed by EDTA treatment. This finding corroborated our previous conclusion that the rapid decay of the transmembrane voltage was attributable to only a few high-conductance channels among many CF0 molecules, typically in the order of one channel/CF1-depleted EDTA vesicle. Inhibition of photophosphorylation in control thylakoids was measured as function of the concentration of CF0 inhibitors. It was compared with the inhibition of proton conduction through exposed CF0 in EDTA vesicles. Photophosphorylation and proton conduction by the high-conductance form of CF0 were inhibited by the same low inhibitor concentrations. This suggested that the high-conducting form of CF0 with a time-averaged single-channel conductance of 1 pS [Lill, H., Althoff, G. & Junge, W. (1987) J. Membrane Biol. 98, 69-78] represented the proton channel in the integral enzyme, which acted as a low- impedance access from the thylakoid lumen to the coupling site in CF0CF1. The slow decay process was attributed to a majority of low- conductance CF0 channels, i.e. about 50 molecules/vesicle. The conductance of these channels was more than 100-fold lower and they did not compete with the very few highly conducting channels for rebinding of added CF1. The low proton conduction of the majority of exposed CF0 molecules, possibly due to a structural rearrangement, may be protecting the thylakoid membrane against rapid energy dissipation caused by accidental loss of CF1. It may also explain the low single- channel conductance of bacterial F0 reported in the literature
RP  - NOT IN FILE
NT  - UI - 89137125LA - engRN - 0 (Ion Channels)RN - 0 (Protons)RN - 60-00-4 (Edetic Acid)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19890403IS - 0014-2956SB - IMCY - GERMANY, WESTJC - EMZ
UR  - PM:2465153
SO  - Eur J Biochem 1989 Feb 1 ;179(2):459-467

915
UI  - 575
AU  - Matsuno-Yagi A
AU  - Hatefi Y
AD  - Department of Basic and Clinical Research, Research Institute of Scripps Clinic, La Jolla, California 92037
TI  - Uncoupling of oxidative phosphorylation: different effects of lipophilic weak acids and electrogenic ionophores on the kinetics of ATP synthesis
AB  - Previous studies from this laboratory have shown that the kinetics of ATP synthesis by bovine heart submitochondrial particles (SMP) are modulated by the coupled rate of respiration between two extremes of Vmax and apparent Km's for ADP and Pi [Matsuno-Yagi, A., & Hatefi, Y. (1986) J. Biol. Chem. 261, 14031-14038; Hekman, C., Matsuno-Yagi, A., & Hatefi, Y. (1988) Biochemistry 27, 7559-7565]. Thus, with ADP as the variable substrate, ATP synthesis occurred with Vmax = 200 nmol of ATP min-1 (mg of protein)-1 at 30 degrees C and an apparent KmADP = 2-4 microM at low rates of respiration, and with Vmax = 11,000 nmol of ATP min-1 (mg of protein)-1 at 30 degrees C and an apparent KmADP = 120-160 microM at high rates of respiration. At intermediate respiration rates, it was necessary to introduce a third intermediate KmADP for best fit of the kinetic data, indicating that transition from one kinetic extreme to the other is not abrupt and involves intermediate kinetic states of the ATP synthase complexes. The present paper shows that uncouplers affect the kinetics of ATP synthesis by SMP in two ways. When used at moderate concentrations, electrogenic ionophores such as gramicidin D or valinomycin plus nigericin decreased the Vmax for ATP synthesis without changing the contributions of the low, intermediate, and high KmADP to the overall rate of ATP synthesis. By contrast, potent lipophilic weak acid uncouplers, such as FCCP, CCCP, S-13, and SF6847, decreased Vmax and converted the kinetics of ATP synthesis toward high KmADP.(ABSTRACT TRUNCATED AT 250 WORDS)
RP  - NOT IN FILE
NT  - UI - 89352492LA - engRN - 0 (Acids)RN - 0 (Ionophores)RN - 0 (Uncoupling Agents)RN - 1405-97-6 (Gramicidin)RN - 370-86-5 (Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone)RN - 56-65-5 (Adenosine Triphosphate)PT - Journal ArticleID - DK08126/DK/NIDDKDA - 19890928IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:2475167
SO  - Biochemistry 1989 May 16 ;28(10):4367-4374

916
UI  - 523
AU  - McEnery MW
AU  - Hullihen J
AU  - Pedersen PL
AD  - Department of Biological Chemistry, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205
TI  - F0 "proton channel" of rat liver mitochondria. Rapid purification of a functional complex and a study of its interaction with the unique probe diethylstilbestrol
AB  - The F0 portion of the rat liver mitochondrial ATP synthase (F0F1- ATPase) has been purified by a rapid, high yield procedure. F0 is selectively extracted from inner membrane vesicles with 3-[(3- cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS) after prior treatment of the vesicles with guanidine HCl to remove F1. The resultant F0 is functional in proton translocation assays and separates in sodium dodecyl sulfate-polyacrylamide gel electrophoresis into four major and three minor Coomassie-stainable bands, all with apparent molecular masses below 30 kDa. This CHAPS-purified F0 preparation was characterized in detail for its capacity to interact with the unique probe diethylstilbestrol (DES) which, depending on conditions, has been shown to interact with rat liver F0F1 to either inhibit or promote ATP hydrolysis (McEnery, M. W., and Pedersen, P.L. (1986) J. Biol. Chem. 261, 1745-1752). DES-inhibitory sensitivity could be conferred on F1- ATPase activity with the same concentration dependence on F0 as conferral of oligomycin sensitivity. DES was shown also to inhibit the magnitude of valinomycin induced proton influx, while initiating proton efflux in asolectin vesicles reconstituted with F0 and loaded with K+. The potency of DES in producing the latter effects was shown to be highly dependent on hydroxyl groups in "para" positions of the two benzene rings within the DES molecule. Finally, in the absence of F0, DES was shown to act as a catalyst of proton influx in K+-loaded asolectin vesicles upon addition of valinomycin. A model based on the structure of DES is presented to account for both the inhibitory and uncoupling properties of this compound
RP  - NOT IN FILE
NT  - UI - 89308611LA - engRN - 0 (Cholic Acids)RN - 0 (Oligomycins)RN - 0 (Protons)RN - 56-53-1 (Diethylstilbestrol)RN - 75621-03-3 (3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - CA 10951/CA/NCIDA - 19890814IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:2545697
SO  - J Biol Chem 1989 Jul 15 ;264(20):12029-12036

917
UI  - 48
AU  - Miller MJ
AU  - Fraga D
AU  - Paule CR
AU  - Fillingame RH
AD  - Department of Physiological Chemistry, University of Wisconsin Medical School, Madison 53706
TI  - Mutations in the conserved proline 43 residue of the uncE protein (subunit c) of Escherichia coli F1F0-ATPase alter the coupling of F1 to F0
AB  - The conserved Pro43 residue of the uncE protein (subunit c) of the Escherichia coli F1F0-ATPase was changed to Ser or Ala by oligonucleotide-directed mutagenesis, and the mutations were incorporated into the chromosome. The resultant mutant strains were capable of oxidative phosphorylation as indicated by their ability to grow on succinate and had growth yields on glucose that were 80-90% of wild type. Membrane vesicles from the mutants were slightly less efficient than wild type vesicles in ATP-driven proton pumping as indicated by ATP-dependent quenching of quinacrine fluorescence. The decreased quenching response was not due to increased H+ leakiness of the mutant membranes or to loss of F1-ATPase activity from the membrane. These results indicate that the mutant F1F0-ATPases are defective in coupling ATP hydrolysis to H+ translocation. The membrane ATPase activity of the mutants was inhibited less by dicyclohexylcarbodiimide than that of wild type. The decrease in sensitivity to inhibition by dicyclohexylcarbodiimide was caused primarily by dissociation of the F1-ATPase from the mutant F0 in the ATPase assay mixture. These results support the idea that Pro43, and neighboring conserved polar residues play an important role in the binding and functional coupling of F1 to F0. Although a Pro residue is found at position 43 in all species of subunit c studied, surprisingly, it is not absolutely essential to function
RP  - NOT IN FILE
NT  - UI - 89079668LA - engRN - 0 (Macromolecular Systems)RN - 0 (Oligonucleotide Probes)RN - 147-85-3 (Proline)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - 5-T32-GM07133/GM/NIGMSID - 5-T32-GM07215/GM/NIGMSID - GM23105/GM/NIGMSID - etcDA - 19890207IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:2521216
SO  - J Biol Chem 1989 Jan 5 ;264(1):305-311

918
UI  - 755
AU  - Miwa K
AU  - Ohtsubo M
AU  - Denda K
AU  - Hisabori T
AU  - Date T
AU  - Yoshida M
AD  - Department of Life Science, Tokyo Institute of Technology, Kanagawa
TI  - Reconstituted F1-ATPase complexes containing one impaired beta subunit are ATPase-active
AB  - Homogeneous populations of hybrid alpha 3 beta 3 gamma complexes of the thermostable F1-ATPase containing one, two, or three copies of the mutationally impaired beta subunits were produced using the solid phase reconstitution method. Two kinds of mutated beta subunits were used for the reconstitution, one of which lacked the ability to bind any adenine nucleotides. The complexes containing one impaired beta and two wild- type beta subunits retained a significant amount of ATPase activity with cooperative kinetics, whereas those containing two or three impaired beta subunits showed very little ATPase activity. These results imply that the catalysis of steady-state ATP hydrolysis can proceed even if one of the three beta subunits in F1-ATPase is not functional
RP  - NOT IN FILE
NT  - UI - 90110069LA - engRN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19900222IS - 0021-924XSB - IMCY - JAPANJC - HIF
UR  - PM:2532649
SO  - J Biochem (Tokyo ) 1989 Oct ;106(4):679-683

919
UI  - 756
AU  - Miwa K
AU  - Yoshida M
AD  - Department of Life Science, Tokyo Institute of Technology, Yokohama, Japan
TI  - The alpha 3 beta 3 complex, the catalytic core of F1-ATPase
AB  - The alpha 3 beta 3 complex was reconstituted from alpha and beta subunits of the thermophilic bacterium PS3 F1-ATPase (TF1) and then isolated. It is less stable at high and low temperatures than TF1, and the complex dissociates into subunits during native polyacrylamide gel electrophoresis. The alpha 3 beta 3 complex has about 20% of the ATPase activity of TF1. Its enzymic properties are similar to those of the native TF1, exhibiting similar cooperative kinetics as a function of ATP concentration, similar substrate specificity for nucleotide triphosphates, and the presence of two peaks in its temperature- activity profile. Differing from TF1, the ATPase activity of the alpha 3 beta 3 complex is insensitive to N3- inhibition, its divalent cation specificity is less stringent, and its optimum pH shifts to the alkaline side. The addition of the gamma subunit to the alpha 3 beta 3 complex leads to the formation of the alpha 3 beta 3 gamma complex, indicating that the alpha 3 beta 3 complex is an intermediate in the process of assembly of the holoenzyme from each subunit. These results definitely show that the essential structure for eliciting the ATPase activity of F1-ATPase is trimeric alpha beta pairs and that the kinetic cooperativity of the F1-ATPase is an inherent property of this trimeric structure but is not due to the presence of single-copy subunits. In this sense, the alpha 3 beta 3 complex is the catalytic core of F1- ATPase
RP  - NOT IN FILE
NT  - UI - 89367273LA - engRN - 0 (Cations, Divalent)RN - 0 (Macromolecular Systems)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19891006IS - 0027-8424SB - IMCY - UNITED STATESJC - PV3
UR  - PM:2528144
SO  - Proc Natl Acad Sci U S A 1989 Sep ;86(17):6484-6487

920
UI  - 361
AU  - Palmer DN
AU  - Fearnley IM
AU  - Medd SM
AU  - Walker JE
AU  - Martinus RD
AU  - Bayliss SL
AU  - Hall NA
AU  - Lake BD
AU  - Wolfe LS
AU  - Jolly RD
AD  - Department of Veterinary Pathology and Public Health, Massey University, Palmerston North, New Zealand
TI  - Lysosomal storage of the DCCD reactive proteolipid subunit of mitochondrial ATP synthase in human and ovine ceroid lipofuscinoses
AB  - The ceroid lipofuscinoses (Batten's disease) are a group of neuro- degenerative lysosomal storage diseases of children and animals that are recessively inherited. In the diseased individuals fluorescent storage bodies accumulate in a wide variety of cells, including neurons. The material stored in the cells of sheep affected with ceroid lipofuscinosis is two-thirds protein. The stored material does not arise from lipid peroxidation or a defect in lipid metabolism, and the lipid content is consistent with a lysosomal origin for the storage bodies. The major protein stains poorly with Coomassie blue dye and is soluble in organic solvents. It has an apparent molecular weight of 3,500 and its amino acids sequence is identical to that of the dicyclohexylcarbodiimide (DCCD) reactive proteolipid, subunit c, of mammalian mitochondrial ATP synthases. Apart from removal of mitochondrial import sequences, it has not been modified post- translationally. At least 50% of the mass of the storage bodies is composed of this protein. A minor protein sequence related to the 17- kDa subunit of vacuolar H(+)-ATPase is also found in storage bodies isolated from pancreas. As in humans and cattle, the ovine protein is the product of two expressed genes named P1 and P2. In normal and diseased animals there are no differences in sequences between P1 cDNAs or P2 cDNAs, nor do levels of mRNAs in liver for P1 or P2 differ substantially between normal and diseased animals. Both normal and diseased sheep also express a spliced pseudogene encoding amino acids 1 to 31 of the mitochondrial import presequence. The peptides they encode differ by one amino acid; arginine-23 is changed to glutamine in the diseased sheep. Storage bodies isolated from brains and pancreas of children affected with the juvenile and late infantile forms of ceroid lipofuscinosis also contain large amounts of material that is identical to subunit c of ATP synthase. However, the protein is not present in storage bodies isolated from brains of patients affected with the infantile form of the disease, and these storage bodies contain other unidentified proteins. It is possible that the cause of ovine, juvenile and late infantile ceroid lipofuscinoses is related to a defect in degradation of the subunit c of mitochondrial ATP synthase
RP  - NOT IN FILE
NT  - UI - 91063242LA - engRN - 0 (Proteins)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialID - NS11238/NS/NINDSDA - 19910107IS - 0065-2598SB - IMCY - UNITED STATESJC - 2LU
UR  - PM:2535017
SO  - Adv Exp Med Biol 1989  ;266():211-222

921
UI  - 391
AU  - Panchenko MV
AU  - Vinogradov AD
TI  - [Kinetics of the interaction of ATPase of submitochondrial fragments and a natural protein-inhibitor]
AB  - Conditions were selected which enable a quantitative assay of the ATPase inhibitor protein in submitochondrial particles. It was found that the isolated soluble inhibitor exhibits a marked pH-dependent hysteretic behaviour, i. e., an instant jump of pH for the inhibitor solution from 4.8 to 8.2 induced a slow alteration of its activity as measured by the inhibition of ATP hydrolysis by submitochondrial particles. In acid media (pH less than 6.8), the inhibitor is in the active, whereas in alkaline media (pH greater than 6.8) in the inactive state; the apparent pKa value for the cooperative active/inactive transition is 6.8. Treatment of the inhibitor protein with diethylpyrocarbonate, a specific reagent for histidine, completely abolishes its inhibitory activity. Two types of the inhibitor protein-- ATPase interaction were revealed, i.e., reversible (ATP-independent) and irreversible (ATP-dependent) ones. Both reactions, i.e., ATP hydrolysis and ATP inhibition by the inhibitor in the presence of Mg2+ are characterized by a hyperbolic dependence of the reaction rate on ATP concentration; however, for both reactions the apparent KmATP values (50 and 5 microM, respectively) differ significantly (pH 8.0). Thus, the inhibitor--ATPase interaction shows that there exists a specific site for ATP in the ATPase which is different from the catalytic one. A model for the inhibitor protein interaction with ATPase which takes account of a slow pH-dependent conformational transformation of the inhibitor protein is proposed
RP  - NOT IN FILE
NT  - UI - 89335841LA - rusRN - 0 (ATPase inhibitory protein)RN - 0 (Proteins)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19890919IS - 0320-9725SB - IMCY - USSRJC - A28
UR  - PM:2527066
SO  - Biokhimiia 1989 Apr ;54(4):569-579

922
UI  - 21168
AU  - Pati S
AU  - Brusilow WS
AD  - Department of Chemistry and Biochemistry, University of Maryland, College Park 20742
TI  - The roles of the alpha and gamma subunits in proton conduction through the Fo sector of the proton-translocating ATPase of Escherichia coli
AB  - Previous genetic and biochemical studies have shown that the Fo sector of the Escherichia coli H+-ATPase is synthesized and assembled in a nonleaky form from plasmid-borne genes. The proton channel then appears to be opened by an interaction of F1 subunits, especially the alpha subunit, with the nonleaky Fo (Brusilow, W. S. A. (1987) J. Bacteriol. 169, 4984-4990; Solomon, K. A., and Brusilow, W. S. A. (1988) J. Biol. Chem. 263, 5402-5407). To study the role of the alpha and gamma subunits in proton conduction, we constructed an inducible alpha plasmid. In an alpha-gamma- background, the induction of alpha synthesis caused lethal proton leakiness, as assayed by the loss of respiration-dependent acridine orange fluorescence quenching of E. coli membranes. The presence of a gamma subunit counteracted the lethal effects as if gamma were blocking the opened channel
MH  - A
MH  - Acridine Orange
MH  - alpha
MH  - ALPHA-SUBUNIT
MH  - ATPase
MH  - Biochemistry
MH  - Chemistry
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1
MH  - fluorescence
MH  - H+-ATPase
MH  - Macromolecular Systems
MH  - membrane
MH  - Membranes
MH  - proton
MH  - Proton-Translocating ATPases
MH  - Protons
MH  - SUBUNIT
MH  - synthesis
MH  - SYSTEM
MH  - SYSTEMS
RP  - NOT IN FILE
NT  - UI - 89123355LA - engRN - 0 (Macromolecular Systems)RN - 0 (Plasmids)RN - 0 (Protons)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleID - AI20010/AI/NIAIDDA - 19890321IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:2536718
SO  - J Biol Chem 1989 Feb 15 ;264(5):2640-2644

923
UI  - 44
AU  - Paule CR
AU  - Fillingame RH
AD  - Department of Physiological Chemistry, University of Wisconsin Medical School, Madison 53706
TI  - Mutations in three of the putative transmembrane helices of subunit a of the Escherichia coli F1F0-ATPase disrupt ATP-driven proton translocation
AB  - Three missense mutants in subunit a of the Escherichia coli F1F0-ATPase were isolated and characterized after hydroxylamine mutagenesis of a plasmid carrying the uncB (subunit a) gene. The mutations resulted in Asp119----His, Ser152----Phe, or Gly197----Arg substitutions in subunit a. Function was not completely abolished by any of the mutations. The F0 membrane sector was assembled in all three cases as judged by restoration of dicyclohexylcarbodiimide sensitivity to the F1F0-ATPase. The H+ translocation capacity of F0 was reduced in all three mutants. ATP-driven H+-translocation was also reduced, with the response in the Gly197----Arg mutant being almost nil and that in the Asp119----His and Ser152----Phe mutants less severely affected. The substituted residues are predicted to lie in the second, third, and fourth transmembrane helices suggested in most models for subunit a. The Gly197----Arg mutation lies in a very conserved region of the protein and the substitution may disrupt a structure that is critical to function. The Asp119----His and Ser152----Phe mutations also lie in areas with sequence conservation. A further analysis of randomly generated mutants may provide more information on regions of the protein that are crucial to function. Heterodiploid transformants, carrying plasmids with either the wild-type uncB gene or mutant uncB genes in an uncB (Trp231---- stop) background, were characterized biochemically. The truncated subunit a was not detected in membranes of the background strain by Western blotting, and the uncB+ plasmid complemented strain showed normal biochemistry. The uncB mutant genes were shown to cause equivalent defects in either the heterodiploid background configuration, or after incorporation into an otherwise wild-type unc operon. The subunit a (Trp231----stop) background strain was shown to bind F1-ATPase nearly normally despite lacking subunit a in its membrane
RP  - NOT IN FILE
NT  - UI - 89372943LA - engRN - 0 (Codon)RN - 0 (Macromolecular Systems)RN - 0 (Plasmids)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - 5-T32-GM07215/GM/NIGMSID - GM23105/GM/NIGMSDA - 19891012IS - 0003-9861SB - IMCY - UNITED STATESJC - 6SK
UR  - PM:2528329
SO  - Arch Biochem Biophys 1989 Oct ;274(1):270-284

924
UI  - 21235
AU  - Podgornik R
TI  - Electrostatic correlation forces between surfaces with surface specific interactions
RP  - IN FILE
SO  - J Chem Phys 1989  ;91():5840-5849

925
UI  - 843
AU  - Richardson DJ
AU  - McEwan AG
AU  - Jackson JB
AU  - Ferguson SJ
AD  - School of Biochemistry, University of Birmingham, England
TI  - Electron transport pathways to nitrous oxide in Rhodobacter species
AB  - 1. Electron transport components involved in nitrous oxide reduction in several strains of Rhodobacter capsulatus and in the denitrifying strain of Rhodobacter sphaeroides (f. sp. denitrificans) have been investigated. Detailed titrations with antimycin A and myxothiazol, inhibitors of the cytochrome bc1 complex, show that part of the electron flow to nitrous oxide passes through this complex. The sensitivity to myxothiazol varies between strains and growth conditions of R. capsulatus; the higher rates of nitrous oxide reduction correlate with the higher sensitivities. Partial inhibition of the nitrous oxide reductase enzyme with azide decreased the sensitivity to myxothiazol of the strains that had the highest nitrous oxide reductase activity. 2. Inhibition of nitrous oxide reduction in cells of R. capsulatus by myxothiazol could be restored under dark conditions by addition of N,N,N',N'-tetramethyl-p-phenylene diamine. The highest activities observed after addition of this electron carrier were found in the strains that had the highest sensitivity to myxothiazol, consistent with the premise that this inhibitor is more effective at the higher flux rates to nitrous oxide. 3. Addition of nitrous oxide to cells of R. capsulatus strain N22DNAR+ under darkness caused oxidation of both b- and c-type cytochromes. The oxidation of b cytochromes was less pronounced in the presence of myxothiazol, consistent with a role for the cytochrome bc1 complex in the electron pathway to nitrous oxide. Ferricyanide, in the absence of myxothiazol, caused a similar extent of oxidation of b cytochromes, but a greater oxidation of c-type, suggesting that there was a pool of c-type cytochrome that was not oxidisable by nitrous oxide. The time course showed that both the b- and c-type cytochromes were oxidised within a few seconds of the addition of nitrous oxide. During the following seconds there was a partial re-reduction of the cytochromes such that after approximately 1 min a lower steady-state of oxidation was attained and this persisted until the nitrous oxide was exhausted. 4. A mutant, MTCBC1, of R. capsulatus that specifically lacked a functional cytochrome bc1 complex reduced nitrous oxide, albeit at 30% of the rate shown by the parent strain MT1131. A reduced minus nitrous-oxide-oxidised difference spectrum for MTCBC1 in the absence of myxothiazol was similar to the corresponding difference spectrum observed for strain N22DNAR+ in the presence of myxothiazol. It is suggested that these difference spectra identify the cytochrome components, including a b-type, involved in a pathway that is alternative to, and independent of, the cytochrome bc1 complex.(ABSTRACT TRUNCATED AT 400 WORDS)
RP  - NOT IN FILE
NT  - UI - 90076187LA - engRN - 0 (Hydroxyquinolines)RN - 0 (Thiazoles)RN - 10024-97-2 (Nitrous Oxide)RN - 341-88-8 (2-(n-heptyl)-4-hydroxyquinoline N-oxide)RN - 642-15-9 (Antimycin A)RN - 76706-55-3 (myxothiazol)RN - EC 1.10.2.2 (Ubiquinol-Cytochrome-c Reductase)PT - Journal ArticleDA - 19900119IS - 0014-2956SB - IMCY - GERMANY, WESTJC - EMZ
UR  - PM:2556273
SO  - Eur J Biochem 1989 Nov 20 ;185(3):659-669

926
UI  - 21088
AU  - Rohde M
AU  - Mayer F
AU  - Hicks DB
AU  - Krulwich TA
AD  - Gesellschaft fur Biotechnologische Forschung, Braunschweig, FRG
TI  - Immunoelectron microscopic localization of the F1F0 ATPase (ATP synthase) on the cytoplasmic membrane of alkalophilic Bacillus firmus RAB
AB  - Evidence that the F1F0 ATPase (ATP synthase) of alkalophilic Bacillus firmus RAB is localized exclusively on the cytoplasmic membrane was obtained by immunogold electron microscopy using a highly specific polyclonal antibody against the beta subunit of Escherichia coli F1F0 ATPase. The energetic problem faced by cells of B. firmus RAB growing oxidatively at pH 10.5 despite a low protonmotive force across the cytoplasmic membrane cannot, therefore, be circumvented by localization of energy transducing functions on hypothetical internal membranes
MH  - A
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Bacillus
MH  - BETA
MH  - BETA-SUBUNIT
MH  - Cells
MH  - electron
MH  - electron microscopy
MH  - ELECTRON-MICROSCOPY
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - function
MH  - membrane
MH  - Membranes
MH  - Microscopy
MH  - pH
MH  - Proton-Translocating ATPases
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 90028331LA - engRN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleID - GM 28454/GM/NIGMSDA - 19891213IS - 0006-3002SB - IMCY - NETHERLANDS
UR  - PM:2529907
SO  - Biochim Biophys Acta 1989 Oct 16 ;985(2):233-235

927
UI  - 389
AU  - Vasil'eva EA
AU  - Panchenko MV
AU  - Vinogradov AD
TI  - [Interaction of ATPase from submitochondrial fragments and a natural inhibitor protein during delta-mu-H+ generation on a membrane]
AB  - An addition of the inhibitor protein (IF1) to submitochondrial particles (SMP) essentially free of endogenous IF1 (AS-SMP) results in a synchroneous inhibition of ATP hydrolysis and ATP-dependent reduction of NAD+ by succinate without any effect on the oxidative phosphorylation rate. The binding of IF1 to the membrane-bound ATPase leads to the loss of the inhibitor protein sensitivity to trypsin despite the delta mu H+ generation. The data obtained are consistent with a model according to which there exist the hydrolase and synthetase forms of F1 and contradict the generally accepted concepts on the delta mu H+-dependent dissociation of the F1-IF1 complex
RP  - NOT IN FILE
NT  - UI - 90074629LA - rusRN - 0 (ATPase inhibitory protein)RN - 0 (Proteins)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.38 (Ca(2+)-Transporting ATPase)PT - Journal ArticleDA - 19900123IS - 0320-9725SB - IMCY - USSRJC - A28
UR  - PM:2531616
SO  - Biokhimiia 1989 Sep ;54(9):1490-1498

928
UI  - 357
AU  - Walker JE
AU  - Powell SJ
AU  - Vinas O
AU  - Runswick MJ
AD  - MRC Laboratory of Molecular Biology, Cambridge, UK
TI  - ATP synthase from bovine mitochondria: complementary DNA sequence of the import precursor of a heart isoform of the alpha subunit
AB  - The alpha-subunit of ATP synthase from mitochondria is a major component of the extrinsic membrane sector of the enzyme. It is encoded in nuclear DNA. A family of overlapping complementary DNA clones encoding its precursor has been isolated from a bovine library by using in the first instance a mixture of 128 synthetic oligonucleotides designed on the basis of the known protein sequence, and the sequence of the full-length cDNA has been determined. The deduced protein sequence shows that the alpha-subunit of ATP synthase has a presequence of 43 amino acids that is not present in the mature protein. Presumably it directs the protein into the mitochondrial matrix and is removed during the import process. The encoded protein sequence is also longer by one amino acid at its C-terminal end than the protein isolated from F1-ATPase, but this alanine residue may have been removed artifactually during release of the F1-ATPase particle from the inner mitochondrial membrane. With the exception of one uncertainty caused by an ambiguity at one position in the nucleotide sequence, the mature protein sequence encoded in the cDNA is exactly the same as the sequence determined previously by direct analysis of the protein isolated from bovine heart mitochondria [Walker et al. (1985) J. Mol. Biol. 184, 677-701]. The cDNA sequence differs in 158 nucleotides over a region of alignment of 1097 nucleotides from a partial cDNA for the alpha-subunit that has been isolated from a bovine cDNA derived from liver RNA [Breen (1988) Biochem. Biophys. Res. Commun. 152, 264-269].(ABSTRACT TRUNCATED AT 250 WORDS)
RP  - NOT IN FILE
NT  - UI - 89352541LA - engRN - 0 (Enzyme Precursors)RN - 0 (Oligonucleotides)RN - 0 (RNA, Messenger)RN - 9007-49-2 (DNA)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19890929IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:2527557
SO  - Biochemistry 1989 May 30 ;28(11):4702-4708

929
UI  - 360
AU  - Walker JW
AU  - Reid GP
AU  - Trentham DR
TI  - Synthesis and properties of caged nucleotides
RP  - NOT IN FILE
NT  - UI - 89313422LA - engRN - 0 (Indicators and Reagents)RN - 0 (Nucleotides)RN - 56-65-5 (Adenosine Triphosphate)PT - Journal ArticleID - HL15835/HL/NHLBIDA - 19890825IS - 0076-6879SB - IMCY - UNITED STATESJC - MVA
UR  - PM:2747531
SO  - Methods Enzymol 1989  ;172():288-301

930
UI  - 19892
AU  - Wandlowski T
AU  - Marecek V
AU  - Holub K
AU  - Samec Z
TI  - Ion transfer across liquid-liquid phase boundaries: Electrochemical kinetics by Faradic impendance
MH  - ion
MH  - TRANSFER
MH  - Kinetics
RP  - IN FILE
SO  - J Phys Chem 1989  ;93():8204-8212

931
UI  - 19790
AU  - Xue Z
AU  - Boyer PD
AD  - Department of Chemistry and Biochemistry, University of California, Los Angeles
TI  - Modulation of the GTPase activity of the chloroplast F1-ATPase by ATP binding at noncatalytic sites
AB  - Although the binding of nucleotides at the noncatalytic sites of F1- ATPase has been regarded as probably having some type of regulatory function, only limited observations have been reported that support such a role. We present here results showing that the presence of ATP at noncatalytic sites can give a fivefold enhancement of the rate of GTP hydrolysis by the chloroplast F1-ATPase. Heat-activation of the chloroplast F1-ATPase in the presence of ATP, followed by column separation from the medium nucleotides gives an enzyme with two of the three noncatalytic sites filled with ATP. In contrast, heat-activation in the presence of ADP gives an enzyme with only one noncatalytic site filled with ADP. Such an enzyme with two noncatalytic sites empty catalyzes MgGTP hydrolysis only very slowly. The filling of a second noncatalytic site with ATP by exposure of the enzyme to ATP without Mg2+ present, followed by column separation, markedly increases the rate of GTP hydrolysis. A further increase occurs when a third noncatalytic site is filled by exposure to Mg2+ and ATP. The rate of MgATP hydrolysis is the same for the enzyme heat-activated in the presence of ATP or ADP, probably because MgATP, unlike MgGTP, rapidly binds to both catalytic and noncatalytic sites
MH  - A
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - ADP
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BINDING
MH  - Biochemistry
MH  - Chemistry
MH  - chloroplast
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - Guanosine Triphosphate
MH  - H(+)-Transporting ATP Synthase
MH  - Hydrolysis
MH  - Magnesium
MH  - NONCATALYTIC SITES
MH  - Nucleotides
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 89153105LA - engRN - 0 (Nucleotides)RN - 28141-84-6 (magnesium GTP)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 86-01-1 (Guanosine Triphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM11094/GM/NIGMSDA - 19890417IS - 0014-2956SB - IMCY - GERMANY, WESTJC - EMZ
UR  - PM:2522043
SO  - Eur J Biochem 1989 Feb 15 ;179(3):677-681

932
UI  - 754
AU  - Yokoyama K
AU  - Hisabori T
AU  - Yoshida M
AD  - Department of Life Science, Tokyo Institute of Technology, Midori-ku Yokohama, Japan
TI  - The reconstituted alpha 3 beta 3 delta complex of the thermostable F1- ATPase
AB  - Previously we reported that ATPase activity was recovered when the subunit alpha + beta + gamma or alpha + beta + delta of the F1-ATPase from the thermophilic bacterium PS3 were combined under appropriate conditions. Unlike that of holoenzyme (TF1) and the alpha + beta + gamma mixture, ATPase activity of the alpha + beta + delta mixture was heat labile and insensitive to azide inhibition (Yoshida, M., Sone, N., Hirata, H., and Kagawa, Y. (1977) J. Biol. Chem. 252, 3480-3485). Here, the properties of purified subunit complexes were compared in detail with those of native TF1. The subunit stoichiometries of the complexes were determined to be alpha 3 beta 3 gamma 1 and alpha 3 beta 3 delta 1. In general, the properties of the alpha 3 beta 3 gamma complex are very similar to those of TF1, whereas those of the alpha 3 beta 3 delta complex are significantly different. ATPase activity of the alpha 3 beta 3 delta complex is cold labile. The alpha 3 beta 3 delta complex showed a less stringent specificity for substrate and divalent cation than TF1 and the alpha 3 beta 3 gamma complex. Two Km values for ATP were exhibited by the alpha 3 beta 3 delta complex with the lower one being in the range of 0.1 microM. Equilibrium dialysis experiments revealed that the alpha 3 beta 3 delta complex cannot specifically bind ADP in the absence of Mg2+, while TF1 and the alpha 3 beta 3 gamma complex bind about 1 and 3 mol of ADP/mol of enzyme, respectively. ADP- dependent inactivation of the alpha 3 beta 3 delta complex by dicyclohexylcarbodiimide was not observed. The alpha 3 beta 3 gamma complex was readily formed when the gamma subunit was added to the alpha 3 beta 3 delta complex, suggesting that the alpha 3 beta 3 delta complex is not a "dead-end" complex. The cause of thermolability of the alpha 3 beta 3 delta complex appears to be the low stability of the complex itself at high temperature and not due to an unusually low thermostability of the delta subunit
RP  - NOT IN FILE
NT  - UI - 90094363LA - engRN - 0 (Cations, Divalent)RN - 0 (Macromolecular Systems)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 58-64-0 (Adenosine Diphosphate)RN - 7439-95-4 (Magnesium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19900201IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:2532213
SO  - J Biol Chem 1989 Dec 25 ;264(36):21837-21841

933
UI  - 875
AU  - Zolkiewska A
TI  - [Proton potential on the inner mitochondrial membrane and its role in mitochondrial processes]
MH  - Action Potentials
MH  - Adenosine Triphosphate
MH  - Biological Transport
MH  - biosynthesis
MH  - Cell Membrane
MH  - Energy Metabolism
MH  - Human
MH  - Membrane Proteins
MH  - metabolism
MH  - Mitochondria
MH  - physiology
MH  - Protons
RP  - NOT IN FILE
NT  - Zaklad Biochemii Komorki, Instytut Biologii Doswiadczalnej PAN im M Nenckiego, Warszawa
SO  - Postepy Biochem 1989  ;35(4):427-435

934
UI  - 183
AU  - Aggeler R
AU  - Mendel-Hartvig J
AU  - Capaldi RA
AD  - Institute of Molecular Biology, University of Oregon, Eugene 97403
TI  - Monoclonal antibody modification of the ATPase activity of Escherichia coli F1 ATPase
AB  - Monoclonal antibodies (mAbs) have been made against each of the five subunits of ECF1 (alpha, beta, gamma, delta, and epsilon), and these have been used in topology studies and for examination of the role of individual subunits in the functioning of the enzyme. All of the mAbs obtained reacted with ECF1, while several failed to react with ECF1F0, including three mAbs against the gamma subunit (gamma II, gamma III, and gamma IV), one mAb against delta, and two mAbs against epsilon (epsilon I and epsilon II). These topology data are consistent with the gamma, delta, and epsilon subunits being located at the interface between the F1 and F0 parts of the complex. Two forms of ECF1 were used to study the effects of mAbs on the ATPase activity of the enzyme: ECF1 with the epsilon subunit tightly bound and acting to inhibit activity and ECF1* in which the delta and epsilon subunits had been removed by organic solvent treatment. ECF1* had an ATPase activity under standard conditions of 93 mumol of ATP hydrolyzed min-1 mg-1, cf. an activity of 7.5 units mg-1 for our standard ECF1 preparation and 64 units mg-1 for enzyme in which the epsilon subunit had been removed by trypsin treatment. The protease digestion of ECF1* reduced activity to 64 units mg-1 in a complicated process involving an inhibition of activity by cleavage of the alpha subunit, activation by cleavage of gamma, and inhibition with cleavage of the beta subunit. mAbs to the gamma subunit, gamma II and gamma III, activated ECF1 by 4.4- and 2.4-fold, respectively, by changing the affinity of the enzyme for the epsilon subunit, as evidenced by density gradient centrifugation experiments. The gamma-subunit mAbs did not alter the ATPase activity of ECF1*- or trypsin-treated enzyme. The alpha-subunit mAb (alpha I) activated ECF1 by a factor of 2.5-fold and ECF1F0 by 1.3-fold, but inhibited the ATPase activity of ECF1* by 30%
RP  - NOT IN FILE
NT  - UI - 91084507LA - engRN - 0 (Antibodies, Monoclonal)RN - EC 3.4.21.4 (Trypsin)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - HL24236/HL/NHLBIDA - 19910207IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:2148117
SO  - Biochemistry 1990 Nov 13 ;29(45):10387-10393

935
UI  - 444
AU  - al Shawi MK
AU  - Parsonage D
AU  - Senior AE
AD  - Department of Biochemistry, University of Rochester Medical Center, New York 14642
TI  - Adenosine triphosphatase and nucleotide binding activity of isolated beta-subunit preparations from Escherichia coli F1F0-ATP synthase
AB  - Adenosine triphosphatase activity and nucleotide binding affinity of isolated beta-subunit preparations from Escherichia coli F1F0-ATP synthase were studied. The aim was to find out whether isolated beta- subunit would provide an experimental model in which effects of mutations on catalysis per se, unencumbered by complications due to their effects on positive catalytic cooperativity, could be studied. Three types of purified, isolated beta-subunit preparations were studied. Type I-beta was from a strain lacking all F1F0 subunits except beta and epsilon. Type II-beta was from F1 carrying the alpha S375F mutation which blocks positive catalytic cooperativity. Type III-beta was from normal F1. Type I- and II-beta had very low ATPase activity (less than 10(-4) s-1) which was azide-insensitive, aurovertin- insensitive, and unaffected by anti-beta antibody. Type I-beta activity was EDTA-insensitive. We conclude that isolated beta-subunit from E. coli F1F0 has zero or at most very low intrinsic ATPase activity. Type III-beta had low ATPase activity (8.4 x 10(-5) s-1 to 1.1 x 10(-3) s-1 in seven different preparations). This activity was aurovertin- sensitive, but varied in azide sensitivity from 0 to 34% inhibited. The azide-sensitive component, like F1 and alpha 3 beta 3 gamma oligomer, was inhibited by anti-beta and anti-alpha antibodies. The azide- insensitive component was stimulated by anti-beta and unaffected by anti-alpha. We show here that (alpha beta)-oligomer has ATPase activity which is azide-insensitive, aurovertin-sensitive, stimulated by anti- beta, and unaffected by anti-alpha. The intrinsic ATPase activity of Type III-beta could be due to contaminating (alpha beta)-oligomer plus alpha 3 beta 3 gamma-oligomer. Isolated beta had very low affinity for nucleotide as compared to the first catalytic site on F1. Taken together with the very low ATPase activity of isolated beta (even if real), the work shows that isolated beta is not a good experimental model of F1 catalysis
RP  - NOT IN FILE
NT  - UI - 90202798LA - engRN - 0 (Antibodies)RN - 0 (Aurovertins)RN - 0 (Azides)RN - 0 (Macromolecular Systems)RN - 0 (Nucleotides)RN - 146-91-8 (Guanosine Diphosphate)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 60-00-4 (Edetic Acid)RN - 65256-31-7 (aurovertin D)RN - 7439-95-4 (Magnesium)RN - 86-01-1 (Guanosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM-25349/GM/NIGMSDA - 19900503IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:2156822
SO  - J Biol Chem 1990 Apr 5 ;265(10):5595-5601

936
UI  - 445
AU  - al Shawi MK
AU  - Parsonage D
AU  - Senior AE
AD  - Department of Biochemistry, University of Rochester Medical Center, New York 14642
TI  - Thermodynamic analyses of the catalytic pathway of F1-ATPase from Escherichia coli. Implications regarding the nature of energy coupling by F1-ATPases
AB  - Thermodynamic properties of 12 different F1-ATPase enzymes were analyzed in order to gain insights into the catalytic mechanism and the nature of energy coupling to delta mu H+. The enzymes were normal soluble Escherichia coli F1, a group of nine beta-subunit mutant soluble E. coli F1 enzymes (G142S, K155Q, K155E, E181Q, E192Q, M209I, D242N, D242V, R246C), and both soluble and membrane-bound bovine heart mitochondrial F1. Unisite activity was studied by use of Gibbs free energy diagrams, difference energy diagrams, and derivation of linear free energy relationships. This allowed construction of binding energy diagrams for both the unisite ATP hydrolysis and ATP synthesis reaction pathways, which were in agreement. The binding energy diagrams showed that the step of Pi binding is a major energy-requiring step in ATP synthesis, as is the step of ATP release. It is suggested that there are two major catalytic enzyme conformations, and ATP- and an ADP- binding conformation. The effects of the mutations on the rate-limiting steps of multisite as compared to unisite activity were correlated, suggesting a direct link between the rate-limiting steps of the two types of activity. Multisite activity was analyzed by Arrhenius plots and by study of relative promotion from unisite to multisite rate. Changes in binding energy due to mutation were seen to have direct effects on multisite catalysis. From all the data, a model is derived to describe the mechanism of ATP synthesis. ATP hydrolysis, and energy coupling to delta mu H+ in F1F0-ATPases
RP  - NOT IN FILE
NT  - UI - 90170945LA - engRN - 0 (Phosphates)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSDA - 19900409IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:2137823
SO  - J Biol Chem 1990 Mar 15 ;265(8):4402-4410

937
UI  - 21266
AU  - Antonenko YN
AU  - Yaguzhinsky LS
AD  - AN Belozersky Laboratory, Moscow State University, USSR
TI  - Effect of changes in cation concentration near bilayer lipid membrane on the rate of carrier-mediated cation fluxes and on the carrier apparent selectivity
AB  - A new approach was applied for the measurements of ion transport through bilayer lipid membranes (BLM) induced by electrically neutral cation/H+ exchangers. This is an improved version of the method of the measurements of the cation/H+ exchange rate based on recording pH shifts in the unstirred layers near the BLM. Using this approach, the pH gradient in the unstirred layers induced by the cation/H+ exchanger was reduced by successive addition of the acetate on one side of the BLM until the pH shift reached zero. The difference in acetate concentration across the membrane is a measure of the cation/H+ exchange rate. In the second part of the work we found that the changes in cation concentration in the unstirred layers under the conditions imposed when measuring cation selectivity (according to Antonenko, Yu.N. and Yaguzhinsky, L.S., Biochim. Biophys. Acta 1988; 938, 125-130) can significantly decrease the apparent value of cation selectivity. It was shown that more accurate results can be obtained if low concentrations of the carrier are used. The values of nigericin cation selectivity for the alkali metals were measured (K+/Rb+ 19 +/- 1, Rb+/Na+ 1.9 +/- 0.2, Na+/Cs+ 8 +/- 0.5, Cs+/Li+ 1.8 +/- 0.3)
MH  - A
MH  - acetate
MH  - Cations
MH  - ion
MH  - Ion Transport
MH  - Ionophores
MH  - Lipid Bilayers
MH  - Lipids
MH  - membrane
MH  - Membranes
MH  - method
MH  - pH
MH  - Phosphatidylcholines
MH  - proton
MH  - Protons
MH  - transport
RP  - NOT IN FILE
NT  - UI - 90335254LA - engRN - 0 (Cations)RN - 0 (Ionophores)RN - 0 (Lipid Bilayers)RN - 0 (Membrane Lipids)RN - 0 (Metals)RN - 0 (Phosphatidylcholines)RN - 0 (Protons)RN - 28380-24-7 (Nigericin)RN - 57-88-5 (Cholesterol)PT - Journal ArticleDA - 19900913IS - 0006-3002SB - IMCY - NETHERLANDS
UR  - PM:2165815
SO  - Biochim Biophys Acta 1990 Jul 24 ;1026(2):236-240

938
UI  - 21019
AU  - Bogdanoff P
AU  - Graber P
TI  - Proton efflux and phosphorylation in flash groups
MH  - flash
MH  - Phosphorylation
MH  - proton
T2  - Curr. Res. Photosynth., Proc. Int. Conf. Photosynth., 8th, Meeting Date 1989, Volume 3, 217-20. Edited by: Baltscheffsky, Margareta. Kluwer: Dordrecht, Neth
Y2  - -32676  
RP  - ON REQUEST (03/18/92)
SO  -  1990  ;():

939
UI  - 20953
AU  - Burkovski A
AU  - Deckers-Hebestreit G
AU  - Altendorf K
AD  - Universitat Osnabruck, Arbeitsgruppe Mikrobiologie, Osnabruck, FRG
TI  - Expression of subunit III of the ATP synthase from spinach chloroplasts in Escherichia coli
AB  - Expression of subunit III of the ATP synthase from spinach chloroplasts in Escherichia coli has been achieved. Although the protein is inserted into the bacterial cytoplasmic membrane, formation of a functional Fo complex was not observed
MH  - A
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - chloroplast
MH  - Chloroplasts
MH  - COMPLEX
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - membrane
MH  - protein
MH  - Proteolipids
MH  - Spinach
MH  - SPINACH-CHLOROPLASTS
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 91032074LA - engRN - 0 (Plasmids)RN - 0 (Proteolipids)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19901204IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:2146153
SO  - FEBS Lett 1990 Oct 1 ;271(1-2):227-230

940
UI  - 19869
AU  - Cross RL
AU  - Taiz L
AD  - Department of Biochemistry and Molecular Biology, SUNY Health Science Center, Syracuse 13210
TI  - Gene duplication as a means for altering H+/ATP ratios during the evolution of FoF1 ATPases and synthases
AB  - In the evolution of the FoF1 family of proton-translocating membrane complexes, two reversals in function appear to have occurred, first changing it from an ATPase to an ATP synthase and then back again to an ATPase. Here we suggest that with each change in function, the ratio of protons transported per ATP hydrolyzed or synthesized (H+/ATP) was altered in order for the complex to better adapt to its new role. We propose that this was accomplished by gene duplication with partial loss in the number of functional catalytic sites (to increase H+/ATP) or functional proton channels (to decrease H+/ATP). This method of changing the H+/ATP ratio preserved overall structural features of the complex essential to energy coupling
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Biochemistry
MH  - COMPLEX
MH  - Evolution
MH  - H(+)-Transporting ATP Synthase
MH  - membrane
MH  - method
MH  - proton
MH  - Protons
MH  - review
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 90092551LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialID - GM-23152/GM/NIGMSDA - 19900208IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:2136729
SO  - FEBS Lett 1990 Jan 1 ;259(2):227-229

941
UI  - 21198
AU  - Danshina SV
AU  - Drachev AL
AU  - Drachev LA
AU  - Eremin SV
AU  - Kaulen AD
AU  - Khitrina LV
AU  - Mitsner BI
AD  - A N Belozersky Laboratory of Molecular Biology and Bioorganic Chemistry, Moscow, USSR
TI  - C(13)-substituted bacteriorhodopsin analogs
AB  - 13-Ethyl-, 13-isopropyl-, 13-tert-butyl-, 13-phenyl-, 13-alpha-naphthyl- , and 13-demethyl-retinals were synthesized and incubated with bacterioopsin (bO) to give the corresponding bacteriorhodopsin (bR) analogs. The capability of the 13-tert-butyl- and 13-alpha-naphthyl-bRs to exist and to photocycle shows that apparently around C(13) of the chromophore there lies a large enough cavity. A study of the light- induced conversions of the artificial pigments prepared has shown that the introduction at position 13 of the chromophore of the hydrocarbon substituents bulkier than that of the natural bR diminished the amplitudes of the electric photoresponses. Bulky C(13)-substituents or absence of substitution at that position decelerated the relaxation of the M-intermediates and disturbed the 13-cis-in equilibrium all-trans- isomerization
MH  - A
MH  - ANALOGS
MH  - Bacteriorhodopsin
MH  - Chemistry
MH  - Light
MH  - M-intermediate
MH  - pigments
MH  - relaxation
MH  - Retinaldehyde
RP  - NOT IN FILE
NT  - UI - 90274403LA - engRN - 116-31-4 (Retinaldehyde)RN - 53026-44-1 (Bacteriorhodopsins)PT - Journal ArticleDA - 19900711IS - 0003-9861SB - IMCY - UNITED STATES
UR  - PM:2350173
SO  - Arch Biochem Biophys 1990 Jun ;279(2):225-231

942
UI  - 89
AU  - Dimroth P
AD  - Institut fur Physiologische Chemie, Technischen Universitat, Munchen, FRG
TI  - Bacterial energy transductions coupled to sodium ions
AB  - In Propionigenium modestum, an Na+ cycle couples the exergonic decarboxylation of methylmalonyl-CoA to endergonic ATP synthesis. The ATPase is an F1F0-type enzyme, closely related to the F1F0 ATPase of Escherichia coli. The specificity of the P. modestum ATPase for Na+ is not absolute, as it catalyses proton transport at low Na+ concentrations. The Na(+)-binding site is located on the F0 sector. Therefore, a hybrid composed of F0 from P. modestum and F1 from E. coli, but not F1F0 from E. coli, was a functional Na+ pump. In Klebsiella pneumoniae, the Na+ ions pumped out of the cell by oxaloacetate decarboxylase are taken up again in symport with the growth substrate citrate. The reaction mechanism of oxaloacetate decarboxylase involves carboxylation of the prosthetic biotin group by carboxyltransfer from oxaloacetate, catalysed by the peripheral alpha- subunit. The firmly membrane-bound subunits beta and gamma complete the cycle by decarboxylation of the carboxybiotin intermediate which is coupled to Na+ translocation through the membrane
RP  - NOT IN FILE
NT  - UI - 91126298LA - engRN - 0 (Bacterial Proteins)RN - 0 (Citrates)RN - 0 (Protons)RN - 56-65-5 (Adenosine Triphosphate)RN - 7440-23-5 (Sodium)RN - 77-92-9 (Citric Acid)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)RN - EC 4.1.1. (Carboxy-Lyases)RN - EC 4.1.1.3 (oxaloacetate decarboxylase)RN - EC 4.1.1.41 (propionyl-CoA carboxylase)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 19910308IS - 0923-2508SB - IMCY - FRANCEJC - R6F
UR  - PM:2177912
SO  - Res Microbiol 1990 Mar ;141(3):332-336

943
UI  - 90
AU  - Dimroth P
AD  - Institut fur Physiologische Chemie der Technischen Universitat Munchen, FRG
TI  - Mechanisms of sodium transport in bacteria
AB  - In some bacteria, an Na+ circuit is an important link between exergonic and endergonic membrane reactions. The physiological importance of Na+ ion cycling is described in detail for three different bacteria. Klebsiella pneumoniae fermenting citrate pumps Na+ outwards by oxaloacetate decarboxylase and uses the Na+ ion gradient thus established for citrate uptake. Another possible function of the Na+ gradient may be to drive the endergonic reduction of NAD+ with ubiquinol as electron donor. In Vibrio alginolyticus, an Na+ gradient is established by the NADH: ubiquinone oxidoreductase segment of the respiratory chain; the Na+ gradient drives solute uptake, flagellar motion and possibly ATP synthesis. In Propionigenium modestum, ATP biosynthesis is entirely dependent on the Na+ ion gradient established upon decarboxylation of methylmalonyl-CoA. The three Na(+)- translocating enzymes, oxaloacetate decarboxylase of Klebsiella pneumoniae, NADH: ubiquinone oxidoreductase of Vibrio alginolyticus and ATPase (F1F0) of Propionigenium modestum have been isolated and studied with respect to structure and function. Oxaloacetate decarboxylase consists of a peripheral subunit (alpha), that catalyses the carboxyltransfer from oxaloacetate to enzyme-bound biotin. The subunits beta and gamma are firmly embedded in the membrane and catalyse the decarboxylation of the carboxybiotin enzyme, coupled to Na+ transport. A two-step mechanism has also been demonstrated for the respiratory Na+ pump. Semiquinone radicals are first formed with the electrons from NADH; subsequently, these radicals dismutate in an Na(+)-dependent reaction to quinone and quinol. The ATPase of P. modestum is closely related in its structure to the F1F0 ATPase of E. coli, but uses Na+ as the coupling ion. A specific role of protons in the ATP synthesis mechanism is therefore excluded
RP  - NOT IN FILE
NT  - UI - 90239121LA - engRN - 7440-23-5 (Sodium)RN - EC 4.1.1. (Carboxy-Lyases)RN - EC 4.1.1.3 (oxaloacetate decarboxylase)PT - Journal ArticlePT - ReviewPT - Review LiteratureDA - 19900606IS - 0962-8436SB - IMCY - ENGLANDJC - P5Z
UR  - PM:1970650
SO  - Philos Trans R Soc Lond B Biol Sci 1990 Jan 30 ;326(1236):465-477

944
UI  - 907
AU  - Dmitriev LF
TI  - [Energy transformation in mitochondria. Active role of the membrane]
MH  - Adenosine Triphosphate
MH  - biosynthesis
MH  - Energy Metabolism
MH  - Intracellular Membranes
MH  - metabolism
MH  - Mitochondria
MH  - Models,Biological
RP  - NOT IN FILE
SO  - Biofizika 1990 Jul ;35(4):685-686

945
UI  - 19787
AU  - Du ZY
AU  - Boyer PD
AD  - Department of Chemistry and Biochemistry, University of California, Los Angeles 90024-1570
TI  - On the mechanism of sulfite activation of chloroplast thylakoid ATPase and the relation of ADP tightly bound at a catalytic site to the binding change mechanism
AB  - Washed chloroplast thylakoid membranes upon exposure to [3H]ADP retain a tightly bound [3H]ADP on a catalytic site of the ATP synthase. The presence of sufficient endogenous or added Mg2+ results in an enzyme with essentially no ATPase activity. Sulfite activates the ATPase, and many molecules of ATP per synthase can be hydrolyzed before most of the bound [3H]ADP is released, a result interpreted as indicating that the ADP is not bound at a site participating in catalysis by the sulfite- activated enzyme [Larson, E. M., Umbach, A., & Jagendorf, A. T. (1989) Biochim. Biophys. Acta 973, 75-85]. We present evidence that this is not the case. The Mg2(+)- and ADP-inhibited enzyme when exposed to MgATP and 20-100 mM sulfite shows a lag of about 1 min at 22 degrees C and of about 15 s at 37 degrees C before reaching the same steady-state rate as attained with light-activated ATPase that has not been inhibited by Mg2+ and ADP. The lag is not eliminated if the enzyme is exposed to sulfite prior to MgATP addition, indicating that ATPase turnover is necessary for the activation. The release of most of the bound [3H]ADP parallels the onset of ATPase activity, although some [3H]ADP is not released even with prolonged catalytic turnover and may be on poorly active or inactive enzyme or at noncatalytic sites. The results are consistent with most of the tightly bound [3H]ADP being at a catalytic site and being replaced as this Mg2(+)- and ADP-inhibited site regains equivalent participation with other catalytic sites on the activated enzyme.(ABSTRACT TRUNCATED AT 250 WORDS)
MH  - A
MH  - ACTIVATION
MH  - ACTIVE
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - ADP
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BINDING
MH  - Biochemistry
MH  - Catalysis
MH  - CHANGE MECHANISM
MH  - Chemistry
MH  - chloroplast
MH  - Magnesium
MH  - mechanism
MH  - membrane
MH  - Membranes
MH  - NONCATALYTIC SITES
MH  - Sulfites
MH  - SYNTHASE
MH  - thylakoid
MH  - THYLAKOID ATPASE
MH  - thylakoid membrane
RP  - NOT IN FILE
NT  - UI - 90148965LA - engRN - 0 (Sulfites)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 7439-95-4 (Magnesium)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleID - GM-11094/GM/NIGMSDA - 19900326IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:2137348
SO  - Biochemistry 1990 Jan 16 ;29(2):402-407

946
UI  - 305
AU  - Engelbrecht S
AU  - Junge W
AD  - Universitat Osnabruck, Abt Biophysik, FRG
TI  - Subunit delta of H(+)-ATPases: at the interface between proton flow and ATP synthesis
AB  - The ATP synthases in photophosphorylation and respiration are of the F- type with a membrane-bound proton channel, F0, and an extrinsic catalytic portion, F1. The properties of one particular subunit, delta (in chloroplasts and Escherichia coli) and OSCP (in mitochondria), are reviewed and the role of this subunit at the interface between F0 and F1 is discussed. Delta and OSCP from the three sources have in common the molecular mass (approximately 20 kDa), an elongated shape (axial ratio in solution about 3:1), one high-affinity binding site to F1 (Kd approximately 100 nM) plus probably one or two further low-affinity sites. When isolated delta is added to CF1-depleted thylakoid membranes, it can block proton flow through exposed CF0 channels, as do CF1 or CF1(-delta)+ delta. This identifies delta as part of the proton conductor or, alternatively, conformational energy transducer between F0 (proton flow) and F1 (ATP). Hybrid constructs as CF1(-delta)+ E. coli delta and EF1(-delta)+ chloroplast delta diminish proton flow through CF0.CF1(-delta) + E. coli delta does the same on EF0. Impairment of proton leaks either through CF0 or through EF0 causes "structural reconstitution' of ATP synthesis by remaining intact F0F1. Functional reconstitution (ATP synthesis by fully reconstructed F0F1), however, is absolutely dependent on the presence of subunit delta and is therefore observed only with CF1 or CF1(-delta) + chloroplast delta on CF0 and EF1 or EF1(-delta) + E. coli delta on EF0. The effect of hybrid constructs on F0 channels is surprising in view of the limited sequence homology between chloroplast and E. coli delta (36% conserved residues including conservative replacements). An analysis of the distribution of the conserved residues at present does not allow us to discriminate between the postulated conformational or proton-conductive roles of subunit delta
RP  - NOT IN FILE
NT  - UI - 90148989LA - engRN - 0 (Membrane Proteins)RN - 0 (Protons)RN - 0 (oligomycin sensitivity-conferring protein)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 19900326IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:2154253
SO  - Biochim Biophys Acta 1990 Feb 22 ;1015(3):379-390

947
UI  - 304
AU  - Engelbrecht S
AU  - Althoff G
AU  - Junge W
AD  - Fachbereich Biologie/Chemie, Universitat Osnabruck, Federal Republic of Germany
TI  - Reconstitution of photophosphorylation in EDTA-treated thylakoids by added chloroplast coupling factor 1 (ATPase) and chloroplast coupling factor 1 lacking the delta subunit. Structural or functional?
AB  - Upon EDTA treatment thylakoids lose the chloroplast coupling factor 1 (CF1) part of their ATP synthase, CF0CF1, this exposes the proton channel, CF0. The previously established ability of the CF1 subunit delta to block the proton leak through CF0 prompted us to study (a) the ability of complete CF1 and, for comparison, CF1 lacking the delta subunit to block proton leakage and thereby to reconstitute structurally some photophosphorylation activity of the remaining CF0CF1 molecules and (b) their ability to form functional enzymes (functional reconstitution). In order to discriminate between activities caused by added CF1 or CF1(-delta) and remaining CF0CF1, the former were inhibited by chemical modification of subunit beta by N,N'-dicyclohexyl carbodiimide (DCCD) and the latter by tentoxin. We found that added CF1 acted both structurally and functionally while added DCCD-treated CF1 (DCCD-CF1) acted only structurally. In contrast to previous observations, CF1(-delta) and DCCD-CF1(-delta) also acted structurally although the reduction of proton leakage was smaller than with DCCD- CF1. Hence there was no functional reconstitution without subunit delta present. Previous studies indicated that only a small fraction of exposed CF0 is highly conducting and that this small fraction is distinguished by its high affinity for added CF1. The results of this study point rather to a wider distribution of CF0 conductance states and binding affinities
RP  - NOT IN FILE
NT  - UI - 90235856LA - engRN - 0 (Macromolecular Systems)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 60-00-4 (Edetic Acid)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19900606IS - 0014-2956SB - IMCY - GERMANY, WESTJC - EMZ
UR  - PM:2139608
SO  - Eur J Biochem 1990 Apr 20 ;189(1):193-197

948
UI  - 353
AU  - Fearnley IM
AU  - Walker JE
AU  - Martinus RD
AU  - Jolly RD
AU  - Kirkland KB
AU  - Shaw GJ
AU  - Palmer DN
AD  - MRC Laboratory of Molecular Biology, Cambridge, UK
TI  - The sequence of the major protein stored in ovine ceroid lipofuscinosis is identical with that of the dicyclohexylcarbodiimide-reactive proteolipid of mitochondrial ATP synthase
AB  - The ceroid lipofuscinoses are a group of neurodegenerative lysosomal storage diseases of children and animals that are recessively inherited. In diseased individuals fluorescent storage bodies accumulate in a wide variety of cells, including neurons. Previous studies of these bodies isolated from tissues of affected sheep confirmed that the storage occurs in lysosomes, and showed that the storage body is mostly made of a single protein with an apparent molecular mass of 3500 Da with an N-terminal amino acid sequence that is the same as residues 1-40 of the c-subunit (or dicyclohexylcarbodi- imide-reactive proteolipid) of mitochondrial ATP synthase. In the present work we have shown by direct analysis that the stored protein is identical in sequence with the entire c-subunit of mitochondrial ATP synthase, a very hydrophobic protein of 75 amino acid residues. As far as can be detected by the Edman degradation, the stored protein appears not to have been subject to any post-translational modification other than the correct removal of the mitochondrial import sequences that have been shown in other experiments to be present at the N-terminal of its two different precursors. No other protein accumulates in the storage bodies to any significant extent. Taken with studies of the cDNAs for the c-subunit in normal and diseased sheep, these results indicate that the material that is stored in lysosomes of diseased animals has probably entered mitochondria and has been subjected to the proteolytic processing that is associated with mitochondrial import. This implies that the defect that leads to the lysosomal accumulation concerns the degradative pathway of the c-subunit of ATP synthase. An alternative, but less likely, hypothesis is that for some unknown reason the precursors of subunit c are being directly mis-targeted to lysosomes, where they become processed to yield a protein identical with the protein that is normally found in the mitochondrial ATP synthase assembly, and which then accumulates
RP  - NOT IN FILE
NT  - UI - 90303272LA - engRN - 0 (Amino Acids)RN - 0 (Carbodiimides)RN - 0 (Carrier Proteins)RN - 0 (Macromolecular Systems)RN - 0 (Proteolipids)RN - 0 (dicyclohexylcarbodiimide-binding proteolipid)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - NS 11238/NS/NINDSDA - 19900808IS - 0264-6021SB - IMCY - ENGLANDJC - 9YO
UR  - PM:2141977
SO  - Biochem J 1990 Jun 15 ;268(3):751-758

949
UI  - 21110
AU  - Ferguson SJ
TI  - Periplasm underestimated
MH  - England
MH  - membrane
MH  - Membrane Proteins
MH  - protein
MH  - Proteins
RP  - NOT IN FILE
NT  - UI - 91068294LA - engRN - 0 (Membrane Proteins)PT - CommentPT - LetterDA - 19910115IS - 0968-0004SB - IMCY - ENGLAND
UR  - PM:2251728
SO  - Trends Biochem Sci 1990 Oct ;15(10):377

950
UI  - 21008
AU  - Fromme P
AU  - Graber P
AD  - Max Volmer Institut fur Biophysikalische und Physikalische Chemie, Technische Universitat Berlin, FRG
TI  - Uni-site catalysis in thylakoids. The influence of membrane energization on ATP hydrolysis and ATP-Pi exchange
AB  - ATP-hydrolysis was measured with thylakoid membranes during continuous illumination. The concentrations of free and enzyme-bound ATP, ADP and Pi were measured using either cold ATP, [gamma-32P]ATP or [14C]ATP. The concentration of free ATP was constant, free ADP and enzyme-bound ATP were below the detection limit. Nevertheless, [gamma-32P]ATP was bound, hydrolyzed and 32Pi was released. The ADP was not released from the enzyme but cold Pi was bound from the medium, cold ATP was resynthesized and released. A quantitative analysis gave the following rate constants: ATP-binding kATP = 2 . 10(5) M-1 s-1, ADP-release: kADP less than 10(-2)s-1, Pi-release: kPi = 0.1 s-1. These rate constants are considerably smaller than under deenergized conditions. The rate constant for the release of ATP can be estimated to be at least 0.2 s-1 under energized conditions. Obviously, energization of the membrane, i.e. protonation of the enzyme leads mainly to a decrease of the rate of ATP-binding, to an increase of the rate of ATP release and to a decrease of the rate of ADP-release
MH  - A
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - ADP
MH  - analysis
MH  - atp
MH  - ATPase
MH  - Catalysis
MH  - CONSTANT
MH  - Hydrolysis
MH  - membrane
MH  - Membranes
MH  - protonation
MH  - rate constant
MH  - thylakoid
MH  - thylakoid membrane
MH  - thylakoids
MH  - uni-site catalysis
MH  - UNISITE CATALYSIS
RP  - NOT IN FILE
NT  - UI - 90353585LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19900927IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:2143736
SO  - FEBS Lett 1990 Aug 20 ;269(1):247-251

951
UI  - 21010
AU  - Fromme P
AU  - Graber P
AD  - Max-Volmer-Institut fur Biophysikalische und Physikalische Chemie, Technische Universitat Berlin (Germany)
TI  - Activation/inactivation and uni-site catalysis by the reconstituted ATP- synthase from chloroplasts
AB  - The proton-translocating ATP-synthase of chloroplasts, CF0F1, was isolated and reconstituted into asolectin liposomes. CF0F1 can exist in at least four different states, oxidized or reduced, either inactive or active. These states are characterized by different kinetics of ADP binding: There is no binding of ADP to the inactive, oxidized state, the rate constant for ADP binding to the inactive, reduced states is 7.10(2) M-1.s-1. ADP binding to the active, reduced state occurs under deenergized conditions with 10(5) M-1.s-1 and transforms the enzyme into the inactive, reduced state. Parallel to the ADP-dependent inactivation, the enzyme can also inactivate without ADP binding with a first-order rate constant of 7.10(-3) M-1.s-1. With the active, reduced enzyme ATP-hydrolysis was measured under uni-site conditions as has been carried out with MF1 (Grubmeyer, C., Cross, R.C. and Penefsky, H.S. (1982) J. Biol. Chem. 257, 12092-12100). The rate constant for ATP binding is 10(6) M-1.s-1, the 'equilibrium constant' on the enzyme EADPPi/EATP is 0.4. The rate constants for Pi release and ADP release are 0.2 s-1 and o.1 s-1, respectively. This indicates that the enzyme carries out a complete turnover under uni-site conditions with rates much higher than that reported for MF1
MH  - A
MH  - ACTIVE
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - ADP
MH  - ADP BINDING
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BINDING
MH  - Catalysis
MH  - CF0F1
MH  - chloroplast
MH  - Chloroplasts
MH  - CONSTANT
MH  - Kinetics
MH  - Liposomes
MH  - Phosphates
MH  - rate constant
MH  - SYNTHASE
MH  - uni-site catalysis
MH  - UNISITE CATALYSIS
RP  - NOT IN FILE
NT  - UI - 90181419LA - engRN - 0 (Liposomes)RN - 0 (Phosphates)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19900413IS - 0006-3002SB - IMCY - NETHERLANDS
UR  - PM:2178683
SO  - Biochim Biophys Acta 1990 Mar 15 ;1016(1):29-42

952
UI  - 750
AU  - Fujiyama Y
AU  - Yokoyama K
AU  - Yoshida M
AU  - Wakabayashi T
AD  - Department of Life Science, Tokyo Institute of Technology, Japan
TI  - Electron microscopy of the reconstituted complexes of the F1-ATPase with various subunit constitution revealed the location of the gamma subunit in the central cavity of the molecule
AB  - The subunit structure of the F1-ATPase from the thermophilic bacterium PS3 was probed by comparing electron microscopic images of the subunit complexes reconstituted to contain different subunit compositions. The following structural features were observed. (1) The alpha 3 beta 3 complex has a hexagonal, apparently symmetrical arrangement of masses with a central cavity. (2) The projections of the alpha 3 beta 3 delta complexes are similar to those of the alpha 3 beta 3 complexes. (3) In contrast, the alpha 3 beta 3 gamma complex has an additional mass in the centre which is similar to that found in the native enzyme (alpha 3 beta 3 gamma delta epsilon). From these observations, it is concluded that the central mass in the F1-ATPase is comprised mostly of the gamma subunit
RP  - NOT IN FILE
NT  - UI - 91032044LA - engRN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19901204IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:2146152
SO  - FEBS Lett 1990 Oct 1 ;271(1-2):111-115

953
UI  - 182
AU  - Gogol EP
AU  - Johnston E
AU  - Aggeler R
AU  - Capaldi RA
AD  - Institute of Molecular Biology, University of Oregon, Eugene 97403
TI  - Ligand-dependent structural variations in Escherichia coli F1 ATPase revealed by cryoelectron microscopy
AB  - The Escherichia coli F1 ATPase, ECF1, has been examined by cryoelectron microscopy after reaction with Fab' fragments generated from monoclonal antibodies to the alpha and epsilon subunits. The enzyme-antibody complexes appeared triangular due to the superposition of three anti- alpha Fab' fragments on alternating densities of the hexagonally arranged alpha and beta subunits. The Fab' to the epsilon subunit superimposed on a beta subunit. A density was observed near the center of the structure in the internal cavity. The position of this central density with respect to peripheral sites was not fixed. Sorting of images of ECF1 labeled with the combination of three anti-alpha Fab' fragments plus an Fab' directed to the epsilon subunit gave three classes in each of which the central density was closest to a different beta subunit. The distribution of the central density among the three classes was measured for different ligand-binding conditions. When ATP was present in catalytic sites under conditions where there was no enzyme turnover (i.e., without Mg2+ present), there were approximately equal numbers of images in each of three classes. When ATP and Mg2+ were added and ATP hydrolysis was allowed to proceed, almost two-thirds of the images were in the class in which the central density was closest to the beta subunit superimposed by the epsilon subunit. We conclude that domains within the ECF1 structure, either the central mass or a domain including the epsilon subunit, move in the enzyme in response to ligand binding. We suggest that this movement is involved in coupling catalytic sites to the proton channel in the F0 part of the ATP synthase
RP  - NOT IN FILE
NT  - UI - 91088562LA - engRN - 0 (Ligands)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 60-00-4 (Edetic Acid)RN - 7439-95-4 (Magnesium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM39806/GM/NIGMSID - HL24526/HL/NHLBIDA - 19910207IS - 0027-8424SB - IMCY - UNITED STATESJC - PV3
UR  - PM:2148209
SO  - Proc Natl Acad Sci U S A 1990 Dec ;87(24):9585-9589

954
UI  - 842
AU  - Golby P
AU  - Carver M
AU  - Jackson JB
AD  - School of Biochemistry, University of Birmingham, England
TI  - Membrane ionic currents in Rhodobacter capsulatus. Evidence for electrophoretic transport of K+, Rb+ and NH4+
AB  - 1. The cytoplasmic membrane ionic current of cells of Rhodobacter capsulatus, washed to lower the endogenous K+ concentration, had a non- linear dependence on the membrane potential measured during photosynthetic illumination. Treatment of the cells with venturicidin, an inhibitor of the H(+)-ATP synthase, increased the membrane potential and decreased the membrane ionic current at values of membrane potential below a threshold. 2. The addition of K+ or Rb+, but not of Na+, led to an increase in the membrane ionic current and a decrease in the membrane potential in either the presence or absence of venturicidin. Approximately 0.4 mM K+ or 2.0 mM Rb+ led to a half- maximal response. At saturating concentrations of K+ and Rb+, the membrane ionic currents were similar. The membrane ionic currents due to K+ and Rb+ were not additive. The K(+)-dependent and Rb(+)-dependent ionic currents had a non-linear relationship with membrane potential: the alkali cations only increased the ionic current when the membrane potential lay above a threshold value. The presence of 1 mM Cs+ did not lead to an increase in the membrane ionic current but it had the effect of inhibiting the membrane ionic current due to either K+ or Rb+. 3. Photosynthetic illumination in the presence of either K+ or Rb+, and weak acids such as acetate, led to a decrease in light-scattering by the cells. This was attributed to the uptake of potassium or rubidium acetate and a corresponding increase in osmotic strength in the cytoplasm. 4. The addition of NH4+ also led to an increase in membrane ionic current and to a decrease in membrane potential (half-maximal at 2.0 mM NH4+). The relationship between the NH4(+)-dependent ionic currents and the membrane potential was similar to that for K+. The NH4(+)-dependent and K(+)-dependent ionic current were not additive. However, illumination in the presence of NH4+ and acetate did not lead to significant light-scattering changes. The NH4(+)-dependent membrane ionic current was inhibited by 1 mM Cs+ but not by 50 microM methylamine. 5. It is proposed that the K(+)-dependent membrane ionic current is catalysed by a low-affinity K(+)-transport system such as that described in Rb. capsulatus [Jasper, P. (1978) J. Bacteriol. 133, 1314-1322]. The possibility is considered that, as well as Rb+, this transport system can also operate with NH4+. However, in our experimental conditions NH4+ uptake is followed by NH3 efflux.(ABSTRACT TRUNCATED AT 400 WORDS)
RP  - NOT IN FILE
NT  - UI - 90151663LA - engRN - 0 (Bacterial Proteins)RN - 0 (Photosynthetic Reaction Center, Bacterial)RN - 0 (Venturicidins)RN - 7440-09-7 (Potassium)RN - 7440-17-7 (Rubidium)RN - 7664-41-7 (Ammonia)PT - Journal ArticleDA - 19900329IS - 0014-2956SB - IMCY - GERMANY, WESTJC - EMZ
UR  - PM:2406135
SO  - Eur J Biochem 1990 Feb 14 ;187(3):589-597

955
UI  - 1129
AU  - Graber P
TI  - Kinetics of proton-transport coupled ATP synthesis in chloroplasts
MH  - atp
MH  - ATP synthesis
MH  - chloroplast
MH  - Chloroplasts
MH  - Kinetics
MH  - synthesis
RP  - ON REQUEST (03/18/92)
SO  - Ettore Majorana Int Sci Ser : Phys Sci 1990  ;51():277-309

956
UI  - 10679
AU  - Graber P
AU  - Boettcher B
AU  - BOEKEMA EJ
TI  - The structure of the ATP-synthase from chloroplasts
MH  - atp
MH  - ATP synthase
MH  - chloroplast
MH  - Chloroplasts
MH  - structure
RP  - NOT IN FILE
SO  - Ettore Majorana Int Sci Ser 1990  ;51():247-276

957
UI  - 1136
AU  - Grotjohann I
AU  - Graber P
TI  - Isolation and properties of the membrane-integrated part of the ATP-synthase from chloroplasts, CF0
MH  - ATP synthase
MH  - chloroplast
MH  - Chloroplasts
RP  - ON REQUEST (03/18/92)
SO  - Biochim Biophys Acta 1990  ;1017():177-180

958
UI  - 19785
AU  - Guerrero KJ
AU  - Xue ZX
AU  - Boyer PD
AD  - Molecular Biology Institute, University of California, Los Angeles 90024
TI  - Active/Inactive state transitions of the chloroplast F1 ATPase are induced by a slow binding and release of Mg2+. Relationship to catalysis and control of F1 ATPases
AB  - Mg2+ is known to be a potent inhibitor of F1 ATPases from various sources. Such inhibition requires the presence of a tightly bound ADP at a catalytic site. Results with the spinach chloroplast F1 ATPase (CF1) show that the time delays of up to 1 min or more in the induction or the relief of the inhibition are best explained by a slow binding and slow release of Mg2+ rather than by slow enzyme conformational changes. CF1 is known to have multiple Mg2+ binding sites with Kd values in the micromolar range. The inhibitory Mg2+ and ADP can bind independently to CF1. When Mg2+ and ATP are added to the uninhibited enzyme, a relatively fast rate of hydrolysis attained soon after the addition is followed by a much slower steady-state rate. The inhibited steady-state rate results from a slowly attained equilibrium of binding of medium Mg2+. The Kd for the binding of the inhibitory Mg2+ is in the range of 1-8 microM, in the presence or absence of added ATP, as based on the extent of rate inhibition induced by Mg2+. Assessments from 18O exchange experiments show that the binding of Mg2+ is accompanied by a relatively rapid change to an enzyme form that is incapable of hydrolyzing MgATP. When ATP is added to the Mg2+- and ADP-inhibited enzyme, the resulting reactivation can be explained by MgATP binding to an alternate catalytic site which results in a displacement of the tightly bound ADP after a slow release of Mg2+. Both an increase in temperature (to 50 degrees C) and the presence of activating anions such as bicarbonate or sulfite reduce the extent of the Mg2+ inhibition markedly. The activating anions may bind to CF1 in place of Pi near the ADP. Whether the inhibitory Mg2+ binds at catalytic or noncatalytic nucleotide binding sites or at another location is not known. The Mg2(+)- and ADP-induced inhibition appears to be a general property of F1 ATPases, which show considerable differences in affinity for ADP, Mg2+, and Pi. These differences may reflect physiological control functions
MH  - A
MH  - ADP
MH  - Anions
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BINDING
MH  - Binding Sites
MH  - Catalysis
MH  - chloroplast
MH  - conformational change
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - H(+)-Transporting ATP Synthase
MH  - Hydrolysis
MH  - Magnesium
MH  - nucleotide binding
MH  - nucleotide binding sites
MH  - NUCLEOTIDE-BINDING-SITES
MH  - Spinach
MH  - SYNTHASE
MH  - Temperature
MH  - Time
RP  - NOT IN FILE
NT  - UI - 90375489LA - engRN - 146-91-8 (Guanosine Diphosphate)RN - 7439-95-4 (Magnesium)RN - EC 3.6.1.- (Ca(2+) Mg(2+)-ATPase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM11094/GM/NIGMSDA - 19901018IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:2144528
SO  - J Biol Chem 1990 Sep 25 ;265(27):16280-16287

959
UI  - 19786
AU  - Guerrero KJ
AU  - Ehler LL
AU  - Boyer PD
AD  - Department of Chemistry and Biochemistry and Molecular Biology Institute, University of California, Los Angeles 90024
TI  - Guanosine and formycin triphosphates bind at non-catalytic nucleotide binding sites of CF1 ATPase and inhibit ATP hydrolysis
AB  - Guanosine triphosphate and formycin triphosphate (FTP) in the presence of excess Mg2+ can bind to empty non-catalytic sites of spinach chloroplast ATPase (CF1). This results in a greatly reduced capacity for ATP hydrolysis compared to the enzyme with non-catalytic sites filled with ATP. With two GTP bound at non-catalytic sites the inhibition is about 90%; with two FTP bound about 80% inhibition is obtained. Binding and release of the nucleotides from the non-catalytic sites are relatively slow processes. Exposure of CF1 with one or two empty non-catalytic sites to 5-10 microM FTP or GTP for 15 min suffices for about 50% of the maximum inhibition. Reactivation of CF1 after exposure to higher FTP or GTP concentrations requires long exposure to 2 microM EDTA. The findings show that, contrary to previous assumptions, GTP can bind tightly to non-catalytic sites of CF1. They suggest that the presence of adenine nucleotides at non-catalytic sites might be essential for high catalytic capacity of the F1 ATPases
MH  - A
MH  - Adenine Nucleotides
MH  - ADENINE-NUCLEOTIDES
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BINDING
MH  - Binding Sites
MH  - Biochemistry
MH  - Calcium
MH  - Chemistry
MH  - chloroplast
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - Guanosine Triphosphate
MH  - H(+)-Transporting ATP Synthase
MH  - Hydrolysis
MH  - Magnesium
MH  - non-catalytic site
MH  - NONCATALYTIC SITES
MH  - nucleotide binding
MH  - nucleotide binding sites
MH  - NUCLEOTIDE-BINDING-SITES
MH  - Nucleotides
MH  - Spinach
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 91032009LA - engRN - 0 (Bicarbonates)RN - 0 (Formycins)RN - 0 (Ribonucleotides)RN - 16409-13-5 (formycin triphosphate)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 7439-95-4 (Magnesium)RN - 7440-70-2 (Calcium)RN - 86-01-1 (Guanosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19901210IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:2146148
SO  - FEBS Lett 1990 Sep 17 ;270(1-2):187-190

960
UI  - 21280
AU  - Gutman M
AU  - Nachliel E
TI  - The dynamic aspects of proton transfer processes
MH  - P
MH  - proton
MH  - Proton transfer
MH  - Proton-transfer
MH  - review
MH  - theory
MH  - TRANSFER
MH  - transport
MH  - wox
RP  - IN FILE
NT  - P 30; Ju Rev
SO  - Biochim Biophys Acta 1990  ;1015():391-414

961
UI  - 20805
AU  - Hnggi P
AU  - Talkner P
AU  - Borkovec M
TI  - Reaction-field theory: fifty years after Kramers
MH  - theory
RP  - IN FILE
SO  - Rev Mod Phys 1990  ;62():251-341

962
UI  - 20954
AU  - Hensel M
AU  - Deckers-Hebestreit G
AU  - Schmid R
AU  - Altendorf K
AD  - Universitat Osnabruck, Arbeitsgruppe Mikrobiologie, FRG
TI  - Orientation of subunit c of the ATP synthase of Escherichia coli--a study with peptide-specific antibodies
AB  - Antibodies were raised against a peptide of subunit c of the ATP synthase from Escherichia coli obtained by cleavage with cyanogen bromide. This peptide comprises the amino acid residues Gly-18 to Met- 57 and contains the highly conserved, hydrophilic stretch of subunit c. Several conformation-specific populations of antibodies recognized this region both in isolated subunit c and in the intact F0 complex. In antibody binding studies with membrane vesicles of different orientations, recognition occurred only after incubation with everted membrane vesicles, independent of the presence or absence of F1, although a higher membrane protein concentration was necessary to observe the same antibody binding in the presence of the F1 part. From these results we conclude that the hydrophilic region of subunit c is exposed to the cytoplasmic side of the membrane
MH  - A
MH  - ACID
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BINDING
MH  - COMPLEX
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - F1
MH  - membrane
MH  - membrane vesicles
MH  - Peptide Fragments
MH  - protein
MH  - RESIDUE
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 90181421LA - engRN - 0 (Peptide Fragments)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19900413IS - 0006-3002SB - IMCY - NETHERLANDS
UR  - PM:2178684
SO  - Biochim Biophys Acta 1990 Mar 15 ;1016(1):63-70

963
UI  - 84
AU  - Hoffmann A
AU  - Dimroth P
AD  - Mikrobiologisches Institut der Eidgenossischen Technischen Hochschule, Zurich, Switzerland
TI  - The ATPase of Bacillus alcalophilus. Purification and properties of the enzyme
AB  - The ATPase of Bacillus alcalophilus was extracted from the bacterial membranes with Triton X-100 and purified by hydroxyapatite column chromatography. SDS gel-electrophoresis of the purified protein indicated the typical subunit pattern of an F1F0 structure with five F1 subunits (alpha, beta, gamma, delta, epsilon) and three F0 subunits (a,b,c). The alpha and beta subunits were antigens for an antiserum against the corresponding subunits of the ATPase of Escherichia coli. Subunit c was extracted from the bacterial membranes with chloroform/methanol. Its amino acid composition was in the range of subunits c from other ATPases. Maximal ATPase activity was observed in the presence of 2-5 mM MgCl2, an ATP/Mg2+ ratio of 2:1 and 25% methanol. In the absence of methanol, only about 1% of the maximal activity was observed. The enzyme was also activated by Ca2+ (in the absence of methanol), reaching about 30% of the maximal activity. The dependence of initial velocity versus ATP of the Ca2(+)-activated but not of the Mg2+/methanol-activated enzyme indicted cooperativity with three strongly cooperative binding sites
RP  - NOT IN FILE
NT  - UI - 91099315LA - engRN - 0 (Amino Acids)RN - 0 (Organotin Compounds)RN - 67-56-1 (Methanol)RN - 7439-95-4 (Magnesium)RN - 7440-70-2 (Calcium)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19910220IS - 0014-2956SB - IMCY - GERMANYJC - EMZ
UR  - PM:2148515
SO  - Eur J Biochem 1990 Dec 12 ;194(2):423-430

964
UI  - 20871
AU  - Iwamoto A
AU  - Miki J
AU  - Maeda M
AU  - Futai M
AD  - Department of Organic Chemistry and Biochemistry, Osaka University, Japan
TI  - H(+)-ATPase gamma subunit of Escherichia coli. Role of the conserved carboxyl-terminal region
AB  - Cloned uncG genes (wild-type or in vitro mutagenized) for the Escherichia coli gamma subunit were introduced into the uncG mutant Gln- 14----end), and the functions of the mutant subunits were studied. The F1's with Ala-283----end and Thr-277----end mutant gamma subunits had 63 and 14% of the wild-type ATPase activity, respectively, and mutants with these subunits showed reduced growth by oxidative phosphorylation, indicating that the 10 residues at the carboxyl terminus (286th residue) are important, but dispensable, for catalysis. On the other hand, F1 with a Gln-269----end gamma subunit was inactive. Replacement of conserved residues (Gln-269, Thr-273, or Glu-275) between Gln-269 and Leu-276 gave enzymes with significantly reduced ATPase activity (2- 41% of that of the wild-type) and lower ATP-driven proton conduction. Thus these residues are required for the normal catalytic activity of F1, although they are not absolutely essential. Membranes with amino acid replacements (Thr-277----end, Gln-269----Leu, or Glu-275----Lys) and the frameshift mutation (downstream of Thr-277) had about 15% of the wild-type ATPase activity, but showed different degrees of ATP- dependent H+ translocation and growth yield by oxidative phosphorylation, suggesting that the gamma subunit, especially its carboxyl-terminal region, functions in coupling between catalysis and H+ translocation
MH  - A
MH  - ACID
MH  - atp
MH  - ATPase
MH  - Biochemistry
MH  - Catalysis
MH  - Chemistry
MH  - coupling
MH  - Dicyclohexylcarbodiimide
MH  - Enzymes
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1
MH  - function
MH  - H(+)ATPase
MH  - H+
MH  - In Vitro
MH  - Macromolecular Systems
MH  - membrane
MH  - Membranes
MH  - mutant
MH  - Oxidative Phosphorylation
MH  - Phosphorylation
MH  - protein
MH  - Proteins
MH  - proton
MH  - RESIDUE
MH  - SUBUNIT
MH  - SYSTEM
MH  - SYSTEMS
MH  - translocation
RP  - NOT IN FILE
NT  - UI - 90202983LA - engRN - 0 (Macromolecular Systems)RN - 0 (Plasmids)RN - 0 (Recombinant Proteins)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19900502IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:2138624
SO  - J Biol Chem 1990 Mar 25 ;265(9):5043-5048

965
UI  - 841
AU  - Jones MR
AU  - McEwan AG
AU  - Jackson JB
AD  - School of Biochemistry, University of Birmingham, UK
TI  - The role of c-type cytochromes in the photosynthetic electron transport pathway of Rhodobacter capsulatus
AB  - (1) Short flash excitation of membrane vesicles of a cytochrome-c2- deficient mutant of Rhodobacter capsulatus (strain MT-G4/S4) led to rapid oxidation of a c-type cytochrome. In redox titrations, the photooxidation of c-type cytochrome was attenuated with a midpoint of approx. +360 mV. Vesicles from a control strain, MT1131, gave similar results. These findings are consistent with those of Prince et al. (Prince, R.C., Davidson, E., Haith, L.E. and Daldal, F. (1986) Biochemistry 25, 5208-5214). (2) In anaerobic intact cells the extent of rapid re-reduction of c-type cytochrome oxidised after a flash was less in MT-G/S4 than in MT1131. Cytochrome c reduction in both strains was inhibited by myxothiazol. The myxothiazol-sensitive component of the electrochromic absorbance change in cells indicated that rapid charge separation through the cytochrome bc1 complex was less extensive after a flash in MT-G4/S4 than in MT 1131. (3) In anaerobic intact cells and in chromatophores of Rb. capsulatus strain MT-GS18, a mutant deficient in both cytochrome c1 and cytochrome c2, flash excitation led to the oxidation of c-type cytochrome. Redox titrations and spectra of chromatophores suggested that this is the same cytochrome as was photooxidized in vesicles of MT-G4/S4 and MT1131. This result is in contrast with earlier findings (Prince, R.C. and Daldal, F. (1987) Biochim. Biophys, Acta 894, 370-378) in which it was reported that no photooxidation of c-type cytochrome occurred in the absence of c1 and c2, and argues against the possibility that cytochrome c1 can rapidly and directly donate electrons to the reaction centre. (4) It is proposed that a previously uncharacterized, membrane-bound c-type cytochrome (Em7 approximately +360 mV) is present in Rb-capsulatus MT1131, in the c2-deficient mutant MT-G4/34 and in the c1/c2-deficient mutant MTGS18. This cytochrome and cytochrome c2 are alternative electron donors to the reaction centre in strain MT1131
RP  - NOT IN FILE
NT  - UI - 90373814LA - engRN - 9007-43-6 (Cytochrome c)RN - 9035-43-2 (cytochrome c2)PT - Journal ArticleDA - 19901016IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:2168749
SO  - Biochim Biophys Acta 1990 Aug 9 ;1019(1):59-66

966
UI  - 85
AU  - Kaim G
AU  - Ludwig W
AU  - Dimroth P
AU  - Schleifer KH
AD  - Lehrstuhl fur Mikrobiologie, Technische Universitat, Munich, FRG
TI  - Sequence of subunits a and b of the sodium ion translocating adenosine triphosphate synthase of Propionigenium modestum
RP  - NOT IN FILE
NT  - UI - 91067471LA - engRN - 7440-23-5 (Sodium)RN - EC 3.6.1.37 (Na(+)-K(+)-Exchanging ATPase)PT - Journal ArticleDA - 19910115IS - 0305-1048SB - IMCY - ENGLANDJC - O8L
UR  - PM:2174545
SO  - Nucleic Acids Res 1990 Nov 25 ;18(22):6697

967
UI  - 21087
AU  - Krulwich TA
AU  - Quirk PG
AU  - Guffanti AA
AD  - Department of Biochemistry, Mount Sinai School of Medicine, City University of New York, New York 10029
TI  - Uncoupler-resistant mutants of bacteria
AB  - The chemiosmotic model of energy transduction offers a satisfying and widely confirmed understanding of the action of uncouplers on such processes as oxidative phosphorylation; the uncoupler, by facilitating the transmembrane movement of protons or other compensatory ions, reduces the electrochemical proton gradient that is posited as the energy intermediate for many kinds of bioenergetic work. In connection with this formulation, uncoupler-resistant mutants of bacteria that neither exclude nor inactivate these agents represent a bioenergetic puzzle. Uncoupler-resistant mutants of aerobic Bacillus species are, in fact, membrane lipid mutants with bioenergetic properties that are indeed challenging in connection with the chemiosmotic model. By contrast, uncoupler-resistant mutants of Escherichia coli probably exclude uncouplers, sometimes only under rather specific conditions. Related phenomena in eucaryotic and procaryotic systems, as well as various observations on uncouplers, decouplers, and certain other membrane-active agents, are also briefly considered
MH  - A
MH  - Bacillus
MH  - Bacteria
MH  - Biochemistry
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - intermediate
MH  - ion
MH  - Ions
MH  - membrane
MH  - model
MH  - Movement
MH  - mutant
MH  - Oxidative Phosphorylation
MH  - Phosphorylation
MH  - proton
MH  - Protons
MH  - review
MH  - SYSTEM
MH  - SYSTEMS
MH  - Uncoupling Agents
RP  - NOT IN FILE
NT  - UI - 90205733LA - engRN - 0 (Uncoupling Agents)PT - Journal ArticlePT - ReviewPT - Review, AcademicID - GM28454/GM/NIGMSDA - 19900503IS - 0146-0749SB - IMCY - UNITED STATES
UR  - PM:2181259
SO  - Microbiol Rev 1990 Mar ;54(1):52-65

968
UI  - 21086
AU  - Krulwich TA
AU  - Guffanti AA
AU  - Seto-Young D
AD  - Department of Biochemistry, Mount Sinai School of Medicine, City University of New York, NY
TI  - pH homeostasis and bioenergetic work in alkalophiles
AB  - A Na+/H+ antiporter catalyses coupled Na+ extrusion and H+ uptake across the membranes of extremely alkalophilic bacilli. This exchange is electrogenic, with H+ translocated inward greater than Na+ extruded. It is energized by the delta chi 2 component of the delta mu H+ that is established during primary proton pumping by the alkalophile respiratory chain complexes. These complexes abound in the membranes of extreme alkalophiles. Combined activity of the respiratory chain, the antiporter, and solute transport systems that are coupled to Na+ re- entry, allow the alkalophiles to maintain a cytoplasmic pH that is several pH units more acidic than optimal external pH values for growth. There is no compelling evidence for a specific and necessary role for any ion other than sodium in pH homeostasis, and although there is very high cytoplasmic buffering capacity in the alkaline range, active mechanisms for pH homeostasis are crucial. Energization of the antiporter as well as the proton translocating F1F0-ATPase that catalyses ATP synthesis in the extreme alkalophiles must accommodate the problem of the low net delta mu H+ and the very low concentrations of protons, per se, in the external medium. This problem is by-passed by other bioenergetic work functions, such as solute uptake or motility, that utilize sodium ions for energy-coupling in the place of protons
MH  - A
MH  - ACTIVE
MH  - atp
MH  - ATP synthesis
MH  - Biochemistry
MH  - Carrier Proteins
MH  - COMPLEX
MH  - delta
MH  - electrogenic
MH  - function
MH  - H+
MH  - Homeostasis
MH  - ion
MH  - Ions
MH  - mechanism
MH  - MECHANISMS
MH  - membrane
MH  - Membranes
MH  - pH
MH  - protein
MH  - Proteins
MH  - proton
MH  - Protons
MH  - review
MH  - Sodium
MH  - synthesis
MH  - SYSTEM
MH  - SYSTEMS
MH  - transport
RP  - NOT IN FILE
NT  - UI - 90351625LA - engRN - 0 (Carrier Proteins)RN - 0 (Sodium-Hydrogen Antiporter)RN - 7440-23-5 (Sodium)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 19900924IS - 0168-6445SB - IMCY - NETHERLANDS
UR  - PM:2167108
SO  - FEMS Microbiol Rev 1990 Jun ;6(2-3):271-278

969
UI  - 21020
AU  - Labahn A
AU  - Fromme P
AU  - Graber P
TI  - Detecting precatalytic conformational changes in F1-ATPase with 4-benzoyl(benzoyl)-1-amidofluorescein, a novel fluorescent nucelotide site-specific photoaffinity label
MH  - A
MH  - atp
MH  - conformation
MH  - conformational change
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - fluorescence
MH  - photoaffinity
RP  - NOT IN FILE
SO  - FEBS Lett 1990  ;271():116-118

970
UI  - 21007
AU  - Labahn A
AU  - Fromme P
AU  - Graber P
AD  - Max Volmer Institut fur Biophysikalische und Physikalische Chemie, Technische Universitat Berlin, FRG
TI  - Uni-site ATP synthesis in thylakoids
AB  - Uni-site ATP synthesis was measured with thylakoids. The membrane-bound ATP-synthase, CF0F1, was brought into the active, reduced state by illumination in the presence of thioredoxin, dithiothreitol and phosphate. This enzyme contains two tightly bound ATP per CF0F1. ATP was released from the enzyme when ADP was added in substoichiometric amounts during illumination. Experiments with [14C]ADP indicated that after binding the same nucleotide was phosphorylated and released as [14C]ATP, i.e. only one site is involved in ATP-synthesis ('uni-site ATP-synthesis'). The two tightly bound ATP are not involved in the catalytic turnover. The rate constant for ADP binding was (4 +/- 2) x 10(6) M-1s-1. Compared to deenergized conditions the rate constant for ADP binding and that for ATP-release were drastically increased, i.e. membrane energization increased the rate constants for the ATP- synthesis direction
MH  - ACTIVE
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - ADP
MH  - ADP BINDING
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - ATPase
MH  - BINDING
MH  - CF0F1
MH  - CONSTANT
MH  - Dithiothreitol
MH  - membrane
MH  - rate constant
MH  - Site
MH  - synthesis
MH  - thylakoid
MH  - thylakoids
RP  - NOT IN FILE
NT  - UI - 91032045LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19901204IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:2226796
SO  - FEBS Lett 1990 Oct 1 ;271(1-2):116-118

971
UI  - 1132
AU  - Labahn A
AU  - Graber P
TI  - Uni-site ATP hydrolysis catalyzed by the ATP-synthase from chloroplasts
MH  - atp
MH  - ATP synthase
MH  - chloroplast
MH  - Chloroplasts
MH  - Hydrolysis
MH  - unisite
T2  - Curr. Res. Photosynth., Proc. Int. Conf. Photosynth., 8th, Meeting Date 1989, Volume 3, 37-40. Edited by: Baltscheffsky, Margareta. Kluwer: Dordrecht, Neth
Y2  - -32676  
RP  - ON REQUEST (03/18/92)
SO  -  1990  ;():

972
UI  - 87
AU  - Laubinger W
AU  - Deckers-Hebestreit G
AU  - Altendorf K
AU  - Dimroth P
AD  - Institut fur Physiologische Chemie, Technischen Universitat Munchen, FRG
TI  - A hybrid adenosinetriphosphatase composed of F1 of Escherichia coli and F0 of Propionigenium modestum is a functional sodium ion pump
AB  - Analyses on immunoblots indicated strong binding of the alpha- and beta- subunits of the ATPase of Propionigenium modestum to antibodies raised against the corresponding subunits of the F1F0 ATPase of Escherichia coli. Cross-reactivities of antibodies against the other ATPase subunits were not observed. The use of Na+ or H+ as alternate coupling ions, observed previously for the P. modestum ATPase [Laubinger, W., & Dimroth, P. (1989) Biochemistry 28, 7194-7198], is not found for the F1F0 ATPase of E. coli, which is specific for protons. However, a hybrid consisting of the F1 moiety of the E. coli ATPase and F0 of that from P. modestum performed Na+ or H+ transport in a reconstituted system. As with the homologous ATPase of P. modestum, H+ pumping of the hybrid was abolished at Na+ concentrations of greater than 1 mM. The F0 sector and not F1, therefore, determines the cation specificity of these F1F0 ATPases
RP  - NOT IN FILE
NT  - UI - 90352295LA - engRN - 0 (Proteolipids)RN - 0 (proteoliposomes)RN - 1333-74-0 (Hydrogen)RN - 7440-23-5 (Sodium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19900927IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:2167128
SO  - Biochemistry 1990 Jun 12 ;29(23):5458-5463

973
UI  - 522
AU  - Lee JH
AU  - Garboczi DN
AU  - Thomas PJ
AU  - Pedersen PL
AD  - Department of Biological Chemistry, Johns Hopkins School of Medicine, Baltimore, Maryland 21205
TI  - Mitochondrial ATP synthase. cDNA cloning, amino acid sequence, overexpression, and properties of the rat liver alpha subunit
AB  - The predicted amino acid sequence of the alpha subunit of the rat liver mitochondrial ATP synthase has been obtained by sequencing a cDNA for the alpha subunit. Analysis of the sequence shows that it contains the A and B consensus sequences found in many nucleotide-binding proteins. Twelve amino acids of the rat liver alpha subunit differ from the sequence of the bovine heart alpha subunit; four of these involve differences in charge. The rat liver alpha subunit, from arginine 15 to the C-terminal proline 510, has been overexpressed in Escherichia coli using the alkaline phosphatase promoter (phoA) and leader peptide to direct the export of the expressed protein to the bacterial periplasm. By treating the cells with lysozyme, osmotic shock, and alkaline pH washes, the alpha subunit can be extracted in high yield (greater than 25 mg/liter) and in a high state of purity. The expressed alpha subunit remains soluble at pH 9.5 or greater and precipitates when treated with Mg2+ ions at low millimolar concentration. The bacterially expressed alpha subunit interacts with 2'(3')-O-(2,4,6-trinitrophenyl)adenosine 5'-triphosphate (TNP-ATP), resulting in a marked fluorescence enhancement upon binding. An enhancement of fluorescence is also observed upon the interaction of the alpha subunit with TNP-ADP. Preincubating the alpha subunit with 1.5 mM ATP significantly reduces the fluorescence enhancement seen with TNP-ATP. The alpha subunit binds TNP-ATP with an apparent Kd in the low micromolar range (1-5 microM) and binds TNP-ADP with an affinity at least 10-fold lower. This work shows that the rat liver alpha subunit can be overexpressed in E. coli to yield a large amount of functional protein. With the acquisition of the overexpressed alpha subunit, it is now possible to test the reconstitution of ATPase activity from a mixture of recombinant and rat liver-derived subunits and to test the formation of complexes by the overexpressed alpha and beta subunits of the rat liver F1-ATPase
RP  - NOT IN FILE
NT  - UI - 90170980LA - engRN - 0 (2',3'-O-(2,4,6-trinitrophenyl)adenosine 5'-triphosphate)RN - 56-65-5 (Adenosine Triphosphate)RN - 7439-95-4 (Magnesium)RN - 9007-49-2 (DNA)RN - EC 3.2.1.17 (Muramidase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - CA 10951/CA/NCIDA - 19900409IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:2137825
SO  - J Biol Chem 1990 Mar 15 ;265(8):4664-4669

974
UI  - 21236
AU  - Leikin S
AU  - Kornyshev AA
TI  - Theory of hydration forces. Nonlocal electrostatic interaction of neutral surfaces
RP  - IN FILE
SO  - J Chem Phys 1990  ;92():6890-6898

975
UI  - 184
AU  - Lucken U
AU  - Gogol EP
AU  - Capaldi RA
AD  - Institute of Molecular Biology, University of Oregon, Eugene 97403
TI  - Structure of the ATP synthase complex (ECF1F0) of Escherichia coli from cryoelectron microscopy
AB  - The structural relationship of the catalytic portion (ECF1) of the Escherichia coli F1F0 ATP synthase (ECF1F0) to the intact, membrane- bound complex has been determined by cryoelectron microscopy and image analysis of single, unordered particles. ECF1F0, reconstituted into membrane structures, has been preserved and examined in its native state in a layer of amorphous ice. Side views of the ECF1F0 show the same elongated bilobed and trilobed projection of the ECF1 views shown previously to be normal to the hexagonal projection. The elongated aqueous cavity of the ECF1 is perpendicular to the membrane bilayer profile in the bilobed view. ECF1 is separated from the membrane- embedded F0 by a narrow stalk approximately 40 A long and approximately 25-30 A thick. The F0 part extends from the lipid bilayer by approximately 10 A on the side facing the ECF1. There is no clear extension of the protein on the opposite side of the membrane
RP  - NOT IN FILE
NT  - UI - 90344773LA - engRN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM39806/GM/NIGMSID - HL24526/HL/NHLBIID - RR02756/RR/NCRRDA - 19900920IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:2200506
SO  - Biochemistry 1990 Jun 5 ;29(22):5339-5343

976
UI  - 86
AU  - Ludwig W
AU  - Kaim G
AU  - Laubinger W
AU  - Dimroth P
AU  - Hoppe J
AU  - Schleifer KH
AD  - Lehrstuhl fur Mikrobiologie, Technische Universitat Munchen, Federal Republic of Germany
TI  - Sequence of subunit c of the sodium ion translocating adenosine triphosphate synthase of Propionigenium modestum
AB  - The 30 N-terminal amino acid residues of the purified ATPase c subunit of Propionigenium modestum have been determined. An oligonucleotide mixture was derived from this sequence and used as probe for cloning the corresponding gene in Escherichia coli. The nucleotide sequence of the gene has been determined and compared with those of ATPase c subunits from other bacteria and chloroplasts. Peculiar sequence similarities are found only at the C-terminus between the c subunits of the ATPases from P. modestum and from Vibrio alginolyticus, another putative Na(+)-translocating ATPase
RP  - NOT IN FILE
NT  - UI - 91031485LA - engRN - 0 (Carrier Proteins)RN - 7440-23-5 (Sodium)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19901213IS - 0014-2956SB - IMCY - GERMANYJC - EMZ
UR  - PM:2146118
SO  - Eur J Biochem 1990 Oct 24 ;193(2):395-399

977
UI  - 573
AU  - Matsuno-Yagi A
AU  - Hatefi Y
AD  - Department of Molecular and Experimental Medicine, Research Institute of Scripps Clinic, La Jolla, California 92037
TI  - Studies on the mechanism of oxidative phosphorylation. ADP promotion of GDP phosphorylation
AB  - The process of ATP or GTP synthesis by bovine heart submitochondrial particles involves the binding of ADP or GDP to 3 exchangeable sites I, II, and III, and only upon substrate occupation of site III does rapid ATP or GTP synthesis take place. The dissociation constants determined for ADP were KADPI less than or equal to 10(-8) M, KADPII approximately 10(-7) M, and KADPIII (equivalent to apparent KADPm), approximately 3 x 10(-6) M in the low Km mode and KADPIII approximately 150 x 10(-6) M in the high Km mode. For GDP, these constants were KGDPI approximately 10(- 6)-10(-5) M, KGDPII approximately 10(-4) M, and KGDPIII approximately 10(-3) M when NADH was the respiratory substrate (Matsuno-Yagi, A., and Hatefi, Y. (1990) J. Biol. Chem. 265, 82-88). Because of its low affinity for the above binding sites, GDP at micromolar concentrations does not lead to GTP synthesis. However, as shown in this paper, micromolar [GDP] undergoes phosphorylation in the presence of micromolar concentrations of ADP. Under these conditions, both ATP and GTP are synthesized. GDP inhibits ATP synthesis with KGDPi congruent to 7 microM, while ADP promotes GTP synthesis in a reaction that requires inorganic phosphate (apparent KPim = 2-3 mM) and is inhibited by uncouplers and inhibitors of the ATP synthase complex. The ADP-promoted GTP synthesis exhibited an "apparent" KGDPm = 4 microM and an "apparent" Vmax = 11 nmol of GTP (min.mg of protein)-1. These results were interpreted to mean that (a) micromolar [ADP] occupies sites I and II, allowing site III to bind and phosphorylate GDP, and (b) the KGDPm and Vmax calculated under these conditions represent values for the low Km-low Vmax mode of GTP synthesis, which in the absence of ADP is not detectable because of the positive cooperativity phase of GTP synthesis with the high KGDPII approximately 10(-4) M
RP  - NOT IN FILE
NT  - UI - 91056074LA - engRN - 0 (Phosphates)RN - 146-91-8 (Guanosine Diphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 86-01-1 (Guanosine Triphosphate)PT - Journal ArticleID - DK08126/DK/NIDDKDA - 19901228IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:2243094
SO  - J Biol Chem 1990 Nov 25 ;265(33):20308-20313

978
UI  - 574
AU  - Matsuno-Yagi A
AU  - Hatefi Y
AD  - Department of Molecular and Experimental Medicine, Research Institute of Scripps Clinic, La Jolla, California 92037
TI  - Studies on the mechanism of oxidative phosphorylation. Positive cooperativity in ATP synthesis
AB  - Kinetic and nucleotide binding studies have shown that submitochondrial particles from bovine heart possess three exchangeable binding sites for ADP or GDP. In order of decreasing affinity at neutral pH, these sites will be referred to as sites I, II, and III, and their respective dissociation constants as KI, KII, and KIII. In oxidative phosphorylation experiments in the presence of saturating amounts of inorganic phosphate, rapid ATP (or GTP) synthesis occurred only upon ADP (or GDP) binding to site III. The Eadie-Hofstee plots (v/[S] on the ordinate versus v on the abscissa) of the kinetics of ATP (or GTP) synthesis at variable ADP (or GDP) were, therefore, composed of an initial upward phase, indicating positive cooperativity with respect to substrate concentration, followed by a downward phase where rapid product formation took place. These data allowed calculation of KII from the upward phase and KIII (equivalent to apparent Km) from the downward phase. KI was estimated from Scatchard plots of binding data with radiolabeled ADP or GDP. Thus, together with our previous results, these findings have allowed characterization of the process of ATP or GTP synthesis by bovine-heart submitochondrial particles in terms of KI, KII, KIII, and kcat
RP  - NOT IN FILE
NT  - UI - 90094463LA - engRN - 146-91-8 (Guanosine Diphosphate)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 86-01-1 (Guanosine Triphosphate)PT - Journal ArticleID - DK08126/DK/NIDDKDA - 19900202IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:2294123
SO  - J Biol Chem 1990 Jan 5 ;265(1):82-88

979
UI  - 19783
AU  - Milgrom YM
AU  - Ehler LL
AU  - Boyer PD
AD  - Department of Chemistry and Biochemistry and Molecular Biology Institute, University of California, Los Angeles 90024
TI  - ATP binding at noncatalytic sites of soluble chloroplast F1-ATPase is required for expression of the enzyme activity
AB  - The F1-ATPase from chloroplasts (CF1) lacks catalytic capacity for ATP hydrolysis if ATP is not bound at noncatalytic sites. CF1 heat activated in the presence of ADP, with less than one ADP and no ATP at non-catalytic sites, shows a pronounced lag in the onset of ATP hydrolysis after exposure to 5-20 microM ATP. The onset of activity correlates well with the binding of ATP at the last two of the three noncatalytic sites. The dependence of activity on the presence of ATP at non-catalytic sites is shown at relatively low or high free Mg2+ concentrations, with or without bicarbonate as an activating anion, and when the binding of ATP at noncatalytic sites is slowed 3-4-fold by sulfate. The latent CF1 activated by dithiothreitol also requires ATP at noncatalytic sites for ATPase activity. A similar requirement by other F1-ATPases and by ATP synthases seems plausible
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - ADP
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BINDING
MH  - Biochemistry
MH  - Chemistry
MH  - chloroplast
MH  - Chloroplasts
MH  - DEPENDENCE
MH  - Dithiothreitol
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - H(+)-Transporting ATP Synthase
MH  - Heat
MH  - Hydrolysis
MH  - Macromolecular Systems
MH  - Magnesium
MH  - non-catalytic site
MH  - NONCATALYTIC SITES
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 91035377LA - engRN - 0 (Macromolecular Systems)RN - 56-65-5 (Adenosine Triphosphate)RN - 7439-95-4 (Magnesium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM-11094/GM/NIGMSDA - 19901210IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:2146260
SO  - J Biol Chem 1990 Nov 5 ;265(31):18725-18728

980
UI  - 19784
AU  - Milgrom YM
AU  - Boyer PD
AD  - Department of Chemistry and Biochemistry, University of California, Los Angeles
TI  - The ADP that binds tightly to nucleotide-depleted mitochondrial F1- ATPase and inhibits catalysis is bound at a catalytic site
AB  - Previous studies have shown that the initial complex formed when ADP binds to nucleotide-depleted F1-ATPase is transformed with a half time of 2 to 3 min to form with a much lower rate of ADP release. The ADP binding results in a strong inhibition of ATPase activity. The present paper reports appraisal of where the inhibitory ADP binds by use of the photoreactive ADP analog, 2-N3-ADP. In presence of Mg2+ the 2-N3-ADP like ADP induces reversible inhibition of nucleotide-depleted F1 (ndF1) with a Kd of about 10 nM. Photoirradiation of the inactive 2-N3-[beta- 32P]ADP-ndF1 complex results in labeling of only the beta-subunit. The major labeled peptide isolated from a trypic digest consists of residues from Ala-338 to Arg-356, with Tyr-345 as the site of labeling. This identifies the site of the inhibitory ADP binding as one of the catalytic sites of the enzyme
MH  - A
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - ADP
MH  - ADP BINDING
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Azides
MH  - BETA-SUBUNIT
MH  - BINDING
MH  - Biochemistry
MH  - Catalysis
MH  - Chemistry
MH  - COMPLEX
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - H(+)-Transporting ATP Synthase
MH  - Nadp
MH  - Peptide Fragments
MH  - SYNTHASE
MH  - Time
MH  - Trypsin
RP  - NOT IN FILE
NT  - UI - 91027772LA - engRN - 0 (Affinity Labels)RN - 0 (Azides)RN - 0 (Peptide Fragments)RN - 53-59-8 (NADP)RN - 58-64-0 (Adenosine Diphosphate)RN - 64020-53-7 (2-azidoadenosine 3',5'-diphosphate)RN - EC 3.4.21.4 (Trypsin)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM-I 11094/GM/NIGMSDA - 19901211IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:2145975
SO  - Biochim Biophys Acta 1990 Oct 24 ;1020(1):43-48

981
UI  - 43
AU  - Miller MJ
AU  - Oldenburg M
AU  - Fillingame RH
AD  - Department of Physiological Chemistry, University of Wisconsin Medical School, Madison 53706
TI  - The essential carboxyl group in subunit c of the F1F0 ATP synthase can be moved and H(+)-translocating function retained
AB  - The proteolipid subunit c of F1F0-type H(+)-transporting ATP synthases [ATP phosphohydrolase (H(+)-transporting), EC 3.6.1.34] contains a conserved Asp/Glu residue that is thought to function in H+ translocation. To test the importance of the position of this residue in the Escherichia coli enzyme, we used oligonucleotide-directed mutagenesis to move the carboxyl side chain from position 61 to position 58, 60, or 62. Mutant cells with these changes were incapable of growth via oxidative phosphorylation on succinate. An Asp-61----Glu mutant grew on succinate but at 50% the efficiency of wild type. Hence, even minor changes in the position of the carboxyl group can significantly reduce function. In a second approach, slow-growing revertants to an Asp-61----Gly mutant were isolated. In one such revertant, Ala-24 was changed to Asp, while the original Asp-61----Gly mutation remained unchanged. The Asp-24-Gly-61 double mutant grew on succinate at 60% the efficiency of wild type. Hence the essential carboxyl group of subunit c can function when anchored at either position 24 or position 61, and this supports the idea that these residues may neighbor each other when subunit c is folded in the membrane. The rate of ATP-driven H+ translocation by mutant membrane vesicles was estimated by the quenching of 9-amino-6-chloro-2- methoxyacridine fluorescence and corresponded to actual H+ pumping rates less than 25% that of wild type
RP  - NOT IN FILE
NT  - UI - 90311311LA - engRN - 0 (Macromolecular Systems)RN - 0 (Oligonucleotide Probes)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - F32 GM09942/GM/NIGMSID - GM23105/GM/NIGMSDA - 19900815IS - 0027-8424SB - IMCY - UNITED STATESJC - PV3
UR  - PM:2142302
SO  - Proc Natl Acad Sci U S A 1990 Jul ;87(13):4900-4904

982
UI  - 1007
AU  - Mitchell P
TI  - Osmochemistry of solute translocation
AB  - The main aim of this brief contribution is to suggest that our understanding of the general principles of osmochemistry may provide useful insights into the type of mechanism by which solute-translocating catalysts work. In particular, I would like to encourage a more widespread and explicit recognition of the special merits of the mobile barrier type of mechanism (Mitchell, 1957, 1987), not as a panacea, but to explain the translocation of the characteristically hydrophilic and somewhat bulky solutes that are the main substrates of solute porters and of some osmoenzymes in bacterial membranes
MH  - Bacteria
MH  - Bacterial Proteins
MH  - Biological Transport,Active
MH  - Carrier Proteins
MH  - Cell Compartmentation
MH  - Chemistry,Physical
MH  - Membrane Proteins
MH  - Membranes
MH  - metabolism
MH  - Osmosis
MH  - Solubility
RP  - NOT IN FILE
NT  - Glynn Research Institute, Bodmin, Cornwall, UK
SO  - Res Microbiol 1990 Mar ;141(3):286-289

983
UI  - 752
AU  - Odaka M
AU  - Kobayashi H
AU  - Muneyuki E
AU  - Yoshida M
AD  - Department of Life Science, Tokyo Institute of Technology, Yokohama, Japan
TI  - Aromatic rings of tyrosine residues at adenine nucleotide binding sites of the beta subunits of F1-ATPase are not necessary for ATPase activity
AB  - Using site-directed mutagenesis, Tyr-307, Tyr-341, or Tyr-364, supposedly located at the adenine nucleotide binding site(s) of the beta subunits of F1-ATPase from the thermophilic bacterium PS3, was replaced with Phe or Cys. The alpha 3 beta 3 complexes reconstituted from the alpha subunits and individual mutant beta subunits hydrolyzed ATP. Thus, neither the hydroxyl groups nor the aromatic rings in these positions are required for ATPase activity of F1-ATPase
RP  - NOT IN FILE
NT  - UI - 90226387LA - engRN - 0 (Macromolecular Systems)RN - 0 (Nucleotides)RN - 55520-40-6 (Tyrosine)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19900518IS - 0006-291XSB - IMCY - UNITED STATESJC - 9Y8
UR  - PM:2139333
SO  - Biochem Biophys Res Commun 1990 Apr 16 ;168(1):372-378

984
UI  - 446
AU  - Pagan J
AU  - Senior AE
AD  - Department of Biochemistry, University of Rochester Medical Center, New York 14642
TI  - Mutations in alpha-subunit of Escherichia coli F1-ATPase obtained by hydroxylamine-mutagenesis of plasmids carrying the uncA gene
AB  - In order to generate mutants randomly in the Escherichia coli uncA gene (encoding the alpha-subunit of F1-ATPase), plasmids carrying uncA were treated in vitro with hydroxylamine. Restriction fragments of the mutated uncA gene were then reconstructed into plasmid pDP34, which expresses all of the F1F0 structural genes, and the reconstructed mutant plasmids were expressed in a strain carrying a deletion of chromosomal uncA. Each of the mutations was characterized by DNA sequencing, growth assays, and biochemical assays of membrane preparations. Three nonsense and one frameshift mutation were identified and their properties were studied briefly. Eight new missense mutations were identified and characterization of their properties is described. These eight mutations were R139H, A177V, R210C, R303C, A306V, T343I, G351S, and P370L
RP  - NOT IN FILE
NT  - UI - 90179188LA - engRN - 0 (Hydroxylamines)RN - 0 (Macromolecular Systems)RN - 0 (Plasmids)RN - 7803-49-8 (Hydroxylamine)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSDA - 19900406IS - 0003-9861SB - IMCY - UNITED STATESJC - 6SK
UR  - PM:2138002
SO  - Arch Biochem Biophys 1990 Mar ;277(2):283-289

985
UI  - 443
AU  - Pagan J
AU  - Senior AE
AD  - Department of Biochemistry, University of Rochester Medical Center, NY 14642
TI  - Tight ATP and ADP binding in the noncatalytic sites of Escherichia coli F1-ATPase is not affected by mutation of bulky residues in the 'glycine- rich loop'
AB  - It is shown that ATP dissociates very slowly (koff less than 6.4 x 10(5) s-1, t1/2 greater than 3 h) from the three noncatalytic sites of E. coli F1-ATPase and that ADP dissociates from these three sites in a homogeneous fashion with koff = 1.5 x 10(-4) s-1 (t1/2 = 1.35 h). Mutagenesis of alpha-subunit residues R171 and Q172 in the 'glycine- rich loop' (Homology A) consensus region of the noncatalytic sites was carried out to test the hypothesis that unusually bulky residues at these positions are responsible wholly or partly for the observed tight binding of adenine nucleotides. The mutations alpha Q172G or alpha R171S,Q172G had no effects on ATP or ADP binding to or rates of dissociation from F1 noncatalytic sites. KdATP and KdADP of isolated alpha-subunit were weakened by approximately 1 order of magnitude in both mutants. The results suggest that neither residue alpha R171 nor alpha Q172 interacts directly with bound nucleotide, and show that the presence of bulky residues per se in the glycine-rich loop region of F1- alpha-subunit is not responsible for tight binding in the noncatalytic sites
RP  - NOT IN FILE
NT  - UI - 91032165LA - engRN - 0 (Macromolecular Systems)RN - 56-40-6 (Glycine)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM 25349/GM/NIGMSDA - 19901213IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:2146165
SO  - FEBS Lett 1990 Oct 29 ;273(1-2):147-149

986
UI  - 21293
AU  - Pagan J
AU  - Senior AE
TI  - Structures of the efrapeptins: potent inhibitors of mitochondrial ATPase from the fungus Tolypocladium niveum
MH  - atp
MH  - ATPase
MH  - inhibitor
MH  - inhibitors
MH  - structure
RP  - NOT IN FILE
SO  - FEBS Lett 1990  ;273():147-149

987
UI  - 21109
AU  - Perez JA
AU  - Ferguson SJ
AD  - Department of Biochemistry, University of Oxford, UK
TI  - Kinetics of oxidative phosphorylation in Paracoccus denitrificans. 1. Mechanism of ATP synthesis at the active site(s) of F0F1-ATPase
AB  - (1) The rate of ATP synthesis during NADH-driven aerobic respiration has been measured in plasma membrane vesicles from Paracoccus denitrificans as a function of the concentration of the substrates, ADP and inorganic phosphate (Pi). In both cases, the response of the reaction to changes in the degree of saturation of the F0F1-ATPase generated a perfect Micaelian dependence which allowed the determination of the corresponding Michaelis constants, KmADP and KmPi. (2) These kinetic parameters possess a real mechanistic significance, as concluded from the partial reduction of the rate of phosphorylation by the energy-transfer inhibitor venturicidin and the consequent analysis of the results within the framework of the theory of metabolic control. (3) The same membrane vesicles, which catalyze very high rates of ATP synthesis, have been shown to support much lower rates of the exchange ATP in equilibrium Pi and negligible rates of ATP hydrolysis. Under similar conditions, the preparations are also capable of generating phosphorylation potentials, delta Gp, of 60-61 kJ.mol-1. (4) These properties have allowed analysis of the synthetic reaction in the presence of significant concentrations of the product, ATP, using integrated forms of the Michaelis-Menten rate equations. (5) It has been shown that ATP produces pure competitive product inhibition of the forward reaction with a value of KiATP = 16 +/- 1 microM, thus indicating that the affinity of the nucleotide for the active site(s) of the F0F1-ATPase, during net ATP synthesis, is significantly higher than previously thought. (6) The order of binding of the substrates, ADP and Pi, to the active site(s) has been determined as random. (7) At very low concentrations of ADP, a second and much smaller Michaelis constant for this substrate has been identified, with an estimated value of KmADP approximately equal to 50 nM, associated with a maximal rate of only 2% of that measured at a higher range of concentrations. (8) The results obtained are discussed in relation to the presence of two or three equivalent catalytic sites operating in the cooperative manner explicitly described by the binding change mechanism
MH  - A
MH  - ACTIVE
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - ADP
MH  - analysis
MH  - atp
MH  - ATP synthesis
MH  - ATPase
MH  - BINDING
MH  - Biochemistry
MH  - CHANGE MECHANISM
MH  - CONSTANT
MH  - delta
MH  - DEPENDENCE
MH  - Energy Transfer
MH  - function
MH  - Hydrolysis
MH  - inhibitor
MH  - INORGANIC-PHOSPHATE
MH  - Kinetics
MH  - mechanism
MH  - membrane
MH  - membrane vesicles
MH  - Oxidative Phosphorylation
MH  - Phosphorylation
MH  - Proton-Translocating ATPases
MH  - Respiration
MH  - Site
MH  - synthesis
MH  - theory
MH  - vesicles
RP  - NOT IN FILE
NT  - UI - 91104961LA - engRN - 0 (Venturicidins)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19910226IS - 0006-2960SB - IMCY - UNITED STATES
UR  - PM:2148690
SO  - Biochemistry 1990 Nov 20 ;29(46):10503-10518

988
UI  - 21108
AU  - Perez JA
AU  - Ferguson SJ
AD  - Department of Biochemistry, University of Oxford, UK
TI  - Kinetics of oxidative phosphorylation in Paracoccus denitrificans. 2. Evidence for a kinetic and thermodynamic modulation of F0F1-ATPase by the activity of the respiratory chain
AB  - (1) The affinity of the F0F1-ATPase from Paracoccus denitrificans for ATP during NADH-driven oxidative phosphorylation has been analyzed under different conditions by examining the type and extent of product inhibition. (2) A limited collapse of the protonmotive force (delta p) due to partial uncoupling does not increase the affinity for ATP at the active site(s) of the enzyme; instead, a partial noncompetitive inhibition becomes apparent, compatible with the binding of ATP to a noncatalytic site (or sites) with high affinity. (3) In contrast, partial inhibition of the electron-transport chain increases the extent of pure competitive product inhibition and, therefore, the affinity for ATP at the active site(s). (4) The results are interpreted as indicative of a modulation of the rate of ATP release from the active site(s) of the F0F1-ATPase which is controlled by the activity of the electron-transport chain and not by delta p
MH  - A
MH  - ACTIVE
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - Antibiotics
MH  - atp
MH  - ATPase
MH  - BINDING
MH  - Biochemistry
MH  - Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone
MH  - Cyanides
MH  - delta
MH  - Electron Transport
MH  - Kinetics
MH  - Oxidative Phosphorylation
MH  - P
MH  - Phosphorylation
MH  - Proton-Translocating ATPases
MH  - Rotenone
MH  - Site
MH  - Thiazoles
RP  - NOT IN FILE
NT  - UI - 91104962LA - engRN - 0 (Antibiotics, Antifungal)RN - 0 (Cyanides)RN - 0 (Thiazoles)RN - 370-86-5 (Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone)RN - 56-65-5 (Adenosine Triphosphate)RN - 76706-55-3 (myxothiazol)RN - 83-79-4 (Rotenone)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19910226IS - 0006-2960SB - IMCY - UNITED STATES
UR  - PM:2148691
SO  - Biochemistry 1990 Nov 20 ;29(46):10518-10526

989
UI  - 21021
AU  - Richard P
AU  - Rigaud JL
AU  - Graber P
TI  - A .DELTA.pH clamp method for analysis of steady-state kinetics of photophosphorylation
MH  - A
MH  - analysis
MH  - atp
MH  - Kinetics
MH  - method
MH  - Photophosphorylation
MH  - pmf
RP  - NOT IN FILE
SO  - Eur J Biochem 1990  ;193():921-925

990
UI  - 21006
AU  - Richard P
AU  - Rigaud JL
AU  - Graber P
AD  - Max-Volmer-Institut, Technische Universitat Berlin, Federal Republic of Germany
TI  - Reconstitution of CF0F1 into liposomes using a new reconstitution procedure
AB  - The H(+)-ATPase (ATP synthase) from chloroplasts was isolated, purified and reconstituted into phosphatidylcholine/phosphatidic-acid liposomes. Liposomes prepared by reverse-phase evaporation were treated with various amounts of Triton X-100 and protein incorporation was studied at each step of the solubilization process. After detergent removal by SM2-Biobeads, the activities of the resulting proteoliposomes were measured indicating that the most efficient reconstitution was obtained by insertion of the protein into preformed, detergent-saturated liposomes. The conditions for the reconstitution were optimized with regard to ATP synthesis driven by an artificially generated delta pH/delta psi. An important benefit of the new reconstituted CF0F1 liposomes is the finding that the rate of ATP synthesis remains constant up to 10 s, indicating a low basal membrane permeability
MH  - A
MH  - ACID
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - ATPase
MH  - CF0F1
MH  - chloroplast
MH  - Chloroplasts
MH  - CONSTANT
MH  - Detergents
MH  - H(+)ATPase
MH  - Liposomes
MH  - membrane
MH  - Octoxynol
MH  - Permeability
MH  - Phosphatidic Acids
MH  - Phosphatidylcholines
MH  - Polyethylene Glycols
MH  - protein
MH  - proteoliposome
MH  - PSI
MH  - RECONSTITUTED CF0F1
MH  - reconstitution
MH  - SYNTHASE
MH  - synthesis
RP  - NOT IN FILE
NT  - UI - 91065402LA - engRN - 0 (Detergents)RN - 0 (Liposomes)RN - 0 (Phosphatidic Acids)RN - 0 (Phosphatidylcholines)RN - 0 (Polyethylene Glycols)RN - 56-65-5 (Adenosine Triphosphate)RN - 9002-93-1 (Octoxynol)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19910111IS - 0014-2956SB - IMCY - GERMANY
UR  - PM:2147417
SO  - Eur J Biochem 1990 Nov 13 ;193(3):921-925

991
UI  - 9917
AU  - Rigoulet M
TI  - Control processes in oxidative phosphorylation: kinetic constraints and stoichiometry.
AB  - Control processes in oxidative phosphorylation have been studied in three experimental models. (1) In isolated yeast mitochondria, external ATP is a regulatory effector of cytochrome-c oxidase activity. In phosphorylating or uncoupling states, the relationships between respiratory rate and delta mu H+, and the respiratory rate and cytochrome-c oxidase reduction level are dependent on this kinetic regulation. (2) In rat liver mitochondria, the response of the respiratory rate to uncoupler addition is age-dependent: liver mitochondria isolated from young rats maintain a greater delta mu H+ than liver mitochondria isolated from adults, with the same respiratory rate obtained with the same concentration of uncoupler. This behaviour is linked to redox proton pump properties, i.e., to the degree of intrinsic uncoupling induced by uncoupler addition. (3) The effect of almitrine, a new kind of ATPase/ATPsynthase inhibitor, was studied in mammalian mitochondria. (i) Almitrine inhibits oligomycin-sensitive ATPase - it decreases the ATPase/O value without any change in delta mu H+; (ii) almitrine increased the mechanistic H+/ATP stoichiometry of ATPase/ATPsynthase; (iii) almitrine-induced changes in H+/ATPase stoichiometry depend on the flux magnitude through ATPase. These results are discussed in terms of the following interdependent parameters; flux value, force, pump efficiency and control coefficient.
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - Adult
MH  - Animal
MH  - atp
MH  - ATPase
MH  - Biological Transport
MH  - Cattle
MH  - Cytochrome c
MH  - Cytochrome-c Oxidase
MH  - H+
MH  - Kinetics
MH  - Liver
MH  - metabolism
MH  - Mitochondria
MH  - Mitochondria,Heart
MH  - Mitochondria,Liver
MH  - model
MH  - Oxidative Phosphorylation
MH  - Oxygen Consumption
MH  - Phosphorylation
MH  - proton
MH  - Proton Pump
MH  - Rats
MH  - regulation
MH  - Saccharomyces cerevisiae
RP  - NOT IN FILE
NT  - Institut de Biochimie Cellulaire et Neurochimie du CNRS, Universite de Bordeaux, FrancePMID- 0002144185
SO  - Biochim Biophys Acta 1990 Jul 25 ;1018(2-3):185-189

992
UI  - 9918
AU  - Rigoulet M
AU  - Fraisse L
AU  - Ouhabi R
AU  - Guerin B
AU  - Fontaine E
AU  - Leverve X
TI  - Flux-dependent increase in the stoichiometry of charge translocation by mitochondrial ATPase/ATP synthase induced by almitrine.
AB  - After studying the effects of almitrine, a new kind of ATPase/ATP synthase inhibitor, on two kinds of isolated mammalian mitochondrion, we have observed that: (1) Almitrine inhibits oligomycin-sensitive ATPase; it decreases the ATP/O value of oxidative phosphorylations without any change in the magnitude of delta mu H+. (2) Almitrine increases the mechanistic H+/ATP stoichiometry of ATPase as shown by measuring either (i) the extent of potassium acetate and of potassium phosphate accumulation sustained by ATP utilisation, or (ii) the electrical charge/ATP (K+/ATP) ratio at steady-state of ATPase activity. (3) Rat liver mitochondria are at least 10-times more sensitive to almitrine than beef heart mitochondria. (4) The change in H+/ATP stoichiometry induced by almitrine depends on the magnitude of the flux through ATPase. The inhibitory effect of almitrine on ATPase/ATP synthase complex, as a consequence of such an H+/ATP stoichiometry change, is discussed.
MH  - acetate
MH  - Acetic Acids
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - Almitrine
MH  - Animal
MH  - antagonists & inhibitors
MH  - atp
MH  - ATPase
MH  - Cattle
MH  - Comparative Study
MH  - COMPLEX
MH  - drug effects
MH  - enzymology
MH  - H(+)-Transporting ATP Synthase
MH  - H+
MH  - Liver
MH  - Male
MH  - metabolism
MH  - Mitochondria
MH  - Mitochondria,Heart
MH  - Mitochondria,Liver
MH  - Mitochondrial Swelling
MH  - Oligomycins
MH  - Oxidative Phosphorylation
MH  - pharmacology
MH  - Phosphates
MH  - Phosphorylation
MH  - Potassium
MH  - Protons
MH  - Rats
MH  - Rats,Inbred Strains
MH  - Support,Non-U.S.Gov't
MH  - SYNTHASE
MH  - Valinomycin
RP  - NOT IN FILE
NT  - Institut de Biochimie Cellulaire et Neurochimie du CNRS, Universite de Bordeaux, FrancePMID- 0002165421
SO  - Biochim Biophys Acta 1990 Jul 17 ;1018(1):91-97

993
UI  - 354
AU  - Runswick MJ
AU  - Medd SM
AU  - Walker JE
AD  - MRC Laboratory of Molecular Biology, Cambridge, UK
TI  - The delta-subunit of ATP synthase from bovine heart mitochondria. Complementary DNA sequence of its import precursor cloned with the aid of the polymerase chain reaction
AB  - The delta-subunit of ATP synthase from bovine heart mitochondria is part of the extrinsic membrane domain, F1-ATPase. The mature protein is 146 amino acids in length and its function is obscure. It is encoded by a nuclear gene and is imported into the organelle. Two mixtures of oligonucleotides 17 bases long, designed on the basis of the known protein sequence, have been synthesized and employed as primers on bovine cDNA in the polymerase chain reaction. By this means a segment of bovine cDNA encoding part of the delta-subunit has been amplified, and this DNA segment has been employed to identify related cDNA clones in a library. These clones encode the mitochondrial import precursor of the delta-subunit; the protein sequence of the mature protein deduced from it is exactly the same as that determined earlier by direct sequence analysis. The clones have also been used to show that both the bovine and human genomes seem to contain a single gene for the delta- subunit
RP  - NOT IN FILE
NT  - UI - 90197618LA - engRN - 0 (Macromolecular Systems)RN - 0 (Protein Precursors)RN - 9007-49-2 (DNA)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19900426IS - 0264-6021SB - IMCY - ENGLANDJC - 9YO
UR  - PM:2138455
SO  - Biochem J 1990 Mar 1 ;266(2):421-426

994
UI  - 751
AU  - Sato MH
AU  - Hisabori T
AU  - Yoshida M
AD  - Department of Life Science, Faculty of Science, Tokyo Institute of Technology, Yokohama, Japan
TI  - The 55-kDa polypeptide released from spinach thylakoid membranes with 1 M LiCl is not the beta subunit of chloroplast F1
AB  - It was reported by Frasch et al. (Frasch, W. D., Green, J., Caguiat, J., and Mejia, A. (1989) J. Biol. Chem. 264, 5064-5069) that washing spinach thylakoid membranes with 1 M LiCl caused the release of the beta subunit of chloroplast F1 (CF1) which, existing as 180-kDa complexes of beta 3, retained considerable ATPase activity. We repeated their procedures and confirmed that a CF1 beta-like 55-kDa polypeptide was a major constituent of the 1 M LiCl-washed extract. However, the extract contained another polypeptide of which the Mr was 14,000, and these two polypeptides comprised a complex with approximate Mr 550,000 that had the same mobility in native polyacrylamide gel electrophoresis as that of ribulose-1,5-bisphosphate carboxylase. Only very low ATPase activity, less than 1% of the reported value, was detected for the extract and the purified complex. Antibody against the beta subunit of F1 from a thermophilic bacterium PS3 showed a clear cross-reactivity with the CF1 beta subunit but not with the 55-kDa polypeptide. Analysis of the N-terminal amino acid sequences of the 55- and 14-kDa polypeptides and the whole complex revealed that the complex was ribulose-1,5-bisphosphate carboxylase and that the 55- and 14-kDa polypeptides were its large and small subunits, respectively
RP  - NOT IN FILE
NT  - UI - 90337936LA - engRN - 0 (Macromolecular Systems)RN - 0 (Membrane Proteins)RN - 0 (Plant Proteins)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)RN - EC 4.1.1.39 (Ribulose-Bisphosphate Carboxylase)PT - Journal ArticleDA - 19900913IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:2143183
SO  - J Biol Chem 1990 Aug 15 ;265(23):13419-13422

995
UI  - 20955
AU  - Schneppe B
AU  - Deckers-Hebestreit G
AU  - Altendorf K
AD  - Universitat Osnabruck, Fachbereich Biologie/Chemie, Arbeitsgruppe Mikrobiologie, Federal Republic of Germany
TI  - Overproduction and purification of the uncI gene product of the ATP synthase of Escherichia coli
AB  - The uncI gene, the first gene of the unc operon, has been cloned into an expression vector carrying the lambda PRPL promoters in tandem orientation and the gene cI857 coding for the thermolabile repressor. Linkage of the uncI gene to an efficient ribosome binding site (the translational initiation region of the uncE gene) resulted in 10-20- fold increased gene expression. The i protein has been extracted from overproducing cells using chloroform/methanol and purified to homogeneity by ion exchange chromatography. Analyzing the products of the uncI gene encoded by different plasmids, we provide evidence that, in contrast to the previously reported data (Walker, J. E., Saraste, M., and Gay, N. J. (1984) Biochim. Biophys. Acta 768, 164-200), the chromosome-encoded i protein contains the N-terminal sequence Ser-Val- Ser-Leu-Val-Ser-Arg and has a molecular weight of 13,504
MH  - A
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BINDING
MH  - Cells
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - ion
MH  - ion exchange
MH  - M
MH  - protein
MH  - purification
MH  - Site
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 90094427LA - engRN - 0 (Plasmids)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19900202IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:2136739
SO  - J Biol Chem 1990 Jan 5 ;265(1):389-395

996
UI  - 447
AU  - Senior AE
AD  - Department of Biochemistry, University of Rochester, New York 14642
TI  - The proton-translocating ATPase of Escherichia coli
AB  - The purpose of this review is to provide an up-to-date summary of E. coli proton-translocating F1F0ATPase. From work on this enzyme, new insights have been gained in the areas of bacterial physiology and energy metabolism, mechanism of enzyme action, mechanism of ion transport through membranes, structure of membrane proteins, mechanism of energy coupling, and regulation of complex enzyme expression and assembly. An important and pressing need is for more structural information. High-resolution structural analyses of F1F0 have not progressed far, and this is likely to present a road block unless overcome. One possibility is to crystallize or apply nuclear magnetic resonance spectroscopy to isolated subunits available in native form from E. coli F1F0. In this way, one might incrementally build a structure of the F1F0 complex. Static views, however, are unlikely to provide a complete picture of a dynamic enzyme such as this, in which long-range interactions between F0 and F1 and cooperative interactions between nucleotide-binding sites play such an important role in catalysis. Mutagenesis and reversion analysis are two powerful techniques, which, combined with direct enzymological measurements, can be exploited in the immediate future to study the intriguing dynamic aspects of F1F0 function. Many questions remain to challenge us. Regulation of enzyme activity in the cell is not understood. The role of the noncatalytic nucleotide sites is unknown. The assembly pathway and regulation of expression are not established. The mechanisms of H+ translocation and catalysis seem to be proving amenable to analysis, and further advances in these areas can be expected. Long-range conformational interaction between the H+ conduction machinery in F0 and the catalytic sites in F1 seems basic to energy coupling; a major future goal is to provide a realistic physical explanation to validate this concept
RP  - NOT IN FILE
NT  - UI - 90303438LA - engRN - 0 (Nucleotides)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, AcademicDA - 19900813IS - 0883-9182SB - IMCY - UNITED STATESJC - ABO
UR  - PM:2141983
SO  - Annu Rev Biophys Biophys Chem 1990  ;19():7-41

997
UI  - 874
AU  - Sherratt HS
AU  - Turnbull DM
TI  - Mitochondrial oxidations and ATP synthesis in muscle
MH  - Adenosine Triphosphate
MH  - Animal
MH  - Biological Transport
MH  - biosynthesis
MH  - Electron Transport
MH  - Energy Metabolism
MH  - Human
MH  - metabolism
MH  - Mitochondria,Muscle
MH  - Oxidation-Reduction
MH  - Proteins
MH  - Thermodynamics
MH  - ultrastructure
RP  - NOT IN FILE
SO  - Baillieres Clin Endocrinol Metab 1990 Sep ;4(3):523-560

998
UI  - 826
AU  - Strotmann H
AU  - Thelen R
AU  - Muller W
AU  - Baum W
AD  - Institut fur Biochemie der Pflanzen, Heinrich-Heine-Universitat Dusseldorf, Federal Republic of Germany
TI  - A delta pH clamp method for analysis of steady-state kinetics of photophosphorylation
AB  - An instrumental device is described which allows steady-state kinetic measurements of photophosphorylation at a desired proton gradient which can be maintained throughout the course of the experiment ('delta pH clamp'). This is achieved by electronic regulation of light intensity using the calibrated 9-aminoacridine fluorescence signal as sensor of the gradient. The instrument is suitable for determination of kinetic parameters of the proton-translocating ATPase in isolated envelope-free chloroplasts under defined conditions. At clamped delta pH, phosphorylation as a function of substrate concentration shows Michaelis-Menten kinetics. The true Michaelis constants and the dissociation constants for phosphate and ADP are reported. The Michaelis constants are not affected by the magnitude of the proton gradient in the investigated range. The significance of these results is discussed
RP  - NOT IN FILE
NT  - UI - 91065396LA - engRN - 0 (Protons)RN - 90-45-9 (Aminacrine)PT - Journal ArticleDA - 19910111IS - 0014-2956SB - IMCY - GERMANYJC - EMZ
UR  - PM:2174369
SO  - Eur J Biochem 1990 Nov 13 ;193(3):879-886

999
UI  - 20870
AU  - Takeyama M
AU  - Ihara K
AU  - Moriyama Y
AU  - Noumi T
AU  - Ida K
AU  - Tomioka N
AU  - Itai A
AU  - Maeda M
AU  - Futai M
AD  - Institute of Scientific and Industrial Research, Osaka University, Japan
TI  - The glycine-rich sequence of the beta subunit of Escherichia coli H(+)- ATPase is important for activity
AB  - A short sequence motif rich in glycine residues, Gly-X-X-X-X-Gly-Lys- Thr/Ser, has been found in many nucleotide-binding proteins including the beta subunit of Escherichia coli H(+)-ATPase (Gly-Gly-Ala-Gly-Val- Gly-Lys-Thr, residues 149-156). The following mutations were introduced in this region of the cloned E. coli unc operon carried by a plasmid pBWU1: Ala-151----Pro or Val; insertion of a Gly residue between Lys- 155 and Thr-156; and replacement of the region by the corresponding sequence of adenylate kinase (Gly-Gly-Pro-Gly-Ser-Gly-Lys-Gly-Thr) or p21 ras protein (ras) (Gly-Ala-Gly-Gly-Val-Gly-Lys-Ser). All F0F1 subunits were synthesized in the deletion strain of the unc operon- dependent on pBWU1 with mutations, and essentially the same amounts of H(+)-ATPase with these mutant beta subunits were found in membranes. The adenylate kinase and Gly insertion mutants showed no oxidative phosphorylation or ATPase activity, whereas the Pro-151 mutants had higher ATPase activity than the wild-type, and the Val-151 and ras mutants had significant activity. It is striking that the enzyme with the ras mutation (differing in three amino acids from the beta sequence) had about half the membrane ATPase activity of the wild-type. These results together with the simulated three-dimensional structures of the wild-type and mutant sequences suggest that in mutant beta subunits with no ATPase activity projection of Thr-156 residues was opposite to that in the wild-type, and that the size and direction of projection of residue 151 are important for the enzyme activity
MH  - A
MH  - ACID
MH  - Amino Acids
MH  - ATPase
MH  - BETA-SUBUNIT
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0F1
MH  - H(+)ATPase
MH  - Macromolecular Systems
MH  - membrane
MH  - Membranes
MH  - mutant
MH  - nucleotide binding
MH  - Oxidative Phosphorylation
MH  - Phosphorylation
MH  - protein
MH  - Proteins
MH  - RESIDUE
MH  - SUBUNIT
MH  - SYSTEM
MH  - SYSTEMS
RP  - NOT IN FILE
NT  - UI - 91065946LA - engRN - 0 (Macromolecular Systems)RN - 0 (Oligonucleotide Probes)RN - 0 (Plasmids)RN - 56-40-6 (Glycine)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19910117IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:2147431
SO  - J Biol Chem 1990 Dec 5 ;265(34):21279-21284

1000
UI  - 21024
AU  - Turina MT
AU  - Venturoli G
AU  - Grber P
AU  - Melandri BA
TI  - Quantitative estimation of the H+ -storage capacity of chromatophores and comparison with acid-base induced ATP sythesis
MH  - atp
MH  - chromatophore
MH  - chromatophores
MH  - H+
MH  - pmf
MH  - slip
RP  - IN FILE
SO  - Biochim Biophys Acta 1990  ;1018():134-137

1001
UI  - 356
AU  - Vinas O
AU  - Powell SJ
AU  - Runswick MJ
AU  - Iacobazzi V
AU  - Walker JE
AD  - MRC Laboratory of Molecular Biology, Cambridge, UK
TI  - The epsilon-subunit of ATP synthase from bovine heart mitochondria. Complementary DNA sequence, expression in bovine tissues and evidence of homologous sequences in man and rat
AB  - The epsilon-subunit of ATP synthase from bovine heart mitochondria is assembled into the extrinsic membrane sector, F1-ATPase. The mature protein is 50 amino acid residues in length and its function is unknown. It is a nuclear gene product that is imported into the organelle. A mixture of 64 oligonucleotides 17 bases long, designed on the basis of the known protein sequence, was synthesized and used as a hybridization probe to isolate a cognate cDNA clone from a bovine library. The DNA sequence of this clone was determined, and the protein sequence of the epsilon-subunit deduced from it agrees exactly with that determined by direct sequence analysis of the protein isolated from bovine hearts. The bovine cDNA was used as a hybridization probe to examine the expression of the epsilon-subunit in various bovine tissues. mRNAs related to the cDNA are found in all of these tissues, and no evidence was obtained of the presence of mRNAs for the epsilon- subunit with similar coding sequences and dissimilar 3' non-coding regions. By hybridization experiments with digests of DNA from cow, man and rat it has been shown that sequences related to the bovine cDNA are present in the genomes of all three species. More than one related sequence was detected in all cases, indicating the presence in all three genomes of more than one gene and/or pseudogenes
RP  - NOT IN FILE
NT  - UI - 90147593LA - engRN - 0 (Macromolecular Systems)RN - 0 (Oligonucleotide Probes)RN - 9007-49-2 (DNA)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19900313IS - 0264-6021SB - IMCY - ENGLANDJC - 9YO
UR  - PM:2137333
SO  - Biochem J 1990 Jan 15 ;265(2):321-326

1002
UI  - 21009
AU  - Wach A
AU  - Ahlers J
AU  - Graber P
AD  - Institut fur Biochemie und Molekularbiologie, Freie Universitat Berlin
TI  - The H(+)-ATPase of the plasma membrane from yeast. Kinetics of ATP hydrolysis in native membranes, isolated and reconstituted enzymes
AB  - The H(+)-ATPase of the plasma membrane from Saccharomyces cerevisiae has been isolated, purified and reconstituted into asolectin liposomes. The kinetics of ATP hydrolysis have been compared for the H(+)-ATPase in the plasma membrane, in a protein/lipid/detergent micelle (isolated enzyme) and in asolectin proteoliposomes (reconstituted enzyme). In all three cases the kinetics of ATP hydrolysis can be described by Michaelis-Menten kinetics with Km = 0.2 mM MgATP (plasma membranes), Km = 2.4 mM MgATP (isolated enzyme) and Km = 0.2 mM MgATP (reconstituted enzyme). However, the maximal turnover decreases only by a factor of two during isolation of the enzyme and does not change during reconstitution; the activation of the H(+)-ATPase by free Mg2+ is also only slightly influenced by the detergent. The dissociation constant of the enzyme-Mg2+ complex Ka, does not alter during isolation and the dissociation constant of the enzyme-substrate complex, Ks, increases from Ks = 30 microM (plasma membranes) to Ks = 90 microM (isolated enzyme). ATP binding to the H(+)-ATPase ('single turnover' conditions) for the isolated and the reconstituted enzyme resulted in both cases in a second-order rate constant k1 = 2.6 x 10(4) M-1.s-1. From these observations it is concluded that the detergent used (Zwittergent TM 3- 14) interacts reversibly with the H(+)-ATPase and that practically all H(+)-ATPase molecules are reconstituted into the liposomes with the ATP- binding site being directed to the outside of the vesicle
MH  - A
MH  - ACTIVATION
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATPase
MH  - BINDING
MH  - COMPLEX
MH  - CONSTANT
MH  - Enzymes
MH  - H(+)ATPase
MH  - Hydrolysis
MH  - Kinetics
MH  - Liposomes
MH  - membrane
MH  - Membranes
MH  - Phospholipids
MH  - proteoliposome
MH  - rate constant
MH  - reconstitution
MH  - Saccharomyces cerevisiae
MH  - Site
RP  - NOT IN FILE
NT  - UI - 90276438LA - engRN - 0 (Liposomes)RN - 0 (Phospholipids)RN - 56-65-5 (Adenosine Triphosphate)RN - 69279-91-0 (asolectin)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19900713IS - 0014-2956SB - IMCY - GERMANY, WEST
UR  - PM:2140984
SO  - Eur J Biochem 1990 May 20 ;189(3):675-682

1003
UI  - 355
AU  - Walker JE
AU  - Fearnley IM
AU  - Lutter R
AU  - Todd RJ
AU  - Runswick MJ
AD  - Medical Research Council Laboratory of Molecular Biology, Cambridge, UK
TI  - Structural aspects of proton-pumping ATPases
AB  - ATP synthase is found in bacteria, chloroplasts and mitochondria. The simplest known example of such an enzyme is that in the eubacterium Escherichia coli; it is a membrane-bound assembly of eight different polypeptides assembled with a stoichiometry of alpha 3 beta 3 gamma 1 delta 1 epsilon 1 a1b2c10-12. The first five of these constitute a globular structure, F1-ATPase, which is bound to an intrinsic membrane domain, F0, an assembly of the three remaining subunits. ATP synthases driven by photosynthesis are slightly more complex. In chloroplasts, and probably in photosynthetic bacteria, they have nine subunits, all homologues of the components of the E. coli enzyme; the additional subunit is a duplicated and diverged relation of subunit b. The mammalian mitochondrial enzyme is more complex. It contains 14 different polypeptides, of which 13 have been characterized. Two membrane components, a (or ATPase-6) and A6L, are encoded in the mitochondrial genome in overlapping genes and the remaining subunits are nuclear gene products that are translated on cytoplasmic ribosomes and then imported into the organelle. The sequence of the proteins of ATP-synthase have provided information about amino acids that are important for its function. For example, amino acids contributing to nucleotide binding sites have been identified. Also, they provide the basis of models of secondary structure of membrane components that constitute the transmembrane proton channel. An understanding of the coupling of the transmembrane potential gradient for protons, delta mu H+, to ATP synthesis will probably require the determination of the structure of the entire membrane bound complex. Crystals have been obtained of the globular domain, F1-ATPase. They diffract to a resolution of 3-4 A and data collection is in progress. As a preliminary step towards crystallization of the entire complex, we have purified it from bovine mitochondria and reconstituted it into phospholipid vesicles
RP  - NOT IN FILE
NT  - UI - 90239114LA - engRN - 0 (Macromolecular Systems)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review LiteratureDA - 19900606IS - 0962-8436SB - IMCY - ENGLANDJC - P5Z
UR  - PM:1970643
SO  - Philos Trans R Soc Lond B Biol Sci 1990 Jan 30 ;326(1236):367-378

1004
UI  - 827
AU  - Werner S
AU  - Schumann J
AU  - Strotmann H
AD  - Institut fur Biochemie der Pflanzen, Heinrich-Heine-Universitat Dusseldorf, FRG
TI  - The primary structure of the gamma-subunit of the ATPase from Synechocystis 6803
AB  - The nucleotide sequence of the gene coding for the F0F1-ATPase gamma- subunit (atpC) from the transformable cyanobacterium Synchocystis 6083 has been determined. The deduced translation product consists of 314 amino acid residues and is highly homologous (72% identical residues) to the sequences of other cyanobacterial gamma-subunits. The Synechocystis 6803 sequence is also homologous to the chloroplast gamma- sequence. Like in the other cyanobacterial subunits, only the first of the 3 cysteine residues, which are involved in energy-linked functions of the gamma-subunit in spinach chloroplasts, is conserved in Synechocystis 6803
RP  - NOT IN FILE
NT  - UI - 90169116LA - engRN - EC 3.1.21.- (Deoxyribonuclease EcoRI)RN - EC 3.1.21.- (Deoxyribonuclease HindIII)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19900406IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:2137788
SO  - FEBS Lett 1990 Feb 12 ;261(1):204-208

1005
UI  - 19762
AU  - Wibom R
AU  - Lundin A
AU  - Hultman E
AD  - Department of Clinical Chemistry II, Karolinska Institutet, Huddinge University Hospital, Sweden
TI  - A sensitive method for measuring ATP-formation in rat muscle mitochondria
AB  - A sensitive method for the measurement of the ATP production rate in isolated skeletal muscle mitochondria is presented. Mitochondrial suspensions were prepared by differential centrifugation from approximately 80 mg of soleus muscle. ATP production rates were measured luminometrically, utilizing a reagent based upon firefly luciferase, which emits light proportional to the ATP concentration. In a group of 10 rats the ATP production rates were measured with the following substrate combinations: pyruvate + malate, palmitoyl-L- carnitine + malate, alpha-ketoglutarate, succinate + rotenone and succinate alone. The variance of the method including tissue preparation, protein determination and the luminometric determination of ATP production was estimated to be 10-14% for the various substrates. Compared to values in the literature, the present results show a good agreement for the substrates pyruvate + malate and palmitoyl-L-carnitine + malate, but lower rates were obtained in our study for alpha-ketoglutarate and succinate + rotenone. The advantage of the luminometric method is its high sensitivity. Only 30-40 mg of tissue is required for a complete determination, compared to 1-2 g for a similar assay of oxygen consumption. The method is intended for use in human subjects and will facilitate studies of mitochondrial respiration both in patients of different age groups and in healthy subjects
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP production
MH  - Chemistry
MH  - England
MH  - Human
MH  - Ketoglutaric Acids
MH  - Light
MH  - method
MH  - Mitochondria
MH  - Oxygen
MH  - Oxygen Consumption
MH  - Rats
MH  - Respiration
MH  - Rotenone
MH  - succinate
MH  - Succinates
MH  - Succinic Acid
RP  - NOT IN FILE
NT  - UI - 90251982LA - engRN - 0 (Ketoglutaric Acids)RN - 0 (Malates)RN - 0 (Pyruvates)RN - 0 (Succinates)RN - 110-15-6 (Succinic Acid)RN - 127-17-3 (Pyruvic Acid)RN - 1935-18-8 (Palmitoylcarnitine)RN - 328-50-7 (alpha-ketoglutaric acid)RN - 56-65-5 (Adenosine Triphosphate)RN - 6915-15-7 (malic acid)RN - EC 1.13.12.- (Luciferase)RN - EC 2.7.4.3 (Adenylate Kinase)PT - Journal ArticleDA - 19900620IS - 0036-5513SB - IMCY - ENGLANDJC - UCP
UR  - PM:2339278
SO  - Scand J Clin Lab Invest 1990 Apr ;50(2):143-152

1006
UI  - 753
AU  - Yoshida M
AU  - Allison WS
AD  - Department of Life Sciences, Faculty of Science, Tokyo Institute of Technology, Yokohama, Japan
TI  - The ATPase activity of the alpha 3 beta 3 complex of the F1-ATPase of the thermophilic bacterium PS3 is inactivated on modification of tyrosine 307 in a single beta subunit by 7-chloro-4-nitrobenzofurazan
AB  - The catalytically active alpha 3 beta 3 complex, assembled as described (Miwa, K., and Yoshida, M. (1989) Proc. Natl. Acad. Sci. U. S. A. 86, 6484-6487) from the isolated alpha and beta subunits of the F1-ATPase of the thermophilic bacterium PS3 (TF1), is inactivated by 7-chloro-4- nitrobenzofurazan (Nbf-Cl) with characteristics very similar to those observed when TF1, which has the subunit composition, alpha 3 beta 3 gamma delta epsilon, is inactivated by the reagent under the same conditions. Both native TF1 and the alpha 3 beta 3 complex are inactivated by 200 microM Nbf-Cl with a pseudo-first order rate constant of 3.7 x 10(-2) min-1 in the presence of 0.2 M Na2SO4 at pH 7.6 and 23 degrees C. The rate of increase in absorbance at 385 nm of reaction mixtures containing 200 microM [14C]Nbf-Cl and TF1, the wild- type alpha 3 beta 3 complex, or the mutant alpha 3(beta Y307----F)3 complex, each at 18 microM was also examined. Since the alpha 3(beta y307----F)3 complex is resistant to inactivation by Nbf-Cl, difference spectrophotometry revealed that inactivation of native TF1 and the wild- type alpha 3 beta 3 complex could be correlated with formation of about 1 mol of Nbf-O-Tyr/mol of enzyme or complex. Fractionation of peptic digests of the labeled enzyme and complexes by reversed-phase high performance liquid chromatography resolved a major radioactive peptide that was common to labeled TF1 and the labeled alpha 3 beta 3 complex but was absent in the digest of the labeled alpha 3(beta Y307----F)3 complex. This labeled peptide was shown to contain Tyr-beta 307 derivatized with [14C]Nbf-Cl by automatic amino acid sequence analyses. From these results, it is concluded that one-third of the sites' reactivity of Nbf-Cl with Tyr-beta 307 in TF1 or its equivalent in other F1-ATPases is not influenced by the presence of the gamma, delta, or epsilon subunits. It has also been shown that Tyr-307 is not modified to an appreciable extent when the isolated beta subunit is treated with [14C]Nbf-Cl under conditions in which this residue is nearly completely labeled in a single beta subunit when TF1 or the alpha 3 beta 3 complex is inactivated by the reagent
RP  - NOT IN FILE
NT  - UI - 90153861LA - engRN - 0 (Benzofurans)RN - 0 (Macromolecular Systems)RN - 0 (Peptide Fragments)RN - 115491-60-6 (7-chloro-4-nitrobenzofuran)RN - 55520-40-6 (Tyrosine)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM 16,974/GM/NIGMSDA - 19900323IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:2137446
SO  - J Biol Chem 1990 Feb 15 ;265(5):2483-2487

1007
UI  - 21167
AU  - Angov E
AU  - Ng TC
AU  - Brusilow WS
AD  - Department of Biochemistry, Wayne State University School of Medicine, Detroit, Michigan 48201
TI  - Effect of the delta subunit on assembly and proton permeability of the F0 proton channel of Escherichia coli F1F0 ATPase
AB  - During the assembly of the Escherichia coli proton-translocating ATPase, the subunits of F1 interact with F0 to increase the proton permeability of the transmembrane proton channel. We tested the involvement of the delta subunit in this process by partially and completely deleting uncH (delta subunit) from a plasmid carrying the genes for the F0 subunits and delta and testing the effects of those F0 plasmids on the growth of unc+ and unc mutant E. coli strains. We found that the delta subunit was required for inhibition of growth of unc+ cells. We also tested membranes isolated from unc-deleted cells containing F0 plasmids for F1-binding ability. In unc-deleted cells, these plasmids produced F0 in amounts comparable to those found in normal unc+ E. coli cells, while having only small effects on cell growth. These studies demonstrate that the delta subunit plays an important role in opening the F0 proton channel but that it does not serve as a temporary plug of F0 during assembly, as had been previously speculated (S. Pati and W. S. A. Brusilow, J. Biol. Chem. 264:2640- 2644, 1989)
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - ATPase
MH  - Biochemistry
MH  - Cells
MH  - delta
MH  - DELTA-SUBUNIT
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - F1
MH  - ion
MH  - Ion Channels
MH  - Macromolecular Systems
MH  - membrane
MH  - Membranes
MH  - mutant
MH  - Permeability
MH  - proton
MH  - Proton-Translocating ATPases
MH  - SUBUNIT
MH  - SYSTEM
MH  - SYSTEMS
RP  - NOT IN FILE
NT  - UI - 91100314LA - engRN - 0 (Ion Channels)RN - 0 (Macromolecular Systems)RN - 0 (Oligonucleotide Probes)RN - 0 (Plasmids)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleID - KO4-AI00882/AI/NIAIDDA - 19910220IS - 0021-9193SB - IMCY - UNITED STATES
UR  - PM:1702783
SO  - J Bacteriol 1991 Jan ;173(1):407-411

1008
UI  - 19782
AU  - Berkich DA
AU  - Williams GD
AU  - Masiakos PT
AU  - Smith MB
AU  - Boyer PD
AU  - LaNoue KF
AD  - Department of Cellular and Molecular Physiology, Milton S Hershey Medical Center, Pennsylvania State University, Hershey 17033
TI  - Rates of various reactions catalyzed by ATP synthase as related to the mechanism of ATP synthesis
AB  - The forward and reverse rates of the overall reaction catalyzed by the ATP synthase in intact rat heart mitochondria, as measured with 32P, were compared with the rates of two partial steps, as measured with 18O. Such rates have been measured previously, but their relationship to one another has not been determined, nor have the partial reactions been measured in intact mitochondria. The partial steps measured were the rate of medium Pi formation from bound ATP (in state 4 this also equals the rate of medium Pi into bound ATP) and the rate of formation of bound ATP from bound Pi within the catalytic site. The rates of both partial reactions can be measured by 31P NMR analysis of the 18O distribution in Pi and ATP released from the enzyme during incubation of intact mitochondria with highly labeled [18O]Pi. Data were obtained in state 3 and 4 conditions with variation in substrate concentrations, temperature, and mitochondrial membrane electrical potential gradient (delta psi m). Although neither binding nor release of ATP is necessary for phosphate/H2O exchange, in state 4 the rate of incorporation of at least one water oxygen atom into phosphate is approximately twice the rate of the overall reaction rate under a variety of conditions. This can be explained if the release of Pi or ATP at one catalytic site does not occur, unless ATP or Pi is bound at another catalytic site. Such coupling provides strong support for the previously proposed alternating site mechanism. In state 3 slow reversal of ATP synthesis occurs within the mitochondrial matrix and can be detected as incorporation of water oxygen atoms into medium Pi even though medium [32P]ATP does not give rise to 32Pi in state 3. These data can be explained by lack of translocation of ATP from the medium to the mitochondrial matrix. The rate of bound ATP formation from bound Pi at catalytic sites was over twice the rate of the overall reaction in both states 4 and 3. The rate of reaction at the catalytic site is considerably less sensitive to the decrease in membrane potential and the concentration of medium ADP than is the rate of medium ATP formation. This supports the view that the active catalytic site is occluded and proceeds at a rapid rate which is relatively independent of delta psi m and of media substrates
MH  - A
MH  - ACTIVE
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - ADP
MH  - analysis
MH  - atp
MH  - ATP FORMATION
MH  - ATP synthase
MH  - ATP synthesis
MH  - BINDING
MH  - H(+)-Transporting ATP Synthase
MH  - mechanism
MH  - membrane
MH  - Membrane Potential
MH  - Mitochondria
MH  - Oxygen
MH  - Oxygen Isotopes
MH  - Phosphates
MH  - Phosphorus
MH  - Phosphorus Radioisotopes
MH  - physiology
MH  - PSI
MH  - SYNTHASE
MH  - synthesis
MH  - Temperature
MH  - translocation
MH  - Water
RP  - NOT IN FILE
NT  - UI - 91093113LA - engRN - 0 (Oxygen Isotopes)RN - 0 (Phosphates)RN - 0 (Phosphorus Radioisotopes)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - 5P01-HL18708/HL/NHLBIID - GM11094/GM/NIGMSDA - 19910214IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:1824691
SO  - J Biol Chem 1991 Jan 5 ;266(1):123-129

1009
UI  - 521
AU  - Bianchet M
AU  - Ysern X
AU  - Hullihen J
AU  - Pedersen PL
AU  - Amzel LM
AD  - Department of Biophysics and Biophysical Chemistry, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205
TI  - Mitochondrial ATP synthase. Quaternary structure of the F1 moiety at 3.6 A determined by x-ray diffraction analysis
AB  - The F1 moiety of the mitochondrial ATP synthase is composed of five different subunits with stoichiometry alpha 3 beta 3 gamma delta epsilon and exhibits the capacity to synthesize ATP from ADP and Pi. We have previously crystallized rat liver F1 and described its structure at 9-A resolution (Amzel, L. M., McKinney, M., Narayanan, P., and Pedersen, P. L. (1982) Proc. Natl. Acad. Sci. U. S. A. 79, 5852-5856). Here we present an x-ray map of this complex enzyme at 3.6 A, which provides a much more informative description of its quaternary structure. The overall dimensions of the F1 molecule are 120 A x 120 A x 74 A. The enzyme exhibits 3-fold symmetry relating the three copies of each of the two major subunits, alpha and beta. As the alpha subunits (but not the beta subunits) contain cysteine residues, it has been possible to identify the alpha subunits by heavy atom labeling with mersalyl and to relate their positions in the F1 molecule to the beta subunits. Significantly, the alpha and beta subunits each exist as trimeric arrays which are organized in two slightly offset, interdigitated layers along the 3-fold axis. In one trimeric layer the alpha subunits are located close to the axis with homologous subunits interacting with each other; in the other trimeric layer the beta subunits are far from the axis, and they interact only with alpha subunits and not with one another. At one end of the structure, part of the interface between each alpha and beta subunit encloses a space or "pocket" that is accessible to the solvent; at the other end the interfaces between the subunits are more open and exposed. The present work represents the highest resolution map reported to date for the F1 moiety of an ATP synthase complex
RP  - NOT IN FILE
NT  - UI - 92041997LA - engRN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - CA 10951/CA/NCIID - GM 25432/GM/NIGMSDA - 19911213IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:1834656
SO  - J Biol Chem 1991 Nov 5 ;266(31):21197-21201

1010
UI  - 388
AU  - Bulygin VV
AU  - Vinogradov AD
AD  - Department of Biochemistry, School of Biology, Moscow State University, USSR
TI  - Interaction of Mg2+ with F0.F1 mitochondrial ATPase as related to its slow active/inactive transition
AB  - Bovine heart submitochondrial particles incubated with a low concentration of ADP in the presence of Mg2+ and passed through a Sephadex column equilibrated with EDTA exhibit sensitivity of their initial ATPase activity to preincubation with Mg2+. By using particles thus prepared, several characteristics of a Mg(2+)-specific inhibitory site on F0.F1 ATPase were studied. The inhibition was shown to be both time- and Mg(2+)-concentration-dependent, with an equilibrium constant (at infinite time) of 2 x 10(-6) M (25 degrees C, pH 7.5). The dependence of the pseudo-first-order rate constant for the inhibition process on Mg2+ concentration suggests the presence of a single Mg(2+)- binding site with K8 = 1.1 x 10(-4) M. The data obtained are consistent with a two-step mechanism of Mg(2+)-F0.F1 interaction which results in a loss of the ATPase activity; it includes rapid pH-dependent binding of Mg2+ at the site with K8 = 1.1 x 10(-4) M, followed by a slow interconversion of the Mg(2+)-F1 complex into inactive ATPase (kin. = 0.65 min-1, kact. = 0.01 min-1). The Mg(2+)-inhibited ATPase is very slowly (t1/2 approximately 90 min) re-activated in the presence of EDTA. The rate of EDTA-induced re-activation is pH-independent and can be dramatically increased by added ATP, Pi and sulphite. The dissociation constants for free ATP and P1 (5 x 10(-7) M and 1 x 10(-3) M respectively) and the maximal activation rates were determined by measuring the hyperbolic dependencies of the EDTA-induced re-activation of Mg(2+)-de-activated ATPase on the concentrations of the accelerating ligands. Taken together, the data obtained show two functionally detectable free nucleotide-specific binding sites, one site for Pi and one Mg(2+)-specific ATPase-inhibitory site on the F0.F1 mitochondrial ATP synthase complex
RP  - NOT IN FILE
NT  - UI - 91248139LA - engRN - 58-64-0 (Adenosine Diphosphate)RN - 60-00-4 (Edetic Acid)RN - 7439-95-4 (Magnesium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19910703IS - 0264-6021SB - IMCY - ENGLANDJC - 9YO
UR  - PM:1828147
SO  - Biochem J 1991 May 15 ;276 ( Pt 1)():149-156

1011
UI  - 8160
AU  - Capozza G
AU  - Dmitriev OY
AU  - Krasnoselskaya IA
AU  - Papa S
AU  - Skulachev VP
TI  - The effect of F0 inhibitors on the Vibrio alginolyticus membrane ATPase
MH  - atp
MH  - ATPase
MH  - Bacteria
MH  - F0
MH  - FO
MH  - inhibitor
MH  - inhibitors
MH  - membrane
MH  - proteolipid
RP  - IN FILE
NT  - katrin inhibitoren, vent-effect ist reversible
SO  - FEBS Lett 1991  ;280(2)():274-276

1012
UI  - 21272
AU  - Cramer WA
AU  - Knaff DB
TI  - Energy transduction in biological membranes: a textbook of bioenergetics.
MH  - Membranes
MH  - membrane
MH  - A
PB  - New York: Springer
RP  - NOT IN FILE
SO  -  1991  ;():

1013
UI  - 9898
AU  - Davis EJ
AU  - Davis-van TW
TI  - An assessment of the role of proton leaks in the mechanistic stoichiometry of oxidative phosphorylation.
AB  - Rat liver mitochondria were incubated in the presence of varying concentrations of ATP, followed by ADP to initiate phosphorylation. Analysis of phosphorylation to oxygen ratios (P/O) was carried out with varied initial phosphorylation potentials (or ATP/ADP ratios). Rates of phosphorylation and respiration and magnitude of membrane potential (delta psi) were measured. The results are discussed in the framework of P/total O and P/"extra" O ratios in determination of the mechanistic P/O ratio. It is concluded that the former underestimates, and the latter overestimates the mechanistic P/O ratio.
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - ADP
MH  - analysis
MH  - Animal
MH  - atp
MH  - Biochemistry
MH  - drug effects
MH  - Liver
MH  - Membrane Potentials
MH  - metabolism
MH  - Mitochondria
MH  - Mitochondria,Liver
MH  - Oxidative Phosphorylation
MH  - Oxygen
MH  - Oxygen Consumption
MH  - pharmacology
MH  - Phosphorylation
MH  - proton
MH  - Protons
MH  - PSI
MH  - Rats
MH  - Respiration
MH  - Support,Non-U.S.Gov't
MH  - Support,U.S.Gov't,P.H.S.
RP  - NOT IN FILE
NT  - Indiana University School of Medicine, Department of Biochemistry and Molecular Biology, Indianapolis 46202-5122PMID- 0001654845
SO  - Arch Biochem Biophys 1991 Aug 15 ;289(1):184-186

1014
UI  - 81
AU  - Dimroth P
AD  - Mikrobiologisches Institut, Eidgenossische Technische Hochschule, ETH- Zentrum, Zurich, Switzerland
TI  - Na(+)-coupled alternative to H(+)-coupled primary transport systems in bacteria
AB  - Protons are the most common coupling ions in bacterial energy conversions. However, while many organisms, such as the alkaliphilic Bacilli, employ H(+)-bioenergetics for electron transport phosphorylation, they use Na+ as the coupling ion for transport and flagellar movement. The Na+ gradient required for these bioenergetic functions is established by the secondary Na+/H+ antiporter. In contrast, Vibrio alginolyticus and methanogenic bacteria have primary pumps for both H+ and Na+. They use the proton gradient for ATP synthesis while other, less energy-consuming membrane reactions are powered by the Na+ gradient. In a third mode, some anaerobic bacteria possess decarboxylases acting as primary Na+ pumps. For instance, in Klebsiella pneumoniae, the Na+ gradient established by oxaloacetate decarboxylase is used for the uptake of the growth substrate citrate, and Propionigenium modestum consumes the energy of the Na+ gradient formed by methylmalonyl-CoA decarboxylase directly for ATP synthesis
RP  - NOT IN FILE
NT  - UI - 92181420LA - engRN - 0 (Bacterial Proteins)RN - 0 (Carrier Proteins)RN - 0 (Membrane Proteins)RN - 0 (Oxidative Phosphorylation Coupling Factors)RN - 0 (Protons)RN - 0 (Sodium-Hydrogen Antiporter)RN - 56-65-5 (Adenosine Triphosphate)RN - 7440-09-7 (Potassium)RN - 7440-23-5 (Sodium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)RN - EC 3.6.1.37 (Na(+)-K(+)-Exchanging ATPase)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 19920406IS - 0265-9247SB - IMCY - ENGLANDJC - 9YY
UR  - PM:1665692
SO  - Bioessays 1991 Sep ;13(9):463-468

1015
UI  - 303
AU  - Engelbrecht S
AU  - Reed J
AU  - Penin F
AU  - Gautheron DC
AU  - Junge W
AD  - Biophysik, Fachbereich Biologie/Chemie, Universitat Osnabruck
TI  - Subunit delta of chloroplast F0F1-ATPase and OSCP of mitochondrial F0F1- ATPase: a comparison by CD-spectroscopy
AB  - CD spectra have been recorded with subunit delta from chloroplast CF0CF1 and with OSCP from mitochondrial MF0MF1. These subunits are supposed to act similarly at the interface between proton transport through the F0-portion and ATP-synthesis in the F1-portion of their respective F0F1-ATPase. Evaluation of the data for both proteins revealed a very high alpha-helix content of approximately 85% and practically no beta-sheets. Despite their low homology on the primary structure level (23% identity) and their different electrostatic properties (pI-values differ by 3 units), spinach delta and porcine OSCP are indistinguishable with respect to their secondary structure as measured by CD. Prediction and analysis of consensual alpha-helices even in poorly conserved regions indicate a high degree of structural similarity between chloroplast delta and OSCP. In view of the topology and function of delta and OSCP in intact F0F1 these findings are interpreted to indicate the dominance of secondary and tertiary structure over the primary structure in their supposed function between proton flow and ATP-synthesis
RP  - NOT IN FILE
NT  - UI - 92088422LA - engRN - 0 (Carrier Proteins)RN - 0 (Macromolecular Systems)RN - 0 (Membrane Proteins)RN - 0 (oligomycin sensitivity-conferring protein)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19920124IS - 0341-0382SB - IMCY - GERMANYJC - ACL
UR  - PM:1836327
SO  - Z Naturforsch [C] 1991 Sep ;46(9-10):759-764

1016
UI  - 20869
AU  - Eya S
AU  - Maeda M
AU  - Futai M
AD  - Department of Organic Chemistry and Biochemistry, Osaka University, Japan
TI  - Role of the carboxyl terminal region of H(+)-ATPase (F0F1) a subunit from Escherichia coli
AB  - The effects of amino acid substitutions in the carboxyl terminal region of the H(+)-ATPase a subunit (271 amino acid residues) of Escherichia coli were studied using a defined expression system for uncB genes coded by recombinant plasmids. The a subunits with the mutations, Tyr- 263----end, Trp-231----end, Glu-219----Gln, and Arg-210----Lys (or Gln) were fully defective in ATP-dependent proton translocation, and those with Gln-252----Glu (or Leu), His-245----Glu, Pro-230----Leu, and Glu- 219----His were partially defective. On the other hand, the phenotypes of the Glu-269----end, Ser-265----Ala (or end), and Tyr-263----Phe mutants were essentially similar to that of the wild-type. These results suggested that seven amino acid residues between Ser-265 and the carboxyl terminus were not required for the functional proton pathway but that all the other residues except Arg-210, Glu-219, and His-245 were required for maintaining the correct conformation of the proton pathway. The results were consistent with a report that Arg-210 is directly involved in proton translocation
MH  - A
MH  - ACID
MH  - ATPase
MH  - Biochemistry
MH  - Chemistry
MH  - conformation
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0F1
MH  - H(+)ATPase
MH  - mutant
MH  - proton
MH  - RESIDUE
MH  - SUBUNIT
MH  - SYSTEM
MH  - translocation
RP  - NOT IN FILE
NT  - UI - 91112808LA - engRN - 0 (Oligonucleotides)RN - 74-79-3 (Arginine)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19910228IS - 0003-9861SB - IMCY - UNITED STATES
UR  - PM:1824913
SO  - Arch Biochem Biophys 1991 Jan ;284(1):71-77

1017
UI  - 976
AU  - Ferguson SJ
TI  - The protonmotive force and the problem of the P/O ratio; some teaching considerations
MH  - Bacteria
MH  - Biochemistry
MH  - education
MH  - Electron Transport
MH  - Energy Metabolism
MH  - metabolism
MH  - Nad
MH  - Oxygen
MH  - Phosphorus
MH  - Protons
RP  - NOT IN FILE
NT  - Department of Biochemistry, University of Oxford, UK
SO  - Biochem Soc Trans 1991 Nov ;19(4):982-983

1018
UI  - 116
AU  - Fillingame RH
AU  - Oldenburg M
AU  - Fraga D
AD  - Department of Physiological Chemistry, University of Wisconsin Medical School, Madison 53706
TI  - Mutation of alanine 24 to serine in subunit c of the Escherichia coli F1F0-ATP synthase reduces reactivity of aspartyl 61 with dicyclohexylcarbodiimide
AB  - Dicyclohexylcarbodiimide (DCCD) inhibits the activity of the F1F0-H+ ATP synthase of Escherichia coli by reacting with aspartyl 61 in subunit c of the FO sector to form a stable N-acylurea. The segment of chromosomal DNA which codes the subunits of the FO was cloned from four independently isolated DCCD-resistant mutants, and the sequence of the subunit c gene (uncE) was determined. An Ala24 to serine (A24S) substitution was found in the subunit c gene of each mutant. The A24S uncE gene was cloned into the BamHI site of a mutant derivative of plasmid pBR322. The A24S subunit c conferred DCCD resistance to a variety of recipient E. coli strains when it was overexpressed from this plasmid. A 7-base pair deletion beginning at position 132 of the plasmid vector was responsible for the observed overexpression. Hoppe et al. (Hoppe, J., Schairer, H. U., and Sebald, W. (1980) Eur. J. Biochem. 112, 17-24) had previously shown that mutation of subunit c Ile28 to threonine or valine resulted in DCCD resistance. The DCCD sensitivities of the membrane ATPase of these mutants and the A24S mutant were compared. DCCD sensitivity decreased in the order: wild- type much greater than I27V greater than I28T = A24S. The venturicidin sensitivities of wild-type and mutant membranes were also examined. The membrane ATPase of the I28T and I28V mutants was venturicidin resistant whereas the A24S substitution resulted in a hypersensitivity to inhibition by venturicidin. These results support a model in which subunit c folds in the membrane like a hairpin, where the region of residues 24-28 in transmembrane helix-1 is close to that of aspartyl 61 in transmembrane helix-2
RP  - NOT IN FILE
NT  - UI - 92041957LA - engRN - 0 (DNA, Bacterial)RN - 0 (Genetic Vectors)RN - 0 (Venturicidins)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-45-1 (Serine)RN - 6898-94-8 (Alanine)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - 5-T32-GM07133/GM/NIGMSID - GM23105/GM/NIGMSDA - 19911213IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:1834653
SO  - J Biol Chem 1991 Nov 5 ;266(31):20934-20939

1019
UI  - 42
AU  - Fraga D
AU  - Fillingame RH
AD  - Department of Physiological Chemistry, University of Wisconsin Medical School, Madison 53706
TI  - Essential residues in the polar loop region of subunit c of Escherichia coli F1F0 ATP synthase defined by random oligonucleotide-primed mutagenesis
AB  - The conserved, polar loop region of subunit c of the Escherichia coli F1F0 ATP synthase is postulated to function in the coupling of proton translocation through F0 to ATP synthesis in F1. We have used a random mutagenesis procedure to define the essential residues in the region. Oligonucleotide-directed mutagenesis was carried out with a random mixture of mutant oligonucleotides, the oligonucleotide mixture being generated by chemical synthesis by using phosphoramidite nucleotide stocks that were contaminated with the other three nucleotides. Thirty mutant genes coding single-amino-acid substitutions in the region between Glu-37 and Leu-45 of subunit c were tested for function by analyzing the capacity of plasmids carrying the mutant genes to complement a Leu-4----amber subunit c mutant. All substitutions at the conserved Arg-41 residue resulted in loss of oxidative phosphorylation, i.e., transformants could not grow on a succinate carbon source. The other conserved residues were more tolerant to substitution, although most substitutions did result in impaired growth on succinate. We conclude that Arg-41 is essential in the function of the polar loop and that the ensemble of other conserved residues collectively maintain an optimal environment required for that function
RP  - NOT IN FILE
NT  - UI - 91193224LA - engRN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM-23105/GM/NIGMSDA - 19910514IS - 0021-9193SB - IMCY - UNITED STATESJC - HH3
UR  - PM:2013577
SO  - J Bacteriol 1991 Apr ;173(8):2639-2643

1020
UI  - 9903
AU  - Hafner RP
AU  - Brand MD
TI  - Effect of protonmotive force on the relative proton stoichiometries of the mitochondrial proton pumps.
AB  - The rate of phosphorylation of ADP by isolated mitochondria respiring on succinate was set by addition of ATP, ADP or ADP plus malonate. We measured the rates of phosphorylation and respiration and the protonmotive force under each of these conditions. We measured the oxygen consumption required to drive the proton leak at the protonmotive force reached under each condition and subtracted it from the respiration rate during phosphorylation to determine the oxygen consumption driving phosphorylation. By dividing the rate of phosphorylation by the rate of respiration driving phosphorylation we calculated the mechanistic P/O ratio (number of molecules of ADP phosphorylated per oxygen atom reduced). This ratio was the same at high, intermediate and low values of protonmotive force, indicating that the relative stoichiometries of the mitochondrial protonmotive-force-producing and protonmotive-force-consuming pumps (i.e. H+/O:H+/ATP) are independent of the protonmotive force. This greatly weakens the case for a decrease in stoichiometry, or 'slip', in the mitochondrial proton pumps at high protonmotive force.
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - ADP
MH  - Animal
MH  - atp
MH  - Biochemistry
MH  - Electrophysiology
MH  - Female
MH  - Ion Channels
MH  - Kinetics
MH  - Membrane Potentials
MH  - metabolism
MH  - Mitochondria
MH  - Onium Compounds
MH  - Oxygen
MH  - Oxygen Consumption
MH  - Phosphorylation
MH  - physiology
MH  - proton
MH  - Proton Pump
MH  - Protons
MH  - Rats
MH  - Rats,Inbred Strains
MH  - Respiration
MH  - succinate
MH  - Support,Non-U.S.Gov't
MH  - Trityl Compounds
RP  - NOT IN FILE
NT  - Department of Biochemistry, University of Cambridge, UKPMID- 0001708235
SO  - Biochem J 1991 Apr 1 ;275(Pt 1):75-80

1021
UI  - 18549
AU  - Heberle J
TI  - Zeitauflsende Untersuchung der Protonentranslokationsschritte von bakteriorhodopsin mittels chemisch-gekoppelter pH-Indikatoren
PB  - Freien Universitt Berlin
RP  - NOT IN FILE
SO  -  1991  ;():

1022
UI  - 572
AU  - Hekman C
AU  - Hatefi Y
AD  - Department of Molecular and Experimental Medicine, Research Institute of Scripps Clinic, La Jolla, California 92037
TI  - The F0 subunits of bovine mitochondrial ATP synthase complex: purification, antibody production, and interspecies cross- immunoreactivity
AB  - The known subunits of the membrane sector F0 of the bovine mitochondrial ATP synthase complex are subunits b, d, 6, F6, OSCP (oligomycin sensitivity-conferring protein), the DCCD (dicyclohexylcarbodiimide) binding proteolipid, and A6L. The first six subunits were purified from SMP or preparations of the ATP synthase complex, and monospecific antibodies were raised against each. The antisera were shown to be competent for immuno-blotting, and each antiserum recognized a single polypeptide of the expected Mr in preparations of the ATP synthase complex. Immunoblots utilizing antibodies to OSCP and subunits d and 6, which exhibit the same Mr on dodecyl sulfate-polyacrylamide gels, showed clearly that these polypeptides are immunologically distinct. Immunological cross- reactivity was demonstrated between bovine, human, rat, Saccharomyces cerevisiae, Paracoccus denitrificans, and Escherichia coli for subunit 6; between bovine, human, and rat for subunits b, d, OSCP, and F6; and between bovine and rat for the DCCD binding proteolipid. Anti-subunit 6 antiserum, before or after immunopurification against the ATP synthase complex, recognized a single polypeptide in the bovine ATP synthase complex and S. cerevisiae mitochondria, but two polypeptides of different Mr in bovine SMP, human, and rat mitochondria, and Paracoccus and E. coli membranes
RP  - NOT IN FILE
NT  - UI - 91112812LA - engRN - 0 (Macromolecular Systems)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - DK08126/DK/NIDDKDA - 19910228IS - 0003-9861SB - IMCY - UNITED STATESJC - 6SK
UR  - PM:1824914
SO  - Arch Biochem Biophys 1991 Jan ;284(1):90-97

1023
UI  - 571
AU  - Hekman C
AU  - Tomich JM
AU  - Hatefi Y
AD  - Department of Molecular and Experimental Medicine, Research Institute of Scripps Clinic, La Jolla, California 92037
TI  - Mitochondrial ATP synthase complex. Membrane topography and stoichiometry of the F0 subunits
AB  - The topography of the subunits of the membrane sector F0 of the ATP synthase complex in the bovine mitochondrial inner membrane was studied with the help of subunit-specific antibodies raised to the F0 subunits b, d, 6, F6, A6L, OSCP (oligomycin-sensitivity-conferring protein), and N,N' -dicyclohexylcarbodiimide (DCCD)-binding proteolipid and to the ATPase inhibitor protein (IF1) as an internal control. Exposure of F0 subunits in inverted and right-side-out inner membranes was investigated by direct antibody binding as well as by susceptibility of these subunits to degradation by various proteases as monitored by gel electrophoresis of the membrane digests and immunoblotting with the subunit-specific antibodies. Results show that subunits b, d, F6, A6L (including its C-terminal end) and OSCP were exposed on the matrix side. Sufficient masses of these subunits to recognize antibodies or undergo proteolysis were not exposed on the cytosolic side. This was also the case for subunit 6 and the DCCD-binding proteolipid on either side of the inner membrane. Quantitative immunoblotting in which bound radio-activity from [125I]protein A was employed to estimate the concentration of an antigen in a sample allowed the determination of the stoichiometry of several F0 subunits and IF1 relative to F1-ATPase. Results showed that per mol of F1 there are in bovine heart mitochondria 1 mol each of d, OSCP, and IF1, and 2 mol each of b and F6. Subunit 6 and the DCCD-binding proteolipid could not be quantitated, because the former transferred poorly to nitrocellulose and the latter's antibody did not bind [125I]protein A
RP  - NOT IN FILE
NT  - UI - 91310623LA - engRN - 0 (Macromolecular Systems)RN - 0 (Membrane Proteins)RN - 0 (Peptide Fragments)RN - EC 3.4.21.1 (Chymotrypsin)RN - EC 3.4.21.4 (Trypsin)RN - EC 3.4.24.- (Pronase)RN - EC 3.4.24.27 (Thermolysin)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - DK08126/DK/NIDDKDA - 19910823IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:1830306
SO  - J Biol Chem 1991 Jul 25 ;266(21):13564-13571

1024
UI  - 20950
AU  - Hensel M
AU  - Deckers-Hebestreit G
AU  - Altendorf K
AD  - Arbeitsgruppe Mikrobiologie, Fachbereich Biologie/Chemie, Universitat Osnabruck, Federal Republic of Germany
TI  - Purification and characterization of the F1 portion of the ATP synthase (F1Fo) of Streptomyces lividans
AB  - The F1 complex of the ATP synthase of Streptomyces lividans was isolated and purified. The procedure involved the solubilization of F1 from membranes with buffer of low ionic strength in the presence of EDTA, ion-exchange chromatography and gel filtration. The purified F1 complex from S. lividans (SLF1) consists of five subunits alpha, beta, gamma, delta and epsilon with molecular masses of 58,000, 50,000, 36,000, 28,000 and 13,000, respectively and exhibits immunological cross-reactivity with the F1 portion purified from Escherichia coli (ECF1). The enzymatic properties of SLF1 were determined by the use of microtiter-plate-based assay and compared with data obtained for ECF1. ATPase activity of SLF1 (specific activity: 20-30 U/mg) was only observed in the presence of high concentrations of Ca2+ (10mM). Stimulation of the ATPase activity by Mg2+ was not detectable; quite to the contrary, Mg2+ inhibited the Ca(2+)-stimulated activity of SLF1. SLF1 was re-bound to F1-stripped membranes of S. lividans, but not to F1-stripped membrane vesicles of E. coli. In contrast, ECF1 could be cross-reconstituted with F1-stripped membranes of S. lividans; however, a structural but not a functional reconstitution of the hybrid F1Fo complex was observed
MH  - A
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - buffer
MH  - COMPLEX
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1
MH  - ion exchange
MH  - Macromolecular Systems
MH  - membrane
MH  - membrane vesicles
MH  - Membranes
MH  - purification
MH  - reconstitution
MH  - SUBUNIT
MH  - SYNTHASE
MH  - SYSTEM
MH  - SYSTEMS
RP  - NOT IN FILE
NT  - UI - 92111511LA - engRN - 0 (Deoxyribonucleotides)RN - 0 (Macromolecular Systems)RN - 0 (Ribonucleotides)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19920214IS - 0014-2956SB - IMCY - GERMANY
UR  - PM:1837270
SO  - Eur J Biochem 1991 Dec 18 ;202(3):1313-1319

1025
UI  - 20868
AU  - Ida K
AU  - Noumi T
AU  - Maeda M
AU  - Fukui T
AU  - Futai M
AD  - Department of Organic Chemistry and Biochemistry, Osaka University, Japan
TI  - Catalytic site of F1-ATPase of Escherichia coli. Lys-155 and Lys-201 of the beta subunit are located near the gamma-phosphate group of ATP in the presence of Mg2+
AB  - The catalytic site of Escherichia coli F1 was probed using a reactive ATP analogue, adenosine triphosphopyridoxal (AP3-PL). For complete loss of enzyme activity, about 1 mol of AP3-PL bound to 1 mol of F1 was estimated to be required in the presence or absence of Mg2+. About 70% of the label was bound to the alpha subunit and the rest to the beta subunit in the absence of Mg2+, and the alpha Lys-201 and beta Lys-155 residues, respectively, were the major target residues (Tagaya, M., Noumi, T., Nakano, K., Futai, M., and Fukui, T. (1988) FEBS Lett. 233, 347-351). Addition of Mg2+ decreased the AP3-PL concentration required for half-maximal inhibition, and predominant labeling of the beta subunit (beta Lys-155 and beta Lys-201) with the reagent. ATP and ADP were protective ligands in the presence and absence of Mg2+. The alpha subunit mutation (alpha Lys-201----Gln or alpha Lys-201 deletion) were active in oxidative phosphorylation. However, purified mutant F1s showed impaired low multi-site activity, although their uni-site catalyses were essentially normal. Thus alpha Lys-201 is not a catalytic residue, but may be important for catalytic cooperativity. Mutant F1s were inhibited less by AP3-PL in the absence of Mg2+, and consistent with this, modifications of their alpha subunits by AP3-PL were reduced. AP3-PL was more inhibitory to the mutant enzymes in the presence of Mg2+, and bound to the beta Lys-155 and beta Lys-201 residues of mutant F1 (alpha Lys-201----Gln). These results strongly suggest that alpha Lys-201, beta Lys-155, and beta Lys-201 are located close together near the gamma-phosphate group of ATP bound to the catalytic site, and that the two beta residues and the gamma-phosphate group become closer to each other in the presence of Mg2+
MH  - A
MH  - ACTIVE
MH  - Adenosine
MH  - ADP
MH  - ALPHA-SUBUNIT
MH  - atp
MH  - ATPase
MH  - BETA-SUBUNIT
MH  - Biochemistry
MH  - Chemistry
MH  - Enzymes
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - Ligands
MH  - M
MH  - Magnesium
MH  - mutant
MH  - Oxidative Phosphorylation
MH  - Phosphorylation
MH  - RESIDUE
MH  - Site
MH  - SUBUNIT
RP  - NOT IN FILE
NT  - UI - 91170203LA - engRN - 0 (Plasmids)RN - 56-87-1 (Lysine)RN - 7439-95-4 (Magnesium)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19910423IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:1825998
SO  - J Biol Chem 1991 Mar 25 ;266(9):5424-5429

1026
UI  - 840
AU  - Jackson JB
AD  - School of Biochemistry, University of Birmingham, Edgbaston, UK
TI  - Phototropic bacteria--useful organisms for class experiments
RP  - NOT IN FILE
NT  - UI - 92175352LA - engPT - Journal ArticleDA - 19920406IS - 0300-5127SB - IMCY - ENGLANDJC - E48
UR  - PM:1794595
SO  - Biochem Soc Trans 1991 Nov ;19(4):976

1027
UI  - 21003
AU  - Junesch U
AU  - Graber P
AD  - Biologisches Institut, Universitat Stuttgart, Germany
TI  - The rate of ATP-synthesis as a function of delta pH and delta psi catalyzed by the active, reduced H(+)-ATPase from chloroplasts
AB  - The H(+)-ATPase from chloroplasts was brought into the active, reduced state. Then, an electrochemical potential difference of protons across the thylakoid membranes was generated by an acid-base transition, delta pH, combined with a K+/valinomycin diffusion potential, delta psi. The initial rate of ATP synthesis was measured with a rapid-mixing quenched- flow apparatus in the time-range between 20-150 ms. The rate of ATP synthesis depends in a sigmoidal way on delta pH. Increasing diffusion potentials shifts the delta pH-dependencies to lower delta pH values. Analysis of the data indicate that the rate of ATP synthesis depends on the electrochemical potential difference of protons irrespective of the relative contribution of delta pH and delta psi
MH  - A
MH  - ACTIVE
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - analysis
MH  - atp
MH  - ATP synthesis
MH  - ATPase
MH  - chloroplast
MH  - Chloroplasts
MH  - DELTA-PH
MH  - Diffusion
MH  - diffusion potential
MH  - function
MH  - H(+)ATPase
MH  - membrane
MH  - Membranes
MH  - pH
MH  - proton
MH  - Protons
MH  - PSI
MH  - synthesis
MH  - thylakoid
MH  - thylakoid membrane
RP  - NOT IN FILE
NT  - UI - 92097787LA - engRN - 0 (Protons)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19920205IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:1661688
SO  - FEBS Lett 1991 Dec 9 ;294(3):275-278

1028
UI  - 20951
AU  - Kauffer S
AU  - Deckers-Hebestreit G
AU  - Altendorf K
AD  - Universitat Osnabruck, Fachbereich Biologie/Chemie, Arbeitsgruppe Mikrobiologie, Federal Republic of Germany
TI  - Substitution of the cysteinyl residue (Cys21) of subunit b of the ATP synthase from Escherichia coli
AB  - The Fo complex of the ATP synthase (F1Fo) of Escherichia coli contains only two cysteinyl residues, Cys21, of the two copies of subunit b. Modification of Cys21 with the hydrophobic maleimide N-(7-dimethylamino- 4-methyl-coumarinyl)maleimide resulted in impairment of Fo functions [Schneider, E. & Altendorf, K. (1985) Eur. J. Biochim. 153, 105-109]. We replaced this residue (via cassette mutagenesis) by Ser, Gly, Ala, Thr, Asp and Pro. None of the replacements resulted in detectable alterations of the function of the ATP synthase, making a functional role for these sulfhydryl residues unlikely. Due to its high tolerance towards amino acid substitutions, the region around Cys21 seems not to be a protein-protein contact area
MH  - A
MH  - ACID
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - COMPLEX
MH  - Cysteine
MH  - Dicyclohexylcarbodiimide
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - function
MH  - Macromolecular Systems
MH  - mutagenesis
MH  - protein
MH  - Proteins
MH  - RESIDUE
MH  - SUBUNIT
MH  - SYNTHASE
MH  - SYSTEM
MH  - SYSTEMS
RP  - NOT IN FILE
NT  - UI - 92111510LA - engRN - 0 (Macromolecular Systems)RN - 0 (Oligodeoxyribonucleotides)RN - 0 (Plasmids)RN - 0 (Recombinant Proteins)RN - 52-90-4 (Cysteine)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19920214IS - 0014-2956SB - IMCY - GERMANY
UR  - PM:1837269
SO  - Eur J Biochem 1991 Dec 18 ;202(3):1307-1312

1029
UI  - 442
AU  - Lee RS
AU  - Pagan J
AU  - Wilke-Mounts S
AU  - Senior AE
AD  - Department of Biochemistry, University of Rochester School of Medicine and Dentistry, New York 14642
TI  - Characterization of Escherichia coli ATP synthase beta-subunit mutations using a chromosomal deletion strain
AB  - (1) We constructed Escherichia coli strain JP17 with a deletion in the ATP synthase beta-subunit gene. JP17 is completely deficient in ATP synthase activity and expresses no beta-subunit. Expression of normal beta-subunit from a plasmid restores haploid levels of ATP synthase in membranes. JP17 was shown to be efficacious for studies of beta-subunit mutations. Site-directed mutants were studied directly in JP17. Randomly generated chromosomal mutants were identified by PCR and DNA sequencing, cloned, and expressed in JP17. (2) Eight novel mutations occurring within the putative catalytic nucleotide-binding domain were characterized with respect to their effects on catalysis and structure. The mutations beta C137S, beta G152D, beta G152R, beta E161Q, beta E161R, and beta G251D each impaired catalysis without affecting enzyme assembly or oligomeric structure and are of interest for future studies of catalytic mechanism. The mutations beta D301V and beta D302V, involving strongly conserved carboxyl residues, caused oligomeric instability of F1. However, growth characteristics of these mutants suggested that neither carboxyl side chain is critical for catalysis. (3) The mutations beta R398C and beta R398W rendered ATP synthase resistant to aurovertin, giving strong support to the view that beta R398 is a key residue in the aurovertin-binding site. Neither beta R398C or beta R398W impaired catalysis significantly
RP  - NOT IN FILE
NT  - UI - 91299736LA - engRN - 0 (Aurovertins)RN - 0 (Receptors, Drug)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSDA - 19910822IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:1829962
SO  - Biochemistry 1991 Jul 16 ;30(28):6842-6847

1030
UI  - 20819
AU  - McCormick KA
AU  - Cain BD
AD  - Department of Biochemistry and Molecular Biology, University of Florida, Gainesville 32610-0245
TI  - Targeted mutagenesis of the b subunit of F1F0 ATP synthase in Escherichia coli: Glu-77 through Gln-85
AB  - Subunit b of Escherichia coli F1F0 ATP synthase contains a large hydrophilic region thought to be involved in the interaction between F1 and F0. Oligonucleotide-directed mutagenesis was used to evaluate the functional importance of a segment of this region from Glu-77 through Gln-85. The mutagenesis procedure employed a phagemid DNA template and a doped oligonucleotide primer designed to generate a predetermined collection of missense mutations in the target segment. Sixty-one mutant phagemids were identified and shown to contain nucleotide substitutions encoding 37 novel missense mutations. Mutations were isolated singly or in combinations of up to four mutations per recombinant phagemid. F1F0 ATP synthase function was studied by mutant phagemid complementation of a novel E. coli strain in which the uncF (b) gene was deleted. Complementation was assessed by observing growth on solid succinate minimal medium. Many phagemid-encoded uncF (b) gene mutations in the targeted segment resulted in growth phenotypes indistinguishable from those of strains expressing the native b subunit, suggesting abundant F1F0 ATP synthase activity. In contrast, several specific mutations were associated with a loss of enzyme function. Phagemids specifying the Ala-79----Pro, Arg-82----Pro, Arg-83- ---Pro, or Gln-85----Pro mutation failed to complement uncF (b) gene- deficient E. coli. F1F0 ATP synthase displayed the greatest sensitivity to mutations altering a single site in the target segment, Ala-79. The evidence suggests that Ala-79 occupies a restricted position in the enzyme complex
MH  - A
MH  - ACID
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Biochemistry
MH  - COMPLEX
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - F1
MH  - function
MH  - glutamic acid
MH  - Glutamine
MH  - Macromolecular Systems
MH  - mutagenesis
MH  - mutant
MH  - Site
MH  - SUBUNIT
MH  - succinate
MH  - SYNTHASE
MH  - SYSTEM
MH  - SYSTEMS
RP  - NOT IN FILE
NT  - UI - 92041622LA - engRN - 0 (Glutamates)RN - 0 (Macromolecular Systems)RN - 0 (Oligodeoxyribonucleotides)RN - 0 (Plasmids)RN - 56-85-9 (Glutamine)RN - 56-86-0 (Glutamic Acid)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleID - GM43495/GM/NIGMSDA - 19911220IS - 0021-9193SB - IMCY - UNITED STATES
UR  - PM:1682301
SO  - J Bacteriol 1991 Nov ;173(22):7240-7248

1031
UI  - 181
AU  - Mendel-Hartvig J
AU  - Capaldi RA
AD  - Institute of Molecular Biology, University of Oregon, Eugene 97403
TI  - Catalytic site nucleotide and inorganic phosphate dependence of the conformation of the epsilon subunit in Escherichia coli adenosinetriphosphatase
AB  - The rate of trypsin cleavage of the epsilon subunit of Escherichia coli F1 (ECF1) has been found to be ligand-dependent, as measured indirectly by the activation of the enzyme that occurs on protease digestion, or when followed directly by monitoring the cleavage of this subunit using monoclonal antibodies. The cleavage of the epsilon subunit was fast in the presence of ADP alone, ADP + MG2+, ATP + EDTA, or AMP-PNP, but slow when Pi was added along with ADP + Mg2+ or when ATP + Mg2+ was added to generate ADP + Pi (+Mg2+) in the catalytic site(s). The half-maximal concentration of Pi required in the presence of ADP + Mg2+ to protect the epsilon subunit from cleavage by trypsin was 50 microM, which is in the range measured for the high-affinity binding of Pi to F1. The ligand-dependent conformational changes in the epsilon subunit were also examined in cross-linking experiments using the water-soluble carbodiimide 1-ethyl-3-[3-(dimethylamino)propyl]carbodiimide (EDC). In the presence of ATP + Mg2+ or ADP + Mg2+ + Pi, the epsilon subunit cross-linked to beta in high yield. With ATP + EDTA or ADP + Mg2+ (no Pi), the yield of the beta-epsilon cross-linked product was much reduced. We conclude that the epsilon subunit undergoes a conformational change dependent on the presence of Pi. It has been found previously that binding of the epsilon subunit to ECF1 inhibits ATPase activity by decreasing the off rate of Pi [Dunn, S. D., Zadorozny, V. D., Tozer, R. G., & Orr, L. E. (1987) Biochemistry 26, 4488-4493]. This reciprocal relationship between Pi binding and epsilon- subunit conformation has important implications for energy transduction by the E. coli ATP synthase
RP  - NOT IN FILE
NT  - UI - 91120772LA - engRN - 0 (Adenine Nucleotides)RN - 0 (Macromolecular Systems)RN - 0 (Phosphates)RN - EC 3.4.21.4 (Trypsin)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - HL24526/HL/NHLBIDA - 19910314IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:1825019
SO  - Biochemistry 1991 Feb 5 ;30(5):1278-1284

1032
UI  - 180
AU  - Mendel-Hartvig J
AU  - Capaldi RA
AD  - Institute of Molecular Biology, University of Oregon, Eugene 97403
TI  - Structure-function relationships of domains of the delta subunit in Escherichia coli adenosine triphosphatase
AB  - The topology of the and subunit of the Escherichia coli adenosinetriphosphatase (ECF1) has been explored by proteinase digestion and chemical labeling methods. The delta subunit of ECF1 could be cleaved selectively by reaction of the enzyme complex with very low amounts of trypsin (1:5000, w/w). Cleavage of the delta subunit occurred serially from the C-terminus. The N-terminal fragments of the delta subunit remained bound to the core ECF1 complex through sucrose gradient centrifugation, indicating that part of the binding of this subunit involves the N-terminal segment. ECF1, in which around 20 amino acids had been removed from the C-terminus of delta, still bound to ECF0 but DCCD sensitivity of the ATPase activity was lost. When ECF1 was reacted with N-ethyl[14C]maleimide ([14C]NEM) in the native state, only one of the two Cys residues on the delta subunit was modified. This residue, Cys-140, was also labeled in ECF1F0. Cys-140 was shown to be involved in the disulfide bridge between alpha and delta subunits that is generated when ECF1 is treated with CuCl2. Thus, the C-terminal part of the delta subunit around Cys-140 can interact with the core ECF1 complex. These results suggest a model for the delta subunit in which the central part of polypeptide is a part of the stalk, with both N- and C-termini associated with ECF1
RP  - NOT IN FILE
NT  - UI - 92002100LA - engRN - 52-90-4 (Cysteine)RN - EC 3.4.21.4 (Trypsin)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - HL24526/HL/NHLBIDA - 19911118IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:1655028
SO  - Biochim Biophys Acta 1991 Sep 27 ;1060(1):115-124

1033
UI  - 179
AU  - Mendel-Hartvig J
AU  - Capaldi RA
AD  - Institute of Molecular Biology, University of Oregon, Eugene 97403
TI  - Nucleotide-dependent and dicyclohexylcarbodiimide-sensitive conformational changes in the epsilon subunit of Escherichia coli ATP synthase
AB  - The rate of trypsin cleavage of the epsilon subunit of Escherichia coli F1F0 (ECF1F0) is shown to be ligand-dependent as measured by Western analysis using monoclonal antibodies. The cleavage of the epsilon subunit was rapid in the presence of ADP alone, ATP + EDTA, or AMP-PNP + Mg2+, but slow when Pi was added along with ADP + Mg2+ or when ATP + Mg2+ was added to generate ADP + Pi (+Mg2+) in the catalytic site. Trypsin treatment of ECF1Fo was also shown to increase enzymic activity on a time scale corresponding to that of the cleavage of the epsilon subunit, indicating that the epsilon subunit inhibits ATPase activity in ECF1Fo. The ligand-dependent conformational changes in the epsilon subunit were also examined in cross-linking experiments using the water- soluble carbodiimide 1-ethyl-3-[3-(dimethylamino)propyl]-carbodiimide (EDC). In the presence of ATP + Mg2+ or ADP + Pi + Mg2+, the epsilon subunit cross-linked product was much reduced. Prior reaction of ECF1Fo with dicyclohexylcarbodiimide (DCCD), under conditions in which only the Fo part was modified, blocked the conformational changes induced by ligand binding. When the enzyme complex was reacted with DCCD in ATP + EDTA, the cleavage of the epsilon subunit was rapid and yield of cross- linking of beta to epsilon subunit low, whether trypsin cleavage was conducted in ATP + EDTA or ATP + Mg2+. When enzyme was reacted with DCCD in ATP + Mg2+, cleavage of the epsilon subunit was slow and yield of cross-linking of beta to epsilon high, under all nucleotide conditions for proteolysis.(ABSTRACT TRUNCATED AT 250 WORDS)
RP  - NOT IN FILE
NT  - UI - 92031559LA - engRN - 0 (Macromolecular Systems)RN - 1892-57-5 (Ethyldimethylaminopropyl Carbodiimide)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 67-42-5 (Egtazic Acid)RN - EC 3.4.21.4 (Trypsin)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - HL24536/HL/NHLBIDA - 19911211IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:1834172
SO  - Biochemistry 1991 Nov 12 ;30(45):10987-10991

1034
UI  - 19781
AU  - Milgrom YM
AU  - Ehler LL
AU  - Boyer PD
AD  - Department of Chemistry and Biochemistry, University of California, Los Angeles 90024
TI  - The characteristics and effect on catalysis of nucleotide binding to noncatalytic sites of chloroplast F1-ATPase
AB  - The recent finding that the presence of ATP at non-catalytic sites of chloroplast F1-ATPase (CF1) is necessary for ATPase activity (Milgrom, Y. M., Ehler, L. L., and Boyer, P. D. (1990) J. Biol. Chem. 265,18725- 18728) prompted more detailed studies of the effect of noncatalytic site nucleotides on catalysis. CF1 containing at noncatalytic sites less than one ADP or about two ATP was prepared by heat activation in the absence of Mg2+ and in the presence of ADP or ATP, respectively. After removal of medium nucleotides, the CF1 preparations were used for measurement of the time course of nucleotide binding from 10 to 100 microM concentrations of 3H-labeled ADP, ATP, or GTP. The presence of Mg2+ strongly promotes the tight binding of ADP and ATP at noncatalytic sites. For example, the ADP-heat-activated enzyme in presence of 1 mM Mg2+ binds ADP with a rate constant of 0.5 x 10(6) M-1 min-1 to give an enzyme with two ADP at noncatalytic sites with a Kd of about 0.1 microM. Upon exposure to Mg2+ and ATP the vacant noncatalytic site binds an ATP rapidly and, as an ADP slowly dissociates, a second ATP binds. The binding correlates with an increase in the ATPase activity. In contrast the tight binding of [3H]GTP to noncatalytic sites gives an enzyme with no ATPase activity. The three noncatalytic sites differ in their binding properties. The noncatalytic site that remains vacant after the ADP-heat-activated CF1 is exposed to Mg2+ and ADP and that can bind ATP rapidly is designated as site A; the site that fills with ATP as ADP dissociates when this enzyme is exposed to Mg2+ and ATP is called site B, and the site to which ADP remains bound is called site C. Procedures are given for attaining CF1 with ADP at sites B and C, with GTP at sites A and/or B, and with ATP at sites A, B, and/or C, and catalytic activities of such preparations are measured. For example, little or no ATPase activity is found unless ATP is at site A, but ADP can remain at site C with no effect on ATPase. Maximal GTPase activity requires ATP at site A but about one-fifth of maximal GTPase is attained when GTP is at sites A and B and ATP at site C. Noncatalytic site occupancy can thus have profound effects on the ATPase and GTPase activities of CF1
MH  - A
MH  - ACTIVATION
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - ADP
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BINDING
MH  - Biochemistry
MH  - Catalysis
MH  - Chemistry
MH  - chloroplast
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - Guanosine Triphosphate
MH  - H(+)-Transporting ATP Synthase
MH  - Heat
MH  - non-catalytic site
MH  - NONCATALYTIC SITES
MH  - nucleotide binding
MH  - Nucleotides
MH  - P
MH  - SYNTHASE
MH  - Time
RP  - NOT IN FILE
NT  - UI - 91268015LA - engRN - 0 (Chlorides)RN - 0 (Nucleotides)RN - 0 (Sulfuric Acids)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 86-01-1 (Guanosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM-11094/GM/NIGMSDA - 19910724IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:1828802
SO  - J Biol Chem 1991 Jun 25 ;266(18):11551-11558

1035
UI  - 1006
AU  - Mitchell P
TI  - Foundations of vectorial metabolism and osmochemistry
AB  - Chemical transformations, like osmotic translocations, are transport processes when looked at in detail. In chemiosmotic systems, the pathways of specific ligand conduction are spatially orientated through osmoenzymes and porters in which the actions of chemical group, electron and solute transfer occur as vectorial (or higher tensorial order) diffusion processes down gradients of total potential energy that represent real spatially-directed fields of force. Thus, it has been possible to describe classical bag-of-enzymes biochemistry as well as membrane biochemistry in terms of transport. But it would not have been possible to explain biological transport in terms of classical transformational biochemistry or chemistry. The recognition of this conceptual asymmetry in favour of transport has seemed to be upsetting to some biochemists and chemists; and they have resisted the shift towards thinking primarily in terms of the vectorial forces and co-linear displacements of ligands in place of their much less informative scalar products that correspond to the conventional scalar energies. Nevertheless, considerable progress has been made in establishing vectorial metabolism and osmochemistry as acceptable biochemical disciplines embracing transport and metabolism, and bioenergetics has been fundamentally transformed as a result
MH  - Biochemistry
MH  - Biological Transport
MH  - Biological Transport,Active
MH  - Chemistry
MH  - Diffusion
MH  - Ligands
MH  - metabolism
MH  - Models,Chemical
MH  - Osmosis
MH  - Support,U.S.Gov't,P.H.S.
RP  - NOT IN FILE
NT  - Glynn Research Institute, Bodmin, Cornwall, United Kingdom
SO  - Biosci Rep 1991 Dec ;11(6):297-344

1036
UI  - 20867
AU  - Moriyama Y
AU  - Iwamoto A
AU  - Hanada H
AU  - Maeda M
AU  - Futai M
AD  - Department of Organic Chemistry and Biochemistry, Osaka University, Japan
TI  - One-step purification of Escherichia coli H(+)-ATPase (F0F1) and its reconstitution into liposomes with neurotransmitter transporters
AB  - About 30% of the protein in the inner membrane of Escherichia coli strain DK8/pBWU13 is H(+)-ATPase (F0F1), and practically homogeneous F0F1 could be obtained by gradient centrifugation after solubilization of these membranes. The recombinant plasmid pBWU13 carries the unc operon for F0F1. When reconstituted into liposomes, F0F1 formed an ATP- dependent proton gradient and membrane potential. Proteoliposomes reconstituted with F0F1 and solubilized transporters from chromaffin granules or synaptic vesicle membranes could transport serotonin, dopamine, and norepinephrine dependent on ATP hydrolysis. F0F1 can be obtained rapidly from DK8/pBWU13, and its reconstitution into liposomes with transporters may be useful for monitoring these transporters during their purification
MH  - atp
MH  - ATPase
MH  - Biochemistry
MH  - Carrier Proteins
MH  - Chemistry
MH  - Chromaffin Granules
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0F1
MH  - H(+)ATPase
MH  - Hydrolysis
MH  - Liposomes
MH  - membrane
MH  - Membrane Potential
MH  - Membranes
MH  - protein
MH  - Proteins
MH  - Proteolipids
MH  - proteoliposome
MH  - proton
MH  - purification
MH  - reconstitution
MH  - transport
RP  - NOT IN FILE
NT  - UI - 92042140LA - engRN - 0 (Biogenic Monoamines)RN - 0 (Carrier Proteins)RN - 0 (Liposomes)RN - 0 (Plasmids)RN - 0 (Proteolipids)RN - 0 (Recombinant Proteins)RN - 0 (proteoliposomes)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19911226IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:1834667
SO  - J Biol Chem 1991 Nov 25 ;266(33):22141-22146

1037
UI  - 748
AU  - Muneyuki E
AU  - Yoshida M
AU  - Bullough DA
AU  - Allison WS
AD  - Research Laboratory of Resources Utilization, Tokyo Institute of Technology, Yokohoma, Japan
TI  - Heterogeneous hydrolysis of substoichiometric ATP by the F1-ATPase from Escherichia coli
AB  - The hydrolysis of 0.3 microM [alpha,gamma-32P]ATP by 1 microM F1-ATPase isolated from the plasma membranes of Escherichia coli has been examined in the presence and absence of inorganic phosphate. The rate of binding of substoichiometric substrate to the ATPase is attenuated by 2 mM phosphate and further attenuated by 50 mM phosphate. Under all conditions examined, only 10-20% of the [alpha,gamma-32P]ATP that bound to the enzyme was hydrolyzed sufficiently slowly to be examined in cold chase experiments with physiological concentrations of non-radioactive ATP. These features differ from those observed with the mitochondrial F1-ATPase. The amount of bound substrate in equilibrium with bound products observed in the slow phase which was subject to promoted hydrolysis by excess ATP was not affected by the presence of phosphate. Comparison of the fluxes of enzyme-bound species detected experimentally in the presence of 2 mM phosphate with those predicted by computer simulation of published rate constants determined for uni- site catalysis (Al-Shawi, M.D., Parsonage, D. and Senior, A.E. (1989) J. Biol. Chem. 264, 15376-15383) showed that hydrolysis of substoichiometric ATP observed experimentally was clearly biphasic. Less than 20% of the substoichiometric ATP added to the enzyme was hydrolyzed according to the published rate constants which were calculated from the slow phase of product release in the presence of 1 mM phosphate. The majority of the substoichiometric ATP added to the enzyme was hydrolyzed with product release that was too rapid to be detected by the methods employed in this study, indicating again that the F1-ATPase from E. coli and bovine heart mitochondria hydrolyze substoichiometric ATP differently
RP  - NOT IN FILE
NT  - UI - 91265525LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM 16,974/GM/NIGMSDA - 19910724IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:1828699
SO  - Biochim Biophys Acta 1991 Jun 17 ;1058(2):304-311

1038
UI  - 19780
AU  - Murataliev MB
AU  - Milgrom YM
AU  - Boyer PD
AD  - Molecular Biology Institute, University of California, Los Angeles 90024-1570
TI  - Characteristics of the combination of inhibitory Mg2+ and azide with the F1 ATPase from chloroplasts
AB  - The interactions between ADP, Mg2+, and azide that result in the inhibition of the chloroplast F1 ATPase (CF1) have been explored further. The binding of the inhibitory Mg2+ with low Kd is shown to occur only when tightly bound ADP is present at a catalytic site. Either the tightly bound ADP forms part of the Mg(2+)-binding site or it induces conformational changes creating the high-affinity site for inhibitory Mg2+. Kinetic studies show that CF1 forms two catalytically inactive complexes with Mg2+. The first complex results from Mg2+ binding with a Kd for Mg2+ dissociation of about 10-15 microM, followed by a slow conversion to a complex with a Kd of about 4 microM. The rate- limiting step of the CF1 inactivation by Mg2+ is the initial Mg2+ binding. When medium Mg2+ is chelated with EDTA, the two complexes dissociate with half-times of about 1 and 7 min, respectively. Azide enhances the extent of Mg(2+)-dependent inactivation by increasing the affinity of the enzyme for Mg2+ 3-4 times and prevents the reactivation of both complexes of CF1 with ADP and Mg2+. This results from decreasing the rate of Mg2+ release; neither the rate of Mg2+ binding to CF1 nor the rate of isomerization of the first inactive complex to the more stable form is affected by azide. This suggests that the tight- binding site for the inhibitory azide requires prior binding of both ADP and Mg2+
MH  - A
MH  - ADP
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Azides
MH  - BINDING
MH  - chloroplast
MH  - Chloroplasts
MH  - COMPLEX
MH  - conformational change
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - H(+)-Transporting ATP Synthase
MH  - Magnesium
MH  - SYNTHASE
MH  - Time
RP  - NOT IN FILE
NT  - UI - 91355181LA - engRN - 0 (Azides)RN - 7439-95-4 (Magnesium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM-11094/GM/NIGMSDA - 19911004IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:1832012
SO  - Biochemistry 1991 Aug 27 ;30(34):8305-8310

1039
UI  - 825
AU  - Pancic PG
AU  - Strotmann H
AU  - Kowallik KV
AD  - Institut fur Biochemie, Pflanzen, Dusseldorf, Germany
TI  - The delta subunit of the chloroplast ATPase is plastid-encoded in the diatom Odontella sinensis
AB  - A 5.2 kb PstI restriction fragment containing the atpA gene cluster of the plastic genome of the centric diatom Odontella sinensis was cloned. Sequencing revealed a reading frame of 561 bp separating the genes atpF and atpA, which is preceded by a putative ribosome binding site. The third nucleotide of the codon for the last amino acid of atpF is the first nucleotide of the initiation codon of the 561 bp reading frame. The amino acid sequence deduced from the nucleotide sequence of this gene (atpD) is colinear with delta subunits of different F0F1-ATPases and shows an overall sequence homology of up to 35% when compared with the sequences of cyanobacteria and Cyanophora paradoxa. The results are discussed in context with the evolution of chloroplasts of the chlorophyll-a + b and -a + c lineages, respectively
RP  - NOT IN FILE
NT  - UI - 91192176LA - engRN - 0 (Codon)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19910513IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:1826484
SO  - FEBS Lett 1991 Mar 25 ;280(2):387-392

1040
UI  - 20952
AU  - Pati S
AU  - Brusilow WS
AU  - Deckers-Hebestreit G
AU  - Altendorf K
AD  - Department of Biochemistry, Wayne State University School of Medicine, Detroit, Michigan 48201
TI  - Assembly of the F0 proton channel of the Escherichia coli F1F0 ATPase: low proton conductance of reconstituted Fo sectors synthesized and assembled in the absence of F1
AB  - We have previously proposed that during assembly of the Escherichia coli F1F0 ATPase, the proton permeability of the Fo sector of the E. coli F1F0 ATPase is increased significantly by interactions with F1 subunits [Pati, S., & Brusilow, W.S.A. (1989) J. Biol. Chem 264, 2640- 2644]. To test this model for Fo assembly, we purified F0 sectors synthesized in the presence and absence of F1 subunits and measured the abilities of these different preparations to bind purified F1 ATPase and to conduct protons when reconstituted into liposomes. The results of these studies demonstrated significant differences in proton- conducting abilities of the different Fo preparations. Fo sectors synthesized in the presence of F1 subunits were more permeable to protons than those synthesized in the absence of F1 subunits
MH  - ATPase
MH  - Biochemistry
MH  - conductance
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - ion
MH  - Ion Channels
MH  - Liposomes
MH  - model
MH  - Permeability
MH  - proton
MH  - Protons
MH  - SUBUNIT
RP  - NOT IN FILE
NT  - UI - 91230106LA - engRN - 0 (Ion Channels)RN - 0 (Liposomes)RN - 0 (Plasmids)RN - 0 (Protons)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleID - KO4-AI00882/AI/NIAIDDA - 19910620IS - 0006-2960SB - IMCY - UNITED STATES
UR  - PM:1709362
SO  - Biochemistry 1991 May 14 ;30(19):4710-4714

1041
UI  - 636
AU  - Penefsky HS
AU  - Cross RL
AD  - Department of Biochemistry and Molecular Biology, SUNY Health Science Center, Syracuse
TI  - Structure and mechanism of FoF1-type ATP synthases and ATPases
RP  - NOT IN FILE
NT  - UI - 91272902LA - engRN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, AcademicID - GM-21737/GM/NIGMSID - GM-23152/GM/NIGMSDA - 19910724IS - 0065-258XSB - IMCY - UNITED STATESJC - 2LM
UR  - PM:1828930
SO  - Adv Enzymol Relat Areas Mol Biol 1991  ;64():173-214

1042
UI  - 21166
AU  - Scarpetta MA
AU  - Hawthorne CA
AU  - Brusilow WS
AD  - Department of Biochemistry, Wayne State University School of Medicine, Detroit, Michigan 48201
TI  - Characterization of semi-uncoupled hybrid Escherichia coli-Bacillus megaterium F1F0 proton-translocating ATPases
AB  - Cloned atp genes for the proton-translocating ATPase of the obligate aerobe Bacillus megaterium have been demonstrated to be capable of complementing Escherichia coli ATPase (unc) mutants (Hawthorne, C. A., and Brusilow, W. S. A. (1986) J. Biol. Chem. 261, 5245-5248). To determine the minimum subunit requirements for cross-species complementation, we constructed all combinations of B. megaterium atpA, G, D, and C genes (coding for the alpha, gamma, beta, and epsilon subunits, respectively) and tested their abilities to complement two uncA (alpha subunit) and two uncD (beta subunit) mutants of E. coli. The results indicated that complementation of either uncD mutant required atpD (beta) only. Complementation of one of the uncA (alpha) mutants required atpA, G, and D (alpha, gamma, and beta) and possibly atpE (epsilon) as well. The other uncA mutant was not complemented by any combination of B. megaterium ATPase genes. Complementation of a beta mutant by atpD (beta) or atpD and C (beta epsilon) produced cells which could grow aerobically on a nonfermentable carbon source (succinate) but not anaerobically on rich medium containing glucose. These E. coli therefore had become obligate aerobes. The ability to grow anaerobically could be restored to the mutant complemented by atpD alone by growth at pH 7.5 or pH 8 in the presence of 0.1 M potassium
MH  - A
MH  - alpha
MH  - ALPHA-SUBUNIT
MH  - atp
MH  - ATPase
MH  - Bacillus
MH  - BETA
MH  - BETA-SUBUNIT
MH  - Biochemistry
MH  - Cells
MH  - EPSILON-SUBUNIT
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - Glucose
MH  - M
MH  - mutant
MH  - pH
MH  - Potassium
MH  - protein
MH  - Proteins
MH  - Proton-Translocating ATPases
MH  - SUBUNIT
MH  - succinate
RP  - NOT IN FILE
NT  - UI - 92011610LA - engRN - 0 (DNA, Bacterial)RN - 0 (Plasmids)RN - 0 (Recombinant Fusion Proteins)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleID - KO4-AI00882/AI/NIAIDDA - 19911108IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:1655755
SO  - J Biol Chem 1991 Oct 5 ;266(28):18567-18572

1043
UI  - 749
AU  - Shirakihara Y
AU  - Yohda M
AU  - Kagawa Y
AU  - Yokoyama K
AU  - Yoshida M
AD  - Department of Physics, Hyogo University of Education
TI  - Purification by dye-ligand chromatography and a crystallization study of the F1-ATPase and its major subunits, beta and alpha, from a thermophilic bacterium, PS3
AB  - For a crystallization study, purification methods for F1-ATPase from a thermophilic bacterium, PS3, and its major subunits, beta and alpha, have been improved. The improvement depended on the introduction of dye- ligand chromatography columns to the previously adopted array of chromatography columns: a Blue-B (a blue dye bound to agarose) column was introduced for the F1 preparation, a Green-A column (a green dye attached to agarose) for the beta subunit, and a Blue-A (another blue dye, Cibacron Blue 3GA, bound to agarose) column for the alpha subunit. The improved preparations of all the proteins had purities of nearly 99%. Using the highly purified preparations of the proteins, crystallization conditions were searched for in a systematic way. Large plate crystals (0.2 X 0.5 X 0.5 mm) of F1 were grown from a polyethylene glycol solution. However, neither of the subunits was crystallized, in spite of extensive search for crystallization conditions
RP  - NOT IN FILE
NT  - UI - 91349189LA - engRN - 0 (Dyes)RN - 0 (Ligands)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19910927IS - 0021-924XSB - IMCY - JAPANJC - HIF
UR  - PM:1831812
SO  - J Biochem (Tokyo ) 1991 Mar ;109(3):466-471

1044
UI  - 747
AU  - Tsutsumi S
AU  - Denda K
AU  - Yokoyama K
AU  - Oshima T
AU  - Date T
AU  - Yoshida M
AD  - Department of Life Science, Tokyo Institute of Technology, Yokohama, Japan
TI  - Molecular cloning of genes encoding major two subunits of a eubacterial V-type ATPase from Thermus thermophilus
AB  - The atpAB genes which encode the alpha and beta subunits of membrane ATPase from a thermophilic eubacterium, Thermus thermophilus HB8, were cloned. The deduced amino-acid sequences of the alpha subunit (583 amino acids) and the beta subunit (478 amino acids) are only moderately similar to the alpha beta subunits of the F0F1-ATPases, while they are highly similar to the major two subunits of the V-type ATPases, a family of ATPases which have been so far found in eukaryotic endomembrane vacuolar vesicles and archaebacterial plasma membranes. Thus, T. thermophilus ATPase belongs to the V-type ATPase family, even though this bacterium is a eubacterium. The hypothesis that the differentiation of an ancestral ATPase into V-type and F0F1-ATPase occurred after the evolution of a primordial cell into archaebacteria and eubacteria should be modified accordingly
RP  - NOT IN FILE
NT  - UI - 92089171LA - engRN - 0 (Macromolecular Systems)RN - 0 (Oligodeoxyribonucleotides)RN - EC 3.6.1.- (V-type ATPase)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19920127IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:1836357
SO  - Biochim Biophys Acta 1991 Dec 3 ;1098(1):13-20

1045
UI  - 611
AU  - Turina P
AU  - Melandri BA
AU  - Graber P
AD  - Department of Biology, University of Bologna, Italy
TI  - ATP synthesis in chromatophores driven by artificially induced ion gradients
AB  - An electrochemical potential difference for protons (delta mu H+) across the membrane of bacterial chromatophores was induced by an artificially generated pH difference (delta pH) and a K+/valinomycin diffusion potential, delta phi. The initial rate of ATP synthesis was measured with a rapid-mixing quenched-flow apparatus in the time range between 70 ms and 30 s after the acid-base transition. The rate of ATP synthesis depends exponentially on delta pH. Increasing diffusion potentials shift the delta pH dependency to lower delta pH values. Diffusion potentials were calculated from the Goldman equation. Using estimated permeability coefficients, the rate of ATP synthesis depends only on the electrochemical potential difference of protons irrespective of the relative contribution of delta pH and delta phi
RP  - NOT IN FILE
NT  - UI - 91160558LA - engRN - 56-65-5 (Adenosine Triphosphate)PT - Journal ArticleDA - 19910415IS - 0014-2956SB - IMCY - GERMANYJC - EMZ
UR  - PM:2001702
SO  - Eur J Biochem 1991 Feb 26 ;196(1):225-229

1046
UI  - 19868
AU  - Vogel PD
AU  - Cross RL
AD  - Department of Biochemistry and Molecular Biology, State University of New York, Syracuse 13210
TI  - Adenine nucleotide-binding sites on mitochondrial F1-ATPase. Evidence for an adenylate kinase-like orientation of catalytic and noncatalytic sites
AB  - Nucleotide-depleted mitochondrial F1-ATPase (F1[0,0]) is inhibited by the diadenosine oligophosphate compounds, AP4A, AP5A, and AP6A (where APxA stands for 5',5'-diadenosine oligophosphates having a chain of x phosphoryl groups linking the two adenosine moieties). When F1[0,0] is preincubated with these compounds and then assayed for ATP hydrolysis activity under conditions that normally allow turnover at all three catalytic sites, the maximal level of inhibition observed is 80%. However, when assayed at lower ATP concentrations under conditions that allow simultaneous turnover at only two of the three sites, no inhibition is observed. A decrease in the number of phosphoryl groups that links the adenosine moieties to less than 4 (AP3A, AP2A) converts the compound to an activator of ATP hydrolysis, similar in effect to that obtained when one mol of ADP or 2-azido-ADP binds at a catalytic site on F1[0,0]. Inhibition by the compounds requires the presence of at least one vacant noncatalytic site. Evidence is provided that the probes also interact with a catalytic site. The stoichiometry for maximal inhibition by AP4A is 0.94 mol/mol of F1. The data presented support a model for the structure of nucleotide-binding sites on F1 that places catalytic and noncatalytic sites in close proximity in an orientation analogous to the ATP and AMP binding sites on adenylate kinase. Inhibition of the enzyme by the dinucleotide compounds can be explained by the cross-bridging of one of the catalytic sites to a noncatalytic site in analogy to the inhibition of adenylate kinase by AP5A. The residual capacity for bi-site catalysis indicates that the second and third catalytic sites remain catalytically active
MH  - A
MH  - ACTIVE
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - ADP
MH  - atp
MH  - ATP synthase
MH  - BINDING
MH  - Binding Sites
MH  - Biochemistry
MH  - Catalysis
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - H(+)-Transporting ATP Synthase
MH  - Hydrolysis
MH  - MITOCHONDRIAL F1-ATPASE
MH  - model
MH  - NONCATALYTIC SITES
MH  - nucleotide binding
MH  - nucleotide binding sites
MH  - NUCLEOTIDE-BINDING-SITES
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 91177852LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 73-24-5 (Adenine)RN - EC 2.7.4.3 (Adenylate Kinase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM 23152/GM/NIGMSDA - 19910501IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:1826104
SO  - J Biol Chem 1991 Apr 5 ;266(10):6101-6105

1047
UI  - 21004
AU  - Wach A
AU  - Graber P
AD  - Biologisches Institut, Universitat Stuttgart, Federal Republic of Germany
TI  - The plasma membrane H(+)-ATPase from yeast. Effects of pH, vanadate and erythrosine B on ATP hydrolysis and ATP binding
AB  - The H(+)-ATPase from the plasma membrane of Saccharomyces cerevisiae was isolated and purified. The rate of ATP hydrolysis and ATP binding was measured as a function of pH and the effect of the vanadate and erythrosine B inhibitors was investigated. The pH dependence of the rate of ATP hydrolysis forms a bell-shaped curve with a maximum at pH 6 and half-maximal rates at pH 5.0 and 7.4. Only the pH dependence between pH 6 and pH 7.6 is reversible. Above pH 7.6 and below pH 5.5, denaturation of the isolated enzyme is observed. The rate of ATP binding shows the same pH dependency as that of ATP hydrolysis. Both pH dependencies can be described by the dissociation of a monovalent acidic group with a pK of 7.4. It is concluded that the enzyme must be protonated before ATP binding. Vanadate does not inhibit ATP binding, ADP release or Pi release at concentrations where complete inhibition of ATP hydrolysis is observed. It is concluded that vanadate inhibits a step of the reaction cycle which occurs after Pi release. In contrast, erythrosine B inhibits ATP binding and thus affects the first step of the reaction cycle
MH  - A
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - ADP
MH  - atp
MH  - ATPase
MH  - BINDING
MH  - DEPENDENCE
MH  - function
MH  - H(+)ATPase
MH  - Hydrolysis
MH  - membrane
MH  - pH
MH  - Saccharomyces cerevisiae
RP  - NOT IN FILE
NT  - UI - 92007896LA - engRN - 0 (Vanadates)RN - 16423-68-0 (Erythrosine)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19911112IS - 0014-2956SB - IMCY - GERMANY
UR  - PM:1655431
SO  - Eur J Biochem 1991 Oct 1 ;201(1):91-97

1048
UI  - 352
AU  - Walker JE
AU  - Lutter R
AU  - Dupuis A
AU  - Runswick MJ
AD  - Medical Research Council, Laboratory of Molecular Biology, Cambridge, UK
TI  - Identification of the subunits of F1F0-ATPase from bovine heart mitochondria
AB  - An oligomycin-sensitive F1F0-ATPase isolated from bovine heart mitochondria has been reconstituted into phospholipid vesicles and pumps protons. this preparation of F1F0-ATPase contains 14 different polypeptides that are resolved by polyacrylamide gel electrophoresis under denaturing conditions, and so it is more complex than bacterial and chloroplast enzymes, which have eight or nine different subunits. The 14 bovine subunits have been characterized by protein sequence analysis. They have been fractionated on polyacrylamide gels and transferred to poly(vinylidene difluoride) membranes, and N-terminal sequences have been determined in nine of them. By comparison with known sequences, eight of these have been identified as subunits beta, gamma, delta, and epsilon, which together with the alpha subunit form the F1 domain, as the b and c (or DCCD-reactive) subunits, both components of the membrane sector of the enzyme, and as the oligomycin sensitivity conferral protein (OSCP) and factor 6 (F6), both of which are required for attachment of F1 to the membrane sector. The sequence of the ninth, named subunit e, has been determined and is not related to any reported protein sequence. The N-terminal sequence of a tenth subunit, the membrane component A6L, could be determined after a mild acid treatment to remove an alpha-N-formyl group. Similar experiments with another membrane component, the a or ATPase-6 subunit, caused the protein to degrade, but the protein has been isolated from the enzyme complex and its position on gels has been unambiguously assigned. No N- terminal sequence could be derived from three other proteins. The largest of these is the alpha subunit, which previously has been shown to have pyrrolidonecarboxylic acid at the N terminus of the majority of its chains. The other two have been isolated from the enzyme complex; one of them is the membrane-associated protein, subunit d, which has an alpha-N-acetyl group, and the second, surprisingly, is the ATPase inhibitor protein. When it is isolated directly from mitochondrial membranes, the inhibitor protein has a frayed N terminus, with chains starting at residues 1, 2, and 3, but when it is isolated from the purified enzyme complex, its chains are not frayed and the N terminus is modified. Previously, the sequences at the N terminals of the alpha, beta, and delta subunits isolated from F1-ATPase had been shown to be frayed also, but in the F1F0 complex they each have unique N-terminal sequences.(ABSTRACT TRUNCATED AT 400 WORDS)
RP  - NOT IN FILE
NT  - UI - 91242449LA - engRN - 0 (Oligomycins)RN - 0 (Polyvinyls)RN - 24937-79-9 (polyvinylidene fluoride)RN - 9007-49-2 (DNA)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19910702IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:1827992
SO  - Biochemistry 1991 Jun 4 ;30(22):5369-5378

1049
UI  - 9893
AU  - Williams RJP
TI  - Uncoupled and Coupled Electron Transfer Reactions
MH  - bioenergetics
MH  - electron
MH  - proton
MH  - TRANSFER
RP  - ON REQUEST (08/14/91)
SO  - Biochim Biophys Acta 1991  ;1058(1)():71-74

1050
UI  - 8612
AU  - Yamasaki H
AU  - Furuya S
AU  - Kawamura A
AU  - Ito A
AU  - Okayama S
AU  - Nishimura M
TI  - Induction of the H+ Release from Thylakoid Membranes by Illumination in the Presence of Protonophores at High Concentrations
MH  - H+
MH  - localized coupling
MH  - membrane
MH  - Membranes
MH  - proton
MH  - protonophore
MH  - thylakoid
RP  - ON REQUEST (03/31/92)
SO  - Plant Cell Physiol 1991  ;32():925-934

1051
UI  - 178
AU  - Aggeler R
AU  - Chicas-Cruz K
AU  - Cai SX
AU  - Keana JF
AU  - Capaldi RA
AD  - Institute of Molecular Biology, University of Oregon, Eugene 97403
TI  - Introduction of reactive cysteine residues in the epsilon subunit of Escherichia coli F1 ATPase, modification of these sites with tetrafluorophenyl azide-maleimides, and examination of changes in the binding of the epsilon subunit when different nucleotides are in catalytic sites
AB  - Cysteine residues have been exchanged for serine residues at positions 10 and 108 in the epsilon subunit of the Escherichia coli F1 ATPase by site-directed mutagenesis to create two mutants, epsilon-S10C and epsilon-S108C. These two mutants and wild-type enzyme were reacted with [14C]N-ethylmaleimide (NEM) to examine the solvent accessibility of Cys residues and with novel photoactivated cross-linkers, tetrafluorophenyl azide-maleimides (TFPAM's), to examine near-neighbor relationships of subunits. In native wild-type F1 ATPase, NEM reacted with alpha subunits at a maximal level of 1 mol/mol of enzyme (1 mol/3 alpha subunits) and with the delta subunit at 1 mol/mol of enzyme; other subunits were not labeled by the reagent. In the mutants epsilon-S10C and epsilon-S108C, Cys10 and Cys108, respectively, were also labeled by NEM, indicating that these are surface residues. Reaction of wild-type enzyme with TFPAM's gave cross-linking of the delta subunit to both alpha and beta subunits. Reaction of the mutants with TFPAM's also cross-linked delta to alpha and beta and in addition formed covalent links between Cys10 of the epsilon subunit and the gamma subunit and between Cys108 of the epsilon subunit and the alpha subunit. The yield of cross-linking between sites on epsilon and other subunits depended on the nucleotide conditions used; this was not the case for delta- alpha or delta-beta cross-linked products. In the presence of ATP+EDTA the yield of cross-linking between epsilon-Cys10 and gamma was high (close to 50%) while the yield of epsilon-Cys108 and alpha was low (around 10%).(ABSTRACT TRUNCATED AT 250 WORDS)
RP  - NOT IN FILE
NT  - UI - 92198911LA - engRN - 0 (Azides)RN - 0 (Cross-Linking Reagents)RN - 0 (Maleimides)RN - 128-53-0 (Ethylmaleimide)RN - 139428-48-1 (N-(4-azido-2,3,5,6-tetrafluorobenzyl)-3- maleimidopropionamide)RN - 139428-49-2 (N-(4-azido-2,3,5,6-tetrafluorobenzyl)-6- maleimidohexanamide)RN - 52-90-4 (Cysteine)RN - 56-65-5 (Adenosine Triphosphate)RN - 60-00-4 (Edetic Acid)RN - 7439-95-4 (Magnesium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM27137/GM/NIGMSID - HL24526/HL/NHLBIDA - 19920424IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:1532326
SO  - Biochemistry 1992 Mar 24 ;31(11):2956-2961

1052
UI  - 177
AU  - Aggeler R
AU  - Capaldi RA
AU  - Dunn S
AU  - Gogol EP
AD  - Institute of Molecular Biology, University of Oregon, Eugene 97403
TI  - Epitope mapping of monoclonal antibodies to the Escherichia coli F1 ATPase alpha subunit in relation to activity effects and location in the enzyme complex based on cryoelectron microscopy
AB  - The interaction of Escherichia coli F1 ATPase (ECF1) with several different monoclonal antibodies (mAbs) specific for the alpha subunit has been examined. The epitopes for each of the mAbs have been localized by using molecular biological approaches to generate fragments of the alpha subunit. The binding of several of the mAbs has also been examined by cryoelectron microscopy of ECF1 Fab complexes. One of the mAbs, alpha II, bound in the region Asn 109-Val 153 without affecting ATPase activity. Most of the mAbs bound in the C-terminal third of the alpha subunit. MAb alpha 1 bound between residues Gln 443 and Trp 513. This mAb activated ATPase activity and was visualized in cryoelectron microscopy, superimposed on the alpha subunit, indicating that the epitope was on the top or bottom of ECF1 in the hexagonal projection. Other mAbs to the C-terminus, including alpha D which also activated the enzyme, reacted between Gly 371 and Trp 513 but failed to bind to small overlapping fragments within this sequence. The epitopes for these mAbs are probably formed by the folded polypeptide which occurs only in Western analysis when long stretches of the alpha subunit are present, suggesting that the C-terminus of alpha is a self- folding domain. In cryoelectron microscopy, Fab fragments for alpha D were seen extending from the sides of the ECF1 complex in hexagonal projection
RP  - NOT IN FILE
NT  - UI - 92337437LA - engRN - 0 (Antibodies, Monoclonal)RN - 0 (Binding Sites, Antibody)RN - 0 (Epitopes)RN - 0 (Immunoglobulins, Fab)RN - 0 (Recombinant Fusion Proteins)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - HL24236/HL/NHLBIDA - 19920820IS - 0003-9861SB - IMCY - UNITED STATESJC - 6SK
UR  - PM:1378717
SO  - Arch Biochem Biophys 1992 Aug 1 ;296(2):685-690

1053
UI  - 175
AU  - Aggeler R
AU  - Capaldi RA
AD  - Institute of Molecular Biology, University of Oregon, Eugene 97403
TI  - Cross-linking of the gamma subunit of the Escherichia coli ATPase (ECF1) via cysteines introduced by site-directed mutagenesis
AB  - The gamma subunit of the Escherichia coli F1 ATPase (ECF1) has been altered by site-directed mutagenesis to create five different mutants, gamma-S8C, gamma-S81C, gamma-T106C, gamma-S179C, and gamma-V286C, respectively. ECF1 isolated from four of these mutants had ATPase activities similar to that of a wild-type isogenic strain used as a control, the exception was enzyme isolated from mutant gamma-S81C, which had an ATPase activity of around 70-80% of the wild type. ECF1 isolated from each of the various mutants was reacted with N-(4-(7- (diethylamino)-4-methylcoumarin-3-yl))maleimide (CM). The fluorescent reagent was incorporated into Cys residues placed at positions 8, 106, 179, and 286, but not at 81, indicating which of these Cys residues are on the surface of the gamma subunit in the enzyme complex. Modification of the Cys at position 106 with CM activated the enzyme, and modification of the Cys at position 8 inhibited ATPase activity a small amount; however, modification of Cys at 179 or 286 had no effect on enzyme activity. The four mutants with a reactive Cys were reacted with tetrafluorophenylazide maleimides (TFPAMs), novel photoactivatable cross-linkers. In the mutant gamma-S8C, cross-links were formed between the introduced Cys on the gamma subunit and sites on the beta subunit. This cross-linking between gamma and beta depended on nucleotide conditions under which the photolysis was carried out, with differently migrating cross-linked products being obtained in ATP + EDTA compared with ATP + Mg2+ or ATP + Mg2+ Pi. Cross-linking between beta and gamma inhibited ATPase activity in proportion to the yield of cross-linked product. In the mutant gamma-V286C, cross-links were formed between the introduced Cys on gamma and the alpha subunit which were the same in all nucleotide conditions and which led to inhibition of ATPase activity
RP  - NOT IN FILE
NT  - UI - 93016072LA - engRN - 0 (Coumarins)RN - 0 (Cross-Linking Reagents)RN - 0 (Fluorescent Dyes)RN - 0 (Maleimides)RN - 0 (Oligodeoxyribonucleotides)RN - 0 (Plasmids)RN - 52-90-4 (Cysteine)RN - 93111-28-5 (N-(4-(7-(diethylamino)-4-methylcoumarin-3-yl))maleimide)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleID - HL24536/HL/NHLBIDA - 19921125IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:1400447
SO  - J Biol Chem 1992 Oct 25 ;267(30):21355-21359

1054
UI  - 440
AU  - al Shawi MK
AU  - Senior AE
AD  - Department of Biochemistry, University of Rochester, School of Medicine and Dentistry, New York 14642
TI  - Catalytic sites of Escherichia coli F1-ATPase. Characterization of unisite catalysis at varied pH
AB  - Using manual rapid-mixing procedures in which small, equal volumes of Escherichia coli F1-ATPase and [gamma-32P]ATP were combined at final concentrations of 2 and 0.2 microM, respectively (i.e., unisite catalysis conditions), it was shown that greater than or equal to 66% of the 32P became bound to the enzyme, with the ratio of bound ATP/bound Pi equal to 0.4 and the rate of dissociation of bound [32P]Pi equal to 3.5 x 10(-3) s-1, similar to previously published values. Azide is known to inhibit cooperative but not unisite catalysis in F1- ATPase [Noumi, T., Maeda, M., & Futai, M. (1987) FEBS Lett. 213, 381- 384]. In the presence of 1 mM sodium azide, 99% of the 32P became bound to the enzyme, with the ratio of bound ATP/bound Pi being 0.57. These experiments demonstrated that when conditions are used which minimize cooperative catalysis, most or all of the F1 molecules bind substoichiometric ATP tightly, hydrolyze it with retention of bound ATP and Pi, and release the products slowly. The data justify the validity of previously published rate constants for unisite catalysis. Unisite catalysis in E. coli F1-ATPase was studied at varied pH from 5.5 to 9.5 using buffers devoid of phosphate. Rate constants for ATP binding/release, ATP hydrolysis/resynthesis, Pi release, and ADP binding/release were measured; the Pi binding rate constant was inferred from the delta G for ATP hydrolysis. ATP binding was pH- independent; ATP release accelerated at higher pH. The highest KaATP (4.4 x 10(9) M-1) was seen at physiological pH 7.5.(ABSTRACT TRUNCATED AT 250 WORDS)
RP  - NOT IN FILE
NT  - UI - 92118898LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSDA - 19920225IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:1531027
SO  - Biochemistry 1992 Jan 28 ;31(3):878-885

1055
UI  - 439
AU  - al Shawi MK
AU  - Senior AE
AD  - Department of Biochemistry, University of Rochester Medical Center, New York 14642
TI  - Effects of dimethyl sulfoxide on catalysis in Escherichia coli F1- ATPase
AB  - (1) Dimethyl sulfoxide (DMSO) markedly inhibited the Vmax of multisite ATPase activity in Escherichia coli F1-ATPase at concentrations greater than 30% (v/v). Vmax/KM was reduced by 2 orders of magnitude in 40% (v/v) DMSO at pH 7.5, primarily due to reduction of Vmax. The inhibition was rapidly reversed on dilution into aqueous buffer. (2) KdATP at the first, high-affinity catalytic site was increased 1500- fold from 2.3 x 10(-10) to 3.4 x 10(-7) M in 40% DMSO at pH 7.5, whereas KdADP was increased 3.2-fold from 8.8 to 28 microM. This suggests that the high-affinity catalytic site presents a hydrophobic environment for ATP binding in native enzyme, that there is a significant difference between the conformation for ADP binding as opposed to ATP binding, and that the ADP-binding conformation is more hydrophilic. (3) Rate constants for hydrolysis and resynthesis of bound ATP in unisite catalysis were slowed approximately 10-fold by 40% DMSO; however, the equilibrium between bound Pi/bound ATP was little changed. The reduction in catalysis rates may well be related to the large increase in KdATP (less constrained site). (4) Significant Pi binding to E. coli F1 could not be detected either in 40% DMSO or in aqueous buffer using a centrifuge column procedure. (5) We infer, on the basis of the measured constants KaATP, K2 (hydrolysis/resynthesis of ATP), k+3 (Pi release), and KdADP and from estimates of k-3 (Pi binding) that delta G for ATP hydrolysis in 40% DMSO-containing pH 7.5 buffer is between -9.2 and -16.8 kJ/mol
RP  - NOT IN FILE
NT  - UI - 92118899LA - engRN - 0 (Phosphates)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 67-68-5 (Dimethyl Sulfoxide)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSDA - 19920225IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:1531028
SO  - Biochemistry 1992 Jan 28 ;31(3):886-891

1056
UI  - 516
AU  - Amzel LM
AU  - Bianchet MA
AU  - Pedersen PL
AD  - Department of Biophysics and Biophysical Chemistry, Johns Hopkins School of Medicine, Baltimore, Maryland 21205
TI  - Quaternary structure of ATP synthases: symmetry and asymmetry in the F1 moiety
AB  - It has been proposed that during ATP synthesis/hydrolysis F1 ATPases experience a complex pattern of nucleotide binding and release during the catalytic cycle (binding change mechanism). This type of mechanism has implications that can be correlated with the structure of the enzyme. F1-ATPases (stoichiometry alpha 3 beta 3 gamma delta epsilon) are essentially a symmetrical trimer of pairs of the major subunits (alpha and beta); the minor subunits (gamma, delta and epsilon) are in single copies and interact with the trimer in an asymmetrical fashion. The asymmetry introduced by the minor subunits has important structural and functional consequences: (1) it introduces differences between the potentially equivalent binding and catalytic sites in the major subunits, (2) it restricts the ways in which a binding change mechanism can occur, and (3) it governs the way in which the F1 interacts with the (asymmetrical) F0 sector
RP  - NOT IN FILE
NT  - UI - 93054451LA - engRN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialID - CA 10951/CA/NCIID - GM 25432/GM/NIGMSDA - 19921222IS - 0145-479XSB - IMCY - UNITED STATESJC - HIO
UR  - PM:1429535
SO  - J Bioenerg Biomembr 1992 Oct ;24(5):429-433

1057
UI  - 823
AU  - Beckers G
AU  - Berzborn RJ
AU  - Strotmann H
AD  - Institut fur Biochemie der Pflanzen, Heinrich Heine Universitat Dusseldorf, Germany
TI  - Zero-length crosslinking between subunits delta and I of the H(+)- translocating ATPase of chloroplasts
AB  - Treatment of spinach thylakoids with 1-ethyl-3-(dimethylaminopropyl)- carbodiimide (EDC)/N-hydroxysulfosuccinimide (sulfo-NHS) induced formation of a zero-length crosslink of an apparent molecular mass of 38 kDa. This product was shown, by immunodetection, to consist of subunit delta of CF1 and subunit I of CF0. The crosslink was isolated by preparative SDS gel electrophoresis and subjected to cyanogen bromide cleavage. Electrophoretic and immunological analysis of the resulting peptides suggested that the crosslink was formed between a glutamyl or aspartyl residue at the C-terminal end of subunit I and a basic amino acid of subunit delta in the range between Val-1 to Met- 165. Treatment of thylakoids with EDC/Sulfo-NHS resulted in inhibition of photophosphorylation and CF0CF1-catalyzed ATP hydrolysis without affecting formation of a proton gradient related to phenazine methosulfate-mediated cyclic electron transport. Inhibition of H+ transport-coupled ATP hydrolysis was more pronounced than non-coupled methanol-stimulated ATP hydrolysis. The results suggest that subunits delta and I form a connection between the partial complexes CF1 and CF0 in situ. Crosslinking of the two subunits may impede the translocation of protons through CF0CF1
RP  - NOT IN FILE
NT  - UI - 92338186LA - engRN - 0 (Cross-Linking Reagents)RN - 1892-57-5 (Ethyldimethylaminopropyl Carbodiimide)RN - 506-68-3 (Cyanogen Bromide)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19920825IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:1385976
SO  - Biochim Biophys Acta 1992 Jul 6 ;1101(1):97-104

1058
UI  - 10669
AU  - BOEKEMA EJ
AU  - Harris D
AU  - Boettcher B
AU  - Graber P
TI  - The structure of the ATP-synthase from chloroplasts
MH  - atp
MH  - ATP synthase
MH  - chloroplast
MH  - Chloroplasts
MH  - structure
T2  - Res.Photosynth
Y2  - -32676  
PB  - Dordrecht: Kluwer
RP  - NOT IN FILE
M2  - 2
SO  -  1992  ;.(1992):645-652

1059
UI  - 10675
AU  - Boettcher B
AU  - Graber P
AU  - BOEKEMA EJ
TI  - The structure of photosystem I from the thermophilic cyanobacterium Synechococcus sp. determined by electron microscopy of two-dimensional crystals
MH  - atp
MH  - cyano
MH  - Cyanobacteria
MH  - electron
MH  - electron microscopy
MH  - ELECTRON-MICROSCOPY
MH  - Microscopy
MH  - structure
MH  - Synechococcus
RP  - NOT IN FILE
SO  - Biochim Biophys Acta 1992  ;1100(2):125-136

1060
UI  - 176
AU  - Capaldi RA
AU  - Aggeler R
AU  - Gogol EP
AU  - Wilkens S
AD  - Institute of Molecular Biology, University of Oregon, Eugene 97403
TI  - Structure of the Escherichia coli ATP synthase and role of the gamma and epsilon subunits in coupling catalytic site and proton channeling functions
AB  - The structure of the Escherichia coli ATP synthase has been studied by electron microscopy and a model developed in which the alpha and beta subunits of the F1 part are arranged hexagonally (in top view) alternating with one another and surrounding a central cavity of around 35 A at its widest point. The alpha and beta subunits are interdigitated in side view for around 60 A of the 90 A length of the molecule. The F1 narrows and has three-fold symmetry at the end furthest from the F0 part. The F1 is linked to F0 by a stalk approximately 45 A long and 25-30 A in diameter. The F0 part is mostly buried in the lipid bilayer. The gamma subunit provides a domain that extends into the central cavity of the F1 part. The gamma and epsilon subunits are in a different conformation when ATP + Mg2+ are present in catalytic sites than when ATP + EDTA are present. This is consistent with these two small subunits switching conformations as a function of whether or not phosphate is bound to the enzyme at the position of the gamma phosphate of ATP. We suggest that this switching is the key to the coupling of catalytic site events with proton translocation in the F0 part of the complex
RP  - NOT IN FILE
NT  - UI - 93054452LA - engRN - 0 (Proton Pump)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 19921222IS - 0145-479XSB - IMCY - UNITED STATESJC - HIO
UR  - PM:1429536
SO  - J Bioenerg Biomembr 1992 Oct ;24(5):435-439

1061
UI  - 19867
AU  - Chernyak BV
AU  - Cross RL
AD  - Department of Biochemistry and Molecular Biology, State University of New York, Syracuse 13210
TI  - Adenine nucleotide-binding sites on mitochondrial F1-ATPase: studies of the inactive complex formed upon binding ADP at a catalytic site
AB  - ADP-induced inhibition of mitochondrial F1-ATPase has been studied. It is shown that in the presence of magnesium and the absence of light, the photoaffinity ADP analog, 2-azido-ADP, induces a reversible inhibition of native F1 that is indistinguishable from that obtained with ADP. Photolysis of the inactive complex results in the predominant labeling of a catalytic-site peptide identified previously (Cross et al., 1987, Proc. Natl. Acad. Sci. USA 84, 5715-5719). Dissociation of the inactive complex formed between F1 and ADP is biphasic with a rapid azide-insensitive phase followed by a slow azide-sensitive phase (k approximately 3 x 10(-3) s-1). It is also shown that incubation of the ADP-inhibited enzyme with EDTA or phosphate does not result in release or migration of ADP from the catalytic site. However, it does convert the complex to a form that reactivates in the presence of 100 microM ATP at a rate too rapid to observe using manual mixing
MH  - A
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - ADP
MH  - atp
MH  - ATP synthase
MH  - Azides
MH  - BINDING
MH  - Biochemistry
MH  - Cations
MH  - COMPLEX
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - H(+)-Transporting ATP Synthase
MH  - Light
MH  - Magnesium
MH  - MITOCHONDRIAL F1-ATPASE
MH  - nucleotide binding
MH  - nucleotide binding sites
MH  - NUCLEOTIDE-BINDING-SITES
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 92264720LA - engRN - 0 (Azides)RN - 0 (Cations, Divalent)RN - 0 (Organophosphorus Compounds)RN - 58-64-0 (Adenosine Diphosphate)RN - 64020-53-7 (2-azidoadenosine 3',5'-diphosphate)RN - 7439-95-4 (Magnesium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM-23152/GM/NIGMSDA - 19920616IS - 0003-9861SB - IMCY - UNITED STATESJC - 6SK
UR  - PM:1534000
SO  - Arch Biochem Biophys 1992 Jun ;295(2):247-252

1062
UI  - 518
AU  - Chuang WJ
AU  - Abeygunawardana C
AU  - Pedersen PL
AU  - Mildvan AS
AD  - Department of Biological Chemistry, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205
TI  - Two-dimensional NMR, circular dichroism, and fluorescence studies of PP- 50, a synthetic ATP-binding peptide from the beta-subunit of mitochondrial ATP synthase
AB  - PP-50, a peptide based on residues 141-190 of the beta-subunit of mitochondrial F1-ATPase, contains the GX4GKT consensus region for nucleoside triphosphate binding and has been shown to bind ATP [Garboczi, D.N., Shenbagamurthi, W.K., Hullihen, J., & Pedersen, P.L. (1988) J. Biol. Chem. 263, 812-816]. At pH 4.0, appropriate for NMR studies, PP-50 retains the ability to bind ATP tightly (KD = 17.5 microM) with a 1:1 stoichiometry as shown by titrations measuring the partial quenching of ATP fluorescence by PP-50. CD spectra of PP-50 at pH 4.0 and at low ionic strength show 5.8% helix, 30.2% beta-structure, and 64% coil. ATP binding increases the structure of PP-50, changing the CD to 7.5% helix, 44.5% beta-structure, and 48% coil. Increasing the ionic strength to 50 mM KCl also increases the structure, changing the CD to 7.4% helix, 64.4% beta-structure, and 28.2% coil. The 600-MHz proton NMR spectrum of PP-50, at pH 4.0 and low ionic strength, has been assigned by 2D methods (TOCSY, DQF-COSY, and NOESY with jump- return water suppression). Based on strong d alpha N NOEs, J alpha N values, and NH chemical shifts differing from random coil values, regions of extended structure are detected from residues 1-7 and 43-48. Based on dNN, dNN(i,i+2), and d alpha N(i,i+2) NOEs and 3J alpha N values, possible type I' and type I turns are found from residues 11-14 and 31-34, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)
RP  - NOT IN FILE
NT  - UI - 92379034LA - engRN - 0 (Peptide Fragments)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - CA10951/CA/NCIID - DK28616/DK/NIDDKDA - 19920930IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:1387322
SO  - Biochemistry 1992 Sep 1 ;31(34):7915-7921

1063
UI  - 21197
AU  - Danshina SV
AU  - Drachev LA
AU  - Kaulen AD
AU  - Korana K
AU  - Marti T
AU  - Mogi T
AU  - Skulachev VI
TI  - [Study of intermediate N using mutant forms of bacteriorhodopsin at Asp- 96]
AB  - It has been found that the N(P, R)-type intermediate of the photocycle is formed in the Asp-96-->Asn mutant at acidic pH. Azide, which strongly activates the M decay in this mutant, allows the N intermediate to be shown also at neutral pH. Under these conditions mutant N decays in a pH-independent fashion. In the presence of azide, the H+ uptake by Asp-96-->Asn mutant bacteriorhodopsin follows the M decay, whereas the N decay occurs at a much slower rate. Two electrogenic stages have been shown to be associated with the M--->bR step in the Asp-96--->Asn mutant photocycle. The faster and slower stages correlate with the M--->N and N--->bR transitions, respectively. In the Asp-96--->Asn mutant, high concentrations of azide are found to increase the M decay rate up to the values higher than those in the wild-type protein, both with or without azide. Such an effect is absent for the Asp-96-->Glu mutant. The activation energies for M--->N and N--- >bR transitions in the wild-type protein are equal to 18 and 19 kcal x mole-1, respectively. In the Asp-96-->Asn mutant without azide, the activation energy of the M decay is only 5 kcal x mole-1, whereas in the presence of azide in this mutant the activation energies for M and N decays are 8 and 9 kcal x mole-1, respectively. A scheme of events accompanying the Schiff base reprotonation during the photocycle is discussed
MH  - A
MH  - ACID
MH  - ACTIVATION
MH  - Bacteriorhodopsin
MH  - BASE
MH  - electrogenic
MH  - H+
MH  - intermediate
MH  - M
MH  - mutant
MH  - pH
MH  - protein
MH  - proton
MH  - Protons
MH  - Schiff base
MH  - Schiff-base
RP  - NOT IN FILE
NT  - UI - 93090994LA - rusRN - 0 (Protons)RN - 53026-44-1 (Bacteriorhodopsins)RN - 56-84-8 (Aspartic Acid)PT - Journal ArticleDA - 19930112IS - 0320-9725SB - IMCY - RUSSIA
UR  - PM:1333821
SO  - Biokhimiia 1992 Oct ;57(10):1574-1585

1064
UI  - 20948
AU  - Deckers-Hebestreit G
AU  - Altendorf K
AD  - Universitat Osnabruck, Fachbereich Biologie/Chemie, Arbeitsgruppe Mikrobiologie, FRG
TI  - The Fo complex of the proton-translocating F-type ATPase of Escherichia coli
AB  - The ATP synthase (F1Fo) of Escherichia coli consists of two structurally and functionally distinct entities. The F1 part is composed of five subunits alpha, beta, gamma, delta and epsilon (3:3:1:1:1) and carries the catalytic centres of the enzyme. The membrane-bound Fo complex functions as a proton channel and consists of the three subunits a, b and c (1:2:10 +/- 1). Subunit c (8288 M(r)) exhibits a hairpin-like structure within the membrane. A conserved acidic residue (Asp-61) in the C-terminal hydrophobic segment is absolutely required for proton translocation through Fo, whereas the hydrophilic loop region is necessary for F1 binding. Expression of the chloroplast proteolipid together with subunits a and b of E. coli did not produce an active Fo hybrid complex. Therefore, the construction of hybrid c subunits consisting of parts of the proteolipid from both organisms is in progress to determine those parts of subunit c that are essential for a functional interplay with subunits a and b. Subunit a (30,276 M(r)), which is also involved in proton translocation, is an extremely hydrophobic protein with 5-8 membrane-spanning helices. Studies with alkaline phosphatase fusion proteins resulted in controversial conclusions about the localization of the N and C termini of the protein. A foreign epitope (13 amino acids) has been inserted into the N- or C-terminal region of subunit a without affecting the function of Fo. Binding studies with a monoclonal antibody against this epitope are now under investigation to determine the orientation of subunit a.(ABSTRACT TRUNCATED AT 250 WORDS)
MH  - A
MH  - ACID
MH  - ACTIVE
MH  - Amino Acids
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BINDING
MH  - chloroplast
MH  - COMPLEX
MH  - England
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1
MH  - function
MH  - membrane
MH  - MONOCLONAL-ANTIBODIES
MH  - phosphatase
MH  - protein
MH  - Proteins
MH  - proton
MH  - Protons
MH  - RESIDUE
MH  - review
MH  - SUBUNIT
MH  - SYNTHASE
MH  - translocation
RP  - NOT IN FILE
NT  - UI - 93147708LA - engRN - 0 (Protons)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 19930303IS - 0022-0949SB - IMCY - ENGLAND
UR  - PM:1337099
SO  - J Exp Biol 1992 Nov ;172():451-459

1065
UI  - 20949
AU  - Deckers-Hebestreit G
AU  - Altendorf K
AD  - Universitat Osnabruck, Fachbereich Biologie/Chemie, Federal Republic of Germany
TI  - Influence of subunit-specific antibodies on the activity of the F0 complex of the ATP synthase of Escherichia coli. II. Effects of subunit c-specific polyclonal antibodies
AB  - After incubation of F1-stripped everted membrane vesicles with antibodies against subunit c of the ATP synthase of Escherichia coli the proton translocation through the open F0 channel was blocked. Rebinding of F1 to those vesicles is affected by the antibody concentration used. In general, the use of F(ab')2 or Fab fragments prepared from anti-c antibodies gave similar results. However, using Fab fragments a higher amount of antigenic binding sites was necessary to block the F0 complex completely, whereas extremely low amounts of Fab fragments were necessary to inhibit the binding of F1. This can be explained by an antigenic determinant of subunit c, which is only accessible to the smaller Fab fragments with a molecular mass of approximately 50,000. Incubation of F1-containing everted membranes with anti-c antibodies showed that the binding of the antibodies resulted in a displacement of F1, while simultaneously the proton translocation through F0 has been blocked. Such a displacement can only be observed after incubation with IgG molecules or F(ab')2 fragments. Fab fragments were not able to displace the F1 part, indicating that the ability of antibodies and F(ab')2 fragments to produce cross-links is responsible for the loss of F1 from the membranes
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BINDING
MH  - Binding Sites
MH  - COMPLEX
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - F1
MH  - membrane
MH  - membrane vesicles
MH  - Membranes
MH  - proton
MH  - Site
MH  - SUBUNIT
MH  - SYNTHASE
MH  - translocation
RP  - NOT IN FILE
NT  - UI - 92291128LA - engRN - 0 (Antibodies)RN - 0 (Epitopes)RN - 0 (Immunoglobulins, Fab)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19920716IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:1376323
SO  - J Biol Chem 1992 Jun 15 ;267(17):12370-12374

1066
UI  - 70
AU  - Dimroth P
AU  - Laubinger W
AU  - Kluge C
AU  - Kaim G
AU  - Ludwig W
AU  - Schleifer KH
AD  - Mikrobiologisches Institut, Eidgenossische Technische Hochschule, ETH- Zentrum, Zurich, Switzerland
TI  - Sodium-translocating adenosinetriphosphatase of Propionigenium modestum
RP  - NOT IN FILE
NT  - UI - 93167631LA - engRN - EC 3.6.1.- (sodium ATPase)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 19930312IS - 0077-8923SB - IMCY - UNITED STATESJC - 5NM
UR  - PM:1288327
SO  - Ann N Y Acad Sci 1992 Nov 30 ;671():310-321

1067
UI  - 74
AU  - Dimroth P
AD  - Mikrobiologisches Institut, Eidgenossische Technische Hochschule, ETH- Zentrum, Zurich, Switzerland
TI  - The ATPases of Propionigenium modestum and Bacillus alcalophilus. Strategies for ATP synthesis under low energy conditions
AB  - In Propionigenium modestum, ATP synthesis is coupled via delta mu Na+ to the decarboxylation of (S)-methylmalonyl-CoA. The low energy yield of this reaction implies that approx. 4 decarboxylation cycles are necessary to synthesize 1 molecule of ATP. Theoretical considerations in accord with experimental results suggest ATP synthesis in P. modestum at delta mu Na+ = -110 mV. Other anaerobic bacteria synthesize ATP at a delta mu H+ of similar size and alkaliphilic bacteria at pH 10.3 have a delta mu H+ of only -103 mV. In these cases, the H+(Na+) to ATP stoichiometry must be at least 4
RP  - NOT IN FILE
NT  - UI - 92338209LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 19920825IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:1385980
SO  - Biochim Biophys Acta 1992 Jul 17 ;1101(2):236-239

1068
UI  - 79
AU  - Dimroth P
AD  - Mikrobiologisches Institut, Eidgenossische Technische Hochschule, Zurich, Switzerland
TI  - Structure and function of the Na(+)-translocating ATPase of Propionigenium modestum
AB  - The strict anaerobic bacterium Propionigenium modestum performs a Na+ cycle over the membrane to couple ATP synthesis to the decarboxylation of methyl-malonyl-CoA. The responsible ATPase has a typical F1F0 structure, the water-soluble F1 moiety being composed of five different subunits and the more firmly membrane-bound F0 part consisting of three different subunits. The F1F0 ATPase but not F1 alone was specifically activated by Na+ ions, suggesting that the Na+ binding site is located on the F0 moiety. The ATPase reconstituted into proteoliposomes catalyzed an ATP-dependent Na+ accumulation that was stimulated to the same extent by dissipating the membrane potential with valinomycin or with the uncoupler carbonylcyanide-m-chloro phenylhydrazone. The transport of Na+ is therefore a primary event, not a secondary event involving the intermediate formation of a proton gradient. The ATPase also catalyzed H+ translocation at Na+ concentrations below 1 mM. Our results indicate a common mechanism of the ATPase for Na+ and H+ (H3O+) translocation and a switch from H+ to Na+ translocation by increasing the Na+ concentration. A hybrid ATPase consisting of F1 from E. coli and F0 from P. modestum had the same specificity with respect to the translocated cations as the homologous F1F0 ATPase of P. modestum, indicating again that the Na+ (H+) binding site is located on the F0 part. Also in accord with this supposition is a diffusion potential- induced translocation of Na+ or H+ through the F0 part of the enzyme complex. The phylogenetic relationship between the Na(+)-translocating ATPase of P. modestum and other F1F0 ATPases has been clearly demonstrated by sequencing studies
RP  - NOT IN FILE
NT  - UI - 93080096LA - engRN - 7440-23-5 (Sodium)RN - EC 3.6.1.- (sodium ATPase)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19921230IS - 0302-2994SB - IMCY - ENGLANDJC - 1UF
UR  - PM:1449075
SO  - Acta Physiol Scand Suppl 1992  ;607():97-103

1069
UI  - 21196
AU  - Drachev LA
AU  - Kaulen AD
AU  - Komrakov AY
AD  - AN Belozersky Institute of Physico-Chemical Biology, Moscow State University, Russia
TI  - Interrelations of M-intermediates in bacteriorhodopsin photocycle
AB  - The photocycles of the wild-type bacteriorhodopsin and the D96N mutant were investigated by the flash-photolysis technique. The M-intermediate formation (400 nm) and the L-intermediate decay (520 nm) were found to be well described by a sum of two exponents (time constants, tau 1 = 65 and tau 2 = 250 microseconds) for the wild-type bR and three exponents (tau 1 = 55 microseconds, tau 2 = 220 microseconds and tau 3 = 1 ms) for the D96N mutant of bR. A component with tau = 1 ms was found to be present in the photocycle of the wild-type bacteriorhodopsin as a lag- phase in the relaxation of photoresponses at 400 and 520 nm. In the presence of Lu3+ ions or 80% glycerol this component was clearly seen as an additional phase of M-formation. The azide effect on the D96N mutant of bR suggests that the 1-ms component is associated with an irreversible conformational change switching the Schiff base from the outward to the inward proton channel. The maximum of the difference spectrum of the 1-ms component of D96N bR is located at 404 nm as compared to 412 nm for the first two components. We suggest that this effect is a result of the alteration of the inward proton channel due to the Asp96-->Asn substitution. Proton release measured with pyranine in the absence of pH buffers was identical for the wild-type bR and D96N mutant and matched the M-->M' conformational transition. A model for M rise in the bR photocycle is proposed
MH  - A
MH  - Azides
MH  - Bacteriorhodopsin
MH  - BASE
MH  - buffer
MH  - Buffers
MH  - conformational change
MH  - CONSTANT
MH  - ion
MH  - Ions
MH  - M
MH  - M-intermediate
MH  - microsecond
MH  - model
MH  - mutant
MH  - pH
MH  - proton
MH  - proton release
MH  - relaxation
MH  - Schiff base
MH  - Schiff-base
MH  - spectra
MH  - Time
RP  - NOT IN FILE
NT  - UI - 93076906LA - engRN - 0 (Azides)RN - 53026-44-1 (Bacteriorhodopsins)PT - Journal ArticleDA - 19921224IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:1446744
SO  - FEBS Lett 1992 Nov 30 ;313(3):248-250

1070
UI  - 19865
AU  - Duncan TM
AU  - Cross RL
AD  - Department of Biochemistry and Molecular Biology, SUNY Health Science Center, Syracuse 13210
TI  - A model for the catalytic site of F1-ATPase based on analogies to nucleotide-binding domains of known structure
AB  - An updated topological model is constructed for the catalytic nucleotide-binding site of the F1-ATPase. The model is based on analogies to the known structures of the MgATP site on adenylate kinase and the guanine nucleotide sites on elongation factor Tu (Ef-Tu) and the ras p21 protein. Recent studies of these known nucleotide-binding domains have revealed several common functional features and similar alignment of nucleotide in their binding folds, and these are used as a framework for evaluating results of affinity labeling and mutagenesis studies of the beta subunit of F1. Several potentially important residues on beta are noted that have not yet been studied by mutagenesis or affinity labeling
MH  - A
MH  - atp
MH  - ATP synthase
MH  - BETA-SUBUNIT
MH  - BINDING
MH  - Biochemistry
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - H(+)-Transporting ATP Synthase
MH  - model
MH  - nucleotide binding
MH  - Nucleotides
MH  - review
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 93054455LA - engRN - 0 (Nucleotides)RN - EC 2.7.4.3 (Adenylate Kinase)RN - EC 3.6.1.- (Peptide Elongation Factor Tu)RN - EC 3.6.1.- (Proto-Oncogene Protein p21(ras))RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialID - GM 23152/GM/NIGMSDA - 19921222IS - 0145-479XSB - IMCY - UNITED STATESJC - HIO
UR  - PM:1429539
SO  - J Bioenerg Biomembr 1992 Oct ;24(5):453-461

1071
UI  - 302
AU  - Engelbrecht S
AU  - Junge W
AD  - Universitat Osnabruck, Germany
TI  - Added subunit beta of CF1 as well as gamma/delta/epsilon restore photophosphorylation in partially CF1-depleted thylakoids
AB  - We investigated the ability of subunits beta, gamma, delta, and epsilon of CF1, the F1-ATPase of chloroplasts, to interact with exposed CF0 in EDTA-treated, partially CF1-depleted thylakoid membranes. We measured the ability of subunits beta, gamma, delta, and epsilon to stimulate the rate of photophosphorylation under continuous light and, for subunit beta, also the ability to diminish the proton leakage through exposed CF0 by deceleration of the decay of electrochromic absorption transients under flashing light. The greatest effect was caused by subunit beta, followed by gamma/delta/epsilon. Pairwise combinations of gamma, delta, and epsilon or each of these subunits alone were only marginally effective. Subunit gamma from the thermophilic bacterium PS 3 in combination with chloroplast delta and epsilon was as effective as chloroplast gamma. The finding that the small CF1 subunits in concert and the beta subunit by itself specifically interacted with the exposed proton channel CF0, qualifies the previous concept of subunit delta acting particularly as a plug to the open CF0 channel. The interactions between the channel and the catalytic portion of the enzyme seem to involve most of the small, and at least beta of the large subunits
RP  - NOT IN FILE
NT  - UI - 93075831LA - engRN - 0 (Plant Extracts)RN - 60-00-4 (Edetic Acid)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19921228IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:1445938
SO  - Biochim Biophys Acta 1992 Dec 7 ;1140(2):157-162

1072
UI  - 40
AU  - Fillingame RH
AD  - Department of Biomolecular Chemistry, University of Wisconsin Medical School, Madison 53706
TI  - Subunit c of F1F0 ATP synthase: structure and role in transmembrane energy transduction
RP  - NOT IN FILE
NT  - UI - 92338210LA - engRN - 56-84-8 (Aspartic Acid)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialID - GM-23105/GM/NIGMSDA - 19920825IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:1385981
SO  - Biochim Biophys Acta 1992 Jul 17 ;1101(2):240-243

1073
UI  - 39
AU  - Fillingame RH
AD  - Department of Biomolecular Chemistry, University of Wisconsin Medical School, Madison 53706
TI  - H+ transport and coupling by the F0 sector of the ATP synthase: insights into the molecular mechanism of function
AB  - The F0 sector of the ATP synthase complex facilitates proton translocation through the membrane, and via interaction with the F1 sector, couples proton transport to ATP synthesis. The molecular mechanism of function is being probed by a combination of mutant analysis and structural biochemistry, and recent progress on the Escherichia coli F0 sector is reviewed here. The E. coli F0 is composed of three types of subunits (a, b, and c) and current information on their folding and organization in F0 is reviewed. The structure of purified subunit c in chloroform-methanol-H2O resembles that in native F0, and progress in determining the structure by NMR methods is reviewed. Genetic experiments suggest that the two helices of subunit c must interact as a functional unit around an essential carboxyl group as protons are transported. In addition, a unique class of suppressor mutations identify a transmembrane helix of subunit a that is proposed to interact with the bihelical unit of subunit c during proton transport. The role of multiple units of subunit c in coupling proton translocation to ATP synthesis is considered. The special roles of Asp61 of subunit c and Arg210 of subunit a in proton translocation are also discussed
RP  - NOT IN FILE
NT  - UI - 93054459LA - engRN - 0 (Protons)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialID - GM-23105/GM/NIGMSDA - 19921222IS - 0145-479XSB - IMCY - UNITED STATESJC - HIO
UR  - PM:1331039
SO  - J Bioenerg Biomembr 1992 Oct ;24(5):485-491

1074
UI  - 38
AU  - Fillingame RH
AU  - Girvin ME
AU  - Fraga D
AU  - Zhang Y
AD  - Department of Biomolecular Chemistry, University of Wisconsin Medical School, Madison 53706
TI  - Correlations of structure and function in H+ translocating subunit c of F1F0 ATP synthase
RP  - NOT IN FILE
NT  - UI - 93167633LA - engRN - 0 (Macromolecular Systems)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialID - GM-23105/GM/NIGMSDA - 19930312IS - 0077-8923SB - IMCY - UNITED STATESJC - 5NM
UR  - PM:1288329
SO  - Ann N Y Acad Sci 1992 Nov 30 ;671():323-333

1075
UI  - 659
AU  - Freedman JC
AU  - Novak TS
AU  - Penefsky HS
AU  - Stein WD
AD  - Department of Physiology, State University of New York Health Science Center, Syracuse 13210
TI  - Quantitative analysis of oxonol V fluorescence in submitochondrial particles
RP  - NOT IN FILE
NT  - UI - 93167667LA - engRN - 0 (Fluorescent Dyes)RN - 0 (Isoxazoles)RN - 0 (Succinates)RN - 61389-30-8 (oxonol V)RN - EC 3.6.1.37 (Na(+)-K(+)-Exchanging ATPase)PT - Journal ArticleID - GM21737/GM/NIGMSDA - 19930312IS - 0077-8923SB - IMCY - UNITED STATESJC - 5NM
UR  - PM:1337685
SO  - Ann N Y Acad Sci 1992 Nov 30 ;671():493-496

1076
UI  - 9193
AU  - Fromme P
AU  - Dahse I
AU  - Graber P
TI  - Effect of Tentoxin on the Activation and on the Catalytic Reaction of Reconstituted H(+)-ATPase from Chloroplasts
MH  - ACTIVATION
MH  - alpha
MH  - atp
MH  - BETA
MH  - Catalysis
MH  - chloroplast
MH  - Chloroplasts
MH  - H(+)ATPase
MH  - inhibitor
MH  - pmf
MH  - regulation
MH  - tentoxin
MH  - unisite
MH  - UNISITE CATALYSIS
RP  - IN FILE
NT  - tentoxin does not act on pmf driven AdN release and unisite catalysis. also Lit. for "binds to alpha or more probably to beta
SO  - Z Naturforsch 1992  ;47():239-244

1077
UI  - 20862
AU  - Futai M
AU  - Iwamoto A
AU  - Omote H
AU  - Orita Y
AU  - Shin K
AU  - Nakamoto RK
AU  - Maeda M
AD  - Department of Organic Chemistry and Biochemistry, Osaka University, Japan
TI  - Escherichia coli ATP synthase (F-ATPase): catalytic site and regulation of H+ translocation
AB  - We discuss our recent results on the Escherichia coli F-ATPase, in particular its catalytic site in the beta subunit and regulation of H+ transport by the gamma subunit. Affinity labelling experiments suggest that beta Lys-155 in the glycine-rich sequence is near the gamma- phosphate moiety of ATP bound at the catalytic site. The enzyme loses activity upon introduction of missense mutations in beta Lys-155 or beta Thr-156 and changes catalytic properties upon introduction of other mutations. By analysis of mutations and their pseudo revertants, residues beta Ser-174, beta Glu-192 and beta Val-198 were found to be located near the glycine-rich sequence. The combined approaches of chemical labelling and genetics have been fruitful in visualizing the structure of the catalytic site. Analysis of mutations in the gamma subunit suggests that this subunit has an essential role in coupling catalysis with proton translocation
MH  - analysis
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BETA-SUBUNIT
MH  - Biochemistry
MH  - Catalysis
MH  - Chemistry
MH  - COLI ATP SYNTHASE
MH  - coupling
MH  - England
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F-ATPASE
MH  - genetics
MH  - H+
MH  - proton
MH  - Protons
MH  - regulation
MH  - RESIDUE
MH  - Site
MH  - SUBUNIT
MH  - SYNTHASE
MH  - translocation
MH  - transport
RP  - NOT IN FILE
NT  - UI - 93147707LA - engRN - 0 (Protons)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19930303IS - 0022-0949SB - IMCY - ENGLAND
UR  - PM:1337098
SO  - J Exp Biol 1992 Nov ;172():443-449

1078
UI  - 20866
AU  - Futai M
AU  - Iwamoto A
AU  - Maeda M
AD  - Department of Organic Chemistry and Biochemistry, Osaka University, Japan
TI  - Catalytic site in the beta subunit of H(+)-ATPase from Escherichia coli:the glycine-rich sequence and its interaction with Ser-174
MH  - Adenosinetriphosphatase
MH  - ATPase
MH  - BETA-SUBUNIT
MH  - Biochemistry
MH  - Chemistry
MH  - England
MH  - H(+)ATPase
MH  - Site
MH  - SUBUNIT
RP  - NOT IN FILE
NT  - UI - 93080061LA - engRN - 56-40-6 (Glycine)RN - 56-45-1 (Serine)RN - EC 3.6.1.- (sodium-translocating ATPase)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 4.6.1.1 (Adenylate Cyclase)PT - Journal ArticleDA - 19921230IS - 0302-2994SB - IMCY - ENGLAND
UR  - PM:1449057
SO  - Acta Physiol Scand Suppl 1992  ;607():125-130

1079
UI  - 21000
AU  - Graber P
AU  - Labahn A
AD  - Biologisches Institut, Universitat Stuttgart, Germany
TI  - Proton transport-coupled unisite catalysis by the H(+)-ATPase from chloroplasts
AB  - Proton transport-coupled unisite catalysis was measured with the H(+)- ATPase from chloroplasts. The reaction was measured in the ATP hydrolysis direction under deenergized conditions and in the ATP synthesis direction under energized conditions. The equilibrium constant of the enzyme does not change upon energization, whereas the dissociation constants of substrates and products change by orders of magnitude. This indicates that the Gibbs free enthalpy derived from proton translocation is used to change binding affinities of substrates and products, and this results in synthesis of free ATP
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthesis
MH  - ATPase
MH  - BINDING
MH  - Catalysis
MH  - chloroplast
MH  - Chloroplasts
MH  - CONSTANT
MH  - H(+)ATPase
MH  - Hydrolysis
MH  - proton
MH  - Protons
MH  - review
MH  - synthesis
MH  - translocation
MH  - UNISITE CATALYSIS
RP  - NOT IN FILE
NT  - UI - 93054460LA - engRN - 0 (Protons)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 19921222IS - 0145-479XSB - IMCY - UNITED STATES
UR  - PM:1331040
SO  - J Bioenerg Biomembr 1992 Oct ;24(5):493-497

1080
UI  - 21085
AU  - Guffanti AA
AU  - Krulwich TA
AD  - Department of Biochemistry, Mount Sinai School of Medicine, City University of New York, New York 10029
TI  - Features of apparent nonchemiosmotic energization of oxidative phosphorylation by alkaliphilic Bacillus firmus OF4
AB  - Oxidative phosphorylation by extremely alkaliphilic Bacillus species violates two major predictions of the chemiosmotic hypothesis: the magnitude of the chemiosmotic driving force, the delta p (electrochemical proton gradient), is too low to account for the phosphorylation potentials observed during growth at pH 10.5 without using a much higher H+/ATP stoichiometry than used during growth at pH 7.5, and artificially imposed diffusion potentials fail to energize ATP synthesis above about pH 9.5 (Guffanti, A. A., and Krulwich, T. A. (1989) Annu. Rev. Microbiol. 43, 435-463). To further examine the latter observation, large valinomycin-mediated potassium diffusion potentials were imposed across starved cells of Bacillus firmus OF4 at various pH values from pH 7.5 to 10.5. As the external pH increased above pH 8, there was a sharp decrease in the rate of ATP synthesis in response to an imposed diffusion potential. The rate of ATP synthesis fell to zero by pH 9.2 and 9.4, respectively, in the presence and absence of a small inwardly directed Na+ gradient. Electrogenic Na+/H+ antiport and Na+/alpha-aminoisobutyric acid symport proceeded at substantial rates throughout. When synthesis was energized by an electron donor, cells under comparable conditions synthesized ATP at rapid rates up to pH 10.5. The proton transfers that occur during respiration-dependent oxidative phosphorylation at pH 10.5 may depend upon specific complexes. Cells grown at pH 7.5, which have one-third the levels of the caa3-type terminal oxidase, and slightly lower levels of certain other respiratory chain complexes than pH 10.5-grown cells, support only low rates of ATP synthesis at pH 10.5, although energy- dependent symport and antiport rates are comparable with those in pH 10.5-grown cells. A model is presented for oxidative phosphorylation by the alkaliphilic Bacillus that involves a nonchemiosmotic direct intramembrane transfer of protons from specific respiratory chain complexes to the F0 sector of the ATPase, whereas remaining respiratory chain complexes extrude protons into the bulk to generate the bulk potential required both for ATP synthesis and other bioenergetic work. A pK-regulated gate or a delocalized proton pathway that fails to work above pH 9.5 are suggested as possible features that account for the loss of efficacy of a bulk-imposed diffusion potential in energizing ATP synthesis above pH 9.4
MH  - A
MH  - ACID
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthesis
MH  - ATPase
MH  - Bacillus
MH  - Biochemistry
MH  - Cells
MH  - COMPLEX
MH  - delta
MH  - Diffusion
MH  - diffusion potential
MH  - electrogenic
MH  - electron
MH  - F0
MH  - model
MH  - Oxidative Phosphorylation
MH  - P
MH  - pH
MH  - Phosphorylation
MH  - Potassium
MH  - proton
MH  - Proton transfer
MH  - Proton-transfer
MH  - Protons
MH  - synthesis
MH  - TRANSFER
MH  - Valinomycin
RP  - NOT IN FILE
NT  - UI - 92250606LA - engRN - 2001-95-8 (Valinomycin)RN - 56-65-5 (Adenosine Triphosphate)PT - Journal ArticleID - GM28454/GM/NIGMSDA - 19920609IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:1577797
SO  - J Biol Chem 1992 May 15 ;267(14):9580-9588

1081
UI  - 570
AU  - Hatefi Y
AU  - Matsuno-Yagi A
AD  - Department of Molecular and Experimental Medicine, Scripps Research Institute, La Jolla, California 92037
TI  - Unisite and multisite ATP hydrolysis and synthesis by bovine submitochondrial particles
RP  - NOT IN FILE
NT  - UI - 93167638LA - engRN - 0 (Macromolecular Systems)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialID - DK08126/DK/NIDDKDA - 19930312IS - 0077-8923SB - IMCY - UNITED STATESJC - 5NM
UR  - PM:1288334
SO  - Ann N Y Acad Sci 1992 Nov 30 ;671():377-384

1082
UI  - 21143
AU  - Heberle J
AU  - Dencher NA
AD  - Hahn-Meitner-Institut, BENSC-N1, W-1000 Berlin 39, Germany
TI  - Surface-bound optical probes monitor protein translocation and surface potential changes during the bacteriorhodopsin photocycle
AB  - Light-induced H+ release and reuptake as well as surface potential changes inherent in the bacterio-rhodopsin reaction cycle were measured between 10 degrees C and 50 degrees C. Signals of optical pH indicators covalently bound to Lys-129 at the extracellular surface of bacteriorhodopsin were compared with absorbance changes of probes residing in the aqueous bulk phase. Only surface-bound indicators monitor the kinetics of H+ ejection from bacteriorhodopsin and allow the correlation of the photocycle with the pumping cycle. During the L550----M412 transition the H+ appears at the extracellular surface of bacteriorhodopsin. Surface potential changes detected by bound fluorescein or by the potentiometric probe 4-[2-(di-n-butylamino)-6- naphthyl]vinyl-1-(3-sulfopropyl)pyridinium betaine (di-4-ANEPPS) occur in milliseconds concomitantly with the formation and decay of the N intermediate. pH indicators residing in the aqueous bulk phase reflect the transfer of H+ from the membrane surface into the bulk but do not probe the early events of H+ pumping. The observed retardation of H+ at the membrane surface for several hundred microseconds is of relevance for energy conversion of biological membranes powered by electrochemical H+ gradients
MH  - Bacteriorhodopsin
MH  - fluorescein
MH  - Fluoresceins
MH  - H+
MH  - indicator
MH  - intermediate
MH  - Kinetics
MH  - membrane
MH  - Membranes
MH  - microsecond
MH  - pH
MH  - pH-indicator
MH  - protein
MH  - SURFACE
MH  - TRANSFER
MH  - translocation
RP  - NOT IN FILE
NT  - UI - 92335224LA - engRN - 0 (Fluoresceins)RN - 2321-07-5 (Fluorescein)RN - 53026-44-1 (Bacteriorhodopsins)PT - Journal ArticleDA - 19920814IS - 0027-8424SB - IMCY - UNITED STATES
UR  - PM:1497755
SO  - Proc Natl Acad Sci U S A 1992 Jul 1 ;89(13):5996-6000

1083
UI  - 746
AU  - Hisabori T
AU  - Muneyuki E
AU  - Odaka M
AU  - Yokoyama K
AU  - Mochizuki K
AU  - Yoshida M
AD  - Department of Biology, Yokohama City University, Japan
TI  - Single site hydrolysis of 2',3'-O-(2,4,6-trinitrophenyl)-ATP by the F1- ATPase from thermophilic bacterium PS3 is accelerated by the chase- addition of excess ATP
AB  - The interaction of 2',3'-O-(2,4,6-trinitrophenyl)-adenosine 5'- triphosphate (TNP-ATP) and TNP-ADP to F1-ATPase from a thermophilic bacterium PS3 (TF1) was investigated. When TNP-ADP or TNP-ATP was added to the isolated alpha or beta subunit of TF1, characteristic difference spectra were generated for each subunit. Difference spectra generated on addition of these analogs to TF1 resembled those observed for the beta subunit, indicating TNP analogs bind to the beta subunits in the molecule of TF1. Results of equilibrium dialysis showed that TNP-ADP binds to a single high affinity site on TF1 in the presence of Mg2+ with a dissociation constant of 2.2 nM. When TNP-ATP was added to TF1 in a substoichiometric molar ratio, it rapidly bound to TF1 and was slowly hydrolyzed. The hydrolysis proceeded nearly to completion without showing stable equilibrium between bound species of TNP-ATP and TNP-ADP. Similar to beef heart mitochondrial F1, this hydrolysis was greatly accelerated by the chase-addition of 100 microM ATP. However, the hydrolyzed product, TNP-ADP, remained bound on the beta subunit even after the chase
RP  - NOT IN FILE
NT  - UI - 92165809LA - engRN - 0 (2',3'-O-(2,4,6-trinitrophenyl)adenosine 5'-triphosphate)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19920402IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:1531655
SO  - J Biol Chem 1992 Mar 5 ;267(7):4551-4556

1084
UI  - 21284
AU  - Israelachvili JN
TI  - Intermolecular and Surface Forces
MH  - SURFACE
PB  - London: Academic Press
RP  - IN FILE
SO  -  1992  ;():

1085
UI  - 21084
AU  - Ivey DM
AU  - Krulwich TA
AD  - Department of Biochemistry, Mount Sinai School of Medicine of CUNY, NY 10029
TI  - Two unrelated alkaliphilic Bacillus species possess identical deviations in sequence from those of other prokaryotes in regions of F0 proposed to be involved in proton translocation through the ATP synthase
AB  - The a and c subunits of two unrelated alkaliphilic Bacillus species contain unusual motifs in regions previously implicated by others in H(+)-coupled oxidative phosphorylation. The facultative alkaliphile B. firmus OF4 apparently does not contain genes encoding an alternative F0, supporting other evidence that a single species of proton- translocating F1F0-ATPase catalyses oxidative phosphorylation both at low and high pH. The unusual F0 sequence motifs may be part of the adaptation of the extreme alkaliphiles to growth at very high pH
MH  - A
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Bacillus
MH  - Biochemistry
MH  - F0
MH  - Oxidative Phosphorylation
MH  - pH
MH  - Phosphorylation
MH  - proton
MH  - Proton-Translocating ATPases
MH  - SUBUNIT
MH  - SYNTHASE
MH  - translocation
RP  - NOT IN FILE
NT  - UI - 93079447LA - engRN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleID - GM28454/GM/NIGMSDA - 19921229IS - 0923-2508SB - IMCY - FRANCE
UR  - PM:1448623
SO  - Res Microbiol 1992 Jun ;143(5):467-470

1086
UI  - 20865
AU  - Jounouchi M
AU  - Takeyama M
AU  - Noumi T
AU  - Moriyama Y
AU  - Maeda M
AU  - Futai M
AD  - Department of Organic Chemistry and Biochemistry, Osaka University, Japan
TI  - Role of the amino terminal region of the epsilon subunit of Escherichia coli H(+)-ATPase (F0F1)
AB  - Escherichia coli strain KF148(SD-) defective in translation of the uncC gene for the epsilon subunit of H(+)-ATPase could not support growth by oxidative phosphorylation due to lack of F1 binding to Fo (M. Kuki, T. Noumi, M. Maeda, A. Amemura, and M. Futai, 1988, J. Biol. Chem. 263, 17, 437-17, 442). Mutant uncC genes for epsilon subunits lacking different lengths from the amino terminus were constructed and introduced into strain KF148(SD-). F1 with an epsilon subunit lacking the 15 amino-terminal residues could bind to F0 in a functionally competent manner, indicating that these amino acid residues are not absolutely necessary for formation of a functional enzyme. However, mutant F1 in which the epsilon subunit lacked 16 amino-terminal residues showed defective coupling between ATP hydrolysis (synthesis) and H(+)-translocation, although the mutant F1 showed partial binding to Fo. These findings suggest that the epsilon subunit is essential for binding of F1 to F0 and for normal H(+)-translocation. Previously, Kuki et al. (cited above) reported that 60 residues were not necessary for a functional enzyme. However, the mutant with an epsilon subunit lacking 15 residues from the amino terminus and 4 residues from the carboxyl terminus was defective in oxidative phosphorylation, suggesting that both terminal regions affect the conformation of the region essential for a functional enzyme
MH  - A
MH  - ACID
MH  - atp
MH  - ATPase
MH  - BINDING
MH  - Biochemistry
MH  - Chemistry
MH  - conformation
MH  - coupling
MH  - COUPLING FACTOR
MH  - EPSILON-SUBUNIT
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - F0F1
MH  - F1
MH  - H(+)ATPase
MH  - Hydrolysis
MH  - M
MH  - membrane
MH  - Membrane Proteins
MH  - mutant
MH  - Oxidative Phosphorylation
MH  - Oxidative Phosphorylation Coupling Factors
MH  - Phosphorylation
MH  - protein
MH  - Proteins
MH  - RESIDUE
MH  - SUBUNIT
MH  - synthesis
RP  - NOT IN FILE
NT  - UI - 92088271LA - engRN - 0 (Membrane Proteins)RN - 0 (Oxidative Phosphorylation Coupling Factors)RN - 0 (Plasmids)RN - 0 (Recombinant Proteins)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19920123IS - 0003-9861SB - IMCY - UNITED STATES
UR  - PM:1530778
SO  - Arch Biochem Biophys 1992 Jan ;292(1):87-94

1087
UI  - 20864
AU  - Jounouchi M
AU  - Takeyama M
AU  - Chaiprasert P
AU  - Noumi T
AU  - Moriyama Y
AU  - Maeda M
AU  - Futai M
AD  - Department of Organic Chemistry and Biochemistry, Osaka University, Japan
TI  - Escherichia coli H(+)-ATPase: role of the delta subunit in binding Fl to the Fo sector
AB  - The roles of the Escherichia coli H(+)-ATPase (FoFl) delta subunit (177 amino acid residues) was studied by analyzing mutants. The membranes of nonsense (Gln-23----end, Gln-29----end, Gln-74----end) and missense (Gly-150----Asp) mutants had very low ATPase activities, indicating that the delta subunit is essential for the binding of the Fl portion to Fo. The Gln-176----end mutant had essentially the same membrane- bound activity as the wild type, whereas in the Val-174----end mutant most of the ATPase activity was in the cytoplasm. Thus Val-174 (and possibly Leu-175 also) was essential for maintaining the structure of the subunit, whereas the two carboxyl terminal residues Gln-176 and Ser- 177 were dispensable. Substitutions were introduced at various residues (Thr-11, Glu-26, Asp-30, Glu-42, Glu-82, Arg-85, Asp-144, Arg-154, Asp- 161, Ser-163), including apparently conserved hydrophilic ones. The resulting mutants had essentially the same phenotypes as the wild type, indicating that these residues do not have any significant functional role(s). Analysis of mutations (Gly-150----Asp, Pro, or Ala) indicated that Gly-150 itself was not essential, but that the mutations might affect the structure of the subunit. These results suggest that the overall structure of the delta subunit is necessary, but that individual residues may not have strict functional roles
MH  - ACID
MH  - analysis
MH  - ATPase
MH  - BINDING
MH  - Biochemistry
MH  - Chemistry
MH  - Cytoplasm
MH  - DELTA-SUBUNIT
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - H(+)ATPase
MH  - Macromolecular Systems
MH  - membrane
MH  - Membranes
MH  - mutant
MH  - RESIDUE
MH  - SUBUNIT
MH  - SYSTEM
MH  - SYSTEMS
RP  - NOT IN FILE
NT  - UI - 92117644LA - engRN - 0 (Codon)RN - 0 (Macromolecular Systems)RN - 0 (Oligodeoxyribonucleotides)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19920218IS - 0003-9861SB - IMCY - UNITED STATES
UR  - PM:1530999
SO  - Arch Biochem Biophys 1992 Feb 1 ;292(2):376-381

1088
UI  - 286
AU  - Junge W
AU  - Engelbrecht S
AU  - Griwatz C
AU  - Groth G
TI  - THE CHLOROPLAST H+-ATPase: PARTIAL REACTIONS OF THE PROTON
AB  - This article reviews proton intake, charge transfer and proton release by F-ATPases, based in part on flash spectrophotometric studies on the chloroplast ATPase in thylakoid membranes, CF1Fo. The synthesis-coupled translocation of charges by CF1Fo (maximum rate <1500 s-1) and the dissipative flow through its exposed channel portion, CFo (rate >10 000 s-1), are extremely proton-specific (selectivity H+:K+>10(7):1). The proton-specific filter is located in CFo. Proton flow through exposed CFo can be throttled by adding subunit (&dgr;) or subunit &bgr; of CF1. These subunits thus may provide energy-transducing contacts between CF1 and CFo. Recently, we characterized two conditions where, in contrast to the above situation, proton intake by CF1Fo was decoupled from proton transfer across the main dielectric barrier: (a) CF1Fo structurally distorted by low ionic strength transiently trapped protons in a highly cooperative manner, but remained proton tight. This result has been interpreted in terms of Mitchell's proton well. (b) In the absence of nucleotides there is a proton slip. Addition of nucleotides (100 nmol l-1 ADP) abolished proton conduction but not proton intake by CF1Fo. These experiments functionally tag proton binding groups on CF1Fo that are located before the main dielectric barrier
RP  - NOT IN FILE
NT  - UI - 0LA - ENGPT - JOURNAL ARTICLEDA - 19990105IS - 0022-0949SB - IMJC - I2F
UR  - PM:9874756
SO  - J Exp Biol 1992 Nov 1 ;172(Pt 1):461-474

1089
UI  - 8603
AU  - Junge W
AU  - Althoff G
AU  - Jahns P
AU  - Engelbrecht S
AU  - Lill H
AU  - Schnknecht G
TI  - Proton pumps, proton flow and proton ATP synthases in photosynthesis of green plants
MH  - atp
MH  - ATP synthase
MH  - Chemistry
MH  - electron
MH  - electron transfer
MH  - Photosynthesis
MH  - plant
MH  - Plants
MH  - proton
MH  - Proton Pump
MH  - Proton transfer
MH  - Proton-transfer
MH  - SYNTHASE
MH  - TRANSFER
MH  - transport
T2  - Electron and proton transfer in chemistry and biology
A2  - Mller A
A2  - Ratajczak H
A2  - Junge W
A2  - Diemann E
Y2  - -32676  
PB  - Amsterdam, Netherlands: Elsevier
RP  - IN FILE
M2  - 78
SO  -  1992  ;():253-272

1090
UI  - 75
AU  - Kaim G
AU  - Ludwig W
AU  - Dimroth P
AU  - Schleifer KH
AD  - Lehrstuhl fur Mikrobiologie, Technische Universitat Munchen, Federal Republic of Germany
TI  - Cloning, sequencing and in vivo expression of genes encoding the F0 part of the sodium-ion-dependent ATP synthase of Propionigenium modestum in Escherichia coli
AB  - A DNA fragment containing the genes encoding subunits of the F0 part of the sodium-translocating ATPase of Propionigenium modestum was cloned in Escherichia coli and sequenced. The predicted amino acid sequences of subunits a, b and c of the P. modestum ATPase were compared with those of the corresponding subunits of proton-translocating ATPases from other bacteria and chloroplasts. Deletion mutants of E. coli, lacking different genes for ATPase subunits, were transformed with a recombinant plasmid, containing the genes for the subunits a, c, b, delta and part of alpha of the ATPase of P. modestum. Functionally reconstituted ATPase activity could be demonstrated for the transformants. The identity of the vector containing P. modestum genes was verified by restriction analysis of plasmid DNA
RP  - NOT IN FILE
NT  - UI - 92339434LA - engRN - 0 (Bacterial Proteins)RN - 0 (Macromolecular Systems)RN - 0 (Oligodeoxyribonucleotides)RN - 56-65-5 (Adenosine Triphosphate)RN - 7440-23-5 (Sodium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19920825IS - 0014-2956SB - IMCY - GERMANYJC - EMZ
UR  - PM:1386022
SO  - Eur J Biochem 1992 Jul 15 ;207(2):463-470

1091
UI  - 69
AU  - Kluge C
AU  - Dimroth P
AD  - Mikrobiologisches Institut, Eidgenossische Technische Hochschule, Zurich, Switzerland
TI  - Studies on Na+ and H+ translocation through the Fo part of the Na(+)- translocating F1Fo ATPase from Propionigenium modestum: discovery of a membrane potential dependent step
AB  - The purified ATPase of Propionigenium modestum (F1Fo) was incorporated into liposomes, and the F1 part was dissociated. The Fo-liposomes catalyzed proton uptake in response to a potassium diffusion potential (inside negative). Proton translocation was abolished by rebinding F1 to the Fo-liposomes or after incubation with the c-subunit-specific inhibitor dicyclohexylcarbodiimide (DCCD). Proton uptake was also sensitive to the presence of external Na+ or Li+ ions and was completely abolished at 2 mM NaCl or 150 mM LiCl, respectively. However, the same concentrations of these salts in the internal volume of the Fo-liposomes were without effect, suggesting that the cation binding site is not accessible from both sides of the membrane simultaneously. An open channel-type of transport through Fo from P. modestum is therefore excluded. The Fo-liposomes also catalyzed Na+ influx or efflux in response to a K+ diffusion potential that was negative on the inside or outside, respectively. These Na+ fluxes could not be created, however, by delta pNa+ of about 60-180 mV. The initial rate of Na+ uptake depended strongly on the size of the membrane potential with no significant conductivity below -40 mV, followed by a proportional increase up to about -115 mV. In the absence of a membrane potential, the Fo-liposomes catalyzed 22Na+ counterflow against a 28- fold concentration gradient. Uptake of 22Na+ into Fo-liposomes against delta pNa+ (counterflow) was completely prevented by imposing an inside- positive potassium diffusion potential of 90 mV. The catalysis of 22Na+ counterflow by Fo from P. modestum is a clear indication of a carrier (transporter)-type mechanism and excludes a channel mechanism.(ABSTRACT TRUNCATED AT 250 WORDS)
RP  - NOT IN FILE
NT  - UI - 93112609LA - engRN - 0 (Liposomes)RN - 0 (Proteolipids)RN - 0 (proteoliposomes)RN - 1333-74-0 (Hydrogen)RN - 7440-09-7 (Potassium)RN - 7440-23-5 (Sodium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19930204IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:1472503
SO  - Biochemistry 1992 Dec 22 ;31(50):12665-12672

1092
UI  - 73
AU  - Kluge C
AU  - Laubinger W
AU  - Dimroth P
AD  - Mikrobiologisches Institute, Eidgenossische Technische Hochschule, ETH- Zentrum, Zurich, Switzerland
TI  - The Na(+)-translocating ATPase of Propionigenium modestum
RP  - NOT IN FILE
NT  - UI - 93050735LA - engRN - 0 (Macromolecular Systems)RN - 7440-23-5 (Sodium)RN - EC 3.6.1.- (sodium ATPase)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 19921201IS - 0300-5127SB - IMCY - ENGLANDJC - E48
UR  - PM:1426593
SO  - Biochem Soc Trans 1992 Aug ;20(3):572-577

1093
UI  - 21083
AU  - Krulwich TA
AU  - Guffanti AA
AD  - Department of Biochemistry, Mount Sinai School of Medicine, City University of New York, New York 10029
TI  - Proton-coupled bioenergetic processes in extremely alkaliphilic bacteria
AB  - Oxidative phosphorylation, which involves an exclusively proton-coupled ATP synthase, and pH homeostasis, which depends upon electrogenic antiport of cytoplasmic Na+ in exchange for H+, are the two known bioenergetic processes that require inward proton translocation in extremely alkaliphilic bacteria. Energy coupling to oxidative phosphorylation is particularly difficult to fit to a strictly chemiosmotic model because of the low bulk electrochemical proton gradient that follows from the maintenance of a cytoplasmic pH just above 8 during growth at pH 10.5 and higher. A large quantitative and variable discrepancy between the putative chemiosmotic driving force and the phosphorylation potential results. This is compounded by a nonequivalence between respiration-dependent bulk gradients and artificially imposed ones in energizing ATP synthesis, and by an apparent requirement for specific respiratory chain complexes that do not relate solely to their role in generation of bulk gradients. Special features of the synthase may contribute to the mode of energization, just as novel features of the Na+ cycle may relate to the extraordinary capacity of the extreme alkaliphiles to achieve pH homeostasis during growth at, or sudden shifts to, an external pH of 10.5 and above
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - Bacteria
MH  - Biochemistry
MH  - Carrier Proteins
MH  - COMPLEX
MH  - coupling
MH  - electrogenic
MH  - H+
MH  - Homeostasis
MH  - model
MH  - Oxidative Phosphorylation
MH  - pH
MH  - Phosphorylation
MH  - protein
MH  - Proteins
MH  - proton
MH  - review
MH  - SYNTHASE
MH  - synthesis
MH  - translocation
RP  - NOT IN FILE
NT  - UI - 93094178LA - engRN - 0 (Carrier Proteins)RN - 0 (Sodium-Hydrogen Antiporter)RN - 56-65-5 (Adenosine Triphosphate)PT - Journal ArticlePT - ReviewPT - Review, TutorialID - GM28454/GM/NIGMSDA - 19930112IS - 0145-479XSB - IMCY - UNITED STATES
UR  - PM:1334072
SO  - J Bioenerg Biomembr 1992 Dec ;24(6):587-599

1094
UI  - 20998
AU  - Labahn A
AU  - Graber P
AD  - Biologisches Institut, Universitat Stuttgart, Germany
TI  - Transport protons do not participate in ATP synthesis/hydrolysis at the nucleotide binding site of the H(+)-ATPase from chloroplasts
AB  - The H(+)-ATPase from chloroplasts CFoF1, was brought into the active, reduced state by illumination of thylakoids in the presence of thioredoxin and dithiothreitol. Uni-site ATP synthesis was initiated by the addition of 20 nM [alpha-32P]ADP, and enzyme-bound and free nucleotides were separated by a pressure column. The ratio of enzyme- bound ADP to ATP was 0.55 +/- 0.05. In a second experiment, uni-site ATP hydrolysis under energized conditions was initiated by the addition of 36 nM [alpha-32P]ATP; enzyme-bound and free nucleotides were separated by a pressure column. Both procedures were carried out under continuous illumination. The ratio of enzyme-bound ADP to ATP was 0.46 +/- 0.04. In a third experiment, uni-site ATP hydrolysis under de- energized conditions was initiated by the addition of 39 nM [alpha- 32P]ATP and NH4Cl/valinomycin in the absence of illumination. Free and enzyme-bound nucleotides were separated also by a pressure column. The ratio of enzyme-bound ADP to ATP was 0.43 +/- 0.02. This ratio was always the same irrespective of whether the reaction runs in the synthesis or the hydrolysis direction. Furthermore, the ratio does not depend on the membrane energization. We conclude, therefore, that the protons are not directly involved in the reaction at the catalytic site
MH  - A
MH  - ACTIVE
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - ADP
MH  - atp
MH  - ATP synthesis
MH  - ATPase
MH  - BINDING
MH  - chloroplast
MH  - Chloroplasts
MH  - Dithiothreitol
MH  - H(+)ATPase
MH  - Hydrolysis
MH  - membrane
MH  - nucleotide binding
MH  - Nucleotides
MH  - proton
MH  - Protons
MH  - Site
MH  - synthesis
MH  - thylakoid
MH  - thylakoids
MH  - transport
RP  - NOT IN FILE
NT  - UI - 93050246LA - engRN - 0 (Protons)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19921222IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:1330704
SO  - FEBS Lett 1992 Nov 23 ;313(2):177-180

1095
UI  - 21002
AU  - Labahn A
AU  - Graber P
AD  - Biologisches Institut, Universitat Stuttgart
TI  - Numerical simulation of uni-site and bi-site ATP-hydrolysis catalyzed by the membrane-bound H(+)-ATPase from chloroplasts
AB  - The kinetics of ATP-hydrolysis catalyzed by the H(+)-ATPase from chloroplasts was described by an enzyme kinetic model where either one (uni-site catalysis) or two (bi-site catalysis) sites are operating. Numerical simulations are carried out with the software package LARKIN using measured rate constants as a basis. When the ATP-concentration is increased up to 20 nM (at 20 nM enzyme concentration) a change from uni- site to bi-site catalysis occurs at about 7 nM. The measured effective rate constants change by a factor two. However, the calculated rate constants for the second site are drastically increased
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - ATPase
MH  - Catalysis
MH  - chloroplast
MH  - Chloroplasts
MH  - CONSTANT
MH  - England
MH  - H(+)ATPase
MH  - Kinetics
MH  - model
MH  - rate constant
MH  - SIMULATION
MH  - SIMULATIONS
MH  - Site
MH  - Software
MH  - uni-site catalysis
MH  - UNISITE CATALYSIS
RP  - NOT IN FILE
NT  - UI - 93080079LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19921230IS - 0302-2994SB - IMCY - ENGLAND
UR  - PM:1449067
SO  - Acta Physiol Scand Suppl 1992  ;607():241-244

1096
UI  - 437
AU  - Lee RS
AU  - Wilke-Mounts S
AU  - Senior AE
AD  - Department of Biochemistry, University of Rochester Medical Center, New York 14642
TI  - F1-ATPase with cysteine instead of serine at residue 373 of the alpha subunit
AB  - Escherichia coli strain AN718 contains the alpha S373F mutation in F1F0- ATP synthase which blocks ATP synthesis (oxidative phosphorylation) and steady-state F1-ATPase activity. The revertant strain AN718SS2 containing the mutation alpha C373 was isolated and shown to confer a phenotype of higher growth yield than that of the wild type in liquid medium containing limiting glucose, succinate, or LB. Purified F1 from strain AN718SS2 was found to have 30% of wild-type steady-state ATPase activity and 60% of wild-type oxidative phosphorylation activity. Azide sensitivity of ATPase activity and ADP-induced enhancement of bound aurovertin fluorescence, both of which are lost in alpha S373F mutant F1, were regained in alpha C373 F1. N-Ethylmaleimide (NEM) inactivated alpha C373 F1 steady-state ATPase potently but had no effect on unisite ATPase. Complete inactivation of alpha C373 F1 steady-state ATPase corresponded to incorporation of one NEM per F1 (mol/mol), in just one of the three alpha subunits. NEM-inactivated enzyme showed azide- insensitive residual ATPase activity and loss of ADP-induced enhancement of bound aurovertin fluorescence. The data confirm the view that placement at residue alpha 373 of a bulky amino acid side-chain (phenylalanyl or NEM-derivatized cysteinyl) blocks positive catalytic cooperativity in F1. The fact that NEM inhibits steady-state ATPase when only one alpha subunit of three is reacted suggests a cyclical catalytic mechanism
RP  - NOT IN FILE
NT  - UI - 92359547LA - engRN - 0 (Macromolecular Systems)RN - 128-53-0 (Ethylmaleimide)RN - 52-90-4 (Cysteine)RN - 56-45-1 (Serine)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM-25349/GM/NIGMSDA - 19920910IS - 0003-9861SB - IMCY - UNITED STATESJC - 6SK
UR  - PM:1386722
SO  - Arch Biochem Biophys 1992 Sep ;297(2):334-339

1097
UI  - 21165
AU  - Monticello RA
AU  - Angov E
AU  - Brusilow WS
AD  - Department of Biochemistry, Wayne State University School of Medicine, Detroit, Michigan 48201
TI  - Effects of inducing expression of cloned genes for the F0 proton channel of the Escherichia coli F1F0 ATPase
AB  - To evaluate whether expression of cloned genes for the F0 proton channel of the Escherichia coli F1F0 ATPase is sufficient to cause membrane proton permeability, plasmids carrying different combinations of the uncB, E, and F genes, encoding the a, c, and b subunits of the F0 sector, cloned behind the inducible lac promoter in pUC9 or pUC18, were constructed. The effects of inducing F0 synthesis in an unc deletion strain were monitored by measuring cell growth rate, quantitating F0 subunits by immunoblotting, and measuring the ability of membranes to maintain a respiration-induced proton gradient and to bind F1 and carry out energy-coupling reactions. The levels of functional reconstitutable F0 in membranes could be increased four- to sixfold with no change in cellular growth rate or membrane proton permeability (assayed by fluorescence quenching). These results were obtained in uninduced cultures, so the F0 genes were presumably being transcribed from some promoter besides lac. Induction of transcription of all three F0 genes produced increased amounts of F0 subunits in membranes as determined by immunoblot and F1-binding assays, but, when reconstituted with F1, the F0 in membranes isolated from induced cultures was significantly less functional than the F0 in membranes isolated from uninduced cultures. Such induction did result in growth inhibition, but there was no correlation between growth inhibition and either increased membrane proton permeability or the presence of functional, reconstitutable F0
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - ATPase
MH  - Bacterial Proteins
MH  - Biochemistry
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - F1
MH  - fluorescence
MH  - ion
MH  - Ion Channels
MH  - membrane
MH  - Membranes
MH  - Nad
MH  - Permeability
MH  - protein
MH  - Proteins
MH  - proton
MH  - Proton-Translocating ATPases
MH  - Protons
MH  - SUBUNIT
MH  - synthesis
RP  - NOT IN FILE
NT  - UI - 92250434LA - engRN - 0 (Bacterial Proteins)RN - 0 (Ion Channels)RN - 0 (Protons)RN - 0 (Recombinant Proteins)RN - 0 (uncB protein, E coli)RN - 0 (uncE protein, E coli)RN - 0 (uncF protein, Escherichia coli)RN - 53-84-9 (NAD)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleID - K04-AI00882/AI/NIAIDDA - 19920609IS - 0021-9193SB - IMCY - UNITED STATES
UR  - PM:1374378
SO  - J Bacteriol 1992 May ;174(10):3370-3376

1098
UI  - 19778
AU  - Murataliev MB
AU  - Boyer PD
AD  - Molecular Biology Institute, University of California Los Angeles 90024- 1570
TI  - The mechanism of stimulation of MgATPase activity of chloroplast F1- ATPase by non-catalytic adenine-nucleotide binding. Acceleration of the ATP-dependent release of inhibitory ADP from a catalytic site
AB  - The presence of ATP at non-catalytic sites of the chloroplast F1-ATPase (CF1) eliminates a considerable lag in onset of enzyme activity that otherwise occurs in the presence of bicarbonate [Milgrom, Y. M., Ehler, L. & Boyer, P. D. (1991) J. Biol. Chem. 266, 11551-11558]. Sulfite is known to be much more effective than bicarbonate in stimulating ATPase activity CF1. Results reported here show that when assayed in the presence of sulfite, CF1, with some non-catalytic sites empty or filled with GT(D)P, is able to hydrolyze both ATP and GTP. Thus, the presence of adenine nucleotides at non-catalytic sites is not necessary for catalytic turnover of CF1. However, even though CF1 with empty non- catalytic sites shows a significant initial activity, the prior binding of adenine nucleotides at non-catalytic site(s) results in further activation of MgATPase and MgGTPase activities, even at relatively high sulfite and substrate concentrations. Although extensive activation of CF1 results from the presence of sulfite, with or without nucleotide binding at non-catalytic sites, the Km remains constant, at about 50 microM for MgATP and 400 microM for MgGTP. The results obtained show that the ATPase activity of CF1 is determined by the fraction of the active enzyme. The inactive CF1.ADP.Mg2+ formed during MgATP hydrolysis can be rapidly trapped by azide to provide a measure of the fraction of inactive enzyme. Increasing the concentration of sulfite increases the fraction of active CF1 in the assay medium. Measurements with radioactively labeled nucleotides show that the presence of ATP at non- catalytic sites promotes the ATP-dependent release of inhibitory ADP from a catalytic site. The activating effect of ATP binding at non- catalytic sites results from increasing the portion of CF1 in an active state during steady-state ATP hydrolysis
MH  - A
MH  - ACTIVATION
MH  - ACTIVE
MH  - Adenine Nucleotides
MH  - ADENINE-NUCLEOTIDES
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - ADP
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BINDING
MH  - chloroplast
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - Guanosine Triphosphate
MH  - H(+)-Transporting ATP Synthase
MH  - Hydrolysis
MH  - mechanism
MH  - non-catalytic site
MH  - NONCATALYTIC SITES
MH  - nucleotide binding
MH  - Nucleotides
MH  - P
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 93049314LA - engRN - 146-91-8 (Guanosine Diphosphate)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 86-01-1 (Guanosine Triphosphate)RN - EC 3.6.1.- (Ca(2+) Mg(2+)-ATPase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM11094/GM/NIGMSDA - 19921203IS - 0014-2956SB - IMCY - GERMANYJC - EMZ
UR  - PM:1425675
SO  - Eur J Biochem 1992 Oct 15 ;209(2):681-687

1099
UI  - 20861
AU  - Nakamoto RK
AU  - Shin K
AU  - Iwamoto A
AU  - Omote H
AU  - Maeda M
AU  - Futai M
AD  - Department of Organic Chemistry and Biochemistry, Osaka University, Japan
TI  - Escherichia coli F0F1-ATPase. Residues involved in catalysis and coupling
AB  - The molecular biological approach has provided important information toward understanding the complexities of the F0F1 ATPase. This article focuses on our recent results on the ATPase catalytic site contained in the beta subunit and the role of the gamma subunit in regulation of proton transport. We used a combination of affinity labeling and mutagenesis to locate several residues of the alpha and beta subunits in the catalytic site. Adenosine triphosphopyridoxal (AP3-PL) labeled beta Lys-155, beta Lys-201 and alpha Lys-201, suggesting that they are near the gamma-phosphate moiety of ATP. Turning to a mutagenesis approach we demonstrated that the two conserved residues, beta Lys-155 and beta Thr-156 in the glycine-rich sequence, are essential for catalysis. Finally, using pseudorevertant analysis, we positioned residue beta Gly-149 (also in the glycine-rich sequence) in proximity to beta Ser-174, beta Glu-192 (binding site for DCCD), and beta Val-198 (only three residues away from the AP3-PL binding site, beta Lys-201). Genetic studies suggested that the gamma subunit plays a role in regulation of catalysis and its coupling with proton conduction. We found that four mutations in the carboxyl-terminal region (gamma Gln- 269-->Leu, gamma Gly-275-->Lys, gamma Thr-277-->end, or frameshift) had similar membrane ATPase activities but different ATP-dependent proton pumping and growth by oxidative phosphorylation. These results suggested a perturbation in the coupling between catalysis and proton translocation. We were able to clearly define the "uncoupling" by introducing mutations in the amino-terminal region of the gamma subunit. We were led to gamma Met-23-->Lys and Arg which resulted in an enzyme still regulated by delta microH+, but with profoundly inefficient coupling between ATPase catalytic sites and proton translocation in both ATP-dependent proton pumping and delta microH(+)- driven ATP synthesis. Second-site mutations in the carboxyl-terminal region of the gamma subunit reversed this effect
MH  - A
MH  - Adenosine
MH  - analysis
MH  - atp
MH  - ATP synthesis
MH  - ATPase
MH  - BETA-SUBUNIT
MH  - BINDING
MH  - Biochemistry
MH  - Catalysis
MH  - Chemistry
MH  - coupling
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0F1
MH  - Macromolecular Systems
MH  - membrane
MH  - mutagenesis
MH  - Oxidative Phosphorylation
MH  - Phosphorylation
MH  - proton
MH  - regulation
MH  - RESIDUE
MH  - review
MH  - Site
MH  - SUBUNIT
MH  - synthesis
MH  - SYSTEM
MH  - SYSTEMS
MH  - translocation
MH  - transport
RP  - NOT IN FILE
NT  - UI - 93167634LA - engRN - 0 (Macromolecular Systems)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 19930312IS - 0077-8923SB - IMCY - UNITED STATES
UR  - PM:1288330
SO  - Ann N Y Acad Sci 1992 Nov 30 ;671():335-343

1100
UI  - 21273
AU  - Nicholls DG
AU  - Ferguson SJ
TI  - Bioenergetics 2
MH  - Membranes
MH  - membrane
MH  - A
MH  - bioenergetics
PB  - London: Academic Press
RP  - NOT IN FILE
SO  -  1992  ;():

1101
UI  - 351
AU  - Palmer DN
AU  - Fearnley IM
AU  - Walker JE
AU  - Hall NA
AU  - Lake BD
AU  - Wolfe LS
AU  - Haltia M
AU  - Martinus RD
AU  - Jolly RD
AD  - Department of Veterinary Pathology and Public Health, Massey University, Palmerston North, New Zealand
TI  - Mitochondrial ATP synthase subunit c storage in the ceroid- lipofuscinoses (Batten disease)
AB  - The ceroid-lipofuscinoses (Batten disease) are neurodegenerative inherited lysosomal storage diseases of children and animals. A common finding is the occurrence of fluorescent storage bodies (lipopigment) in cells. These have been isolated from tissues of affected sheep. Direct protein sequencing established that the major component is identical to the dicyclohexylcarbodiimide (DCCD) reactive proteolipid, subunit c, of mitochondrial ATP synthase and that this protein accounts for at least 50% of the storage body mass. No other mitochondrial components are stored. Direct sequencing of storage bodies isolated from tissues of children with juvenile and late infantile ceroid- lipofuscinosis established that they also contain large amounts of complete and normal subunit c. It is also stored in the disease in cattle and dogs but is not present in storage bodies from the human infantile form. Subunit c is normally found as part of the mitochondrial ATP synthase complex and accounts for 2-4% of the inner mitochondrial membrane protein. Mitochondria from affected sheep contain normal amounts of this protein. The P1 and P2 genes that code for it are normal as are mRNA levels. Oxidative phosphorylation is also normal. These findings suggest that ovine ceroid-lipofuscinosis is caused by a specific failure in the degradation of subunit c after its normal inclusion into mitochondria, and its consequent abnormal accumulation in lysosomes. This implies a unique pathway for subunit c degradation. It is probable that the human late infantile and juvenile diseases and the disease in cattle and dogs involve lesions in the same pathway
RP  - NOT IN FILE
NT  - UI - 92303621LA - engRN - 0 (Carrier Proteins)RN - 0 (Pigments)RN - 0 (Proteolipids)RN - 0 (dicyclohexylcarbodiimide-binding proteolipid)RN - 0 (lipopigments)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - NS11238/NS/NINDSDA - 19920722IS - 0148-7299SB - IMCY - UNITED STATESJC - 3L4
UR  - PM:1535179
SO  - Am J Med Genet 1992 Feb 15 ;42(4):561-567

1102
UI  - 824
AU  - Pancic PG
AU  - Strotmann H
AU  - Kowallik KV
AD  - Institut fur Biochemie der Pflanzen, Heinrich-Heine-Universitat Dusseldorf, Germany
TI  - Chloroplast ATPase genes in the diatom Odontella sinensis reflect cyanobacterial characters in structure and arrangement
AB  - We have cloned and sequenced a 5200 base restriction fragment and an overlapping 3100 base fragment of the large single copy region of the chloroplast genome of the diatom Odontella sinensis, which hybridized to several ATPase gene probes. These fragments contain six closely linked reading frames that were identified as atpI, atpH, atpG, atpF, atpD, and atpA, coding for subunits IV, III, II, I, delta, and alpha, respectively. Remarkably, the genes atpG and atpD, which are nucleus- encoded in chlorophyll a + b plants, are present in the Odontella chloroplast gene cluster. They map at the same positions as in cyanobacteria. The genes atpD and atpF overlap by four base-pairs as in certain photosynthetic and heterotrophic eubacteria. Upstream from the atpA gene cluster an open reading frame coding for 251 amino acid residues was found, which shows sequence similarity to ATP-binding subunits of periplasmic prokaryotic and eukaryotic transport systems. No similar reading frame is present in the land plant chloroplast genomes analysed so far. Sequences and arrangement of the genes are discussed with respect to the peculiar evolution of the chlorophyll a + c-containing chromophytic plastids
RP  - NOT IN FILE
NT  - UI - 92219274LA - engRN - 9007-49-2 (DNA)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19920512IS - 0022-2836SB - IMCY - ENGLANDJC - J6V
UR  - PM:1532839
SO  - J Mol Biol 1992 Mar 20 ;224(2):529-536

1103
UI  - 20947
AU  - Papa S
AU  - Guerrieri F
AU  - Zanotti F
AU  - Capozza G
AU  - Fiermonte M
AU  - Cocco T
AU  - Altendorf K
AU  - Deckers-Hebersteit G
AD  - Institute of Medical Biochemistry and Chemistry, University of Bari, Italy
TI  - F0 and F1 subunits involved in the gate and coupling function of mitochondrial H+ ATP synthase
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Biochemistry
MH  - Chemistry
MH  - coupling
MH  - F0
MH  - F1
MH  - function
MH  - H+
MH  - Macromolecular Systems
MH  - review
MH  - SUBUNIT
MH  - SYNTHASE
MH  - SYSTEM
MH  - SYSTEMS
RP  - NOT IN FILE
NT  - UI - 93167635LA - engRN - 0 (Macromolecular Systems)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 19930312IS - 0077-8923SB - IMCY - UNITED STATES
UR  - PM:1288331
SO  - Ann N Y Acad Sci 1992 Nov 30 ;671():345-358

1104
UI  - 517
AU  - Pedersen PL
AU  - Amzel LM
AD  - Department of Biological Chemistry, John Hopkins University, School of Medicine, Baltimore, Maryland
TI  - F-type ATPases. Introduction
RP  - NOT IN FILE
NT  - UI - 93054450LA - engRN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 19921222IS - 0145-479XSB - IMCY - UNITED STATESJC - HIO
UR  - PM:1429534
SO  - J Bioenerg Biomembr 1992 Oct ;24(5):427-428

1105
UI  - 514
AU  - Pedersen PL
AU  - Thomas PJ
AU  - Garboczi DN
AU  - Bianchet M
AU  - Amzel LM
AD  - Department of Biological Chemistry, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205
TI  - F-type ATPases: are nucleotide domains in adenylate kinase appropriate models for nucleotide domains in ATP synthase/ATPase complexes?
RP  - NOT IN FILE
NT  - UI - 93167636LA - engRN - 0 (Macromolecular Systems)RN - 0 (Recombinant Proteins)RN - EC 2.7.4.3 (Adenylate Kinase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialID - CA 10951/CA/NCIID - GM 25432/GM/NIGMSDA - 19930312IS - 0077-8923SB - IMCY - UNITED STATESJC - 5NM
UR  - PM:1288332
SO  - Ann N Y Acad Sci 1992 Nov 30 ;671():359-365

1106
UI  - 21295
AU  - Rau DC
AU  - Parsegian VA
AD  - Laboratory of Biochemistry and Metabolism, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland 20892
TI  - Direct measurement of the intermolecular forces between counterion- condensed DNA double helices. Evidence for long range attractive hydration forces
AB  - Rather than acting by modifying van der Waals or electrostatic double layer interactions or by directly bridging neighboring molecules, polyvalent ligands bound to DNA double helices appear to act by reconfiguring the water between macromolecular surfaces to create attractive long range hydration forces. We have reached this conclusion by directly measuring the repulsive forces between parallel B-form DNA double helices pushed together from the separations at which they have self organized into hexagonal arrays of parallel rods. For all of the wide variety of "condensing agents" from divalent Mn to polymeric protamines, the resulting intermolecular force varies exponentially with a decay rate of 1.4-1.5 A, exactly one-half that seen previously for hydration repulsion. Such behavior qualitatively contradicts the predictions of all electrostatic double layer and van der Waals force potentials previously suggested. It fits remarkably well with the idea, developed and tested here, that multivalent counterion adsorption reorganizes the water at discrete sites complementary to unadsorbed sites on the apposing surface. The measured strength and range of these attractive forces together with their apparent specificity suggest the presence of a previously unexpected force in molecular organization
MH  - A
MH  - Biochemistry
MH  - DOUBLE-LAYER
MH  - INTERACTION
MH  - Kidney
MH  - Ligands
MH  - metabolism
MH  - Site
MH  - SURFACE
MH  - united states
MH  - Water
RP  - NOT IN FILE
NT  - UI - 92173087LA - engRN - 0 (Protamines)RN - 7439-96-5 (Manganese)RN - 7440-48-4 (Cobalt)RN - 9007-49-2 (DNA)PT - Journal ArticleDA - 19920403IS - 0006-3495SB - IMCY - UNITED STATES
UR  - PM:1540693
SO  - Biophys J 1992 Jan ;61(1):246-259

1107
UI  - 20999
AU  - Richard P
AU  - Graber P
AD  - Biologisches Institut, Universitat Stuttgart, Federal Republic of Germany
TI  - Kinetics of ATP synthesis catalyzed by the H(+)-ATPase from chloroplasts (CF0F1) reconstituted into liposomes and coreconstituted with bacteriorhodopsin
AB  - The H(+)-ATPase from chloroplasts (CF0F1) was isolated, purified and reconstituted into liposomes from phosphatidylcholine/phosphatidic acid. A transmembrane pH difference, delta pH, and a transmembrane electric potential difference, delta psi, were generated by an acid/base transition. The rate of ATP synthesis was measured at constant delta pH and constant delta psi as a function of temperature between 5 degrees C and 45 degrees C. The activation energy was 55 kJ mol-1. CF0F1 was coreconstituted with bacteriorhodopsin at a molar ratio of approximately 1:170 in the same type of liposomes. Illumination of the proteoliposomes leads to proton transport into the vesicles generating a constant delta pH = 1.8. The dependence of the rate of ATP synthesis on ADP concentration was measured with CF0F1 in the oxidized state, E(ox), and in the reduced state, E(red). The results can be described by Michaelis-Menten kinetics with the following parameters: Vmax = 0.5 s-1, Km = 8 microM for E(ox) and Vmax = 2.0 s-1, Km = 8 microM for E(red)
MH  - A
MH  - ACID
MH  - ACTIVATION
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - ADP
MH  - atp
MH  - ATP synthesis
MH  - ATPase
MH  - Bacteriorhodopsin
MH  - CF0F1
MH  - chloroplast
MH  - Chloroplasts
MH  - CONSTANT
MH  - DELTA-PH
MH  - DEPENDENCE
MH  - function
MH  - H(+)ATPase
MH  - Kinetics
MH  - Liposomes
MH  - pH
MH  - proteoliposome
MH  - proton
MH  - PSI
MH  - synthesis
MH  - Temperature
MH  - transport
RP  - NOT IN FILE
NT  - UI - 93076798LA - engRN - 0 (Liposomes)RN - 53026-44-1 (Bacteriorhodopsins)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19921230IS - 0014-2956SB - IMCY - GERMANY
UR  - PM:1446676
SO  - Eur J Biochem 1992 Nov 15 ;210(1):287-291

1108
UI  - 434
AU  - Senior AE
AU  - al Shawi MK
AD  - Department of Biochemistry, University of Rochester Medical Center, New York 14642
TI  - Further examination of seventeen mutations in Escherichia coli F1- ATPase beta-subunit
AB  - Seventeen mutations in beta-subunit of Escherichia coli F1-ATPase which had previously been characterized in strain AN1272 (Mu-induced mutant) were expressed in strain JP17 (beta-subunit gene deletion). Six showed unchanged behavior, namely: C137Y; G142D; G146S; G207D; Y297F; and Y354F. Five failed to assemble F1F0 correctly, namely: G149I; G154I; G149I,G154I; G223D; and P403S,G415D. Six assembled F1F0 correctly, but with membrane ATPase lower than in AN1272, namely: K155Q; K155E; E181Q; E192Q; D242N; and D242V. AN1272 was shown to unexpectedly produce a small amount of wild-type beta-subunit; F1-ATPase activities reported previously in AN1272 were referable to hybrid enzymes containing both mutant and wild-type beta-subunits. Purified F1 was obtained from K155Q; K155E; E181Q; E192Q; and D242N mutants in JP17. Vmax ATPase values were lower, and unisite catalysis rate and equilibrium constants were perturbed to greater extent, than in AN1272. However, general patterns of perturbation revealed by difference energy diagrams were similar to those seen previously, and the new data correlated well in linear free energy relationships for reaction steps of unisite catalysis. Correlation between multisite and unisite ATPase activity was seen in the new enzymes. Overall, the data give strong support to previously proposed mechanisms of unisite catalysis, steady-state catalysis, and energy coupling in F1-ATPases (Al-Shawi, M. K., Parsonage, D. and Senior, A. E. (1990) J. Biol. Chem. 265, 4402-4410). The K155Q, K155E, D242N, and E181Q mutations caused 5000-fold, 4000- fold, 1800-fold, and 700-fold decrease, respectively, in Vmax ATPase, implying possibly direct roles for these residues in catalysis. Experiments with the D242N mutant suggested a role for residue beta D242 in catalytic site Mg2+ binding
RP  - NOT IN FILE
NT  - UI - 93016088LA - engRN - 0 (Plasmids)RN - 7439-95-4 (Magnesium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSDA - 19921125IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:1400460
SO  - J Biol Chem 1992 Oct 25 ;267(30):21471-21478

1109
UI  - 435
AU  - Senior AE
AD  - Department of Biochemistry, University of Rochester Medical Center, New York 14642
TI  - Catalytic sites of Escherichia coli F1-ATPase
AB  - The catalytic site of Escherichia coli F1-ATPase is reviewed in terms of structure and function. Structural prediction, biochemical analyses, and mutagenesis experiments suggest that the catalytic site is formed primarily by residues 137-335 of beta-subunit. Subdomains of the site involved in phosphate-bond cleavage/synthesis and adenine-ring binding are discussed. Ambiguities inherent in steady-state catalytic measurements due to catalytic site cooperativity are discussed, and the advantages of pre-steady-state ("unisite") techniques are emphasized. The emergence of a single high-affinity catalytic site occurs as a result of F1-oligomer assembly. Measurements of unisite catalysis rate and equilibrium constants, and their modulation by varied pH, dimethylsulfoxide, and mutations, are described and conclusions regarding the nature of the high-affinity catalytic site and mechanism of catalysis are presented
RP  - NOT IN FILE
NT  - UI - 93054458LA - engRN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialID - GM25439/GM/NIGMSDA - 19921222IS - 0145-479XSB - IMCY - UNITED STATESJC - HIO
UR  - PM:1429542
SO  - J Bioenerg Biomembr 1992 Oct ;24(5):479-484

1110
UI  - 436
AU  - Senior AE
AU  - Lee RS
AU  - al Shawi MK
AU  - Weber J
AD  - Department of Biochemistry, University of Rochester Medical Center, New York 14642
TI  - Catalytic properties of Escherichia coli F1-ATPase depleted of endogenous nucleotides
AB  - Nucleotide-depleted Escherichia coli F1 was prepared by the procedure of Wise et al. (1983, Biochem. J. 215, 343-350). This enzyme had high rates of steady-state ATPase and GTPase activity. When "unisite" ATP hydrolysis was measured using an F1/ATP concentration ratio of 10, all of the substoichiometric ATP became bound to the high-affinity catalytic site and none became bound to noncatalytic sites. The association rate constant for ATP binding was 7 x 10(5) M-1 s-1 and the KdATP was 7.9 x 10(-10) M, as compared to values of 3.8 x 10(5) M-1 s-1 and 1.9 x 10(-10) M, respectively, in native (i.e., nucleotide-replete) F1. Rate constants for bound ATP hydrolysis, ATP resynthesis, and P(i) release, and the reaction equilibrium constant, were similar in nucleotide-depleted and native F1. Therefore, we conclude that occupancy of the noncatalytic sites is not required for formation of the high-affinity catalytic site of F1 and has no significant effect on unisite catalysis. In further experiments we looked for the occurrence of inhibitory, catalytic-site-bound MgADP in E. coli F1. Such an entity has been reported for chloroplast and mitochondrial F1. However, our experiments gave no indication for inhibitory MgADP in E. coli F1
RP  - NOT IN FILE
NT  - UI - 92359548LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.1.- (GTP Phosphohydrolases)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSDA - 19920910IS - 0003-9861SB - IMCY - UNITED STATESJC - 6SK
UR  - PM:1386723
SO  - Arch Biochem Biophys 1992 Sep ;297(2):340-344

1111
UI  - 20863
AU  - Shin K
AU  - Nakamoto RK
AU  - Maeda M
AU  - Futai M
AD  - Department of Organic Chemistry and Biochemistry, Osaka University, Japan
TI  - F0F1-ATPase gamma subunit mutations perturb the coupling between catalysis and transport
AB  - We introduced mutations to test the function of the conserved amino- terminal region of the gamma subunit from the Escherichia coli ATP synthase (F0F1-ATPase). Plasmid-borne mutant genes were expressed in an uncG strain which is deficient for the gamma subunit (gamma Gln-14-- >end). Most of the changes, which were between gamma Ile-19 and gamma Lys-33, gamma Asp-83 and gamma Cys-87, or at gamma Asp-165, had little effect on growth by oxidative phosphorylation, membrane ATPase activity, or H+ pumping. Notable exceptions were gamma Met-23-->Arg or Lys mutations. Strains carrying these mutations grew only very slowly by oxidative phosphorylation. Membranes prepared from the strains had substantial levels of ATPase activity, 100% compared with wild type for gamma Arg-23 and 65% for gamma Lys-23, but formed only 32 and 17%, respectively, of the electrochemical gradient of protons. In contrast, other mutant enzymes with similar ATPase activities (including gamma Met-23-->Asp or Glu) formed H+ gradients like the wild type. Membranes from the gamma Arg-23 and gamma Lys-23 mutants were not passively leaky to protons and had functional F0 sectors. These results suggested that substitution by positively charged side chains at position 23 perturbed the energy coupling. The catalytic sites of the mutant enzymes were still regulated by the electrochemical H+ gradient but were inefficiently coupled to H+ translocation in both ATP-dependent H+ pumping and delta mu H+ driven ATP synthesis
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - ATPase
MH  - Biochemistry
MH  - Carbonyl Cyanide m-Chlorophenyl Hydrazone
MH  - Catalysis
MH  - Chemistry
MH  - COLI ATP SYNTHASE
MH  - coupling
MH  - Dicyclohexylcarbodiimide
MH  - Enzymes
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - function
MH  - H+
MH  - Macromolecular Systems
MH  - membrane
MH  - Membranes
MH  - mutant
MH  - Oxidative Phosphorylation
MH  - Phosphorylation
MH  - protein
MH  - Proteins
MH  - proton
MH  - Protons
MH  - Site
MH  - SUBUNIT
MH  - SYNTHASE
MH  - synthesis
MH  - SYSTEM
MH  - SYSTEMS
MH  - translocation
MH  - transport
RP  - NOT IN FILE
NT  - UI - 93015990LA - engRN - 0 (Macromolecular Systems)RN - 0 (Oligodeoxyribonucleotides)RN - 0 (Plasmids)RN - 0 (Recombinant Proteins)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 555-60-2 (Carbonyl Cyanide m-Chlorophenyl Hydrazone)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19921118IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:1400398
SO  - J Biol Chem 1992 Oct 15 ;267(29):20835-20839

1112
UI  - 879
AU  - Skulachev VP
TI  - The laws of cell energetics
AB  - Recent progress in membrane bioenergetics studies has resulted in the important discovery that Na+ can effectively substitute for H+ as the energy coupling ion. This means that living cells can possess three convertible energy currencies, i.e. ATP, protonic and sodium potentials. Analysis of interrelations of these components in various types of living cells allows bioenergetic laws of universal applicability to be inferred
MH  - Adenosine Triphosphate
MH  - Bacteria
MH  - Cell Membrane
MH  - Cells
MH  - Energy Metabolism
MH  - metabolism
MH  - Protons
MH  - Sodium
RP  - NOT IN FILE
NT  - Department of Bioenergetics, A N Belozersky Institute of Physico-Chemical Biology, Moscow State University, Russia
SO  - Eur J Biochem 1992 Sep 1 ;208(2):203-209

1113
UI  - 515
AU  - Thomas PJ
AU  - Garboczi DN
AU  - Pedersen PL
AD  - Department of Biological Chemistry, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205
TI  - Mutational analysis of the consensus nucleotide binding sequences in the rat liver mitochondrial ATP synthase beta-subunit
AB  - The coupling step in the biosynthesis of ATP in biological systems is generally believed to involve an energy-requiring release of ATP bound to the beta-subunit of the ATP synthase complex. A molecular description of the ATP binding site on the beta-subunit is, therefore, critical to understanding the mechanism of coupling in the enzyme. Previously, we reported that a purified, bacterially expressed rat liver beta-subunit binds adenine nucleotides tightly and specifically (Garboczi, D. N., Hullihen, J. H., and Pedersen, P. L. (1988) J. Biol. Chem. 263, 15694-15698). In order to assess the contribution of various regions of the isolated beta-subunit to the ATP binding site we have systematically deleted four different regions: the N-terminal region, the Walker A consensus region, the Walker B consensus region (Walker, J. E., Saraste, M., Runswick, M. J., and Gay, N. (1982) EMBO J. 1, 945- 951), and a "C" region, which, like the A and B regions, bears homology to adenylate kinase. Plasmids directing the expression of double deletions of A and B regions, and B and C regions were also constructed. In addition, 2 residues outside of these regions, His-177 and Tyr-345, which have been predicted to play a central role in nucleotide binding, were mutated. Rabbit antisera to synthetic peptides of the A and C regions verified the identity of the bacterially expressed mutant proteins. Seven of the eight mutant proteins overexpressed in Escherichia coli were resistant to E. coli proteases in the preparative stages, as predicted for compact folded proteins. Furthermore, circular dichroism spectropolarimetry revealed no profound structural alterations in the purified mutant proteins. Relative to the overexpressed full-length beta-subunit, the mutant lacking the A consensus region suffered a 30-fold loss of affinity for ATP and a loss of specificity for 2'(3')-O-(2,4,6-trinitrophenyl)adenosine 5'- triphosphate (TNP-ATP) over 2'(3')-O-(2,4,6-trinitrophenyl)adenosine 5'- monophosphate. The mutant proteins lacking either the N-terminal region or the B region exhibited nucleotide binding properties similar to the full-length beta-subunit, whereas the mutant protein lacking the C region suffered an order of magnitude reduction in affinity for ATP. The affinity of the A and B region double deletion was indistinguishable from the A region deletion in regard to TNP-ATP binding, while the double deletion mutant lacking the B and C regions was not stably expressed in the E. coli SE6004.(ABSTRACT TRUNCATED AT 400 WORDS)
RP  - NOT IN FILE
NT  - UI - 93015911LA - engRN - 0 (Adenine Nucleotides)RN - 0 (Plasmids)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - CA 10951/CA/NCIDA - 19921116IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:1400352
SO  - J Biol Chem 1992 Oct 5 ;267(28):20331-20338

1114
UI  - 520
AU  - Thomas PJ
AU  - Garboczi DN
AU  - Pedersen PL
AD  - Department of Biological Chemistry, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205
TI  - Mitochondrial F-type ATPases: the glycine-rich loop of the beta-subunit is a pyrophosphate binding domain
AB  - The beta-subunit of the mitochondrial ATP synthase complex comprises the bulk, if not all, of the catalytic nucleotide binding site on the enzyme. A region of homologous sequence rich in glycines (G) and containing a basic lysine (K) and a threonine (T) is found in the beta- subunit as well as many other purine nucleotide binding proteins. The consensus sequence of this region is Gx4GKT, where x represents any amino acid, and is called the A region or glycine-rich loop. The related function of these proteins implies that the glycine-rich loop is directly involved in nucleotide binding. Here we directly test the involvement of the beta-subunit's glycine-rich region in adenine nucleotide binding using two independent approaches. A synthetic fifty amino acid peptide, PP-50, containing the glycine-rich region and the surrounding sequence was assessed for secondary structure and interaction with potential ligands. Circular dichroism spectropolarimetry indicates that PP-50 assumes a predominantly beta- sheet conformation in solution. Significantly, the peptide precipitates from solution when ATP, ADP, GTP, ITP, and pyrophosphate are added, but not when AMP or phosphate are included. Magnesium is not required for the interaction with the purine nucleotides. Complimentary to these studies, the sequence around the Gx4GKT motif was deleted from a recombinant rat liver beta-subunit overexpressed in E. coli. While the wild type beta-subunit showed specificity for the tri- and diphosphonucleotides, the deletion mutant bound tri-, di-, and monophosphate nucleotides with equal affinity.(ABSTRACT TRUNCATED AT 250 WORDS)
RP  - NOT IN FILE
NT  - UI - 93080077LA - engRN - 0 (Adenine Nucleotides)RN - 0 (Diphosphates)RN - 0 (Ligands)RN - 0 (Purine Nucleotides)RN - 56-40-6 (Glycine)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19921230IS - 0302-2994SB - IMCY - ENGLANDJC - 1UF
UR  - PM:1333155
SO  - Acta Physiol Scand Suppl 1992  ;607():23-29

1115
UI  - 519
AU  - Thomas PJ
AU  - Bianchet M
AU  - Garboczi DN
AU  - Hullihen J
AU  - Amzel LM
AU  - Pedersen PL
AD  - Department of Biological Chemistry, Johns Hopkins University, School of Medicine, Baltimore, MD
TI  - ATP synthase: structure-function relationships
AB  - Recent work has focused on obtaining a better understanding of the three-dimensional structural relationships between the alpha and beta subunits of the F1 moiety and the location of nucleotide binding domains within these subunits. Four types of approach are currently being pursued: X-ray crystallographic, chemical, molecular biological and biochemical. Here we briefly review some of the major conclusions of these studies, and point out some of the problems that must be resolved before an adequate model that relates structure to function in the ATP synthase molecule can be formulated
RP  - NOT IN FILE
NT  - UI - 92338207LA - engRN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialID - CA 10951/CA/NCIID - GM 25432/GM/NIGMSDA - 19920825IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:1385978
SO  - Biochim Biophys Acta 1992 Jul 17 ;1101(2):228-231

1116
UI  - 21001
AU  - Turina P
AU  - Rumberg B
AU  - Melandri BA
AU  - Graber P
AD  - Max-Volmer Institut, Technische Universitat Berlin, Germany
TI  - Activation of the H(+)-ATP synthase in the photosynthetic bacterium Rhodobacter capsulatus
AB  - The regulation of the membrane-bound H(+)-ATPase from the photosynthetic bacterium Rhodobacter capsulatus was investigated. In the presence of uncouplers the rate of ATP hydrolysis was about 40 mM ATP/M bacteriochlorophyll (Bchl)/s. Without uncouplers this rate increased and if, additionally, the chromatophores were illuminated, it was almost doubled. If uncouplers were added shortly after illumination, the rate increased to 300-350 mM ATP/M Bchl/s. Obviously, energization of the membrane leads to the formation of a metastable, active state of the H(+)-ATPase. The maximal rate of ATP hydrolysis can be measured only when first all H(+)-ATPases are activated by delta mu H+ and when the delta mu H+ is abolished in order to release its back pressure on the hydrolysis rate. The half-life time of the metastable state in the absence of delta mu H+ is about 30 s. It is increased by 3 mM Pi to about 80 s and it is decreased by 1 mM ADP to about 15 s. Quantitatively, the fraction of active H(+)-ATPases shows a sigmoidal dependence on pHin (at constant pHout) and the magnitude of delta psi determines the maximal fraction of enzymes which can be activated: delta pH and delta psi are not equivalent for the activation process
MH  - A
MH  - ACTIVATION
MH  - ACTIVE
MH  - Adenine Nucleotides
MH  - ADENINE-NUCLEOTIDES
MH  - ADP
MH  - atp
MH  - ATPase
MH  - Bacteria
MH  - capsulatus
MH  - chromatophore
MH  - chromatophores
MH  - CONSTANT
MH  - DELTA-PH
MH  - DEPENDENCE
MH  - Enzymes
MH  - H(+)ATPase
MH  - H+
MH  - Hydrolysis
MH  - membrane
MH  - Nucleotides
MH  - pH
MH  - Phosphates
MH  - PSI
MH  - regulation
MH  - Rhodobacter
MH  - rhodobacter capsulatus
MH  - RHODOBACTER-CAPSULATUS
MH  - SYNTHASE
MH  - Time
RP  - NOT IN FILE
NT  - UI - 92283802LA - engRN - 0 (Adenine Nucleotides)RN - 0 (Phosphates)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19920706IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:1534558
SO  - J Biol Chem 1992 Jun 5 ;267(16):11057-11063

1117
UI  - 19866
AU  - Vogel PD
AU  - Nett JH
AU  - Sauer HE
AU  - Schmadel K
AU  - Cross RL
AU  - Trommer WE
AD  - Fachbereich Chemie der Universitat, Kaiserslautern, Federal Republic of Germany
TI  - Nucleotide binding sites on mitochondrial F1-ATPase. Electron spin resonance spectroscopy and photolabeling by azido-spin-labeled adenine nucleotides support an adenylate kinase-like orientation
AB  - A spin-labeled photoaffinity ATP analog, 2-N3-2',3'-SL-ATP (2-N3-SL- ATP) was specifically loaded at catalytic (exchangeable) or noncatalytic (nonexchangeable) nucleotide-binding sites on nucleotide- depleted mitochondrial F1-ATPase. Photolysis of the enzyme complexes resulted in the specific modification of beta-Tyr-345 when the catalytic sites were occupied and beta-Tyr-368 when noncatalytic sites were filled. These are the same amino acid assignments that were made previously using 2-N3ATP. The results demonstrate that the attachment of a spin label moiety to the ribose ring does not prevent proper binding of the analog at both types of nucleotide sites on F1-ATPase and suggest that the probe can be used for investigations of the nucleotide-binding sites using ESR spectroscopy. Enzyme that is in complex with the 2-N3-SL-ATP exhibits an ESR spectrum that is typical for highly immobilized nitroxyl radicals both in the dark or after photolysis. Additional peaks in the high- and low-field regions arise due to dipolar spin interactions most likely involving a pair of catalytic and noncatalytic sites. The two sites are calculated to be approximately 15 A apart. This distance, obtained through ESR spectroscopy, combined with the finding that the 2 labeled amino acids are only 23 residues apart from each other, further supports an adenylate kinase-like arrangement of nucleotide binding sites on F1- ATPase where catalytic and noncatalytic sites are in close proximity (Vogel, P. D., and Cross, R. L. (1991) J. Biol. Chem. 266, 6101-6105)
MH  - A
MH  - Adenine Nucleotides
MH  - ADENINE-NUCLEOTIDES
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - Amino Acids
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Azides
MH  - BINDING
MH  - Binding Sites
MH  - COMPLEX
MH  - electron
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - H(+)-Transporting ATP Synthase
MH  - MITOCHONDRIAL F1-ATPASE
MH  - NONCATALYTIC SITES
MH  - nucleotide binding
MH  - nucleotide binding sites
MH  - NUCLEOTIDE-BINDING-SITES
MH  - Nucleotides
MH  - P
MH  - spectroscopy
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 92291073LA - engRN - 0 (Adenine Nucleotides)RN - 0 (Affinity Labels)RN - 0 (Azides)RN - 0 (Spin Labels)RN - 143796-55-8 (2-azido-2'-3'-(2,2,5,5-tetramethyl-3-pyrroline-1-oxyl-3- carboxylic acid ester)ATP)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM 23152/GM/NIGMSDA - 19920716IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:1318307
SO  - J Biol Chem 1992 Jun 15 ;267(17):11982-11986

1118
UI  - 441
AU  - Weber J
AU  - Lee RS
AU  - Grell E
AU  - Wise JG
AU  - Senior AE
AD  - Department of Biochemistry, University of Rochester Medical Center, New York 14642
TI  - On the location and function of tyrosine beta 331 in the catalytic site of Escherichia coli F1-ATPase
AB  - 1) Using a combination of site-directed mutagenesis and fluorescence spectroscopy we have studied the location and function of residue beta Y331 in the catalytic site of Escherichia coli F1-ATPase. The fluorescent analog lin-benzo-ADP was used as a catalytic-site probe, and was found to bind to three sites in normal F1, with Kd1 = 0.20 microM and Kd2,3 = 5.5 microM. lin-Benzo-ATP was a good substrate for hydrolysis. 2) The mutants investigated were beta Y331F, L, A and E. kcat/KM for ATP hydrolysis in purified F1 was reduced according to the series Y greater than or equal to F greater than L greater than A greater than E, with E being severely impaired; concomitant decreases in binding affinity for lin-benzo-ADP were seen. 3) Fluorescence properties of lin-benzo-ADP bound to F1 differed widely, depending on the residue present at position beta 331. Red shifts of excitation and emission spectra occurred with F and L residues, but not with Y, A, or E. There was strong quenching of fluorescence with wild-type (Y), partial quenching with A, and no quenching with F, L, or E. 4) We conclude that (a) the environment around the bound adenine moiety in the catalytic site is nonpolar, (b) residue beta 331 is part of the adenine-binding subdomain and when tyrosine is the residue, the phenolic hydroxyl makes direct interaction with the fluorophore, (c) an aromatic residue is not absolutely required at position beta 331 for catalytic function, but an increase in polarity leads to functional impairment, and (d) in terms of fluorescence response of bound lin- benzo-ADP all three catalytic sites behaved the same. 5) F1 from mutant beta Y297F bound lin-benzo-ADP with the same fluorescence and binding characteristics as normal F1, and catalytic properties were similar to normal. Therefore, there was no reason to conclude that residue beta Y297 is involved in binding the adenine moiety of ATP
RP  - NOT IN FILE
NT  - UI - 92112892LA - engRN - 0 (Oligodeoxyribonucleotides)RN - 0 (Plasmids)RN - 0 (Recombinant Proteins)RN - 55520-40-6 (Tyrosine)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSDA - 19920218IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:1530942
SO  - J Biol Chem 1992 Jan 25 ;267(3):1712-1718

1119
UI  - 438
AU  - Weber J
AU  - Lee RS
AU  - Grell E
AU  - Senior AE
AD  - Department of Biochemistry, University of Rochester Medical Center, New York 14642
TI  - Investigation of the aurovertin binding site of Escherichia coli F1- ATPase by fluorescence spectroscopy and site-directed mutagenesis
AB  - (1) Previous mutational analyses have shown that residue beta R398 of the beta-subunit is a key residue for binding of the inhibitory antibiotic aurovertin to Escherichia coli F1Fo-ATP synthase. Here, we studied purified F1 from the beta R398C and beta R398W mutants. ATPase activity in both cases was resistant to aurovertin inhibition. The fluorescence spectrum (lambda exc = 278 or 295 nm) of beta R398W F1 showed a significant red-shift as compared to wild-type and beta R398C enzymes, indicating that residue beta R398 lies in a polar environment. On the basis of this and previous evidence, we propose that aurovertin binding to F1-ATPase involves a specific charged donor-acceptor H-bond between residue beta R398 and the 7-hydroxyl group of aurovertin. (2) The fluorescent substrate analog lin-benzo-ADP was shown to bind to beta R398W F1 catalytic sites with the same Kd values as to wild-type F1, and with the same quenching of the fluorescence of the analog. Fluorescence energy transfer was seen between the beta R398W residue and bound lin-benzo-ADP. Analysis of transfer efficiency at varying stoichiometry of bound lin-benzo-ADP showed that interaction occurred between one beta R398W residue and one catalytic-site-bound analog molecule at a distance of approximately 23 A. The relationships of the aurovertin and catalytic sites in the primary and tertiary structure are discussed
RP  - NOT IN FILE
NT  - UI - 92297617LA - engRN - 0 (Aurovertins)RN - 58-64-0 (Adenosine Diphosphate)RN - 61925-59-5 (linear-benzoadenosine diphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSDA - 19920723IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:1534996
SO  - Biochemistry 1992 Jun 9 ;31(22):5112-5116

1120
UI  - 174
AU  - Wilkens S
AU  - Capaldi RA
AD  - Institute of Molecular Biology, University of Oregon, Eugene 97403
TI  - Monomaleimidogold labeling of the gamma subunit of the Escherichia coli F1 ATPase examined by cryoelectron microscopy
AB  - A novel approach for locating sites of interest in a protein complex has been developed using monomaleimidonanogold (MMN). The Escherichia coli F1 ATPase, when prepared without the delta subunit, contains only a single reactive cysteine on one of the three copies of the alpha subunit. This site was reacted with MMN and the gold cluster visualized on the protein complex by cryoelectron microscopy. Additional sites for modification with MMN were added by introducing cysteine residues through site-directed mutagenesis. Labeling of two mutants, gamma S8-C and gamma T106-C, in which Ser8 and Thr106, respectively, had been replaced by a cysteine, placed the gold cluster on the central mass that is seen in the hexagonal projection of the ECF1 complex. The results establish that the central mass contains the N-terminal part of the gamma subunit
RP  - NOT IN FILE
NT  - UI - 93073941LA - engRN - 0 (Immunoglobulins, Fab)RN - 0 (Macromolecular Systems)RN - 0 (Maleimides)RN - 0 (monomaleimido gold)RN - 7440-57-5 (Gold)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - HL24536/HL/NHLBIDA - 19921202IS - 0003-9861SB - IMCY - UNITED STATESJC - 6SK
UR  - PM:1444442
SO  - Arch Biochem Biophys 1992 Nov 15 ;299(1):105-109

1121
UI  - 9950
AU  - Yamasaki H
AU  - Okayama S
AU  - Shibata M
AU  - Nishimura M
AD  - Department of Biology, Faculty of Science, Kyushu University, Fukuoka, Japan
TI  - Inhibition of the light-induced H+ release from uncoupled thylakoid membranes by N-ethylmaleimide
AB  - The light-induced H+ release from thylakoids, which can be observed under completely uncoupled conditions, was inhibited by the SH reagent N-ethylmaleimide (NEM) and its analogs, while the conventional H+ uptake and electron transfer were not affected. The half-inhibiting concentration of NEM for the H+ release was 10 mM and 4 mM in thylakoids in the presence of nigericin and in CF1-depleted thylakoids, respectively. The inhibitory effect increased with the increase in hydrophobicity of the NEM analogs: N-methylmaleimide less than N- ethylmaleimide less than N-phenylmaleimide. It is suggested that SH groups in hydrophobic interior within the membrane are essential to the release of protons
MH  - ANALOGS
MH  - electron
MH  - Ethylmaleimide
MH  - H+
MH  - Membranes
MH  - proton
MH  - Protons
MH  - TRANSFER
RP  - NOT IN FILE
NT  - UI - 92171940LA - engRN - 0 (Maleimides)RN - 128-53-0 (Ethylmaleimide)PT - Journal ArticleDA - 19920327IS - 0006-291XSB - IMCY - UNITED STATESJC - 9Y8
UR  - PM:1540171
SO  - Biochem Biophys Res Commun 1992 Feb 14 ;182(3):1277-1281

1122
UI  - 19779
AU  - Zhou JM
AU  - Boyer PD
AD  - Molecular Biology Institute, University of California, Los Angeles 90024-1570
TI  - MgADP and free Pi as the substrates and the Mg2+ requirement for photophosphorylation
AB  - Previous studies have not provided definitive information about whether ADP or Pi or their complexes with Mg2+ serve as substrates for photophosphorylation and whether free Mg2+ or ADP is required. Results presented show MgADP, MgGDP, or MgUDP are substrates. At variable Mg2+ concentrations, observed velocities are determined by MgADP and not the free ADP concentration. The approximate Km for MgADP with spinach chloroplasts is about 30 microM, for MgGDP 260 microM, and for MgUDP above 5 mM. The apparent Km values for added ADP or Mg2+ are decreased to constant low values near 30 microM as the added Mg2+ or ADP concentrations, respectively, are increased to the millimolar range. With 100 microM added Mg2+, near-maximal velocities can be obtained with excess ADP, but not with excess GDP or UDP. This is explainable by the apparent Km values for MgGDP and MgUDP being well above 100 microM. High phosphorylation rates with excess of either Mg2+ or ADP present show that little or no (less than 2-3 microM) free Mg2+ or ADP is required. In addition, the results show that during rapid photophosphorylation, when one or more catalytic sites are always filled with nucleotide, free ADP does not combine and block the combination of MgADP to catalytic sites that become vacant. This is in contrast to the ability of free ADP to combine tightly with one catalytic site when all catalytic sites are empty. The apparent Km for added ADP above a few micromolar concentration, and with excess Mg2+ present, results from binding of MgADP at a second catalytic site.(ABSTRACT TRUNCATED AT 250 WORDS)
MH  - A
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - ADP
MH  - BINDING
MH  - chloroplast
MH  - Chloroplasts
MH  - COMPLEX
MH  - Magnesium
MH  - membrane
MH  - Membrane Proteins
MH  - Phosphates
MH  - Phosphorylation
MH  - Photophosphorylation
MH  - Proteins
MH  - Spinach
MH  - SPINACH-CHLOROPLASTS
RP  - NOT IN FILE
NT  - UI - 92207930LA - engRN - 0 (Membrane Proteins)RN - 0 (Phosphates)RN - 146-91-8 (Guanosine Diphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 58-98-0 (Uridine Diphosphate)RN - 7439-95-4 (Magnesium)PT - Journal ArticleID - GM-11094/GM/NIGMSDA - 19920501IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:1554702
SO  - Biochemistry 1992 Mar 31 ;31(12):3166-3171

1123
UI  - 349
AU  - Abrahams JP
AU  - Lutter R
AU  - Todd RJ
AU  - van Raaij MJ
AU  - Leslie AG
AU  - Walker JE
AD  - MRC Laboratory of Molecular Biology, Cambridge, UK
TI  - Inherent asymmetry of the structure of F1-ATPase from bovine heart mitochondria at 6.5 A resolution
AB  - ATP synthase, the assembly which makes ATP in mitochondria, chloroplasts and bacteria, uses transmembrane proton gradients generated by respiration or photosynthesis to drive the phosphorylation of ADP. Its membrane domain is joined by a slender stalk to a peripheral catalytic domain, F1-ATPase. This domain is made of five subunits with stoichiometries of 3 alpha: 3 beta: 1 gamma: 1 delta: 1 epsilon, and in bovine mitochondria has a molecular mass of 371,000. We have determined the 3-dimensional structure of bovine mitochondrial F1- ATPase to 6.5 A resolution by X-ray crystallography. It is an approximately spherical globule 110 A in diameter, on a 40 A stem which contains two alpha-helices in a coiled-coil. This stem is presumed to be part of the stalk that connects F1 with the membrane domain in the intact ATP synthase. A pit next to the stem penetrates approximately 35 A into the F1 particle. The stem and the pit are two examples of the many asymmetric features of the structure. The central element in the asymmetry is the longer of the two alpha-helices in the stem, which extends for 90 A through the centre of the assembly and emerges on top into a dimple 15 A deep. Features with threefold and sixfold symmetry, presumed to be parts of homologous alpha and beta subunits, are arranged around the central rod and pit, but the overall structure is asymmetric. The central helix provides a possible mechanism for transmission of conformational changes induced by the proton gradient from the stalk to the catalytic sites of the enzyme
RP  - NOT IN FILE
NT  - UI - 93259118LA - engRN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19930611IS - 0261-4189SB - IMCY - ENGLANDJC - EMB
UR  - PM:8491170
SO  - EMBO J 1993 May ;12(5):1775-1780

1124
UI  - 20806
AU  - Adams J
TI  - MUDPACK-2: Multigrid Software for Elliptic Partial Differential Equations on Uniform Grids with any Resolution
MH  - Software
RP  - IN FILE
SO  - Applied Math Comp 1993  ;53():235-249

1125
UI  - 172
AU  - Aggeler R
AU  - Capaldi RA
AD  - Institute of Molecular Biology, University of Oregon, Eugene 97403
TI  - ATP hydrolysis-linked structural changes in the N-terminal part of the gamma subunit of Escherichia coli F1-ATPase examined by cross-linking studies
AB  - A mutant of Escherichia coli F1-ATPase (ECF1) in which the serine residue in position 8 of the gamma subunit has been replaced by a cysteine residue (gamma S8C) has been used to study nucleotide- dependent cross-linking of the gamma subunit to a beta subunit. When examined in the presence of ADP+Mg2+, either supplied directly or as produced during catalytic turnover of ATP+Mg2+, the main cross-linked product generated using the heterobifunctional, photoactivatable, cross- linker tetrafluorophenylazide maleimide-6 had a M(r)(app) of 108,000. When ATP hydrolysis was inhibited, either by cold or by reaction with sodium azide, or when ATP hydrolysis was prevented by the use of adenyl- 5'-yl beta,gamma-imidodiphosphate, the main cross-linked products were species with M(r)(app) of 102,000 and 84,000. The nucleotide-dependent switching from one cross-linking pattern to another could only be observed when the epsilon subunit was bound to ECF1; it was not seen in ECF1*, an enzyme preparation missing delta and epsilon subunits, but was observed in preparations selectively depleted of the delta subunit. We conclude that the changes detected in these cross-linking experiments are occurring during the hydrolysis of ATP when the beta- gamma phosphate bond is cleaved and that they are related to the coupling of ATP hydrolysis to proton translocation
RP  - NOT IN FILE
NT  - UI - 93315415LA - engRN - 0 (Azides)RN - 0 (Cross-Linking Reagents)RN - 0 (Maleimides)RN - 139428-49-2 (N-(4-azido-2,3,5,6-tetrafluorobenzyl)-6- maleimidohexanamide)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 7439-95-4 (Magnesium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - HL 24236/HL/NHLBIDA - 19930812IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:8392054
SO  - J Biol Chem 1993 Jul 15 ;268(20):14576-14578

1126
UI  - 171
AU  - Aggeler R
AU  - Cai SX
AU  - Keana JF
AU  - Koike T
AU  - Capaldi RA
AD  - Institute of Molecular Biology, University of Oregon, Eugene 97403
TI  - The gamma subunit of the Escherichia coli F1-ATPase can be cross-linked near the glycine-rich loop region of a beta subunit when ADP + Mg2+ occupies catalytic sites but not when ATP + Mg2+ is bound
AB  - A mutant of the Escherichia coli F1-ATPase, gamma S8C, has been reacted with a novel bifunctional reagent, N-maleimido-N'-(4-azido-2,3,5,6- tetrafluorobenzamido) cystamine (TFPAM-SS1). Modification of Cys-8 via the maleimide, followed by photolysis to convert the azido group to a reactive nitrene, led to cross-linking of the gamma subunit to a beta subunit. When this cross-linking was conducted with ADP + Mg2+ in catalytic sites, the predominant cross-linked product had a M(r) of 108,000. If cross-linking was done with uncleaved ATP + Mg2+ in catalytic sites, cross-linked products of 102,000 and 84,000 were formed. Cross-linking under both conditions led to inhibition of ATPase activity. TFPAM-SS1 could be cleaved by using reducing agents to break the disulfide bond that links the malemide and tetrafluorophenylazide moieties. Cleavage of this disulfide bond after formation of 102,000 and 84,000 species led to full recovery of ATPase activity. When the 108-kDa cross-linked product was cleaved, full activity was not restored, presumably because of insertion of the tetrafluorophenylazide into a functionally important site on the beta subunit. After cleavage of the disulfide bond, the free thiols could be reacted with [14C]N- ethylmaleimide, thereby radioactively tagging the sites of insertion of the tetrafluorophenylnitrene moiety. In this way, the site of cross- linking from Cys-8 of gamma to the beta subunit in the presence of ADP + Mg2+ was localized to within the sequence Val 145-Lys-155, which contains the glycine-rich loop. This loop region is a part of the catalytic site of the enzyme
RP  - NOT IN FILE
NT  - UI - 94012621LA - engRN - 0 (Cross-Linking Reagents)RN - 0 (Fluorobenzenes)RN - 0 (Maleimides)RN - 139428-48-1 (N-(4-azido-2,3,5,6-tetrafluorobenzyl)-3- maleimidopropionamide)RN - 56-40-6 (Glycine)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 7439-95-4 (Magnesium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM27137/GM/NIGMSID - HL24236/HL/NHLBIDA - 19931124IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:8407913
SO  - J Biol Chem 1993 Oct 5 ;268(28):20831-20837

1127
UI  - 21265
AU  - Antonenko YN
AU  - Kovbasnjuk ON
AU  - Yaguzhinsky LS
AD  - AN Belozersky Institute of Physico-chemical Biology, Moscow State University, Russian Federation
TI  - Evidence in favor of the existence of a kinetic barrier for proton transfer from a surface of bilayer phospholipid membrane to bulk water
AB  - When the hydrogen-ion flux is induced by nigericin across the planar bilayer lipid membrane (BLM) with bulk pH values being equal at the opposite sides of the BLM, formation of a difference in boundary potentials (delta phi b) on the membrane is observed by the method of inner membrane field compensation. pH gradients are titrated routinely by the addition of sodium acetate at one side of the membrane. The increase in buffer concentration (citrate, phosphate, Mes) leads to a decrease in delta phi b. delta phi b forms in the presence of phosphatidylserine in the membrane-forming solution only. It is concluded that the steady-state difference of the hydrogen ion binding to the opposite surfaces of the membrane (HIBD) is created under the conditions of equal pH values near surfaces of the BLM. The model of the processes implies that nigericin transfers proton predominantly from interface to interface while acetate transfers the proton from bulk phase to bulk phase. In the other series of experiments the monensin-mediated formation of the HIBD leads to the formation of an potassium-ion gradient in the presence of nigericin. Thus, a possibility of performing a work due to the formation of HIBD is demonstrated. Owing to these properties the hydrogen-ion binding difference can be interpreted in a first approximation as a difference of surface hydrogen-ion concentration at the opposite sides of the membrane, arising due to the existence of a kinetic barrier for the proton transfer at the membrane interfaces. These findings can be significant for the mechanism of energy transduction in membrane phosphorylation in mitochondria and chloroplasts
MH  - A
MH  - acetate
MH  - BINDING
MH  - buffer
MH  - Buffers
MH  - chloroplast
MH  - Chloroplasts
MH  - delta
MH  - FIELD
MH  - Hydrogen
MH  - Hydrogen-Ion Concentration
MH  - interfaces
MH  - ion
MH  - Lipid Bilayers
MH  - mechanism
MH  - membrane
MH  - method
MH  - Mitochondria
MH  - model
MH  - pH
MH  - Phospholipids
MH  - Phosphorylation
MH  - proton
MH  - Proton transfer
MH  - Proton-transfer
MH  - Protons
MH  - Sodium
MH  - SURFACE
MH  - TRANSFER
MH  - Valinomycin
MH  - Water
RP  - NOT IN FILE
NT  - UI - 93326602LA - engRN - 0 (Buffers)RN - 0 (Lipid Bilayers)RN - 0 (Phospholipids)RN - 0 (Protons)RN - 2001-95-8 (Valinomycin)RN - 28380-24-7 (Nigericin)RN - 69279-91-0 (asolectin)RN - 7732-18-5 (Water)PT - Journal ArticleDA - 19930823IS - 0006-3002SB - IMCY - NETHERLANDS
UR  - PM:8392870
SO  - Biochim Biophys Acta 1993 Jul 25 ;1150(1):45-50

1128
UI  - 19902
AU  - Benjamin I
TI  - Mechanism and dynamics of ion transfer across a liquid-liquid interface
MH  - mechanism
MH  - ion
MH  - TRANSFER
MH  - A
RP  - IN FILE
SO  - Science 1993  ;261():1558-1560

1129
UI  - 21193
AU  - Bibikov SI
AU  - Grishanin RN
AU  - Kaulen AD
AU  - Marwan W
AU  - Oesterhelt D
AU  - Skulachev VP
AD  - AN Belozersky Institute of Physico-Chemical Biology, Moscow State University, Russia
TI  - Bacteriorhodopsin is involved in halobacterial photoreception
AB  - The bacterio-opsin gene was introduced into a "blind" Halobacterium salinarium mutant that (i) lacked all the four retinal proteins [bacteriorhodopsin (BR), halorhodopsin, and sensory rhodopsins (SRs) I and II] and the transducer protein for SRI and (ii) showed neither attractant response to long wavelength light nor repellent response to short wavelength light. The resulting transformed cells acquired the capability to sense light stimuli. The cells accumulated in a light spot, demonstrating the BR-mediated orientation in spatial light gradients. As in wild-type cells, a decrease in the intensity of long wavelength light caused a repellent response by inducing reversals of swimming direction, but, in contrast to wild-type cells, a decrease in the intensity of short wavelength light also repelled the cells. An increase in light intensity evoked an attractant response (i.e., a transient suppression of spontaneous reversals). Signal processing times and adaptation kinetics were similar to the SRI-mediated reactions. However, compared to SR-mediated photoresponses, higher light intensities were necessary to induce the BR-mediated responses. The light sensitivity of the transformant was increased by adding 1 mM cyanide and decreased by the addition of arginine, agents that respectively reduce and increase the light-independent generation of the electrochemical potential difference of H+ ions (delta mu H+). A decrease in irradiance to an intensity that was still high enough to saturate BR-initiated delta mu H+ changes failed to induce the repellent effect, but the addition of a protonophorous uncoupler sensitized the cell to these light stimuli. The BR D96N mutant (Asp-96 is replaced by Asn) with decreased proton pump activity showed strongly reduced BR-mediated responses. Azide, which increases this mutant's H+ pump efficiency, increased the photosensitivity of the mutant cells. Moreover, azide diminished (i) the membrane potential decreasing and (ii) repellent effects of blue light added to the orange background illumination in this mutant. We conclude that the BR-mediated photoreception is due to the light-dependent generation of delta mu H+. Our data are consistent with the assumption that the H. salinarium cell monitors the membrane energization level with a "protometer" system measuring total delta mu H+ changes or its electric potential difference component
MH  - A
MH  - Bacteriorhodopsin
MH  - Cells
MH  - delta
MH  - H+
MH  - Halobacterium
MH  - Halobacterium salinarium
MH  - ion
MH  - Ions
MH  - Kinetics
MH  - Light
MH  - light-gradient
MH  - membrane
MH  - Membrane Potential
MH  - mutant
MH  - Potassium
MH  - protein
MH  - Proteins
MH  - proton
MH  - Proton Pump
MH  - retinal
MH  - SYSTEM
MH  - Time
RP  - NOT IN FILE
NT  - UI - 94022391LA - engRN - 151-50-8 (Potassium Cyanide)RN - 53026-44-1 (Bacteriorhodopsins)PT - Journal ArticleDA - 19931119IS - 0027-8424SB - IMCY - UNITED STATES
UR  - PM:8415720
SO  - Proc Natl Acad Sci U S A 1993 Oct 15 ;90(20):9446-9450

1130
UI  - 19776
AU  - Boyer PD
AD  - Department of Chemistry and Biochemistry, University of California, Los Angeles 90024-1570
TI  - The binding change mechanism for ATP synthase--some probabilities and possibilities
MH  - atp
MH  - ATP synthase
MH  - BINDING
MH  - Biochemistry
MH  - CHANGE MECHANISM
MH  - Chemistry
MH  - H(+)-Transporting ATP Synthase
MH  - mechanism
MH  - Nucleotides
MH  - review
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 93112640LA - engRN - 0 (Nucleotides)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, AcademicID - GM-11094/GM/NIGMSDA - 19930201IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:8417777
SO  - Biochim Biophys Acta 1993 Jan 8 ;1140(3):215-250

1131
UI  - 21164
AU  - Brusilow WS
AD  - Department of Biochemistry, Wayne State University School of Medicine, Detroit, Michigan 48201
TI  - Assembly of the Escherichia coli F1F0 ATPase, a large multimeric membrane-bound enzyme
AB  - The F1F0 proton translocating ATPase of Escherichia coli is a large membrane-bound enzyme complex consisting of more than 20 polypeptides that are encoded by the unc operon. Besides being a system for analysing the enzymology of ATP synthesis and energy coupling, the ATPase is a model system for determining how large oligomeric membrane- bound proteins are synthesized and assembled. The assembly of the ATPase involves differential gene expression and assembly of the subunits within the membrane and with each other. This review discusses the influence of F1 subunits on the assembly and proton permeability of the F0 proton channel, and the possible advantages to assembly of the particular arrangement of genes in the unc operon
MH  - A
MH  - atp
MH  - ATP synthesis
MH  - ATPase
MH  - Biochemistry
MH  - COMPLEX
MH  - coupling
MH  - England
MH  - enzymology
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - F1
MH  - Macromolecular Systems
MH  - membrane
MH  - Membrane Proteins
MH  - model
MH  - Permeability
MH  - protein
MH  - Proteins
MH  - proton
MH  - Proton-Translocating ATPases
MH  - Protons
MH  - review
MH  - SUBUNIT
MH  - synthesis
MH  - SYSTEM
MH  - SYSTEMS
RP  - NOT IN FILE
NT  - UI - 94018637LA - engRN - 0 (Macromolecular Systems)RN - 0 (Membrane Proteins)RN - 0 (Protons)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticlePT - ReviewPT - Review, TutorialID - AI00882/AI/NIAIDDA - 19931109IS - 0950-382XSB - IMCY - ENGLAND
UR  - PM:8412691
SO  - Mol Microbiol 1993 Aug ;9(3):419-424

1132
UI  - 387
AU  - Bulygin VV
AU  - Syroeshkin AV
AU  - Vinogradov AD
AD  - Department of Biochemistry, School of Biology, Moscow State University, Russian Federation
TI  - Nucleotide/H(+)-dependent change in Mg2+ affinity at the ATPase inhibitory site of the mitochondrial F1-F0 ATP synthase
AB  - The interactions between ADP and Mg2+ that result in the slowly reversible inhibition of the mitochondrial F1-F0 ATPase were studied. The Ki for the inhibitory Mg2+ is shown to be strongly dependent on the occupation of the nucleotide-binding sites. The inhibitory binding site for Mg2+ is not seen unless a stoichiometric amount of ADP is added [Biochem. J. 276 (1991) 149-156]; it appears (Ki = 2.10(-6) M) in the presence of stoichiometric ADP and the affinity for inhibitory Mg2+ decreases to a Ki value of 7.10(-5) M when the second nucleotide binding site with Kd = 5.10(-6) M is loaded with ADP. The binding of the inhibitory Mg2+ is competitively inhibited by H+ ions within the pH interval 6.8-8.2. The nucleotide-dependent affinity transition of the Mg(2+)-specific site suggests that H+/Mg2+ exchange may play an important role in the catalytic mechanism of ATP synthesis/hydrolysis at the active site(s) of F1-F0 ATP synthase
RP  - NOT IN FILE
NT  - UI - 93345686LA - engRN - 0 (Enzyme Inhibitors)RN - 0 (Ions)RN - 1333-74-0 (Hydrogen)RN - 58-64-0 (Adenosine Diphosphate)RN - 7439-95-4 (Magnesium)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19930909IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:8344425
SO  - FEBS Lett 1993 Aug 9 ;328(1-2):193-196

1133
UI  - 20946
AU  - Dmitriev O
AU  - Deckers-Hebestreit G
AU  - Altendorf K
AD  - Universitat Osnabruck, Fachbereich Biologie/Chemie, Arbeitsgruppe Mikrobiologie, Osnabruck, Germany
TI  - ATP synthesis energized by delta pNa and delta psi in proteoliposomes containing the F0F1-ATPase from Propionigenium modestum
AB  - After incorporation of the purified Na(+)-translocating F0F1-ATPase from Propionigenium modestum into preformed phospholipid vesicles the synthesis of ATP from ADP and inorganic phosphate could be observed under conditions where a valinomycin-mediated K+ diffusion potential (delta psi) and/or a Na+ concentration gradient (delta pNa) were imposed. This reaction was not inhibited by the protonophore carbonyl cyanide p-tri-fluoromethoxyphenylhydrazone (FCCP). Furthermore, the delta pNa-driven ATP synthesis was stimulated by FCCP. In contrast, the addition of the Na+/H+ antiporter monensin or of the F0F1 inhibitors N,N'-dicyclohexylcarbodiimide and venturicidin abolished the synthesis of ATP completely. Finally, delta pNa alone was able to elicit ATP synthesis, when a Na+ concentration gradient of sufficient magnitude was applied. In this case ATP synthesis occurred above a threshold level of approximately 120 mV and, furthermore, delta psi and delta pNa appear to be equivalent as driving forces for this process. Therefore, the data provide firm evidence for the concept that delta"mu Na+ is the primary driving force for the synthesis of ATP in P. modestum
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - ADP
MH  - atp
MH  - ATP synthesis
MH  - ATPase
MH  - Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone
MH  - Dicyclohexylcarbodiimide
MH  - Diffusion
MH  - diffusion potential
MH  - F0F1
MH  - INORGANIC-PHOSPHATE
MH  - Liposomes
MH  - P
MH  - Potassium
MH  - Proteolipids
MH  - proteoliposome
MH  - protonophore
MH  - PSI
MH  - Sodium
MH  - synthesis
MH  - Valinomycin
RP  - NOT IN FILE
NT  - UI - 93315447LA - engRN - 0 (Liposomes)RN - 0 (Proteolipids)RN - 0 (Venturicidins)RN - 0 (proteoliposomes)RN - 17090-79-8 (Monensin)RN - 2001-95-8 (Valinomycin)RN - 370-86-5 (Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - 7440-09-7 (Potassium)RN - 7440-23-5 (Sodium)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19930812IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:8325855
SO  - J Biol Chem 1993 Jul 15 ;268(20):14776-14780

1134
UI  - 21194
AU  - Drachev LA
AU  - Dracheva SV
AU  - Kaulen AD
AD  - AN Belozersky Institute of Physico-Chemical Biology, Moscow State University, Russian Federation
TI  - pH dependence of the formation of an M-type intermediate in the photocycle of 13-cis-bacteriorhodopsin
AB  - An M-type intermediate is formed in the 13-cis-bR photocycle in purple membranes at high pH. This is presumably due to deprotonation of the same group whose deprotonation causes a large increase in rate of M formation in the trans-bR photocycle (the 'alkaline transition'). For Triton X-100-solubilized bR, the alkaline transition is shifted to a lower pH value by more than 2 pH units. The alkaline transition in Triton-solubilized preparations changes the efficiency of the M intermediate formation in the 13-cis-sbR photocycle. The M intermediate formation in 13-cis-sbR, as in the case of trans-sbR, is completely inhibited when the blue 'acidic' bR is formed at low pH. The protonation state of the group affecting formation of the M intermediate in 13-cis-bR at high pH and the group which is responsible for the transition to the blue acidic form influence in a similar way the equilibrium between bR isomers in the dark-adapted form as well as the rate of dark adaptation
MH  - A
MH  - Bacteriorhodopsin
MH  - DEPENDENCE
MH  - intermediate
MH  - M
MH  - M-intermediate
MH  - membrane
MH  - Membranes
MH  - pH
MH  - protonation
MH  - purple membrane
RP  - NOT IN FILE
NT  - UI - 94009691LA - engRN - 53026-44-1 (Bacteriorhodopsins)PT - Journal ArticleDA - 19931117IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:8405451
SO  - FEBS Lett 1993 Oct 11 ;332(1-2):67-70

1135
UI  - 21195
AU  - Drachev LA
AU  - Kaulen AD
AU  - Komrakov AY
AD  - Department of New Physical Methods in Biology, AN Belozersky Institute of Physico-Chemical Biology, Moscow State University, Russia
TI  - On the two pathways of the M-intermediate formation in the photocycle of bacteriorhodopsin
AB  - The flash-photolysis technique was used to study the photocycles of the wild-type bacteriorhodopsin (WT bR) and D96N mutant. Kinetics of the L- intermediate decay and M-intermediate formation at pH 7.0, 20 degrees C fit well a sum of two components having time constants, tau (1) = 60 microS and tau (2) = 250 microS, for the WT bR, and a sum of three components having time constants, tau (1) = 55 microS, tau (2) = 220 microS and tau (3) = 1 mS, for the D96N mutant. The fast component with a time constant of 1.4 microS was found in the photoresponse at 400 nm. It constituted 10% of the total amplitude and may be attributed to the K-->L transition. The component with tau = 1 mS was observed in the photocycle of the WT bR as a lag phase in the relaxation of the photoresponse at 400 nm. The difference absorbance minima, corresponding to the first (55-60 microS) and the second (220-260 microS) components of the M-formation, were located at 550 and 530 nm, respectively. The absorbance spectra, corresponding to the 1-mS- component of the M-formation of the D96N bR, may be represented as a superposition of spectra, corresponding to the first and the second components in the region of 460-700 nm. The effect of azide on the D96N bR revealed two azide-independent components in the decay of L- intermediate. Azide was shown to protonate all M-forms simultaneously. This indicates that the Schiff base pK rises almost immediately after deprotonation.(ABSTRACT TRUNCATED AT 250 WORDS)
MH  - A
MH  - Azides
MH  - Bacteriorhodopsin
MH  - BASE
MH  - CONSTANT
MH  - intermediate
MH  - Kinetics
MH  - M-intermediate
MH  - method
MH  - Methods
MH  - mutant
MH  - pH
MH  - relaxation
MH  - Schiff base
MH  - Schiff-base
MH  - Sodium
MH  - spectra
MH  - Time
RP  - NOT IN FILE
NT  - UI - 94004585LA - engRN - 0 (Azides)RN - 26628-22-8 (Sodium Azide)RN - 53026-44-1 (Bacteriorhodopsins)PT - Journal ArticleDA - 19931029IS - 1039-9712SB - IMCY - AUSTRALIA
UR  - PM:8401304
SO  - Biochem Mol Biol Int 1993 Jul ;30(3):461-469

1136
UI  - 348
AU  - Dyer MR
AU  - Walker JE
AD  - Medical Research Council Laboratory of Molecular Biology, Cambridge, UK
TI  - Sequences of members of the human gene family for the c subunit of mitochondrial ATP synthase
AB  - Subunit c is an intrinsic membrane component of ATP synthase, and in mammals it is encoded by two expressed nuclear genes, P1 and P2. Both genes encode the same mature c subunit, but the mitochondrial import pre-sequences in the precursors of subunit c are different. The DNA sequences of the human P1 and P2 genes are described. They occupy about 3.0 and 10.9 kb respectively of the human genome, and both genes are split into five exons. The human genome also contains about 14 related spliced pseudogenes, and the sequence of one such pseudogene related to P2 is described. Sequences flanking the 5' ends of the human P1 and P2 coding sequences each contain a CpG-rich island. Potential promoter elements (TATA and CCAAT boxes) are present in the 5' sequences of the P1 gene, but not that of P2, although there is no direct experimental evidence to show the involvement of these sequences in transcription of the genes
RP  - NOT IN FILE
NT  - UI - 93319529LA - engRN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19930812IS - 0264-6021SB - IMCY - ENGLANDJC - 9YO
UR  - PM:8328972
SO  - Biochem J 1993 Jul 1 ;293 ( Pt 1)():51-64

1137
UI  - 9958
AU  - Gennis RB
AU  - Barquera B
AU  - Hacker B
AU  - Van Doren SR
AU  - Arnaud S
AU  - Crofts AR
AU  - Davidson E
AU  - Gray KA
AU  - Daldal F
AD  - School of Chemical Sciences, University of Illinois, Urbana 61801
TI  - The bc1 complexes of Rhodobacter sphaeroides and Rhodobacter capsulatus
AB  - Photosynthetic bacteria offer excellent experimental opportunities to explore both the structure and function of the ubiquinol-cytochrome c oxidoreductase (bc1 complex). In both Rhodobacter sphaeroides and Rhodobacter capsulatus, the bc1 complex functions in both the aerobic respiratory chain and as an essential component of the photosynthetic electron transport chain. Because the bc1 complex in these organisms can be functionally coupled to the photosynthetic reaction center, flash photolysis can be used to study electron flow through the enzyme and to examine the effects of various amino acid substitutions. During the past several years, numerous mutations have been generated in the cytochrome b subunit, in the Rieske iron-sulfur subunit, and in the cytochrome c1 subunit. Both site-directed and random mutagenesis procedures have been utilized. Studies of these mutations have identified amino acid residues that are metal ligands, as well as those residues that are at or near either the quinol oxidase (Qo) site or the quinol reductase (Qi) site. The postulate that these two Q-sites are located on opposite sides of the membrane is supported by these studies. Current research is directed at exploring the details of the catalytic mechanism, the nature of the subunit interactions, and the assembly of this enzyme
MH  - Bacteria
MH  - Bacterial Proteins
MH  - COMPLEX
MH  - Cytochrome b
MH  - Cytochrome c1
MH  - electron
MH  - Electron Transport
MH  - Ligands
MH  - mechanism
MH  - Proteins
MH  - Quinones
MH  - review
MH  - Rhodobacter sphaeroides
MH  - RHODOBACTER-CAPSULATUS
MH  - SUBUNIT
MH  - transport
MH  - Ubiquinol-Cytochrome-c Reductase
RP  - NOT IN FILE
NT  - UI - 93352442LA - engRN - 0 (Bacterial Proteins)RN - 0 (Fungal Proteins)RN - 0 (Iron-Sulfur Proteins)RN - 0 (Ligands)RN - 0 (Plant Proteins)RN - 0 (Quinones)RN - 0 (Rieske iron-sulfur protein)RN - 9035-42-1 (Cytochrome c1)RN - EC 1.10.2.2 (Ubiquinol-Cytochrome-c Reductase)PT - Journal ArticlePT - ReviewPT - Review, TutorialID - 1FOS TWO 4723-01/TW/FICID - GM35438/GM/NIGMSID - etcDA - 19930916IS - 0145-479XSB - IMCY - UNITED STATESJC - HIO
UR  - PM:8394316
SO  - J Bioenerg Biomembr 1993 Jun ;25(3):195-209

1138
UI  - 68
AU  - Gerike U
AU  - Dimroth P
AD  - Mikrobiologisches Institut der Eidgenossischen Technischen Hochschule, ETH-Zentrum, Zurich, Switzerland
TI  - N-terminal amino acid sequences of the subunits of the Na(+)- translocating F1F0 ATPase from Propionigenium modestum
AB  - We report here the N-terminal protein sequences of the subunits of the ATPase from Propionigenium modestum. Subunits c, b, delta, alpha and beta start with an N-terminal methionine residue, the gamma and epsilon subunits have an alanine N-terminus, from which N-formylmethionine was hydrolyzed by posttranslational modification, and subunit a contains a blocked N-terminus. Each of the N-terminal sequences exactly matches a portion of the DNA sequence in the gene encoding the respective subunit protein on the unc operon. Thus, the exact translational start for each subunit protein can be identified and the primary structures of the protein transcripts can be clearly defined. Based on these data the putative size of the open reading frame that was envisaged from the DNA sequence had to be revised for the alpha and delta subunits
RP  - NOT IN FILE
NT  - UI - 93138123LA - engRN - 0 (Cations, Monovalent)RN - 0 (DNA, Bacterial)RN - 7440-23-5 (Sodium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19930223IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:8422943
SO  - FEBS Lett 1993 Jan 18 ;316(1):89-92

1139
UI  - 115
AU  - Girvin ME
AU  - Fillingame RH
AD  - Department of Biomolecular Chemistry, University of Wisconsin, Madison 53706
TI  - Helical structure and folding of subunit c of F1F0 ATP synthase: 1H NMR resonance assignments and NOE analysis
AB  - Subunit c of the H(+)-transporting F1F0 ATP synthase (EC 3.6.1.34) is thought to fold across the membrane as a hairpin of two alpha-helices and function as a key component of the H(+)-translocase of F0. We report here the initial results of a structural study of purified subunit c in a chloroform-methanol-water (4:4:1) solvent mixture using standard two-dimensional NMR techniques. The spin systems of 78 of the 79 amino acid side chains have been assigned to residue type, and 44 of these have been assigned to specific residues in the sequence. Stretches of alpha-helical secondary structure were observed for Asp7- ILe26 in the first proposed transmembrane helix, and for Arg50-Ile55 and Ala67-Val78 in the second proposed transmembrane helix. Nuclear Overhauser effects (NOEs) were observed between residues at both ends of the predicted transmembrane helices. The intensities of the NOEs between helix-1 and helix-2 were not diminished by mixing of 2H-subunit c with 1H-subunit c, and therefore the NOEs must be due to intramolecular, rather than intermolecular, interactions. Hence the purified protein must fold as a hairpin in this solvent system, just as it is thought to fold in the lipid bilayer of the membrane. In native F0, dicyclohexylcarbodiimide reacts specifically with Asp61 in the second transmembrane helix of subunit c, and the rate of this reaction is reduced by substitution of Ile28 by Thr on the first transmembrane helix. The I28T substitution is shown here to alter the chemical shifts of protons at and around Asp61. This observation provides a further indication that subunit c may fold in chloroform-methanol-water solvent much like it does in the membrane
RP  - NOT IN FILE
NT  - UI - 94032352LA - engRN - 0 (Amino Acids)RN - 0 (Solvents)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - F32-GM11096/GM/NIGMSID - GM23105/GM/NIGMSID - RR02301/RR/NCRRID - etcDA - 19931221IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:8218294
SO  - Biochemistry 1993 Nov 16 ;32(45):12167-12177

1140
UI  - 301
AU  - Groth G
AU  - Junge W
AD  - Universitat Osnabruck, Germany
TI  - Proton slip of the chloroplast ATPase: its nucleotide dependence, energetic threshold, and relation to an alternating site mechanism of catalysis
AB  - The F-ATPase of chloroplasts couples proton flow to ATP synthesis, but is leaky to protons in the absence of nucleotides. This "proton slip" can be blocked by small concentrations of ADP or by inhibitors of the channel portion, CF0. We studied charge flow through the ATPase by flash spectrophotometry and analyzed the inhibition of proton slip by nucleotides, phosphate/arsenate, and insufficient proton motive force. The following inhibition constants (at given background concentrations) were observed: ADP, 0.2 microM (0.5 mM P(i)); ADP, 13.4 microM (no P(i)); P(i), 43 microM (1 microM ADP); GDP, 2.5 microM (0.5 mM P(i)); ATP, 2 microM. ADP and P(i) mutually lowered their respective inhibition constants. Phosphate could be replaced by arsenate. Proton slip occurred only if the proton motive force exceeded a certain threshold, similar to that for ATP synthesis. The inhibition of proton slip by ADP and GDP qualified the respective nucleotide binding sites as belonging to the subset of two (or three) potentially catalytic sites out of the total of six. We interpreted the ADP-induced transition between different conduction states of the ATPase from "slipping" to "closed" to "coupled" as a consequence of the alternating site mechanism of catalysis. Whereas the proton translocator idles in the absence of nucleotides, the high-affinity binding of the first ADP/P(i) couple to one site clutches proton flow to some (conformational) change that can only be executed after the binding of another ADP/P(i) couple to a second site. From there on these sites alternate in the catalytic cycle. An entropic machine is presented which likewise models proton slip, unisite, and multisite ATP synthesis and hydrolysis
RP  - NOT IN FILE
NT  - UI - 93349898LA - engRN - 0 (Nucleotides)RN - 0 (Phosphates)RN - 0 (Protons)RN - 28380-24-7 (Nigericin)RN - 58-64-0 (Adenosine Diphosphate)RN - 7439-95-4 (Magnesium)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19930916IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:8394125
SO  - Biochemistry 1993 Aug 17 ;32(32):8103-8111

1141
UI  - 20817
AU  - Hartzog PE
AU  - Cain BD
AD  - Department of Biochemistry and Molecular Biology, University of Florida, Gainesville 32610
TI  - The aleu207-->arg mutation in F1F0-ATP synthase from Escherichia coli. A model for human mitochondrial disease
AB  - The mitochondrial ATPase 6 gene encodes a subunit of F1F0 adenosine triphosphate (ATP) synthase. A mutation in the ATPase 6 gene has been genetically linked to two maternally inherited genetic diseases: neurological muscle weakness, ataxia, and retinitis pigmentosa (NARP) and certain cases of subacute necrotizing encephalopathy (SNE). Although the severity of both NARP and SNE disease were correlated with the quantity of the ATPase 6leu156-->arg mutation in each patient, the mutation could not be shown to alter F1F0-ATP synthase activity. To investigate the biochemical effects of the ATPase 6leu156-->arg mutation on F1F0-ATP synthase, the aleu207-->arg mutation was constructed in the F1F0-ATP synthase from Escherichia coli to serve as a model for the disease mutation. Characterization of the model bacterial enzyme revealed that the mutation abolishes detectable ATP synthesis via oxidative phosphorylation. The aleu207-->arg mutation results in a structural perturbation blocking proton translocation through F1F0-ATP synthase. The results suggest that a structural defect in human F1F0-ATP synthase is the biochemical basis for NARP and SNE
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthesis
MH  - ATPase
MH  - Biochemistry
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1F0-ATP SYNTHASE
MH  - Human
MH  - Macromolecular Systems
MH  - model
MH  - Oxidative Phosphorylation
MH  - Phosphorylation
MH  - proton
MH  - SUBUNIT
MH  - SYNTHASE
MH  - synthesis
MH  - SYSTEM
MH  - SYSTEMS
MH  - translocation
RP  - NOT IN FILE
NT  - UI - 93286047LA - engRN - 0 (Macromolecular Systems)RN - 0 (Plasmids)RN - 61-90-5 (Leucine)RN - 74-79-3 (Arginine)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleID - GM43495/GM/NIGMSDA - 19930713IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:8509361
SO  - J Biol Chem 1993 Jun 15 ;268(17):12250-12252

1142
UI  - 20818
AU  - Hartzog PE
AU  - Cain BD
AD  - Department of Biochemistry and Molecular Biology, University of Florida, Gainesville 32610
TI  - Mutagenic analysis of the a subunit of the F1F0 ATP synthase in Escherichia coli: Gln-252 through Tyr-263
AB  - The a subunit of F1F0 ATP synthase contains a highly conserved region near its carboxyl terminus which is thought to be important in proton translocation. Cassette site-directed mutagenesis was used to study the roles of four conserved amino acids Gln-252, Phe-256, Leu-259, and Tyr- 263. Substitution of basic amino acids at each of these four sites resulted in marked decreases in enzyme function. Cells carrying a subunit mutations Gln-252-->Lys, Phe-256-->Arg, Leu-259-->Arg, and Tyr- 263-->Arg all displayed growth characteristics suggesting substantial loss of ATP synthase function. Studies of both ATP-driven proton pumping and proton permeability of stripped membranes indicated that proton translocation through F0 was affected by the mutations. Other mutations, such as the Phe-256-->Asp mutation, also resulted in reduced enzyme activity. However, more conservative amino acid substitutions generated at these same four positions produced minimal losses of F1F0 ATP synthase. The effects of mutations and, hence, the relative importance of the amino acids for enzyme function appeared to decrease with proximity to the carboxyl terminus of the a subunit. The data are most consistent with the hypothesis that the region between Gln-252 and Tyr-263 of the a subunit has an important structural role in F1F0 ATP synthase
MH  - A
MH  - ACID
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - Amino Acids
MH  - analysis
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Bacterial Proteins
MH  - Biochemistry
MH  - Cells
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - function
MH  - membrane
MH  - Membranes
MH  - mutagenesis
MH  - Peptide Fragments
MH  - Permeability
MH  - protein
MH  - Proteins
MH  - proton
MH  - Protons
MH  - Site
MH  - site-directed
MH  - SUBUNIT
MH  - SYNTHASE
MH  - translocation
RP  - NOT IN FILE
NT  - UI - 93186700LA - engRN - 0 (Bacterial Proteins)RN - 0 (DNA, Bacterial)RN - 0 (Peptide Fragments)RN - 0 (Protons)RN - 0 (uncB protein, E coli)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleID - GM43495/GM/NIGMSDA - 19930405IS - 0021-9193SB - IMCY - UNITED STATES
UR  - PM:8383111
SO  - J Bacteriol 1993 Mar ;175(5):1337-1343

1143
UI  - 567
AU  - Hatefi Y
AD  - Department of Molecular and Experimental Medicine, Scripps Research Institute, La Jolla, CA 92037
TI  - ATP synthesis in mitochondria
RP  - NOT IN FILE
NT  - UI - 94109374LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialID - DK08126/DK/NIDDKDA - 19940214IS - 0014-2956SB - IMCY - GERMANYJC - EMZ
UR  - PM:8281928
SO  - Eur J Biochem 1993 Dec 15 ;218(3):759-767

1144
UI  - 821
AU  - Hisabori T
AU  - Kothen G
AU  - Strotmann H
AD  - Institut fur Biochemie der Pflanzen, Heinrich-Heine-Universitat Dusseldorf, Germany
TI  - Effect of covalent binding of a derivative of 2',3'-O-(2,4,6- trinitrophenyl)-ADP to the tight binding site of CF1 on the enzyme activity
AB  - Irradiation of isolated chloroplast thylakoids with TNP-ADP results in non-covalent binding and covalent incorporation of a reaction product of TNP-ADP formed by photosynthetic reduction into the so-called "tight" nucleotide binding site of CF1 [Ponse et al. (1992) Z. Naturforsch. 47c, 264-274]. CF1 extracted from thus-loaded thylakoid membranes yielded maximal incorporation of 1 mol/mol of CF1. Almost half had the covalent bond with CF1. In experiments with TNP-[14C]ADP, radioactivity was detected almost equivalently on alpha and beta subunits, suggesting that the binding site may be at the interface between alpha and beta subunits. Enzyme activities of the thylakoid membrane-bound enzyme after covalent labeling were measured. Inhibition, ranging from 20 to 25%, was less than expected from the percentage of CF1 molecules labeled (40-50%). It is suggested that only half of the labeled enzymes, probably those with the nucleotide analog linked to the beta subunit, are inactive
RP  - NOT IN FILE
NT  - UI - 94110251LA - engRN - 299-11-6 (Methylphenazonium Methosulfate)RN - 58-64-0 (Adenosine Diphosphate)RN - 84430-17-1 (2',3'-(O-(2,4,6-trinitrocyclohexadienylidine))adenosine 5'- diphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19940217IS - 0021-924XSB - IMCY - JAPANJC - HIF
UR  - PM:8282720
SO  - J Biochem (Tokyo ) 1993 Sep ;114(3):324-328

1145
UI  - 9935
AU  - Holzenburg A
AU  - Jones PC
AU  - Franklin T
AU  - Pali T
AU  - Heimburg T
AU  - Marsh D
AU  - Findlay JB
AU  - Finbow ME
AD  - Department of Biochemistry & Molecular Biology, University of Leeds, England
TI  - Evidence for a common structure for a class of membrane channels
AB  - Electron microscopic analysis of gap-junction-like structures isolated from an anthropod (Nephrops norvegicus) and composed of a 16-kDa polypeptide, show the functional unit to be a star-shaped hexamer of protein arranged around a central channel which runs perpendicular to the plane of the membrane. Estimations of the molecular volume carried out on an averaged projection are consistent with a subunit mass of 16- 18 kDa. Fourier transform infrared spectroscopy indicates a high alpha- helical content for the protein, supporting secondary-structure predictions of four transmembrane alpha helices/monomer. The averaged projection shows a close resemblance to a hexamer of the 16-kDa protein built on the basis of a four alpha-helical bundle [Finbow, M. E., Eliopoulos, E. E., Jackson, P. J., Keen, J. N., Meagher, L., Thompson, P., Jones, P. C. & Findlay, J. B. C. (1992) Protein Eng. 5, 7-15]. The reconstructed image is also similar to that obtained for gap-junction- like channels isolated from a related arthropod [Homarus americanus; Sikerwar, S. S., Downing, K. H. & Glaeser, R. M. (1991) J. Struct. Biol. 106, 255-263] whose protein content was unknown but which we demonstrate may be composed of a related 16-kDa protein. Previous studies have shown a high sequence identity of the Nephrops 16-kDa protein with the 16-kDa proteolipid subunit c of the vascular H(+)- ATPase, both of which in turn bear similarity to the 8-kDa proteolipid subunit of the F1F0-ATP synthase. Expression of cDNA coding for the Nephrops 16-kDa protein in Saccharomyces cerevisiae, in which the endogenous gene coding for the V-ATPase proteolipid has been inactivated, restores V-ATPase activity and cell growth
MH  - analysis
MH  - ATPase
MH  - Biochemistry
MH  - electron
MH  - England
MH  - F1F0-ATP SYNTHASE
MH  - Ion Channels
MH  - Membrane Proteins
MH  - Proteins
MH  - Saccharomyces cerevisiae
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 93238688LA - engRN - 0 (Ion Channels)RN - 0 (Membrane Proteins)PT - Journal ArticleDA - 19930521IS - 0014-2956SB - IMCY - GERMANYJC - EMZ
UR  - PM:7682941
SO  - Eur J Biochem 1993 Apr 1 ;213(1):21-30

1146
UI  - 745
AU  - Ishii N
AU  - Yoshimura H
AU  - Nagayama K
AU  - Kagawa Y
AU  - Yoshida M
AD  - Research Laboratory for Resources Utilization, Tokyo Institute of Technology, Yokohama
TI  - Three-dimensional structure of F1-ATPase of thermophilic bacterium PS3 obtained by electron crystallography
AB  - The three-dimensional molecular structure of the F1-ATPase from a thermophilic bacterium PS3 (TF1) at 3 nm resolution was reconstructed from a series of tilted, negatively stained electron microscopic images of two-dimensional crystals which were formed on a clean surface of mercury. It was shown that six ellipsoidal columns, each approximately 7 nm in length and approximately 3 nm in diameter, corresponding to the alpha and beta subunits, surrounded a central hollow cavity of approximately 2.8 nm in diameter. The cavity, however, did not penetrate the molecule, and a mass, most likely the gamma subunit, existed in the cavity at a depth of approximately 3.4 nm from the top
RP  - NOT IN FILE
NT  - UI - 93224508LA - engRN - 0 (Organometallic Compounds)RN - 541-09-3 (uranyl acetate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19930513IS - 0021-924XSB - IMCY - JAPANJC - HIF
UR  - PM:7682216
SO  - J Biochem (Tokyo ) 1993 Feb ;113(2):245-250

1147
UI  - 20859
AU  - Iwamoto A
AU  - Park MY
AU  - Maeda M
AU  - Futai M
AD  - Department of Organic Chemistry and Biochemistry, Osaka University, Japan
TI  - Domains near ATP gamma phosphate in the catalytic site of H+-ATPase. Model proposed from mutagenesis and inhibitor studies
AB  - The beta Gly-149 residue is in a glycine-rich sequence (Gly-Gly-Ala-Gly- Val-Gly-Lys-Thr; residues 149-156) of the Escherichia coli H(+)-ATPase (ATP synthase) beta subunit. Substitution of beta Gly-149 by Ser suppressed the effect of the beta Ser-174-->Phe mutation (Iwamoto, A., Omote, H., Hanada, H., Tomioka, N., Itai, A., Maeda, M., and Futai, M. (1991) J. Biol. Chem. 266, 16350-16355), suggesting that beta Gly-149 is located near beta Ser-174. In this study, we introduced different residues at position 149 and found that a single mutant beta Cys-149 was defective. The effect of beta Cys-149 mutation was suppressed by beta Gly-172-->Glu, beta Ser-174-->Phe, beta Glu-192-->Val, or beta Val- 198-->Ala replacement. These results suggest that beta Gly-149, beta Gly-172, beta Ser-174, beta Glu-192, and beta Val-198 residues are located close together in the catalytic site. From these findings we propose a model of the catalytic site of the enzyme near the gamma phosphate moiety of ATP. F1 enzymes with the double mutations beta Cys- 149/beta Glu-172, beta Cys-149/beta Phe-174, beta Cys-149/beta Val-192, and beta Cys-149/beta Ala-198 were less sensitive than wild-type F1 to dicyclohexylcarbodiimide and adenosine triphosphopyridoxal (an affinity analogue of ATP forming a Schiff base with the epsilon-amino group of beta Lys-155 or beta Lys-201), and became sensitive to N-ethylmaleimide in an ATP-protected manner. These results of inhibitor studies are consistent with the proposed model
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BASE
MH  - BETA-SUBUNIT
MH  - Biochemistry
MH  - Chemistry
MH  - Dicyclohexylcarbodiimide
MH  - Enzymes
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - Ethylmaleimide
MH  - F1
MH  - H(+)ATPase
MH  - H+-ATPase
MH  - M
MH  - Macromolecular Systems
MH  - model
MH  - mutagenesis
MH  - mutant
MH  - protein
MH  - Proteins
MH  - RESIDUE
MH  - Schiff base
MH  - Schiff-base
MH  - Site
MH  - SUBUNIT
MH  - SYNTHASE
MH  - SYSTEM
MH  - SYSTEMS
RP  - NOT IN FILE
NT  - UI - 93155151LA - engRN - 0 (Macromolecular Systems)RN - 0 (Oligodeoxyribonucleotides)RN - 0 (Plasmids)RN - 0 (Recombinant Proteins)RN - 101418-63-7 (adenosine triphosphopyridoxal)RN - 128-53-0 (Ethylmaleimide)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19930309IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:8428992
SO  - J Biol Chem 1993 Feb 15 ;268(5):3156-3160

1148
UI  - 20857
AU  - Jounouchi M
AU  - Maeda M
AU  - Futai M
AD  - Department of Organic Chemistry and Biochemistry, Osaka University
TI  - The alpha subunit of ATP synthase (F0F1): the Lys-175 and Thr-176 residues in the conserved sequence (Gly-X-X-X-X-Gly-Lys-Thr/Ser) are located in the domain required for stable subunit-subunit interaction
AB  - The sequence (Gly-X-X-X-X-Gly-Lys-Thr/Ser) is conserved in nucleotide binding proteins including the alpha and beta subunits of the ATP synthase. Various mutations were introduced in the alpha Lys-175 and alpha Thr-176 residues in the sequence (Gly-Asp-Arg-Gln-Thr-Gly-Lys- Thr, residues 169-176) of the Escherichia coli ATP synthase alpha subunit. Surprisingly, single amino acid substitutions drastically affected the subunit assembly of the enzyme. The entire enzyme assembly was lost by alpha Lys-175-->Phe (or Trp) or alpha Thr-176-->Phe (or Tyr) mutation. Other mutants had similar (alpha His-175, alpha Ser-175, alpha Gly-175, alpha Ser-176, and alpha His-176 mutants) or lower (alpha Ala-176, alpha Cys-176, alpha Leu-176, and alpha Val-176 mutants) effects on assembly of the active enzyme compared with that of the wild-type. However, all these mutant enzymes except the alpha Ser- 176 enzyme showed enhanced cold sensitivities and reduced stabilities at high temperature. Mutant enzymes such as alpha Gly-175 and alpha His- 176 showed low multi-site (steady state) catalysis, possibly due to loss of proper subunit-subunit interactions. These results suggest that the alpha Lys-175 and alpha Thr-176 residues are not absolutely essential for catalysis, but that they, or possibly the entire conserved sequence, are located in the key domain for the subunit- subunit interactions essential for enzyme stability and steady state activity.(ABSTRACT TRUNCATED AT 250 WORDS)
MH  - ACID
MH  - ACTIVE
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - ALPHA-SUBUNIT
MH  - Amino Acids
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BETA-SUBUNIT
MH  - BINDING
MH  - Biochemistry
MH  - Catalysis
MH  - Chemistry
MH  - COLI ATP SYNTHASE
MH  - Enzyme Stability
MH  - Enzymes
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0F1
MH  - mutant
MH  - nucleotide binding
MH  - protein
MH  - Proteins
MH  - RESIDUE
MH  - SUBUNIT
MH  - SYNTHASE
MH  - Temperature
RP  - NOT IN FILE
NT  - UI - 94086461LA - engRN - 0 (Amino Acids)RN - 0 (Plasmids)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19940126IS - 0021-924XSB - IMCY - JAPAN
UR  - PM:8262895
SO  - J Biochem (Tokyo ) 1993 Aug ;114(2):171-176

1149
UI  - 10577
AU  - Junge W
AU  - Engelbrecht S
AU  - Lill H
AU  - Groth G
TI  - Protolytic Reactions And The Functioning of F-ATPases
MH  - F-ATPASE
RP  - IN FILE
SO  - Biol Chem Hoppe-Seyler 1993  ;374():741-742

1150
UI  - 744
AU  - Kaibara C
AU  - Odaka M
AU  - Hisabori T
AU  - Yoshida M
AD  - Research Laboratory of Resources Utilization, Tokyo Institute of Technology, Yokohama, Japan
TI  - AT(D)PMg-induced dissociation of the alpha 3 beta 3 complex of the F1- ATPase from a thermophilic Bacillus PS3 into alpha 1 beta 1 heterodimers is prevented by mutation beta (Y341C)
AB  - AT(D)PMg induces dissociation of the alpha 3 beta 3 complex of F1- ATPase from a thermophilic Bacillus strain. PS3, into the alpha 1 beta 1 heterodimers [(1991) Biochim. Biophys. Acta 1056, 279-284] but the location of the AT(D)PMg binding site responsible is not known. From the analysis of AT(D)PMg binding properties of the isolated mutant beta subunit, beta(Y341C), and the stability of the alpha 3 beta(Y341C)3 complex in the presence of AT(D)PMg, we conclude that binding of AT(D)PMg to the Tyr-341 site of the beta subunit(s) in the alpha 3 beta 3 complex triggers the dissociation of the alpha 3 beta 3 complex into the alpha 1 beta 1 heterodimers
RP  - NOT IN FILE
NT  - UI - 93223843LA - engRN - 0 (Macromolecular Systems)RN - 55520-40-6 (Tyrosine)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19930510IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:8467909
SO  - FEBS Lett 1993 Apr 19 ;321(1):46-50

1151
UI  - 63
AU  - Kaim G
AU  - Dimroth P
AD  - Mikrobiologisches Institut, Eidgenossische Technische Hochschule, Switzerland
TI  - Formation of a functionally active sodium-translocating hybrid F1F0 ATPase in Escherichia coli by homologous recombination
AB  - A deletion mutant of Escherichia coli lacking the genes for ATPase subunits a, c, b, delta and part of the alpha subunit was transformed with a plasmid containing the corresponding genes of the sodium- translocating ATPase of Propionigenium modestum. The respective DNA fragment of P. modestum was integrated into the genome of the E. coli deletion mutant by site-specific homologous recombination. The sites of this recombination event were identified by cloning and DNA sequencing. As a consequence of the recombination event, a functionally active hybrid ATPase was obtained by in vivo complementation. The biochemical characterization of this hybrid ATPase revealed high sensitivity to dicyclohexylcarbodiimide as well as strong activation by the addition of sodium ions. After reconstitution into liposomes, the hybrid ATPase catalysed the transport of Na+ upon ATP addition. In the absence of Na+, the ATPase hybrid was able to pump protons, as was shown by the ATP-dependent fluorescence quenching of 9-amino-6-chloro-2- methoxyacridine
RP  - NOT IN FILE
NT  - UI - 94109393LA - engRN - 0 (Proteolipids)RN - 0 (proteoliposomes)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 7440-23-5 (Sodium)RN - EC 3.6.1.- (sodium ATPase)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19940214IS - 0014-2956SB - IMCY - GERMANYJC - EMZ
UR  - PM:8281946
SO  - Eur J Biochem 1993 Dec 15 ;218(3):937-944

1152
UI  - 902
AU  - Kammermeier H
TI  - Efficiency of energy conversion from metabolic substrates to ATP and mechanical and chemiosmotic energy
AB  - The free energy available from substrate oxidation is a largely invariable figure in biological systems. The extent to which this energy is conserved in high-energy intermediates appears to be optimized during evolution to high efficiency. Since energy transformation by enzymatic and transport processes take place largely with fixed stoichiometrics, high efficiency can only be achieved by adapting free energy levels rather than by adapting turnover (number of molecules synthesized/hydrolyzed, etc.) The relatively small steps (equivalent with high efficiency) in free energy of various metabolic steps implicitly mean that changes in free energy leading to abolition of these steps (e.g., reduction of free energy of ATP) abolish the driving force and interrupt the (net) reaction. However, in the myocardium under limited energy supply (hypoxia) various protective mechanisms appear to be involved which keep free energy levels high for surviving of the cell but for the cost of function (contraction). Those mechanisms might play a role in the phenomena of hibernating and stunning
MH  - Adenosine Triphosphate
MH  - Anoxia
MH  - Energy Metabolism
MH  - Evolution
MH  - Human
MH  - metabolism
MH  - Myocardial Contraction
MH  - Myocardium
MH  - physiology
RP  - NOT IN FILE
NT  - Lehrstuhl fur Physiologie, Medizinische Fakultat der RWTH Aachen, FRG
SO  - Basic Res Cardiol 1993  ;88 Suppl 2:15-20.():15-20

1153
UI  - 19777
AU  - Kasho VN
AU  - Allison WS
AU  - Boyer PD
AD  - Department of Chemistry and Biochemistry, University of California, Los Angeles 90024
TI  - Study of the mechanism of MF1 ATPase inhibition by fluorosulfonylbenzoyl inosine, quinacrine mustard, and efrapeptin using intermediate 18O exchange as a probe
AB  - The mitochondrial F1-ATPase (MF1) is known to be largely or totally inhibited by combination or reaction with one fluorosulfonylbenzoyl inosine (FSBI), quinacrine mustard, or efrapeptin per enzyme. Measurements were made with 18O in attempt to ascertain if the weak catalytic activity remaining after exposure to excess of these reagents was due to retention of some activity by the enzyme modified by these inhibitors. Any such activity could have different characteristics that might be revealed by the distribution of [18O]Pi isotopomers formed from [gamma-18O]ATP. The MF1 inhibited by FSBI showed progressive appearance of two new catalytic pathways as inhibition proceeded. Both pathways appeared to be operative in the enzyme after one beta subunit per enzyme had been modified by FSBI. A high exchange pathway showed no detectable change as ATP concentration was lowered. The lower exchange pathway showed an increase in the amount of exchange with lowering of the ATP concentration, similar to the cooperative behavior observed with the unmodified enzyme. With excess ATP more product was formed by the low exchange pathway, showing that compulsory alternation between two catalytic sites was not retained. The behavior can be explained by the ability of the modified beta subunit to undergo binding changes similar to those occurring in catalysis, with the other two beta subunits catalyzing sluggish hydrolysis by different pathways because of the asymmetry introduced by the modification. Inhibition by quinacrine mustard also resulted in the appearance of two new pathways, somewhat similar to those from FSBI inhibition. In contrast, activity remaining with excess efrapeptin present showed only one pathway like that of the native enzyme. This can be attributed to a low equilibrium concentration of free enzyme and total inhibition of MF1 combined with efrapeptin
MH  - A
MH  - Antibiotics
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BETA-SUBUNIT
MH  - BINDING
MH  - Biochemistry
MH  - Catalysis
MH  - Chemistry
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - H(+)-Transporting ATP Synthase
MH  - Hydrolysis
MH  - mechanism
MH  - MITOCHONDRIAL F1-ATPASE
MH  - Oxygen
MH  - Oxygen Isotopes
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 93143329LA - engRN - 0 (Affinity Labels)RN - 0 (Antibiotics)RN - 0 (Oxygen Isotopes)RN - 0 (Peptides)RN - 4213-45-0 (Quinacrine Mustard)RN - 56645-91-1 (efrapeptin)RN - 58-63-9 (Inosine)RN - 83133-70-4 (5'-(4-fluorosulfonylbenzoyl)inosine)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM-16974/GM/NIGMSDA - 19930223IS - 0003-9861SB - IMCY - UNITED STATESJC - 6SK
UR  - PM:8424665
SO  - Arch Biochem Biophys 1993 Jan ;300(1):293-301

1154
UI  - 65
AU  - Kluge C
AU  - Dimroth P
AD  - Mikrobiologisches Institut, Eidenossische Technische Hochschule, ETH- Zentrum, Zurich, Switzerland
TI  - Kinetics of inactivation of the F1Fo ATPase of Propionigenium modestum by dicyclohexylcarbodiimide in relationship to H+ and Na+ concentration: probing the binding site for the coupling ions
AB  - Purified F1Fo ATPase of Propionigenium modestum was rapidly inactivated by dicyclohexylcarbodiimide (DCCD) with k2 = 1.2 x 10(5) M-1 min-1 at pH 5.6 and 0 degree C. Na+ ions provided specific protection from the modification by DCCD while protons stimulated the reaction. Plots of pseudo-first-order rate constants of inactivation (kobs) against pH yielded titration curves with pK(H+) = 7.0 in the absence of Na+ and pK(H+) = 6.2 in the presence of 0.5 mM Na+. From the dependencies of kobs on Na+, pK(Na+) of about 2.5 and 3.3 were obtained at pH 6.5 and 8.0, respectively. These results indicate that DCCD reacts with a protonated group of the enzyme that dissociates with pK(H+) = 7.0 in the absence of Na+, and that Na+ ions promote the dissociation of this group. Additionally, higher Na+ concentrations were required at more acidic pH values to yield half-maximal protection from inactivation. These effects fit a competitive binding model for Na+ or H+ at the DCCD- reactive conserved acidic amino acid of subunit c (Glu-65). The active- site carboxylate could either be protonated and modified by DCCD or bind Na+ which then provides protection. Complementary results were obtained from the effects of Na+ and H+ on ATPase activity. The pH-rate profile of numax (with saturating Na+) indicated an increase of activity with apparent pK = 6.8, an optimum around pH 7.5, and decreasing activity with apparent pK = 8.7.(ABSTRACT TRUNCATED AT 250 WORDS)
RP  - NOT IN FILE
NT  - UI - 94002023LA - engRN - 0 (Protons)RN - 1154-25-2 (ethylisopropylamiloride)RN - 2609-46-3 (Amiloride)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - 7440-23-5 (Sodium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19931109IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:8399181
SO  - Biochemistry 1993 Oct 5 ;32(39):10378-10386

1155
UI  - 66
AU  - Kluge C
AU  - Dimroth P
AD  - Mikrobiologisches Institut, Eidgenossischen Technischen Hochschule, ETH- Zentrum, Zurich, Switzerland
TI  - Specific protection by Na+ or Li+ of the F1F0-ATPase of Propionigenium modestum from the reaction with dicyclohexylcarbodiimide
AB  - Incubation of the purified F1F0-ATPase of Propionigenium modestum with dicyclohexylcarbodiimide (DCCD) led to inactivation of the enzyme in a strongly pH-dependent manner. Rapid inactivation occurred at pH 5-7, while the increase of the pH from 7 to 9 resulted in a continuous reduction of the inactivation rate. In the presence of Na+ ions, the ATPase was specifically protected from inactivation by DCCD. The protective effect of Na+ was most pronounced at pH 9.0 and less significant at pH 7.0. In addition to Na+, Li+ also protected the ATPase from inactivation by DCCD, but approximately 10 times higher concentrations were required for the same effect. Similarly, the Na+ concentration causing half-maximal stimulation of ATPase activity was about 10 times below the Li+ concentration required for the same activation. It is concluded from these results that a binding site is present for Na+ or Li+ on the enzyme with an about 10 times lower affinity for the latter alkali ion, which when occupied stimulates ATPase activity and protects it from inactivation by DCCD. Inactivation of ATPase activity by DCCD correlated well with a specific labeling of subunit c of the enzyme in the presence of the [14C]DCCD derivative. Like ATPase inactivation, the labeling was promoted by more acidic pH values and inhibited by Na+ ions. We suggest from these data that the DCCD-reactive amino acid residue of subunit c (most likely Glu-65) must be protonated for the reaction with the carbodiimide and provides the Na(+)-binding site in its deprotonated state. Dissociation of the carboxylic acid (at high pH) and binding of Na+ ions to the carboxylate thus abolish the reactivity toward DCCD
RP  - NOT IN FILE
NT  - UI - 93315410LA - engRN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 7439-93-2 (Lithium)RN - 7440-23-5 (Sodium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19930812IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:8392053
SO  - J Biol Chem 1993 Jul 15 ;268(20):14557-14560

1156
UI  - 9908
AU  - Krenn BE
AU  - Van WH
AU  - Scholts MJ
AU  - Kraayenhof R
TI  - Modulation of the proton-translocation stoichiometry of H(+)-ATP synthases in two phototrophic prokaryotes by external pH.
AB  - The stoichiometry between proton translocation and ATP synthesis/hydrolysis was studied in two different photosynthetic prokaryotes, the thermophilic cyanobacterium Synechococcus 6716 and the purple bacterium Rhodospirillum rubrum. The H+/ATP ratio was determined by acid-base transitions as a function of the external pH. The H+/ATP ratio of the Synechococcus 6716 ATP synthase was found to increase with increasing pH. In contrast, in R. rubrum this ratio decreased with increasing pH. These results were qualitatively supported by experiments using the fluorescence probe 9-aminoacridine. The degree of coupling between the H+ flux and the ATP synthesis/hydrolysis reaction is apparently modulated by the conditions under which the proton pump has to work. Such modulation of the H+/ATP ratio may be of physiological significance for an organism, for example when ATP synthesis is necessary at low proton-electrochemical potential difference (delta mu H+ levels). The different pH dependencies of the H+/ATP ratios in these organisms are considered in relation to the differences in the charged amino acids that are present in the F0 subunits a and c.
MH  - Amino Acid Sequence
MH  - Amino Acids
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - Bacteria
MH  - Chemistry
MH  - Comparative Study
MH  - Cyanobacteria
MH  - Energy Metabolism
MH  - enzymology
MH  - F0
MH  - fluorescence
MH  - H(+)-Transporting ATP Synthase
MH  - H+
MH  - Hydrogen-Ion Concentration
MH  - Membrane Potentials
MH  - metabolism
MH  - Molecular Sequence Data
MH  - proton
MH  - Proton Pump
MH  - Rhodospirillum
MH  - Rhodospirillum rubrum
MH  - Sequence Alignment
MH  - SUBUNIT
MH  - Support,Non-U.S.Gov't
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - Department of Molecular and Cellular Biology, Vrije Universiteit, Amsterdam, The NetherlandsPMID- 0008379927EDAT- 1993/09/15 00:00MHDA- 1993/09/15 00:00
SO  - Biochem J 1993 Sep 15 ;294(Pt 3):705-709

1157
UI  - 21163
AU  - Ksenzenko SM
AU  - Brusilow WS
AD  - Department of Biochemistry, Wayne State University School of Medicine, Detroit, Michigan 48201
TI  - Protein-lipid interactions of the proteolipid c subunit of the Escherichia coli proton-translocating adenosinetriphosphatase
AB  - Interactions between Escherichia coli membrane phospholipids and the hydrophobic c subunit of the F1F0 proton-translocating ATPase were characterized. Extraction of E. coli membranes with a neutral mixture of chloroform and methanol and subsequent separation steps produced several protein-containing fractions. The protein-containing fraction most soluble in organic solvents contained subunit c and a lipid fraction enriched in phosphatidylglycerol compared to total E. coli membrane phospholipids. Other ATPase subunits and some additional proteins extracted from the membranes by this procedure could be separated from the c subunit by subsequent extraction. The purified and delipidated c subunit contained fatty acids which were released upon treatment with boron trifluoride methanol. Furthermore, deleting and restoring the genes for the F0 subunits changed the composition of extractable membrane phospholipid and fatty acids, indicating that the F0 plays a significant structural role in the membrane
MH  - A
MH  - ACID
MH  - Adenosinetriphosphatase
MH  - ATPase
MH  - Biochemistry
MH  - Chloroform
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - Lipids
MH  - membrane
MH  - Membrane Proteins
MH  - Membranes
MH  - Methanol
MH  - Phospholipids
MH  - protein
MH  - Proteins
MH  - Proton-Translocating ATPases
MH  - SOLVENT
MH  - SUBUNIT
RP  - NOT IN FILE
NT  - UI - 93343638LA - engRN - 0 (Fatty Acids)RN - 0 (Membrane Lipids)RN - 0 (Membrane Proteins)RN - 0 (Phosphatidylethanolamines)RN - 0 (Phosphatidylglycerols)RN - 0 (Phospholipids)RN - 67-56-1 (Methanol)RN - 67-66-3 (Chloroform)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleID - AI00882/AI/NIAIDDA - 19930902IS - 0003-9861SB - IMCY - UNITED STATES
UR  - PM:8342958
SO  - Arch Biochem Biophys 1993 Aug 15 ;305(1):78-83

1158
UI  - 20996
AU  - Labahn A
AU  - Graber P
AD  - Biologisches Institut, Universitat Stuttgart, Germany
TI  - From uni-site to multi-site ATP synthesis in thylakoid membranes
AB  - The membrane-bound H(+)-ATPase from chloroplasts, CF0F1, was brought into the active, reduced state by illumination in the presence of thioredoxin and dithiothreitol. The endogenous nucleotides were removed by a washing procedure so that the active, reduced enzyme contained one tightly bound ATP per CF0F1. When [14C]ADP was added in substoichiometric amounts during continuous illumination, ADP was bound to the enzyme, phosphorylated and released as [14C]ATP, i.e., the tightly bound ATP was not involved in the catalytic turnover ('uni-site ATP-synthesis'). The rate constant for ADP binding was k = (2.0 +/- 0.5) x 10(6) M-1 s-1. The rate of ATP synthesis was measured as a function of the ADP concentration from 8 nM up to 1 mM in the presence of 2 mM phosphate during continuous illumination. A linear increase of the rate was observed up to 100 nM. Above this concentration a supralinear increase was found, indicating the occupation of a second ADP-binding site. A plateau was reached between 1.5 microM and 2.3 microM ADP with a rate of vpl = 3.7 s-1. The half-maximal rate from this plateau was observed at 780 nM. Above 2.3 microM ADP up to 1 mM ADP the data were described by Michaelis-Menten kinetics (vmax = 80 s- 1; apparent KM = 32 microM). These results indicated the participation of at least two different ADP binding sites in ATP synthesis catalyzed by the membrane-bound CF0F1
MH  - A
MH  - ACTIVE
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - ADP
MH  - ADP BINDING
MH  - atp
MH  - ATP synthesis
MH  - ATPase
MH  - BINDING
MH  - Binding Sites
MH  - CF0F1
MH  - chloroplast
MH  - Chloroplasts
MH  - CONSTANT
MH  - Dithiothreitol
MH  - function
MH  - H(+)ATPase
MH  - Kinetics
MH  - membrane
MH  - Membranes
MH  - Nucleotides
MH  - rate constant
MH  - Site
MH  - synthesis
MH  - thylakoid
MH  - thylakoid membrane
RP  - NOT IN FILE
NT  - UI - 93379036LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19931012IS - 0006-3002SB - IMCY - NETHERLANDS
UR  - PM:8369335
SO  - Biochim Biophys Acta 1993 Sep 13 ;1144(2):170-176

1159
UI  - 513
AU  - Lebowitz MS
AU  - Pedersen PL
AD  - Department of Biological Chemistry, Johns Hopkins School of Medicine, Baltimore, Maryland 21205
TI  - Regulation of the mitochondrial ATP synthase/ATPase complex: cDNA cloning, sequence, overexpression, and secondary structural characterization of a functional protein inhibitor
AB  - The ATPase inhibitor protein of the rat liver mitochondrial ATP synthase/ATPase complex has been cloned from a rat liver cDNA library, and its nucleotide sequence determined. The sequence is highly homologous to both the bovine heart (approximately 70%) and the yeast inhibitor proteins (approximately 40%). The deduced protein sequence is 107 amino acids in length, and based on homology to the bovine heart protein, the first 25 N-terminal amino acids encode a putative mitochondrial targeting sequence. The "mature" protein (without the targeting sequence) fused to the maltose binding protein has been overexpressed in Escherichia coli. The maltose binding protein was used as a handle for the development of a rapid one-step purification of the fusion protein by affinity chromatography on an amylose resin. The purified fusion protein was cleaved with Factor Xa protease at the fusion junction, and the resulting ATPase inhibitor protein was purified to > 90% purity. The purified, overexpressed inhibitor protein displays normal inhibitor activity. The protein inhibits ATP hydrolysis catalyzed by the ATP synthase/ATPase complex in submitochondrial particles in a manner kinetically indistinguishable from the same protein purified from rat liver mitochondria, and exhibits a specific activity of approximately 10,000 units/mg. The secondary structure of the inhibitor protein was determined by circular dichroism spectropolarimetry. The experimentally determined structure shows a high content of alpha-helix and is in good agreement with sequence- based structural predictions. As the function of the inhibitor protein is known to exhibit a high dependence on pH, a study of the pH dependence of inhibitor secondary structure was performed. It is shown that as pH is lowered, conditions which activate inhibitory capacity, the protein loses significant alpha-helical structure. This is the first report of the overexpression in E. coli of a functional ATPase inhibitor protein. Secondary structural analysis of this protein indicates that conversion from its active to its inactive form involves a significant conformational change
RP  - NOT IN FILE
NT  - UI - 93183021LA - engRN - 0 (ATPase inhibitory protein)RN - 0 (Proteins)RN - 0 (Recombinant Fusion Proteins)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - CA10951/CA/NCIDA - 19930331IS - 0003-9861SB - IMCY - UNITED STATESJC - 6SK
UR  - PM:8442667
SO  - Arch Biochem Biophys 1993 Feb 15 ;301(1):64-70

1160
UI  - 21237
AU  - Leikin S
AU  - Parsegian VA
AU  - Rau DC
AU  - Rand RP
AD  - Division of Computer Research and Technology, National Institutes of Health, Bethesda, Maryland 20892
TI  - Hydration forces
MH  - Lipid Bilayers
MH  - protein
MH  - Proteins
MH  - review
MH  - Water
RP  - NOT IN FILE
NT  - UI - 94079712LA - engRN - 0 (Aluminum Silicates)RN - 0 (Glass)RN - 0 (Lipid Bilayers)RN - 0 (Minerals)RN - 0 (Polysaccharides)RN - 0 (Proteins)RN - 1302-87-0 (clay)RN - 7732-18-5 (Water)RN - 9007-49-2 (DNA)PT - Journal ArticlePT - ReviewPT - Review, AcademicDA - 19940119IS - 0066-426XSB - IMCY - UNITED STATES
UR  - PM:8257560
SO  - Annu Rev Phys Chem 1993  ;44():369-395

1161
UI  - 300
AU  - Lill H
AU  - Burkovski A
AU  - Altendorf K
AU  - Junge W
AU  - Engelbrecht S
AD  - Abteilung Biophysik, Universitat Osnabruck, Germany
TI  - Complementation of Escherichia coli unc mutant strains by chloroplast and cyanobacterial F1-ATPase subunits
AB  - The genes encoding the five subunits of the F1 portion of the ATPases from both spinach chloroplasts and the cyanobacterium Synechocystis sp. PCC 6803 were cloned into expression vectors and expressed in Escherichia coli. The recombinant subunits formed inclusion bodies within the cells. Each particular subunit was expressed in the respective unc mutant, each unable to grow on non-fermentable carbon sources. The following subunits restored growth under conditions of oxidative phosphorylation: alpha (both sources, cyanobacterial subunit more than spinach subunit), beta (cyanobacterial subunit only), delta (both spinach and Synechocystis), and epsilon (both sources), whereas no growth was achieved with the gamma subunits from both sources. Despite a high degree of sequence homology the large subunits alpha and beta of spinach and cyanobacterial F1 were not as effective in the substitution of their E. coli counterparts. On the other hand, the two smallest subunits of the E. coli ATPase could be more effectively replaced by their cyanobacterial or chloroplast counterparts, although the sequence identity or even similarity is very low. We attribute these findings to the different roles of these subunits in F1: The large alpha and beta subunits contribute to the catalytic centers of the enzyme, a function rendering them very sensitive to even minor changes. For the smaller delta and epsilon subunits it was sufficient to maintain a certain tertiary structure during evolution, with little emphasis on the conservation of particular amino acids
RP  - NOT IN FILE
NT  - UI - 94002139LA - engRN - 0 (Recombinant Proteins)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19931104IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:8399279
SO  - Biochim Biophys Acta 1993 Oct 4 ;1144(3):278-284

1162
UI  - 344
AU  - Lutter R
AU  - Saraste M
AU  - van Walraven HS
AU  - Runswick MJ
AU  - Finel M
AU  - Deatherage JF
AU  - Walker JE
AD  - Medical Research Council, Laboratory of Molecular Biology, Cambridge, UK
TI  - F1F0-ATP synthase from bovine heart mitochondria: development of the purification of a monodisperse oligomycin-sensitive ATPase
AB  - A new procedure for the isolation of ATP synthase from bovine mitochondria has been developed, with the primary objective of producing enzyme suitable for crystallization trials. Proteins were extracted from mitochondrial membranes with dodecyl-beta-D-maltoside, and the ATP synthase was purified from the extract in the presence of the same detergent by a combination of ion-exchange and gel-filtration chromatography and ammonium sulphate precipitation. This simple and rapid procedure yields 20-30 mg of highly pure and monodisperse enzyme, evidently consisting of 14 different subunits, amongst them, in apparently stoichiometric amounts with the established subunits, subunit e, a recently discovered subunit of unknown function. The enzyme preparation has an oligomycin-sensitive ATP hydrolysis activity, and so the F1 domain is functionally associated with the membrane domain, F0. In contrast with the N-termini of some of the subunits of bovine mitochondrial F1-ATPase, those of the F1F0-ATP synthase are not degraded by proteolysis during the isolation procedure. This preparation therefore satisfies prerequisites for crystallization trials
RP  - NOT IN FILE
NT  - UI - 94059037LA - engRN - 0 (Detergents)RN - 0 (Glucosides)RN - 0 (Oligomycins)RN - 56-65-5 (Adenosine Triphosphate)RN - 69227-93-6 (dodecyl maltoside)RN - 7783-20-2 (Ammonium Sulfate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19931214IS - 0264-6021SB - IMCY - ENGLANDJC - 9YO
UR  - PM:8240295
SO  - Biochem J 1993 Nov 1 ;295 ( Pt 3)():799-806

1163
UI  - 350
AU  - Lutter R
AU  - Abrahams JP
AU  - van Raaij MJ
AU  - Todd RJ
AU  - Lundqvist T
AU  - Buchanan SK
AU  - Leslie AG
AU  - Walker JE
AD  - MRC Laboratory of Molecular Biology, Cambridge, UK
TI  - Crystallization of F1-ATPase from bovine heart mitochondria
AB  - Crystals of the F1-ATPase sector of the ATP synthase complex from bovine heart mitochondria have been grown from solutions containing polyethylene glycol 6000. The crystals diffract to 2.9 A resolution on a laboratory X-ray source. They are orthorhombic and belong to the space group P2(1)2(1)2(1). The unit cell axes are a = 285 A, b = 108 A, c = 140 A. There is one molecule of F1-ATPase in the asymmetric unit
RP  - NOT IN FILE
NT  - UI - 93164259LA - engRN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19930316IS - 0022-2836SB - IMCY - ENGLANDJC - J6V
UR  - PM:8433373
SO  - J Mol Biol 1993 Feb 5 ;229(3):787-790

1164
UI  - 568
AU  - Matsuno-Yagi A
AU  - Hatefi Y
AD  - Department of Molecular and Experimental Medicine, Scripps Research Institute, La Jolla, California 92037
TI  - Studies on the mechanism of oxidative phosphorylation. ATP synthesis by submitochondrial particles inhibited at F0 by venturicidin and organotin compounds
AB  - Oligomycin,N,N'-dicyclohexylcarbodiimide (DCCD), venturicidin, and tetracoordinate organotin compounds (R3SnX) are potent inhibitors of the mitochondrial ATP synthase complex, all acting on the membrane sector, F0. Oligomycin and DCCD inhibit proton translocation through F0 and energy transfer between F0 and the catalytic sector, F1, of the ATP synthase complex. Our results have shown that venturicidin and organotin compounds (tributyltin and triphenyltin chloride were used) greatly attenuate these processes, but do not cause complete inhibition. As a result, bovine submitochondrial particles (SMP) treated with venturicidin or tributyltin chloride were shown to be capable of ATP hydrolysis and synthesis, albeit at very slow rates. We had shown previously that in ATP synthesis Vmax and apparent Km for ADP and Pi increase or decrease, respectively, as the steady-state membrane potential is elevated or lowered (Matsuno-Yagi, A., and Hatefi, Y. (1986) J. Biol. Chem. 261, 14031-14038). These changes occurred at constant Vmax/Km, suggesting that the apparent Km changes were due mainly to kcat changes. Results presented here show that, in respiring SMP treated with venturicidin or organotin compounds, the membrane potential is near the static-head level, but the slow rate of ATP synthesis takes place with a low KmADP value of 2-3 microM. In agreement with our previous conclusions, these results indicate that it is not the membrane potential per se that affects KmADP during ATP synthesis, but rather it is the rate of energy transfer from F0 to F1 that influences both Vmax and KmADP. Further conclusions from the above studies have been discussed in relation to the possible mechanism of energy transfer between F0 and F1 and the manner in which venturicidin and organotin compounds might attenuate this process
RP  - NOT IN FILE
NT  - UI - 93203200LA - engRN - 0 (Oligomycins)RN - 0 (Organotin Compounds)RN - 0 (Succinates)RN - 0 (Trialkyltin Compounds)RN - 0 (Venturicidins)RN - 110-15-6 (Succinic Acid)RN - 56-65-5 (Adenosine Triphosphate)RN - 688-73-3 (tributyltin)RN - 892-20-6 (triphenyltin)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - DK08126/DK/NIDDKDA - 19930422IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:8454592
SO  - J Biol Chem 1993 Mar 25 ;268(9):6168-6173

1165
UI  - 569
AU  - Matsuno-Yagi A
AU  - Hatefi Y
AD  - Department of Molecular and Experimental Medicine, Scripps Research Institute, La Jolla, California 92037
TI  - Studies on the mechanism of oxidative phosphorylation. Different effects of F0 inhibitors on unisite and multisite ATP hydrolysis by bovine submitochondrial particles
AB  - Bovine submitochondrial particles prepared in the presence of GTP (G- SMP), as well as G-SMP washed in 150 mM KCl, catalyzed unisite ATP hydrolysis with a first order rate constant of 0.12 s-1. This rate constant remained unchanged at ATP concentrations < 0.06 microM but increased sharply at higher ATP concentrations, presumably because of ATP binding to other catalytic or regulatory sites. Pretreatment of the particles with oligomycin greatly inhibited unisite ATP binding, in agreement with previous findings. Pretreatment of the particles with N,N'-dicyclohexylcarbodiimide had a slight effect on unisite ATP binding, whereas pretreatment with the inhibitors venturicidin and tributyl(or triphenyl)tin chloride had no effect. Titration of unisite ATPase activity with increasing concentrations of oligomycin or efrapeptin resulted in sigmoidal inhibition curves, as though more than a single inhibition site was being titrated by each inhibitor. Venturicidin and organotin compounds had little effect on the ATPase activity of SMP at [ATP] < or = [F1] and did not cause 100% inhibition at [ATP] >> [F1]. By analogy to our previous studies on the inhibition of the ubiquinol-cytochrome c reductase complex by antimycin (Hatefi, Y., and Yagi, T. (1982) Biochemistry 24, 6614-6618), it is proposed that venturicidin and organotin compounds freeze the structure of the F0 sector of the ATP synthase complex in such a manner that prevents the subunit molecular motions required for rapid proton flux but allows a slow proton flux generated by ATPase activity at low ATP concentrations
RP  - NOT IN FILE
NT  - UI - 93131887LA - engRN - 0 (Oligomycins)RN - 0 (Trialkyltin Compounds)RN - 0 (Venturicidins)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - 688-73-3 (tributyltin)RN - 86-01-1 (Guanosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - DK08126/DK/NIDDKDA - 19930218IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:8380571
SO  - J Biol Chem 1993 Jan 25 ;268(3):1539-1545

1166
UI  - 20816
AU  - McCormick KA
AU  - Deckers-Hebestreit G
AU  - Altendorf K
AU  - Cain BD
AD  - Department of Biochemistry and Molecular Biology, University of Florida, Gainesville 32610
TI  - Characterization of mutations in the b subunit of F1F0 ATP synthase in Escherichia coli
AB  - Site-directed mutagenesis was used to investigate the restrictions on Ala-79 of the b subunit in F1F0 adenosine triphosphate synthase. This amino acid had been previously identified as particularly sensitive to mutation (McCormick, K. A., and Cain, B. D. (1991) J. Bacteriol. 173, 7240-7248). Mutant uncF (b) genes were placed under control of the lac promoter and monitored for F1F0 ATP synthase function in an uncF(b) deletion strain. Three deleterious bAla-79 mutations were moved to the unc operon in the chromosome by homologous recombination. Decreases in enzymatic activity in the uncF (b) mutant strains resulted from reduced amounts of enzyme. With the exception of the bAla-79-->Pro mutation, high expression of mutant uncF (b) genes resulted in increases in F1F0 ATP synthase activity which were sufficient to overcome the defects. In addition to the decrease in the amount of enzyme, the bAla-79-->Lys mutation affected ATP synthesis to a much greater extent than ATP- driven proton translocation. The evidence supports our earlier hypothesis, in which bAla-79 was proposed to play an important, but not essential, structural role in F1F0 ATP synthase assembly or stability
MH  - A
MH  - ACID
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - adenosine triphosphate synthase
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - ATPase
MH  - Biochemistry
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - function
MH  - mutagenesis
MH  - mutant
MH  - proton
MH  - site-directed
MH  - SUBUNIT
MH  - SYNTHASE
MH  - synthesis
MH  - translocation
RP  - NOT IN FILE
NT  - UI - 94043324LA - engRN - 0 (DNA Primers)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleID - GM43495/GM/NIGMSDA - 19931220IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:8227028
SO  - J Biol Chem 1993 Nov 25 ;268(33):24683-24691

1167
UI  - 347
AU  - Medd SM
AU  - Walker JE
AU  - Jolly RD
AD  - Medical Research Council Laboratory of Molecular Biology, Cambridge, UK
TI  - Characterization of the expressed genes for subunit c of mitochondrial ATP synthase in sheep with ceroid lipofuscinosis
AB  - The human and bovine genomes each contain two expressed nuclear genes, called P1 and P2, for subunit c, a hydrophobic subunit of the membrane sector, Fo, of mitochondrial ATP synthase. Both P1 and P2 encode the same mature protein, but the associated mitochondrial import sequences are different. In sheep with the neurodegenerative disease ceroid lipofuscinosis, and also in humans with Batten's disease, unmodified subunit c accumulates in lysosome-derived organelles in a variety of tissues. However, the sequences of cDNAs for P1 and P2 from sheep with ceroid lipofuscinosis were identical to those in healthy control animals. Therefore, since there was no mutation in either of the mitochondrial import sequences of subunit c in the diseased animals, ceroid lipofuscinosis does not arise from changes in an import sequence causing mis-targeting of the c subunit to lysosomes. The levels of expression of P1 and P2 genes were approximately the same in diseased and healthy animals, and so the protein is unlikely to accumulate because of excessive transcription of either gene. Transcription of a spliced pseudogene related to P2 was detected in both a control animal and a sheep with ceroid lipofuscinosis. The transcripts encode amino acids 1-31 of the P2 mitochondrial targeting sequence. In the diseased animal, an arginine replaced a glutamine in the control sequence. However, restriction fragment analysis of genomic DNA from a further 12 sheep established that the sequence differences were not linked to ceroid lipofuscinosis
RP  - NOT IN FILE
NT  - UI - 93319530LA - engRN - 9007-49-2 (DNA)RN - EC 2.7.4.- (ATP synthetase complex)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19930812IS - 0264-6021SB - IMCY - ENGLANDJC - 9YO
UR  - PM:8328973
SO  - Biochem J 1993 Jul 1 ;293 ( Pt 1)():65-73

1168
UI  - 19864
AU  - Milgrom YM
AU  - Cross RL
AD  - Department of Biochemistry and Molecular Biology, State University of New York Health Science Center, Syracuse 13210
TI  - Nucleotide binding sites on beef heart mitochondrial F1-ATPase. Cooperative interactions between sites and specificity of noncatalytic sites
AB  - We have studied the properties of beef heart mitochondrial F1 having inhibitory MgADP bound at one of the three catalytic sites and various levels of occupancy of the three noncatalytic nucleotide sites including zero, two, or three ADP/ATPs or two ADP/ATP plus one GTP. The properties examined include the rate of MgATP-dependent reactivation and the rate of increase in the fraction of F1 containing transiently bound intermediates. For each form of the enzyme tested, the rate of reactivation closely paralleled the rate of increase in the level of bound intermediates, indicating that when one catalytic site on F1 is blocked by inhibitory MgADP, the remaining two sites are incapable of residual uni- or bi-site activity. It was also found that the stability of the MgADP-inhibited complex decreases with full occupancy of the noncatalytic sites. This demonstrates that the noncatalytic sites modulate the properties of catalytic sites. Finally, it was found that the noncatalytic sites on mitochondrial F1 do not, as has long been believed, bind adenine nucleotides exclusively. Evidence is presented that both GTP and PPi bind tightly at noncatalytic sites
MH  - Adenine Nucleotides
MH  - ADENINE-NUCLEOTIDES
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthase
MH  - BINDING
MH  - Binding Sites
MH  - Biochemistry
MH  - COMPLEX
MH  - Diphosphates
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - Guanosine Triphosphate
MH  - H(+)-Transporting ATP Synthase
MH  - MITOCHONDRIAL F1-ATPASE
MH  - NONCATALYTIC SITES
MH  - nucleotide binding
MH  - nucleotide binding sites
MH  - NUCLEOTIDE-BINDING-SITES
MH  - Nucleotides
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 94043102LA - engRN - 0 (Diphosphates)RN - 0 (Ribonucleotides)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 86-01-1 (Guanosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM 23152/GM/NIGMSDA - 19931129IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:8226836
SO  - J Biol Chem 1993 Nov 5 ;268(31):23179-23185

1169
UI  - 743
AU  - Muneyuki E
AU  - Makino M
AU  - Kamata H
AU  - Kagawa Y
AU  - Yoshida M
AU  - Hirata H
AD  - Research Laboratory of Resources Utilization, Tokyo Institute of Technology, Japan
TI  - Inhibitory effect of NaN3 on the F0F1 ATPase of submitochondrial particles as related to nucleotide binding
AB  - The inhibitory effects of NaN3 on the F0F1 ATPase of beef heart submitochondrial particles were investigated. It was shown that NaN3 inhibited the ATPase activity only in the presence of ATP or ADP and the inhibition proceeded slowly. Analysis of the time-course of the inhibition process lead to a conclusion that an ATP binding site which has an apparent Kd of 14.0 +/- 8.7 microM is responsible for the increase of NaN3 sensitivity. This value agreed well with the low Km of ATP hydrolysis characterized before (Muneyuki, E., and Hirata, H. (1988) FEBS Lett. 234, 455-458) and in the range of so-called bi-site catalysis. The same conclusion was derived as for isolated F1 ATPase. From similar analysis, the Kd of this site for ADP was deduced to be 1.34 +/- 0.45 microM, which also agreed with that reported by Pedersen (Pedersen, P.L. (1975) Biochem. Biophys. Res. Commun. 64, 610-616) and also in the same range as reported for the low Km of ATP synthesis by activated submitochondrial particles. These results suggest that hydrolysis through the low Km mode of ATPase reaction leads the enzyme NaN3 sensitive form and this reaction cycle corresponds to the low Km mode of ATP synthesis
RP  - NOT IN FILE
NT  - UI - 93349961LA - engRN - 0 (Azides)RN - 0 (Phosphates)RN - 26628-22-8 (Sodium Azide)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19930914IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:8347662
SO  - Biochim Biophys Acta 1993 Aug 16 ;1144(1):62-68

1170
UI  - 20860
AU  - Nakamoto RK
AU  - Maeda M
AU  - Futai M
AD  - Department of Organic Chemistry and Biochemistry, Osaka University, Japan
TI  - The gamma subunit of the Escherichia coli ATP synthase. Mutations in the carboxyl-terminal region restore energy coupling to the amino- terminal mutant gamma Met-23-->Lys
AB  - The gamma subunit mutations, gamma Met-23-->Lys or Arg, in the Escherichia coli ATP synthase were previously reported to cause dramatically inefficient energy coupling between ATPase catalysis and H+ translocation (Shin, K., Nakamoto, R.K., Maeda, M., and Futai, M. (1992) J. Biol. Chem. 267, 20835-20839). In this paper, we report that second-site mutations in the gamma subunit can suppress the effects of gamma Met-23-->Lys. By screening randomly mutagenized uncG (gamma Met- 23-->Lys), eight mutations in the carboxyl-terminal region were identified; strains carrying gamma Arg-242-->Cys, gamma Gln-269-->Arg, gamma Ala-270-->Val, gamma Ile-272-->Thr, gamma Thr-273-->Ser, gamma Glu-278-->Gly, gamma Ile-279-->Thr, or gamma Val-280-->Ala in combination with gamma Met-23-->Lys were able to grow by oxidative phosphorylation. H+ pumping assayed in membranes prepared from double mutation strains demonstrated that efficient ATP-dependent H+ transport was restored. Interestingly, the single mutations, gamma Gln-269-->Arg or gamma Thr-273-->Ser, caused reduced growth by oxidative phosphorylation; however, when these mutations were in combination with gamma Met-23-->Lys, growth was substantially increased. Furthermore, strains carrying gamma Met-23-->Lys, gamma Gln-269-->Arg, or gamma Thr- 273-->Ser as single mutations were temperature sensitive, whereas, strains with the double mutations, gamma Met-23-->Lys/gamma Gln-269-- >Arg or gamma Met-23-->Lys/gamma Thr-273-->Ser, were thermally stable. Taken together, these results strongly suggest that gamma Met-23, gamma Arg-242, and the region between gamma Gln-269 to gamma Val-280 are close to each other and interact to mediate efficient energy coupling
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Biochemistry
MH  - Catalysis
MH  - Chemistry
MH  - COLI ATP SYNTHASE
MH  - coupling
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - H+
MH  - M
MH  - Macromolecular Systems
MH  - membrane
MH  - Membranes
MH  - mutant
MH  - Oxidative Phosphorylation
MH  - Phosphorylation
MH  - protein
MH  - Proteins
MH  - SUBUNIT
MH  - SYNTHASE
MH  - SYSTEM
MH  - SYSTEMS
MH  - Temperature
MH  - translocation
MH  - transport
RP  - NOT IN FILE
NT  - UI - 93123296LA - engRN - 0 (Codon)RN - 0 (Macromolecular Systems)RN - 0 (Oligodeoxyribonucleotides)RN - 0 (Recombinant Proteins)RN - 56-87-1 (Lysine)RN - 63-68-3 (Methionine)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19930205IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:8419364
SO  - J Biol Chem 1993 Jan 15 ;268(2):867-872

1171
UI  - 740
AU  - Odaka M
AU  - Kiribuchi K
AU  - Allison WS
AU  - Yoshida M
AD  - Research Laboratory of Resources Utilization, Tokyo Institute of Technology, Yokohama, Japan
TI  - In vivo affinity label of a protein expressed in Escherichia coli. Coenzyme A occupied the AT(D)P binding site of the mutant F1-ATPase beta subunit (Y307C) through a disulfide bond
AB  - When Tyr-307 of the beta subunit of F1-ATPase from a thermophilic Bacillus strain PS3 is replaced by cysteine and expressed in Escherichia coli cells, about a half population of the mutant beta subunit are labeled by Coenzyme A at Cys-307 through a disulfide bond which is cleavable by reducing treatment. The mutant beta subunit can be reconstituted into the alpha 3 beta 3 complex of which ATPase activity is stimulated two-fold by reducing treatment either prior or after reconstitution. Since Tyr-307 has been supposed to be located at one of subdomains which form the ATP binding site of the beta subunit, Coenzyme A binds to the mutant beta subunit as an AT(D)P analogue in E. coli cells and then covalently attaches to Cys-307
RP  - NOT IN FILE
NT  - UI - 94085611LA - engRN - 0 (Affinity Labels)RN - 0 (Bacterial Proteins)RN - 0 (Disulfides)RN - 0 (Recombinant Proteins)RN - 52-90-4 (Cysteine)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 85-61-0 (Coenzyme A)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19940127IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:8262235
SO  - FEBS Lett 1993 Dec 27 ;336(2):231-235

1172
UI  - 741
AU  - Paik SR
AU  - Yokoyama K
AU  - Yoshida M
AU  - Ohta T
AU  - Kagawa Y
AU  - Allison WS
AD  - Department of Chemistry, University of California at San Diego, La Jolla 92093
TI  - The TF1-ATPase and ATPase activities of assembled alpha 3 beta 3 gamma, alpha 3 beta 3 gamma delta, and alpha 3 beta 3 gamma epsilon complexes are stimulated by low and inhibited by high concentrations of rhodamine 6G whereas the dye only inhibits the alpha 3 beta 3, and alpha 3 beta 3 delta complexes
AB  - The ATPase activity of the F1-ATPase from the thermophilic bacterium PS3 is stimulated at concentrations of rhodamine 6G up to about 10 microM where 70% stimulation is observed at 36 degrees C. Half maximal stimulation is observed at about 3 microM dye. At rhodamine 6G concentrations greater than 10 microM, ATPase activity declines with 50% inhibition observed at about 75 microM dye. The ATPase activities of the alpha 3 beta 3 gamma and alpha 3 beta 3 gamma delta complexes assembled from isolated subunits of TF1 expressed in E. coli deleted of the unc operon respond to increasing concentrations of rhodamine 6G nearly identically to the response of TF1. In contrast, the ATPase activities of the alpha 3 beta 3 and alpha 3 beta 3 delta complexes are only inhibited by rhodamine 6G with 50% inhibition observed, respectively, at 35 and 75 microM dye at 36 degrees C. The ATPase activity of TF1 is stimulated up to 4-fold by the neutral detergent, LDAO. In the presence of stimulating concentrations of LDAO, the ATPase activity of TF1 is no longer stimulated by rhodamine 6G, but rather, it is inhibited with 50% inhibition observed at about 30 microM dye at 30 degrees C. One interpretation of these results is that binding of rhodamine 6G to a high-affinity site on TF1 stimulates ATPase activity and unmasks a low-affinity, inhibitory site for the dye which is also exposed by LDAO
RP  - NOT IN FILE
NT  - UI - 94193578LA - engRN - 0 (Macromolecular Systems)RN - 0 (Rhodamines)RN - 989-38-8 (rhodamine 6G)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM 16974/GM/NIGMSDA - 19940505IS - 0145-479XSB - IMCY - UNITED STATESJC - HIO
UR  - PM:8144495
SO  - J Bioenerg Biomembr 1993 Dec ;25(6):679-684

1173
UI  - 822
AU  - Pancic PG
AU  - Strotmann H
AD  - Institut fur Biochemie der Pflanzen, Heinrich-Heine-Universitat Dusseldorf, Germany
TI  - Structure of the nuclear encoded gamma subunit of CF0CF1 of the diatom Odontella sinensis including its presequence
AB  - Using a PCR-product as homologous probe for screening of a cDNA library of the diatom Odontella sinensis overlapping cDNA clones were obtained which showed homologies to atpC-genes of F0F1-ATPases from different sources. Comparison of the deduced amino acid sequence with the N- terminal sequence of the Odontella gamma subunit obtained by protein sequencing, indicated that the complete 370 amino acid protein is processed to a mature protein of 315 amino acids. The 55 amino acids comprising the presequence consists of two segments, one resembling a signal sequence for cotranslational transport through ER membranes and one showing characteristics of a transit sequence for transport of proteins into chloroplasts of higher plants. This result is discussed with respect to the particular envelope structure of chromophytic plastids consisting of four membranes. The outer membrane contains ribosomes on its cytosolic surface. As in cyanobacterial gamma subunits the regulatory sequence region, which is involved in thiol modulation of chloroplast ATPase of green algae and higher plants, is absent in the Odontella gamma subunit
RP  - NOT IN FILE
NT  - UI - 93215822LA - engRN - 0 (DNA, Single-Stranded)RN - 0 (Protein Precursors)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19930430IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:8462677
SO  - FEBS Lett 1993 Mar 29 ;320(1):61-66

1174
UI  - 512
AU  - Pedersen PL
AU  - Amzel LM
AD  - Department of Biological Chemistry, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205
TI  - ATP synthases. Structure, reaction center, mechanism, and regulation of one of nature's most unique machines
RP  - NOT IN FILE
NT  - UI - 93252965LA - engRN - 0 (Macromolecular Systems)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialID - CA 10951/CA/NCIID - GM 25432/GM/NIGMSDA - 19930608IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:8486720
SO  - J Biol Chem 1993 May 15 ;268(14):9937-9940

1175
UI  - 432
AU  - Senior AE
AU  - Wilke-Mounts S
AU  - al Shawi MK
AD  - Department of Biochemistry, University of Rochester Medical Center, New York 14642
TI  - Lysine 155 in beta-subunit is a catalytic residue of Escherichia coli F1 ATPase
AB  - The "homology A" ("glycine-rich" or "P-loop") consensus sequence occurs in the catalytic sites of F1F0 ATP synthase enzymes. The conserved lysine of this motif is beta-subunit Lys-155 in Escherichia coli F1. The role of this lysine in binding and catalysis at the high affinity ATP binding site was studied with the mutants beta K155Q and beta K155E by measuring the rates of ATP binding/release, ATP hydrolysis/synthesis, and Pi release as a function of pH varied from 5.5 to 9.5. In wild type, protonated beta Lys-155 appears to contribute significantly to high affinity binding of ATP, probably through hydrogen bonding to the gamma-phosphate. ATP hydrolysis and synthesis were impaired strongly in the mutants, and the reaction equilibrium constant, which was pH-independent in wild type, was highly pH- dependent in beta K155Q and beta K155E. Studies of steady-state ATPase turnover showed that positive catalytic cooperativity was virtually absent and the pH-dependent component of positive catalytic cooperativity was abolished or reversed in the mutants. The data demonstrate that residue beta K155 is a critical catalytic residue in F1 ATPase
RP  - NOT IN FILE
NT  - UI - 93216633LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - 56-87-1 (Lysine)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSDA - 19930505IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:8463232
SO  - J Biol Chem 1993 Apr 5 ;268(10):6989-6994

1176
UI  - 802
AU  - Strelow F
AU  - Rumberg B
AD  - Max-Volmer-Institut fur Biophysikalische und Physikalische Chemie, Technische Universitat Berlin, Germany
TI  - Kinetics and energetics of redox regulation of ATP synthase from chloroplasts
AB  - The rate of ATP hydrolysis catalyzed by the membrane-bound CF0F1 ATP synthase from chloroplasts served as a probe for the determination of the reduction grade of the enzyme treated with dithiothreitol (DTT) or thioredoxin. Rate constants for reduction were obtained. It turns out that reduction by thioredoxin is about a factor of 6,000 more effective than DTT reduction. The activation profiles with respect to delta pH were obtained for reduced and oxidized ATPases. The activation curve of reduced enzyme turns out to have its half-maximum degree of activation at delta pH = 1.65, which is considerably lower than reported hitherto. The corresponding value of the oxidized enzyme has been obtained from the rate of ATP hydrolysis in the case of incomplete reduced ATPases, taking into account the aforementioned rate constants, and comes to delta pH = 3.35
RP  - NOT IN FILE
NT  - UI - 93265934LA - engRN - 0 (Sulfhydryl Compounds)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19930624IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:8495739
SO  - FEBS Lett 1993 May 24 ;323(1-2):19-22

1177
UI  - 742
AU  - Tozawa K
AU  - Miyauchi M
AU  - Yoshida M
AD  - Research Laboratory of Resources Utilization, Tokyo Institute of Technology, Yokohama, Japan
TI  - Structure of the alpha subunit of F1-ATPase probed by limited proteolysis
AB  - The structure of the isolated alpha subunit of F1-ATPase from the thermophilic Bacillus strain PS3 was probed using limited proteolysis by four different proteases, and the following results were obtained. 1) Distribution of 21 protease-cleaved sites is similar to that of the beta subunit of F1-ATPase (Tozawa, K., Odaka, M., Date, T., and Yoshida, M. (1992) J. Biol. Chem. 267, 16484-16490), thus providing experimental evidence for similar folding topology of the two subunits, and the locations of 11 water-exposed loop regions in the tertiary structure are predicted. 2) Most proteolytic peptides remain associated to maintain the gross structure of the alpha subunit and can reassociate each other after denaturing urea treatment. 3) However, the carboxyl-terminal peptides comprising approximately 80 residues (C1 peptides) are released from other peptide(s) during proteolysis, and those comprising approximately 105 residues (C2 peptides) are released during native polyacrylamide gel electrophoresis after proteolysis. 4) Inclusion of Mg-ATP in the native electrophoretic system prevents the release of the C2 peptide. Addition of Mg-ATP to the proteolysis mixtures results in an increase of the C2 peptide population and a decrease of the C1 peptide population. Thus, Mg-ATP induces a conformational change at the regions of C1 and C2 peptides of the alpha subunit. 5) Except for the trypsin-treated one, protease-treated alpha subunits are reconstitutable with the native beta subunit into the form of alpha 3 beta 3 complexes, which show significantly higher ATPase activities than the intact alpha 3 beta 3 complex. This activation is attributable to the cleavage of a peptide bond that produces C2 peptides. The carboxyl-terminal region of the alpha subunit is likely to be involved in the regulation of ATPase activity in F1-ATPase
RP  - NOT IN FILE
NT  - UI - 93366828LA - engRN - 0 (Peptide Fragments)RN - 56-65-5 (Adenosine Triphosphate)RN - 57-13-6 (Urea)RN - EC 3.4.- (Endopeptidases)RN - EC 3.4.21 (Serine Endopeptidases)RN - EC 3.4.21.1 (Chymotrypsin)RN - EC 3.4.21.19 (glutamyl endopeptidase)RN - EC 3.4.21.4 (Trypsin)RN - EC 3.4.24.27 (Thermolysin)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19930930IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:8360191
SO  - J Biol Chem 1993 Sep 5 ;268(25):19044-19054

1178
UI  - 173
AU  - Turina P
AU  - Aggeler R
AU  - Lee RS
AU  - Senior AE
AU  - Capaldi RA
AD  - Department of Biology, University of Oregon, Eugene 97403
TI  - The cysteine introduced into the alpha subunit of the Escherichia coli F1-ATPase by the mutation alpha R376C is near the alpha-beta subunit interface and close to a noncatalytic nucleotide binding site
AB  - Mutation of the alpha subunit of the Escherichia coli F1-ATPase to convert Arg-376 to a Cys (alpha R376C) lowers multisite ATPase activity 400-1,000-fold while affecting unisite catalysis only around 6-fold, suggesting that the mutation is in a region important for transmission of conformational changes between catalytic sites (Soga, S., Noumi, T., Takeyama, M., Maeda, M., and Futai, M. (1989) Arch. Biochem. Biophys. 268, 643-648; this study). To learn more of the structural features of the segment of the alpha subunit around Arg-376, mutant enzyme with a Cys at this position was modified with several maleimides. N- [14C]Ethylmaleimide reacted rapidly with this Cys in one of the three alpha subunits/F1 (2,500 M-1 s-1); more slowly with a second alpha subunit (390 M-1 s-1); and the same Cys in the third copy of the alpha subunit was completely unreactive to the reagent, indicating asymmetry of alpha subunits in the ECF1 complex. The photoactivatable cross- linker N-(4-azido-2,3,5,6-tetrafluorobenzyl)-3-maleimidopropionamide++ +, when reacted via its maleimide to alpha Cys-376 of the mutant, covalently linked alpha to beta subunits upon photolysis, indicating that Cys-376 of alpha is close to an interface between the alpha and beta subunits. The EDTA-induced exchangeable noncatalytic site could be filled by TNP-ATP in both wild type and alpha R376C mutant ECF1. Occupancy of this site in the alpha R376C mutant altered the rate of reaction of the second-fastest reacting Cys-376 from 390 M-1 s-1 to below 130 M-1 s-1, suggesting that the two sites are on the same alpha subunit. TNP-ATP in the EDTA-induced exchangeable noncatalytic site was quenched by reacting Cys-376 with 4-maleimido-(2,2,6,6- tetramethylpiperidine-N(oxyl), indicating that the region around Cys- 376, which is involved in transmission of conformational changes between alpha and beta, and noncatalytic sites are maximally 10-12 A from each other
RP  - NOT IN FILE
NT  - UI - 93216631LA - engRN - 128-53-0 (Ethylmaleimide)RN - 52-90-4 (Cysteine)RN - 56-65-5 (Adenosine Triphosphate)RN - 61368-63-6 (2',3'-O-(2,4,6-trinitro-cyclohexadienylidine)adenosine 5'- triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSID - HL24526/HL/NHLBIDA - 19930505IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:8463230
SO  - J Biol Chem 1993 Apr 5 ;268(10):6978-6984

1179
UI  - 346
AU  - van Walraven HS
AU  - Lutter R
AU  - Walker JE
AD  - Medical Research Council, Laboratory of Molecular Biology, Cambridge, UK
TI  - Organization and sequences of genes for the subunits of ATP synthase in the thermophilic cyanobacterium Synechococcus 6716
AB  - The sequences of the genes for the nine subunits of ATP synthase in the thermophilic cyanobacterium Synechococcus 6716 have been determined. The genes were identified by comparison of the encoded proteins with sequences of ATP synthase subunits in other species, and confirmed for subunits alpha, beta, delta and epsilon, by determining their N- terminal sequences. They are arranged at three separate loci. Six of them are in one cluster in the order a: c: b': b: delta: alpha, and those for the beta and epsilon subunits form a second and separate cluster. The gene for the gamma-subunit is at a third site. As in other bacteria, the gene for subunit a is immediately preceded by a gene coding for a small hydrophobic protein of unknown function, known as uncI in Escherichia coli. The gene orders in Synechococcus 6716 are related to the orders of ATP synthase genes in the plastid genomes of higher plants, and particularly of a red alga and a diatom. The sequences of the subunits are similar to those of chloroplast ATP synthase, the alpha, beta and c subunits being particularly well conserved. Differences in the primary structures of the Synechococcus 6716 and chloroplast gamma subunits probably underlie different mechanisms of activation of ATP synthase. The nucleotide sequences that are presented also contain 12 other open reading frames. One of them encodes a protein sequence related to the E. coli DNA repair enzyme, photolyase, and another codes for a protein that contains internal repeats related to sequences in the myosin heavy chain
RP  - NOT IN FILE
NT  - UI - 93371369LA - engRN - 0 (DNA, Bacterial)RN - 0 (Peptide Fragments)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19930924IS - 0264-6021SB - IMCY - ENGLANDJC - 9YO
UR  - PM:8363578
SO  - Biochem J 1993 Aug 15 ;294 ( Pt 1)():239-251

1180
UI  - 20997
AU  - Wach A
AU  - Dencher NA
AU  - Graber P
AD  - Biologisches Institut, Universitat Stuttgart, Germany
TI  - Co-reconstitution of plasma membrane H(+)-ATPase from yeast and bacteriorhodopsin into liposomes. ATP hydrolysis as a function of external and internal pH
AB  - The H(+)-ATPase from the plasma membrane of Saccharomyces cerevisiae was isolated and purified. The enzyme was reconstituted with bacteriorhodopsin into asolectin liposomes by detergent dialysis at a molar ratio of 1 H(+)-ATPase to 50 bacteriorhodopsins. The overall orientation of the proteins is such that proton pumping to the vesicle interior occurs upon illumination and after addition of ATP. All liposomes which contain H(+)-ATPase also contain bacteriorhodopsin. The rate of ATP hydrolysis was measured as function of pH in the dark and during illumination of the proteoliposomes. The pH dependency can be described by the protonation of a monovalent group from the outside with an apparent pK of 7.3 and the deprotonation of a monovalent group at the inside with an apparent pK of 3.7. Inside and outside refer to the orientation of the H(+)-ATPase in the liposomes which is opposite to that occurring in vivo. It is concluded that the first step in the reaction cycle is the binding of a proton from the cytosol which is followed by ATP binding, ATP hydrolysis on the enzyme and the release of ADP and phosphate, and finally the proton is released from the enzyme into the external medium
MH  - A
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - ADP
MH  - atp
MH  - ATPase
MH  - Bacteriorhodopsin
MH  - BINDING
MH  - Dialysis
MH  - function
MH  - H(+)ATPase
MH  - Hydrolysis
MH  - Liposomes
MH  - membrane
MH  - pH
MH  - protein
MH  - Proteins
MH  - proteoliposome
MH  - proton
MH  - protonation
MH  - Saccharomyces cerevisiae
RP  - NOT IN FILE
NT  - UI - 93292519LA - engRN - 0 (Liposomes)RN - 53026-44-1 (Bacteriorhodopsins)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19930719IS - 0014-2956SB - IMCY - GERMANY
UR  - PM:8513805
SO  - Eur J Biochem 1993 Jun 1 ;214(2):563-568

1181
UI  - 345
AU  - Walker JE
AU  - Runswick MJ
AD  - Medical Research Council Laboratory of Molecular Biology, Cambridge, UK
TI  - The mitochondrial transport protein superfamily
AB  - The ADP/ATP, phosphate, and oxoglutarate/malate carrier proteins found in the inner membranes of mitochondria, and the uncoupling protein from mitochondria in mammalian brown adipose tissue, belong to the same protein superfamily. Established members of this superfamily have polypeptide chains approximately 300 amino acids long that consist of three tandem related sequences of about 100 amino acids. The tandem repeats from the different proteins are interrelated, and probably have similar secondary structures. The common features of this superfamily are also present in nine proteins of unknown functions characterized by DNA sequencing in various species, most notably in Caenorhabditis elegans and Saccharomyces cerevisiae. The high level expression in Escherichia coli of the bovine oxoglutarate/malate carrier, and the reconstitution of active carrier from the expressed protein, offers encouragement that the identity of superfamily members of known sequence but unknown function may be uncovered by a similar route
RP  - NOT IN FILE
NT  - UI - 94179126LA - engRN - 0 (Carrier Proteins)RN - 0 (Membrane Proteins)RN - 0 (Recombinant Fusion Proteins)RN - 0 (mitochondrial uncoupling protein)RN - 0 (oxoglutarate translocator)RN - EC 2.7.7.- (Adenine Nucleotide Translocase)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 19940421IS - 0145-479XSB - IMCY - UNITED STATESJC - HIO
UR  - PM:8132484
SO  - J Bioenerg Biomembr 1993 Oct ;25(5):435-446

1182
UI  - 433
AU  - Weber J
AU  - Lee RS
AU  - Wilke-Mounts S
AU  - Grell E
AU  - Senior AE
AD  - Department of Biochemistry, University of Rochester Medical Center, New York 14642
TI  - Combined application of site-directed mutagenesis, 2-azido-ATP labeling, and lin-benzo-ATP binding to study the noncatalytic sites of Escherichia coli F1-ATPase
AB  - Noncatalytic nucleotide sites of Escherichia coli F1-ATPase were studied by site-directed mutagenesis, covalent photolabeling with 2- azido-ATP, and lin-benzo-ATP binding. In wild-type, 89% of 2-azido-ATP label was bound to beta-subunit, whereas in the beta Y354F mutant, 95% of the label was bound to alpha-subunit. In the alpha R365Y mutant, label was seen on both alpha (38%) and beta (62%); whereas in the alpha R365F mutant, 93% was on beta. The fluorescence of noncatalytic site- bound lin-benzo-ATP was quenched markedly in F1 from wild-type (76% quench), alpha R365F (85%), alpha R365Y (90%), and alpha R365Y, beta Y354F (83%), but only by 28% in beta Y354F. These results together demonstrate that residues alpha R365 and beta Y354 lie close to the base moiety of adenine nucleotide bound in F1 noncatalytic sites. From comparison of sequences of alpha- and beta-subunits, it appears that residue alpha R365 in noncatalytic sites is equivalent to residue beta Y331 of the catalytic sites. Two unintended mutants were obtained in which alpha-subunit was increased in length by 17 amino acids due to repeat of residues alpha 361 to alpha 377, with either F or Y in the repeated alpha 365 position. Soluble F1 was obtained from both mutants, with novel properties
RP  - NOT IN FILE
NT  - UI - 93203211LA - engRN - 0 (Affinity Labels)RN - 0 (Azides)RN - 0 (DNA, Bacterial)RN - 56-65-5 (Adenosine Triphosphate)RN - 61925-58-4 (benzo-ATP)RN - 72884-75-4 (2-azidoadenosine 5'-triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSDA - 19930422IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:8454597
SO  - J Biol Chem 1993 Mar 25 ;268(9):6241-6247

1183
UI  - 431
AU  - Weber J
AU  - Wilke-Mounts S
AU  - Lee RS
AU  - Grell E
AU  - Senior AE
AD  - Department of Biochemistry, University of Rochester Medical Center, New York 14642
TI  - Specific placement of tryptophan in the catalytic sites of Escherichia coli F1-ATPase provides a direct probe of nucleotide binding: maximal ATP hydrolysis occurs with three sites occupied
AB  - Residue beta Y331 of Escherichia coli F1-ATPase is known from previous affinity labeling, mutagenesis, and lin-benzo-ADP binding experiments to interact directly with the adenine moiety of substrates bound in catalytic sites. Here we mutagenized beta Y331 to tryptophan. Mutant cells grew well on succinate or limiting glucose; purified mutant F1 had kappa cat/Km and lin-benzo-ADP binding characteristics similar to wild type. Fluorescence from beta W331 residues exhibited a maximum at 349 nm, indicating a polar environment in unoccupied sites. ATP, ADP, or AMPPNP caused virtually complete quenching of beta W331 fluorescence, so that the fluorescence of mutant F1 with occupied catalytic sites resembled that of wild-type enzyme. Therefore the beta W331 fluorescence provided a direct probe of nucleotide binding to catalytic sites under true equilibrium conditions. We measured ATP binding and hydrolysis in parallel experiments and found that occupancy of one or two catalytic sites per F1 molecule did not yield significant rates of hydrolysis while occupancy of all three sites yielded Vmax rates. Km(ATP) was similar to Kd3, the Kd for ATP binding to the third catalytic site. We also measured AMPPNP and ADP binding parameters. For ADP, the "on" rate at the first catalytic site was much faster (> or = 5 x 10(5) M-1 s-1) than seen previously by centrifuge column procedures, although the Kd was not much changed. For AMPPNP, the "on" rate at the first site was 2 orders of magnitude less than for ADP or ATP, and the Kd was similar to that for ADP
RP  - NOT IN FILE
NT  - UI - 93388577LA - engRN - 0 (Oligodeoxyribonucleotides)RN - 0 (Recombinant Proteins)RN - 25612-73-1 (Adenylyl Imidodiphosphate)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 61925-59-5 (linear-benzoadenosine diphosphate)RN - 73-22-3 (Tryptophan)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSDA - 19931020IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:8376371
SO  - J Biol Chem 1993 Sep 25 ;268(27):20126-20133

1184
UI  - 21329
AU  - Wetzel CM
AU  - McCarty RE
TI  - Aspects of Subunit Interactions in the Chloroplast ATP Synthase (II. Characterization of a Chloroplast Coupling Factor 1-Subunit III Complex from Spinach Thylakoids)
AB  - A complex between chloroplast-coupling factor 1 (CF1) and subunit III of the membrane-spanning portion of the chloroplast ATP synthase (CF0), isolated as described in the accompanying paper (C.M. Wetzel and R.E. McCarty [1993] Plant Physiol 102: 241-249), has been further characterized. A comparison of the ATPase activities of CF1, CF1-subunit III, and the chloroplast ATP synthase (CF1-CF0) holoenzyme revealed that the properties of CF1-subunit III more closely resemble those of CF1-CF0 than those of CF1. In particular, the Ca2+-ATPase activity after reduction of the enzyme with dithiothreitol was much lower in CF1-subunit III and CF1-CF0 than in CF1, suggesting that the association of the inhibitory [epsilon] subunit is tightened by the presence of either CF0 or subunit III. Cold stability is a property of CF1-CF0 in thylakoid membranes. The ATPase activity of CF1 incubated in the cold in the presence of asolectin liposomes was lost more rapidly than that of either CF1-subunit III or CF1-CF0 incorporated into liposomes. Removal of the [epsilon] subunit from all three preparations resulted in marked stimulation of their ATPase activity. Although subunit III was also removed during depletion of the [epsilon] subunit, it is not known whether the two subunits interact directly. CF1 deficient in the [epsilon] subunit binds to liposomes containing either subunit III or CF0. Taken together, these results provide evidence that the association of CF1 and subunit III of CFo is specific and may play a role in enzyme regulation
MH  - A
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - chloroplast
MH  - COMPLEX
MH  - coupling
MH  - COUPLING FACTOR
MH  - Dithiothreitol
MH  - enzyme
MH  - FIELD
MH  - INTERACTION
MH  - liposome
MH  - Liposomes
MH  - membrane
MH  - Membranes
MH  - plant
MH  - REDUCTION
MH  - regulation
MH  - Spinach
MH  - SUBUNIT
MH  - SYNTHASE
MH  - thylakoid
MH  - thylakoid membrane
MH  - thylakoids
MH  - universities
RP  - NOT IN FILE
NT  - Field of Botany, Cornell University, Ithaca, New York 14853 (CMW)
SO  - Plant Physiol 1993 May ;102(1):251-259

1185
UI  - 21330
AU  - Wetzel CM
AU  - McCarty RE
TI  - Aspects of Subunit Interactions in the Chloroplast ATP Synthase (I. Isolation of a Chloroplast Coupling Factor 1-Subunit III Complex from Spinach Thylakoids)
AB  - A chloroplast ATP synthase complex (CF1 [chloroplast-coupling factor 1]-CF0 [membrane-spanning portion of chloroplast ATP synthase]) depleted of all CF0 subunits except subunit III (also known as the proteolipid subunit) was purified to study the interaction between CF1 and subunit III. Subunit III has a putative role in proton translocation across the thylakoid membrane during photophosphorylation; therefore, an accurate model of subunit inter-actions involving subunit III will be valuable for elucidating the mechanism and regulation of energy coupling. Purification of the complex from a crude CF1-CF0 preparation from spinach (Spinacia oleracea) thylakoids was accomplished by detergent treatment during anion-exchange chromatography. Subunit III in the complex was positively identified by amino acid analysis and N-terminal sequencing. The association of subunit III with CF1 was verified by linear sucrose gradient centrifugation, immunoprecipitation, and incorporation of the complex into asolectin liposomes. After incorporation into liposomes, CF1 was removed from the CF1-III complex by ethylenediaminetetracetate treatment. The subunit III-proteoliposomes were competent to rebind purified CF1. These results indicate that subunit III directly interacts with CF1 in spinach thylakoids
MH  - A
MH  - ACID
MH  - analysis
MH  - atp
MH  - ATP synthase
MH  - chloroplast
MH  - COMPLEX
MH  - coupling
MH  - COUPLING FACTOR
MH  - energy
MH  - FIELD
MH  - INTERACTION
MH  - liposome
MH  - Liposomes
MH  - mechanism
MH  - membrane
MH  - model
MH  - Photophosphorylation
MH  - proteolipid
MH  - proton
MH  - purification
MH  - regulation
MH  - Spinach
MH  - SUBUNIT
MH  - Sucrose
MH  - SYNTHASE
MH  - thylakoid
MH  - thylakoid membrane
MH  - thylakoids
MH  - translocation
MH  - universities
RP  - NOT IN FILE
NT  - Field of Botany, Cornell University, Ithaca, New York 14853 (CMW)
SO  - Plant Physiol 1993 May ;102(1):241-249

1186
UI  - 965
AU  - Williams RJP
TI  - The history of proton-driven ATP formation
AB  - This article sets down the beginnings of some thoughts in bio-energetics. It illustrates how difficult it is in science as elsewhere to know how a new idea is generated. The literature needs very careful examination and separation from personalities
MH  - Adenosine Triphosphate
MH  - Autobiography
MH  - Biochemistry
MH  - Biological Transport
MH  - biosynthesis
MH  - Chemistry
MH  - Energy Metabolism
MH  - H(+)-Transporting ATP Synthase
MH  - history
MH  - History of Medicine,20th Cent.
MH  - metabolism
MH  - Models,Molecular
MH  - Protons
RP  - NOT IN FILE
NT  - University of Oxford, Inorganic Chemistry Laboratory, UK
SO  - Biosci Rep 1993 Aug ;13(4):191-212

1187
UI  - 20858
AU  - Xie DL
AU  - Lill H
AU  - Hauska G
AU  - Maeda M
AU  - Futai M
AU  - Nelson N
AD  - Roche Institute of Molecular Biology, Roche Research Center, Nutley, NJ 07110
TI  - The atp2 operon of the green bacterium Chlorobium limicola
AB  - The operon (atp2) encoding the beta and epsilon subunits of F-ATPase from Chlorobium limicola was cloned and sequenced. In contrast with purple bacteria these genes are arranged in a separate operon similar to the cyanobacteria. The operon terminates with a pronounced stem-loop structure. About 0.8 kb upstream of the beta subunit a gene encoding the enzyme phospho enol pyruvate carboxykinase was identified. This gene is transcribed in the opposite direction of the atp2 operon and also ends with a stem-loop structure. These genes of green bacteria are among the first to be sequenced, and therefore the genetic distance between these genes and corresponding genes from other bacteria and eukaryotes was studied. Even though the operon structure resembles that of cyanobacteria, the evolutionary tree compiled from these data places the chlorobium gene close to purple bacteria. Chlorobium limicola beta and epsilon subunits complemented Escherichia coli mutants defective in the corresponding subunits, indicating that the hybrid enzyme formed from subunits of the two bacteria is active in ATP synthesis
MH  - A
MH  - ACTIVE
MH  - atp
MH  - ATP synthesis
MH  - ATPase
MH  - Bacteria
MH  - BETA-SUBUNIT
MH  - Cyanobacteria
MH  - EPSILON-SUBUNIT
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F-ATPASE
MH  - mutant
MH  - SUBUNIT
MH  - synthesis
RP  - NOT IN FILE
NT  - UI - 93192317LA - engRN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19930414IS - 0006-3002SB - IMCY - NETHERLANDS
UR  - PM:8448205
SO  - Biochim Biophys Acta 1993 Mar 20 ;1172(3):267-273

1188
UI  - 21192
AU  - Zaslavsky D
AU  - Kaulen AD
AU  - Smirnova IA
AU  - Vygodina T
AU  - Konstantinov AA
AD  - AN Belozersky Institute of Physico-Chemical Biology, Moscow State University, Russian Federation
TI  - Flash-induced membrane potential generation by cytochrome c oxidase
AB  - Flash-induced single-electron reduction of cytochrome c oxidase. Compound F (oxoferryl state) by RuII(2,2'-bipyridyl)3(2+) [Nilsson (1992) Proc. Natl. Acad. Sci. USA 89, 6497-6501] gives rise to three phases of membrane potential generation in proteoliposomes with tau values and contributions of ca. 45 microsecond (20%), 1 ms (20%) and 5 ms (60%). The rapid phase is not sensitive to the binuclear centre ligands, such as cyanide or peroxide, and is assigned to vectorial electron transfer from CuA to heme a. The two slow phases kinetically match reoxidation of heme a, require added H2O2 or methyl peroxide for full development, and are completely inhibited by cyanide; evidently, they are associated with the reduction of Compound F to the Ox state by heme a. The charge transfer steps associated with the F to Ox conversion are likely to comprise (i) electrogenic uptake of a 'chemical' proton from the N phase required for protonation of the reduced oxygen atom and (ii) electrogenic H+ pumping across the membrane linked to the F to Ox transition. Assuming heme a 'electrical location' in the middle of the dielectric barrier, the ratio of the rapid to slow electrogenic phase amplitudes indicates that the F to Ox transition is linked to transmembrane translocation of 1.5 charges (protons) in addition to an electrogenic uptake of one 'chemical' proton required to form Fe(3+)-OH- from Fe4+ = O2-. The shortfall in the number of pumped protons and the biphasic kinetics of the millisecond part of the electric response matching biphasic reoxidation of heme a may indicate the presence of 2 forms of Compound F, reduction of only one of which being linked to full proton pumping
MH  - A
MH  - cytochrome
MH  - Cytochrome c
MH  - Cytochrome-c Oxidase
MH  - development
MH  - electrogenic
MH  - electron
MH  - electron transfer
MH  - Electron-transfer
MH  - H+
MH  - Kinetics
MH  - Ligands
MH  - liposome
MH  - Liposomes
MH  - membrane
MH  - Membrane Potential
MH  - microsecond
MH  - Oxygen
MH  - Phosphatidylcholines
MH  - Phospholipids
MH  - Potassium
MH  - Proteolipids
MH  - proteoliposome
MH  - proton
MH  - protonation
MH  - Protons
MH  - TRANSFER
MH  - translocation
RP  - NOT IN FILE
NT  - UI - 94109574LA - engRN - 0 (Liposomes)RN - 0 (Phosphatidylcholines)RN - 0 (Phospholipids)RN - 0 (Proteolipids)RN - 0 (proteoliposomes)RN - 151-50-8 (Potassium Cyanide)RN - 69279-91-0 (asolectin)RN - EC 1.11.1.6 (Catalase)RN - EC 1.9.3.1 (Cytochrome-c Oxidase)PT - Journal ArticleDA - 19940214IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:8282099
SO  - FEBS Lett 1993 Dec 28 ;336(3):389-393

1189
UI  - 19775
AU  - Zhou JM
AU  - Boyer PD
AD  - Molecular Biology Institute, University of California, Los Angeles 90024-1570
TI  - Evidence that energization of the chloroplast ATP synthase favors ATP formation at the tight binding catalytic site and increases the affinity for ADP at another catalytic site
AB  - Previous results have not established whether the attainment of a rapid photophosphorylation rate as ADP concentration is increased in the micromolar range (apparent Km = approximately 30 microM) results from the filling of a second or a third catalytic site. Measurements reported here show that the ATP synthase of chloroplast thylakoids, with 2-4 microM medium ADP present during steady-state photophosphorylation, has one catalytic site filled with tightly bound nucleotides, but other catalytic sites are largely empty. Thus, the rapid increase in the photophosphorylation rate with higher ADP concentrations results from the filling of a second catalytic site. Even with 30 microM added ADP in the dark, the binding of more than one ADP per synthase was not detectable. The sensitivity of the assay was such that the Kd for binding of ADP at a second catalytic site of the de-energized synthase is > 150 microM, considerably above the apparent Km for rapid photophosphorylation. This result can be explained by an increase in the affinity of a second catalytic site for ADP upon energization. Other experiments assessed the effect of ADP binding at a second catalytic site on the equilibrium between bound ATP and ADP and P(i) at the tight catalytic site. When the rate of photophosphorylation is limited by a low ADP concentration, about equal amounts of ATP and ADP are bound at one catalytic site on the synthase. In contrast, when the rate is limited by a low P(i) concentration with 100 microM ADP present, the equilibrium of bound reactants is shifted so that close to one ATP per synthase is present. This is as expected if the binding of ADP at a second catalytic site allows the protonmotive force to promote ATP formation from ADP and P(i) at a tight binding catalytic site. A scheme for the binding change mechanism incorporating these results is presented
MH  - A
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Monophosphate
MH  - Adenosine Triphosphate
MH  - ADP
MH  - ADP BINDING
MH  - atp
MH  - ATP FORMATION
MH  - ATP synthase
MH  - BINDING
MH  - BOUND NUCLEOTIDES
MH  - CHANGE MECHANISM
MH  - chloroplast
MH  - H(+)-Transporting ATP Synthase
MH  - mechanism
MH  - Nucleotides
MH  - Phosphates
MH  - Photophosphorylation
MH  - SYNTHASE
MH  - thylakoid
RP  - NOT IN FILE
NT  - UI - 93131886LA - engRN - 0 (Phosphates)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 61-19-8 (Adenosine Monophosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM-11094/GM/NIGMSDA - 19930218IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:8420929
SO  - J Biol Chem 1993 Jan 25 ;268(3):1531-1538

1190
UI  - 635
AU  - Ziegler M
AU  - Penefsky HS
AD  - Department of Biochemistry and Molecular Biology, State University of New York, Health Science Center, Syracuse 13210
TI  - The adenine nucleotide translocase modulates oligomycin-induced quenching of pyranine fluorescence in submitochondrial particles
AB  - Incorporation of the fluorescent, nonpermeant pH indicator pyranine into submitochondrial particles (pyranine-SMP) permitted monitoring of intravesicular pH changes brought about by proton translocation due to oxidation of respiratory chain substrates or to hydrolysis of ATP. Addition of oligomycin to beef heart pyranine-SMP was followed by a pH- independent quenching of pyranine fluorescence. Quenching was influenced by the presence of adenine nucleotides both inside and outside the submitochondrial particles. The nature of the nucleotides required for quenching resembled the specificity of the adenine nucleotide translocase rather than F1-ATPase. Removal of F1 from pyranine-SMP by treatment of the particles with urea did not alter oligomycin-induced quenching. Atractyloside, a specific inhibitor of the adenine nucleotide translocase, prevented oligomycin-induced quenching when the inhibitor was coincorporated into submitochondrial particles with pyranine. Bongkrekic acid prevented or reversed the oligomycin-dependent quenching when added to pyranine-SMP either before or after oligomycin, respectively, but only when ATP was present within the particles. A mutant of Saccharomyces cerevisiae, lacking translocase genes, exhibited oligomycin-dependent fluorescence quenching which was not inhibited by bongkrekic acid. The results support the interpretation that oligomycin promotes sequestration of the fluorescent probe in a region of the submitochondrial particle, probably the F0F1 complex, that leads to a quenching of fluorescence. The observed quenching can be modulated in a way that suggests an interaction between the translocase and F0
RP  - NOT IN FILE
NT  - UI - 94064591LA - engRN - 0 (Arylsulfonates)RN - 0 (Fluorescent Dyes)RN - 0 (Oligomycins)RN - 0 (Ribonucleotides)RN - 11076-19-0 (Bongkrekic Acid)RN - 17754-44-8 (Atractyloside)RN - 370-86-5 (Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone)RN - 56-65-5 (Adenosine Triphosphate)RN - 6358-69-6 (pyranine)RN - EC 2.7.7.- (Adenine Nucleotide Translocase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM21737/GM/NIGMSDA - 19940104IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:8244963
SO  - J Biol Chem 1993 Dec 5 ;268(34):25320-25328

1191
UI  - 341
AU  - Abrahams JP
AU  - Leslie AG
AU  - Lutter R
AU  - Walker JE
AD  - Medical Research Council Laboratory of Molecular Biology, Cambridge, UK
TI  - Structure at 2.8 A resolution of F1-ATPase from bovine heart mitochondria
AB  - In the crystal structure of bovine mitochondrial F1-ATPase determined at 2.8 A resolution, the three catalytic beta-subunits differ in conformation and in the bound nucleotide. The structure supports a catalytic mechanism in intact ATP synthase in which the three catalytic subunits are in different states of the catalytic cycle at any instant. Interconversion of the states may be achieved by rotation of the alpha 3 beta 3 subassembly relative to an alpha-helical domain of the gamma- subunit
RP  - NOT IN FILE
NT  - UI - 94344236LA - engRN - 0 (Nucleotides)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19940916IS - 0028-0836SB - IMCY - ENGLANDJC - NSC
UR  - PM:8065448
SO  - Nature 1994 Aug 25 ;370(6491):621-628

1192
UI  - 21150
AU  - Alexiev U
AU  - Marti T
AU  - Heyn MP
AU  - Khorana HG
AU  - Scherrer P
AD  - Department of Physics, Freie Universitat Berlin, Germany
TI  - Surface charge of bacteriorhodopsin detected with covalently bound pH indicators at selected extracellular and cytoplasmic sites
AB  - We present a method that allows the detection of the surface charge density of bacteriorhodopsin (bR) at any selected protein surface site. The optical pH indicator fluorescein was covalently bound to the sulfhydryl groups of single cysteine residues, which were introduced at selected positions in bR by site-directed mutagenesis. On the extracellular side, the positions were in the BC loop (72) and in the DE loop (129-134). On the cytoplasmic side, one position in each loop was labeled: 35 (AB), 101 (CD), 160 (EF), and 231 (carboxy tail). The apparent pKs of fluorescein in these positions were determined for various salt concentrations. The local surface charge density was calculated from the dependence of the apparent pK of the dye on the ionic strength using the Gouy-Chapman equation. The surface charge density at pH 6.6 is more negative on the cytoplasmic side (averaged over all positions, -2.5 +/- 0.2 elementary charges per bR) than on the extracellular side (average, -1.8 +/- 0.2 elementary charges per bR) with little variation along the surface. Since the experiments were performed with electrically neutral CHAPS/DMPC micelles, these values represent the charge present on bR itself.(ABSTRACT TRUNCATED AT 250 WORDS)
MH  - A
MH  - Bacteriorhodopsin
MH  - Cysteine
MH  - DEPENDENCE
MH  - DYE
MH  - fluorescein
MH  - indicator
MH  - method
MH  - Micelles
MH  - mutagenesis
MH  - pH
MH  - pH-indicator
MH  - protein
MH  - Proteins
MH  - RESIDUE
MH  - Site
MH  - site-directed
MH  - SURFACE
RP  - NOT IN FILE
NT  - UI - 94114500LA - engRN - 0 (Recombinant Proteins)RN - 53026-44-1 (Bacteriorhodopsins)PT - Journal ArticleID - AI 11479-18/AI/NIAIDDA - 19940224IS - 0006-2960SB - IMCY - UNITED STATES
UR  - PM:8286351
SO  - Biochemistry 1994 Jan 11 ;33(1):298-306

1193
UI  - 21147
AU  - Alexiev U
AU  - Marti T
AU  - Heyn MP
AU  - Khorana HG
AU  - Scherrer P
AD  - Department of Physics, Freie Universitat Berlin, Germany
TI  - Covalently bound pH-indicator dyes at selected extracellular or cytoplasmic sites in bacteriorhodopsin. 2. Rotational orientation of helices D and E and kinetic correlation between M formation and proton release in bacteriorhodopsin micelles
AB  - The kinetics of the light-induced proton release in bacteriorhodopsin/lipid/detergent micelles was monitored with the optical pH-indicator fluorescein bound covalently to positions 127-134 (helices D and E and the DE loop) on the extracellular side of the protein (the proton release side). Single cysteine residues were introduced in these positions by site-directed mutagenesis, and fluorescein was attached to the sulfhydryl group by reaction with (iodoacetamido)fluorescein. Two characteristic proton release times (approximately 20 and 70 microseconds) were observed. The faster time constant was recorded when fluorescein was attached to positions 127, 130, 131, 132, and 134, while the slower time was observed with the indicator bound to positions 128, 129, and 133. The results are rationalized by assuming specific helical wheel orientations for helics D and E and by making a choice for the residues in the DE loop: (i) The fast time constants occur with fluorescein either attached to residues 130, 131, and 132 that form the DE loop or when pointing toward the interior of the protein with its aqueous proton channel [residues 127 (helix D) and 134 (helix E)]. (ii) The slower time constants are detected with fluorescein exposed to the exterior lipid/detergent phase when bound to residues 128, 129 (both helix D), and 133 (helix E). This interpretation is supported by measurements of the polarity of the label environment which indicate for fluorescein in group i a more hydrophilic environment and for group ii a more hydrophobic environment. The fastest proton release time (10 microseconds) was observed with fluorescein bound to position 127.(ABSTRACT TRUNCATED AT 250 WORDS)
MH  - A
MH  - Bacteriorhodopsin
MH  - CONSTANT
MH  - Cysteine
MH  - DYE
MH  - dyes
MH  - fluorescein
MH  - Fluoresceins
MH  - indicator
MH  - Kinetics
MH  - M
MH  - Micelles
MH  - microsecond
MH  - mutagenesis
MH  - pH-indicator
MH  - protein
MH  - Proteins
MH  - proton
MH  - proton release
MH  - Protons
MH  - RESIDUE
MH  - Site
MH  - site-directed
MH  - Time
RP  - NOT IN FILE
NT  - UI - 95034804LA - engRN - 0 (Fluoresceins)RN - 0 (Protons)RN - 0 (Recombinant Proteins)RN - 2321-07-5 (Fluorescein)RN - 52-90-4 (Cysteine)RN - 53026-44-1 (Bacteriorhodopsins)PT - Journal ArticleID - GM 28289/GM/NIGMSDA - 19941221IS - 0006-2960SB - IMCY - UNITED STATES
UR  - PM:7947778
SO  - Biochemistry 1994 Nov 22 ;33(46):13693-13699

1194
UI  - 733
AU  - Amano T
AU  - Yoshida M
AU  - Matsuo Y
AU  - Nishikawa K
AD  - Research Laboratory of Resources Utilization, R-1, Tokyo Institute of Technology, Yokohama, Japan
TI  - Structural model of the ATP-binding domain of the F1-beta subunit based on analogy to the RecA protein
AB  - In contrast to the previous topological model of the ATP binding domain of the F1-ATPase beta subunit based on analogies to those of ras p21 and adenylate kinase, a more consistent model can be constructed with the known structure of the recA protein as a reference. The secondary structure of the F1-ATPase beta subunit predicted from the primary structure agrees well with that of the recA protein. The topology includes a repetitive beta alpha c beta alpha beta alpha beta alpha beta structure where all beta strands are parallel and surround the central alpha c helix above which bound ATP is located. Several residues thought to be located at catalytic site as reported in genetic and chemical labeling work can be consistently positioned in this model
RP  - NOT IN FILE
NT  - UI - 94357258LA - engRN - 0 (Rec A Protein)RN - 55520-40-6 (Tyrosine)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.- (Oncogene Protein p21(ras))RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19941003IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:8076673
SO  - FEBS Lett 1994 Aug 29 ;351(1):1-5

1195
UI  - 734
AU  - Amano T
AU  - Tozawa K
AU  - Yoshida M
AU  - Murakami H
AD  - Research Laboratory of Resources Utilization, Tokyo Institute of Technology, Yokohama, Japan
TI  - Spatial precision of a catalytic carboxylate of F1-ATPase beta subunit probed by introducing different carboxylate-containing side chains
AB  - Combining mutation and chemical modification, we have introduced Asp, Gln, Cys, S-carboxymethylcysteine (Cax) and S-carbamoylmethylcysteine (Cam) into the positions of Glu190 and Glu201 of the beta subunit of F1- ATPase from the thermophilic Bacillus PS3. The steady-state ATPase activities of alpha 3 beta 3 gamma complexes containing these changed beta subunits were 12% (E190Cax), 7% (E190D), 3% (E190Cam), < 1% (E190C), < 1% (E190Q), and 73% (E201D), 40% (E201Cax), 25% (E201C), 20% (E201Q), 4% (E201Cam), of that of that of the wild-type alpha 3 beta 3 gamma complex. For the complexes containing E190C or E190Q, even the ability of single-site catalysis was lost. Thus, the presence of a carboxylate at 190 (but not at 201) is absolutely required for catalysis and its spatial precision is very strict. Analysis of inactivation of the complexes by dicyclohexylcarbodiimide suggests that Glu190 and Glu201 are interacting in the F1-ATPase
RP  - NOT IN FILE
NT  - UI - 94298962LA - engRN - 0 (Amides)RN - 0 (Carboxylic Acids)RN - 0 (Glutamates)RN - 0 (Sulfhydryl Reagents)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-86-0 (Glutamic Acid)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19940809IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:7913050
SO  - FEBS Lett 1994 Jul 4 ;348(1):93-98

1196
UI  - 21162
AU  - Angov E
AU  - Brusilow WS
AD  - Department of Biochemistry, Wayne State University School of Medicine, Detroit, MI 48201
TI  - Effects of deletions in the uncA-uncG intergenic regions on expression of uncG, the gene for the gamma subunit of the Escherichia coli F1Fo- ATPase
AB  - The gamma subunit of the E. coli F1Fo-ATPase is coded for by uncG. This gene is poorly expressed compared to uncA (alpha subunit), which precedes uncG in the unc operon. The genes are separated by a 50- nucleotide intergenic region. We examined the effects of a set of deletions in this region on the relative expression of uncA'-'lacZ and uncG'-'lacZ translational fusion genes located either in the chromosomal unc operon or at the chromosomal lambda att site. The gene for the alpha subunit was expressed 3-6-times better than the gene for the gamma subunit, depending upon chromosomal location. Deletion analysis revealed that the uncA-uncG intergenic region significantly affects expression of uncG, but the Shine-Dalgarno region is not absolutely required for expression of uncG. Different deletions resulted in either increased or decreased expression of uncG
MH  - A
MH  - alpha
MH  - ALPHA-SUBUNIT
MH  - analysis
MH  - ATPase
MH  - Biochemistry
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - Proton-Translocating ATPases
MH  - Site
MH  - SUBUNIT
RP  - NOT IN FILE
NT  - UI - 94114553LA - engRN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleID - KO4-AI00882/AI/NIAIDDA - 19940224IS - 0006-3002SB - IMCY - NETHERLANDS
UR  - PM:8286398
SO  - Biochim Biophys Acta 1994 Jan 4 ;1183(3):499-503

1197
UI  - 511
AU  - Bianchet M
AU  - Medjahed D
AU  - Hulihen J
AU  - Pedersen PL
AU  - Amzel LM
AD  - Department of Biophysics and Biophysical Chemistry, Johns Hopkins University, School of Medicine, Baltimore, MD 21205
TI  - The three-dimensional structure of rat liver mitochodria F1-ATPase: X- ray diffraction studies
RP  - NOT IN FILE
NT  - UI - 94355333LA - engRN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19941003IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:8075110
SO  - Biochim Biophys Acta 1994 Aug 30 ;1187(2):163-164

1198
UI  - 21160
AU  - Brusilow WS
AU  - Monticello RA
AD  - Department of Biochemistry, Wayne State University School of Medicine, Detroit, Michigan 48201
TI  - Synthesis and assembly of the F0 proton channel from F0 genes cloned into bacteriophage lambda and integrated into the Escherichia coli chromosome
AB  - The promoter region and the first four genes of the Escherichia coli proton-translocating ATPase (unc) operon, uncIBEF, were cloned into bacteriophage lambda, enabling this region to be recombined into an unc- deleted E. coli chromosome at the lambda att site. The resultant E. coli strain, carrying single-copy F0 genes, was tested for synthesis and assembly of functional F0 proton channels. Membranes isolated from this strain contained all three F0 subunits and were capable of binding purified F1 and reconstituting F1F0-dependent energy coupling activities. The presence of these F0 sectors did not affect cell growth or membrane proton permeability assayed by fluorescence quenching. When compared with wild type membranes, membranes from the single-copy F0 strain contained less a and b subunits. When the single-copy lambda F0 strain was transformed with an F1 plasmid, the cells became phenotypically and biochemically Unc+, with membrane-bound ATPase and ATP synthase activities that were 50-60% of wild type. The results demonstrate that F0 produced from single-copy genes in the absence of F1 is membrane-bound and functional (i.e. reconstitutable) but not freely permeable to protons. The presence of F1 genes and/or subunits during F0 synthesis and assembly both increases the relative amounts of membrane-bound a and b subunits and produces an F0 sector more like that found in wild type cells than is produced from the single-copy F0 genes alone
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BINDING
MH  - Biochemistry
MH  - Cells
MH  - coupling
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - F1
MH  - fluorescence
MH  - membrane
MH  - Membranes
MH  - Permeability
MH  - proton
MH  - Proton-Translocating ATPases
MH  - Protons
MH  - Site
MH  - SUBUNIT
MH  - SYNTHASE
MH  - synthesis
RP  - NOT IN FILE
NT  - UI - 94171743LA - engRN - 0 (Plasmids)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleID - KO4-AI00882/AI/NIAIDDA - 19940412IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:8125942
SO  - J Biol Chem 1994 Mar 11 ;269(10):7285-7289

1199
UI  - 20943
AU  - Burkovski A
AU  - Deckers-Hebestreit G
AU  - Altendorf K
AD  - Universitat Osnabruck, Fachbereich Biologie/Chemie, Germany
TI  - Hybrid Fo complexes of the ATP synthases of spinach chloroplasts and Escherichia coli. Immunoprecipitation and mutant analyses
AB  - Hybrid Fo complexes of the ATP synthases of spinach chloroplast (CFo) and Escherichia coli (EFo) were investigated. Immunoprecipitations with polyclonal antibodies against the different Fo subunits clearly revealed that hybrid Fo complexes derived from CFo subunit III and EFo subunits a and b were formed in vivo. In addition, the ATPase activities of the hybrid ATP synthase, measured in everted cytoplasmic membranes of an atpE mutant strain transformed with the atpH gene coding for CFo III, were comparable to activities obtained for the same mutant strain complemented with the atpE gene (EFo c). Nevertheless, CFo III was not able to replace EFo c functionally, since the strain containing the hybrid ATP synthase was not able to grow on succinate. In order to investigate the reason for this lack of function, hybrid proteolipids of CFo III and EFo c were constructed. Only a chimaeric protein comprising the seven N-terminal amino acid residues from CFo III and the remaining part of EFo c was able to replace wild-type EFo c, whereas hybrid proteins with 13 and 33 N-terminal amino acids of CFo III were not functional. The results suggested that a network of interactions between the subunits essential for proton translocation and/or coupling of the F1 part exists, which was optimized for each species during evolution, although the overall structure of FoF1 complexes has been conserved
MH  - A
MH  - ACID
MH  - Amino Acids
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - chloroplast
MH  - Chloroplasts
MH  - COMPLEX
MH  - coupling
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - Evolution
MH  - F1
MH  - function
MH  - membrane
MH  - Membranes
MH  - mutant
MH  - protein
MH  - Proteins
MH  - Proteolipids
MH  - proton
MH  - RESIDUE
MH  - Spinach
MH  - SPINACH-CHLOROPLASTS
MH  - SUBUNIT
MH  - succinate
MH  - SYNTHASE
MH  - translocation
RP  - NOT IN FILE
NT  - UI - 95045552LA - engRN - 0 (Chimeric Proteins)RN - 0 (Genetic Vectors)RN - 0 (Proteolipids)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19941221IS - 0014-2956SB - IMCY - GERMANY
UR  - PM:7957212
SO  - Eur J Biochem 1994 Nov 1 ;225(3):1221-1228

1200
UI  - 166
AU  - Capaldi DC
AU  - Reese CB
AD  - Department of Chemistry, King's College London, UK
TI  - Use of the 1-(2-fluorophenyl)-4-methoxypiperidin-4-yl (Fpmp) and related protecting groups in oligoribonucleotide synthesis: stability of internucleotide linkages to aqueous acid
AB  - The internucleotide linkage of uridylyl-(3'-->5')-uridine (r[UpU]) does not undergo detectable hydrolytic cleavage or migration in ca. 24 hr in 0.01 mol dm-3 hydrochloric acid (pH 2.0) at 25 degrees C. However, unlike r[UpU] and previously examined relatively high molecular weight oligoribonucleotides, oligouridylic acids are very sensitive to aqueous acid under the latter conditions (pH 2.0, 25 degrees C). Thus when the 1-(2-fluorophenyl)-4-methoxypiperidin-4-yl (Fpmp) group is used to protect the 2'-hydroxy functions in the synthesis of r[(Up)9U] and r[(Up)19U], the final unblocking process must be carried out above pH 3 if hydrolytic cleavage and migration are to be avoided. It is demonstrated that the rate of acid-catalyzed hydrolysis of the internucleotide linkages of oligoribonucleotides is sequence dependent. As Fpmp groups may be virtually completely removed from average partially-protected oligoribonucleotides within ca. 24 hr at pH 3 and 25 degrees C, it is concluded that Fpmp is a suitable 2'-protecting group even in the synthesis of particularly acid-sensitive sequences
RP  - NOT IN FILE
NT  - UI - 94310045LA - engRN - 0 (Acids)RN - 0 (Oligoribonucleotides)RN - 0 (Piperidines)RN - 7732-18-5 (Water)PT - Journal ArticleDA - 19940815IS - 0305-1048SB - IMCY - ENGLANDJC - O8L
UR  - PM:8036146
SO  - Nucleic Acids Res 1994 Jun 25 ;22(12):2209-2216

1201
UI  - 164
AU  - Capaldi RA
TI  - F1-ATPase in a spin
RP  - NOT IN FILE
NT  - UI - 95360705LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - NewsDA - 19950914IS - 1072-8368SB - IMCY - UNITED STATESJC - B98
UR  - PM:7634064
SO  - Nat Struct Biol 1994 Oct ;1(10):660-663

1202
UI  - 165
AU  - Capaldi RA
AU  - Aggeler R
AU  - Turina P
AU  - Wilkens S
AD  - Institute of Molecular Biology, University of Oregon, Eugene 97403
TI  - Coupling between catalytic sites and the proton channel in F1F0-type ATPases
AB  - F1F0-type ATPases catalyse both ATP-driven proton translocation and proton-gradient-driven ATP synthesis. Recent cryoelectronmicroscopy and low-resolution X-ray studies provide a first glimpse at the structure of this complicated membrane-bound enzyme. The F1 part is roughly globular and linked to the membrane-intercalated F0 part by a narrow stalk domain, which contains the gamma-, delta- and epsilon-subunits along with domains of the b-subunit of the F0 part. Here, we review evidence that conformational and positional changes in the gamma- and epsilon-subunits provide the coupling between catalytic sites and proton translocation within the F1F0 complex
RP  - NOT IN FILE
NT  - UI - 94324085LA - engRN - 0 (Nucleotides)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialID - HL 24526/HL/NHLBIDA - 19940831IS - 0968-0004SB - IMCY - ENGLANDJC - WEF
UR  - PM:8048168
SO  - Trends Biochem Sci 1994 Jul ;19(7):284-289

1203
UI  - 342
AU  - Collinson IR
AU  - Runswick MJ
AU  - Buchanan SK
AU  - Fearnley IM
AU  - Skehel JM
AU  - van Raaij MJ
AU  - Griffiths DE
AU  - Walker JE
AD  - MRC Laboratory of Molecular Biology, Cambridge, UK
TI  - Fo membrane domain of ATP synthase from bovine heart mitochondria: purification, subunit composition, and reconstitution with F1-ATPase
AB  - The Fo membrane domain of the F1Fo-ATP synthase complex has been purified from bovine heart mitochondria. The purification procedure involves the removal of peripheral membrane proteins, including F1- ATPase, from submitochondrial particles with guanidine hydrochloride, followed by extraction of Fo and other membrane proteins from the stripped membranes in the presence of the detergent n-dodecyl beta-D- maltoside. Fo was then purified by ion-exchange and dye ligand chromatography in the presence of the same detergent. Approximately 15 mg of pure Fo was recovered from 1.8 g of mitochondrial membrane protein. The purified Fo is a complex of nine different polypeptides. They are subunits a, b, c, d, e, F6, and A6L characterized before in F1Fo-ATPase preparations, and two new hitherto undetected subunits, named f and g. The sequences of subunits f and g have been determined. They are not related significantly to any known protein, but subunit f appears to contain a membrane-spanning alpha-helix. Proteins f and g are also present in approximately stoichiometric amounts in a highly purified preparation of intact F1Fo-ATPase, and so it is concluded that they are authentic subunits of the bovine enzyme with unknown functions. Dibutyltin 3-hydroxyflavone, an inhibitor of F1Fo-ATPase, also binds to the purified Fo in detergent and competes for binding with venturicidin. In the presence of F1 and OSCP, the purified Fo was reassembled into the intact F1Fo-ATPase complex. Therefore, this procedure provides a relatively abundant source of pure and functional Fo that is suitable for structural analysis
RP  - NOT IN FILE
NT  - UI - 94281230LA - engRN - 0 (Macromolecular Systems)RN - 0 (Membrane Proteins)RN - 0 (oligomycin sensitivity-conferring protein)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19940726IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:8011660
SO  - Biochemistry 1994 Jun 28 ;33(25):7971-7978

1204
UI  - 340
AU  - Collinson IR
AU  - van Raaij MJ
AU  - Runswick MJ
AU  - Fearnley IM
AU  - Skehel JM
AU  - Orriss GL
AU  - Miroux B
AU  - Walker JE
AD  - Medical Research Council Laboratory of Molecular Biology, Cambridge, UK
TI  - ATP synthase from bovine heart mitochondria. In vitro assembly of a stalk complex in the presence of F1-ATPase and in its absence
AB  - Four subunits of the F1F0-ATPase from bovine heart mitochondria have been produced by heterologous over-expression in Escherichia coli. They are the oligomycin sensitivity conferral protein (OSCP), coupling factor 6 (F6) and subunits b and d. Likewise, fragments b', bI, bC, and bM (amino acid residues 79 to 214, 121 to 214, 165 to 214 and 79 to 164, respectively, of subunit b), and fragment d' (subunit d lacking residue 1 to 14) have been produced in abundant quantities by bacterial expression. These subunits, and the fragments of subunits b and d, have been assayed singly and in various combinations by gel-filtration chromatography for their abilities to bind to bovine heart F1-ATPase. Only the OSCP was found to be capable of forming a stable binary complex with F1-ATPase. When fragments b', bI or bC were added to F1- ATPase together with the OSCP, the ternary complexes F1.OSCP.b', F1.OSCP.bI or F1.OSCP.bC were formed, but b', bI and bC appeared to be present in sub-stoichiometric amounts. When F6 was added also, then the stoichiometric quaternary complexes F1.OSCP.b'.F6 and F1.OSCP.bI.F6 were obtained, as was a fourth quaternary complex containing approximately equivalent amounts of F1 and OSCP, and sub-stoichiometric quantities of bC and F6. Finally, three pentameric complexes F1.OSCP.b'.F6.d, F1.OSCP.b'.F6.d' and F1.OSCP.b.F6.d were isolated. In a further series of reconstitution experiments, the binary complexes b'.OSCP and b'.d, the ternary complex b'.d'.F6, and the quaternary complex OSCP.b'.F6.d were obtained. The pre-formed quaternary complex produced a stoichiometric pentameric complex with F1-ATPase. It was shown by S-carboxymethylation of cysteine residues with iodo-[2- 14C]acetic acid that bovine F1F0-ATPase and the reconstituted F1.stalk complex, F1.OSCP.b'.d.F6, each contained one copy per complex of subunits b (or b'), OSCP and d, and that the separate stalk complex contained the same three subunits in the approximate molar ratio 1:1:1. The ratio of b to d in purified F0 was 1:1. Finally, it was demonstrated that the binding of the various subunits to F1-ATPase increases the ATP hydrolase activity and diminishes its inactivation by exposure to cold. These assembly experiments help to define some of the inter-subunit interactions in the stalk region of the F1F0-ATPase complex, and they are an essential step forward towards the goal of extending the high-resolution structure of bovine F1-ATPase into the stalk
RP  - NOT IN FILE
NT  - UI - 95018208LA - engRN - 0 (DNA Primers)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19941102IS - 0022-2836SB - IMCY - ENGLANDJC - J6V
UR  - PM:7932700
SO  - J Mol Biol 1994 Sep 30 ;242(4):408-421

1205
UI  - 339
AU  - Collinson IR
AU  - Fearnley IM
AU  - Skehel JM
AU  - Runswick MJ
AU  - Walker JE
AD  - Medical Research Council Laboratory of Molecular Biology, Cambridge, UK
TI  - ATP synthase from bovine heart mitochondria: identification by proteolysis of sites in F0 exposed by removal of F1 and the oligomycin- sensitivity conferral protein
AB  - The exposure to trypsinolysis of subunits of F1F0-ATPase and of its F0 domain have been compared in everted inner membrane vesicles (submitochondrial particles) made from bovine mitochondria. Treatment of submitochondrial particles with guanidine hydrochloride removed the subunits of F1-ATPase and the oligomycin-sensitivity conferral protein (OSCP), and exposed sites that were occluded in the intact F1F0-ATPase complex. These sites were identified by purifying the subunits from the isolated F0 and F1F0-ATPase complexes before and after proteolysis of the vesicles, and by characterizing them by N-terminal sequencing and electrospray-ionization mass spectrometry. In the stripped vesicles, subunit F6 was completely digested away by either trypsin or chymotrypsin. Trypsin also cleaved subunit b, first at the bond arginine-166-glutamine-167, and then at the consecutive linkages, lysine-120-arginine-121 and arginine-121-histidine-122. Chymotrypsin- sensitive sites were observed after the adjacent methionines 164 and 165. Trypsin also removed amino acids 1-3 of subunit d, and minor cleavage sites were observed in subunit d between amino acids 24 and 25, in subunit g between amino acids 5 and 6, and after amino acid 40 in subunit e. The other subunits remained protected from proteolysis. In membrane-bound F1F0-ATPase, the N-terminus of subunit d was also accessible to trypsin, and subunit e was more susceptible to proteolysis than in F0. Otherwise the F0 subunits and the OSCP were protected. Subunits alpha and beta were cleaved by trypsin at the same sites in their N-terminal regions as in purified F1-ATPase. The trypsinized F0 was incapable of binding F1-ATPase in the presence of the OSCP. These experiments and in vitro re-assembly experiments described elsewehere, that were guided by the results of the proteolysis experiments, have helped to establish a central role for subunit b in the formation of the stalk connecting the F1 and F0 domains of the F1F0-ATPase complex
RP  - NOT IN FILE
NT  - UI - 95071270LA - engRN - 0 (Guanidines)RN - 0 (Membrane Proteins)RN - 0 (Oligomycins)RN - 0 (oligomycin sensitivity-conferring protein)RN - 113-00-8 (Guanidine)RN - EC 3.4.21.1 (Chymotrypsin)RN - EC 3.4.21.4 (Trypsin)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19941130IS - 0264-6021SB - IMCY - ENGLANDJC - 9YO
UR  - PM:7980427
SO  - Biochem J 1994 Oct 15 ;303 ( Pt 2)():639-645

1206
UI  - 20994
AU  - Creczynski-Pasa TB
AU  - Graber P
AD  - Biologisches Institut, Universitat Stuttgart, Germany
TI  - ADP binding and ATP synthesis by reconstituted H(+)-ATPase from chloroplasts
AB  - The H(+)-ATPase from chloroplasts, CF0F1, was isolated, purified and reconstituted into asolectin liposomes. The enzyme was brought either into the oxidized state or into the reduced state, and the rate of ATP synthesis was measured after energisation of the proteoliposomes with an acid-base transition delta pH (pHin = 5.0, pHout = 8.5) and a K+/valinomycin diffusion potential, delta phi (Kin+ = 0.6 mM, Kout+ = 60 mM). A rate of 250 s-1 was observed with the reduced enzyme (85 s-1 in the absence of delta phi). A rate of 50 s-1 was observed with the oxidized enzyme under the same conditions (15 s-1 in the absence of delta phi). The reconstituted enzyme contained 2 ATPbound per CF0F1 and 1 ADPbound per CF0F1. Upon energisation the enzyme was activated and 0.9 ADP per CF0F1 was released. Binding of ADP to the active reduced enzyme was observed under different conditions. In the absence of phosphate the rate constant for ADP binding was 10(5) M-1.s-1 under energized and de-energized conditions. In the presence of phosphate the rate of ADP binding drastically increased under energized conditions, and strongly decreased under de-energized conditions
MH  - A
MH  - ACTIVE
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - ADP
MH  - ADP BINDING
MH  - atp
MH  - ATP synthesis
MH  - ATPase
MH  - BINDING
MH  - CF0F1
MH  - chloroplast
MH  - Chloroplasts
MH  - CONSTANT
MH  - DELTA-PH
MH  - Diffusion
MH  - diffusion potential
MH  - H(+)ATPase
MH  - Liposomes
MH  - pH
MH  - Proteolipids
MH  - proteoliposome
MH  - rate constant
MH  - synthesis
RP  - NOT IN FILE
NT  - UI - 94350097LA - engRN - 0 (Proteolipids)RN - 0 (proteoliposomes)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19940926IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:8070563
SO  - FEBS Lett 1994 Aug 22 ;350(2-3):195-198

1207
UI  - 19863
AU  - Cross RL
TI  - Enzyme structure. Our primary source of ATP
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthase
MH  - England
MH  - H(+)-Transporting ATP Synthase
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 94344231LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - CommentPT - NewsDA - 19940916IS - 0028-0836SB - IMCY - ENGLANDJC - NSC
UR  - PM:8065443
SO  - Nature 1994 Aug 25 ;370(6491):594-595

1208
UI  - 61
AU  - Dimroth P
AD  - Mikrobiologisches Institut, Eidgenossische Technische Hochschule, ETH- Zentrum, Zurich, Switzerland
TI  - Bacterial sodium ion-coupled energetics
AB  - For many bacteria Na+ bioenergetics is important as a link between exergonic and endergonic reactions in the membrane. This article focusses on two primary Na+ pumps in bacteria, the Na(+)-translocating oxaloacetate decarboxylase of Klebsiella pneumoniae and the Na(+)- translocating F1Fo ATPase of Propionigenium modestum. Oxaloacetate decarboxylase is an essential enzyme of the citrate fermentation pathway and has the additional function to conserve the free energy of decarboxylation by conversion into a Na+ gradient. Oxaloacetate decarboxylase is composed of three different subunits and the related methylmalonyl-CoA decarboxylase consists of five different subunits. The genes encoding these enzymes have been cloned and sequenced. Remarkable are large areas of complete sequence identity in the integral membrane-bound beta-subunits including two conserved aspartates that may be important for Na+ translocation. The coupling ratio of the decarboxylase Na+ pumps depended on delta muNa+ and decreased from two to zero Na+ uptake per decarboxylation event as delta mu Na+ increased from zero to the steady state level. In P. modestum, delta mu Na+ is generated in the course of succinate fermentation to propionate and CO2. This delta mu Na+ is used by a unique Na(+)-translocating F1Fo ATPase for ATP synthesis. The enzyme is related to H(+)-translocating F1Fo ATPases. The Fo part is entirely responsible for the coupling of ion specificity. A hybrid ATPase formed by in vivo complementation of an Escherichia coli deletion mutant was completely functional as a Na(+)-ATP synthase conferring the E. coli strain the ability of Na(+)-dependent growth on succinate. The hybrid consisted of subunits a, c, b, delta and part of alpha from P. modestum and of the remaining subunits from E. coli. Studies on Na+ translocation through the Fo part of the P. modestum ATPase revealed typical transporter-like properties. Sodium ions specifically protected the ATPase from the modification of glutamate-65 in subunit c by dicyclohexylcarbodiimide in a pH-dependent manner indicating that the Na+ binding site is at this highly conserved acidic amino acid residue of subunit c within the middle of the membrane
RP  - NOT IN FILE
NT  - UI - 95134027LA - engRN - 0 (Citrates)RN - 56-65-5 (Adenosine Triphosphate)RN - 7440-23-5 (Sodium)RN - 77-92-9 (Citric Acid)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)RN - EC 3.6.1.37 (Na(+)-K(+)-Exchanging ATPase)RN - EC 4.1.1. (Carboxy-Lyases)RN - EC 4.1.1.3 (oxaloacetate decarboxylase)RN - EC 4.1.1.41 (propionyl-CoA carboxylase)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 19950217IS - 0003-6072SB - IMCY - NETHERLANDSJC - 6JE
UR  - PM:7832594
SO  - Antonie Van Leeuwenhoek 1994  ;65(4):381-395

1209
UI  - 20993
AU  - Fischer S
AU  - Etzold C
AU  - Turina P
AU  - Deckers-Hebestreit G
AU  - Altendorf K
AU  - Graber P
AD  - Biologisches Institut, Universitat Stuttgart, Germany
TI  - ATP synthesis catalyzed by the ATP synthase of Escherichia coli reconstituted into liposomes
AB  - The H(+)-translocating F0F1-ATPase from Escherichia coli (EF0F1) was purified and reconstituted into preformed reverse-phase liposomes prepared from egg yolk phosphatidylcholine/phosphatidic acid. The EF0F1 liposomes were energized by an acid/base transition (pHout = 8.3; pHin = 5.0) and a superimposed K+/valinomycin diffusion potential ([K+]out = 100 mM; [K+]in = 0.6 mM) yielding a maximum rate (turnover number) of ATP synthesis of 27 +/- 8 mol ATP . mol EF0F1(-1) . s-1), i.e. 27 +/- 8 s-1. This reaction was inhibited by NH4Cl or by addition of the F0F1 inhibitor N,N'-dicyclohexylcarbodiimide. The rate of ATP synthesis measured as a function of the phosphate and ADP concentrations, can be described by Michaelis-Menten kinetics with a Km of 0.7 +/- 0.2 mM for phosphate ([ADP] = 200 microM) and a Km of 27 +/- 7 microM for ADP ([phosphate] = 5 mM), respectively
MH  - A
MH  - ACID
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - ADP
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - ATPase
MH  - Diffusion
MH  - diffusion potential
MH  - EF0F1
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0F1
MH  - function
MH  - Kinetics
MH  - Liposomes
MH  - Phosphates
MH  - Phosphatidic Acids
MH  - Phosphatidylcholines
MH  - SYNTHASE
MH  - synthesis
RP  - NOT IN FILE
NT  - UI - 95010102LA - engRN - 0 (Liposomes)RN - 0 (Phosphates)RN - 0 (Phosphatidic Acids)RN - 0 (Phosphatidylcholines)RN - 12125-02-9 (Ammonium Chloride)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19941123IS - 0014-2956SB - IMCY - GERMANY
UR  - PM:7925434
SO  - Eur J Biochem 1994 Oct 1 ;225(1):167-172

1210
UI  - 35
AU  - Fraga D
AU  - Hermolin J
AU  - Fillingame RH
AD  - Department of Biomolecular Chemistry, University of Wisconsin Medical School, Madison 53706
TI  - Transmembrane helix-helix interactions in F0 suggested by suppressor mutations to Ala24-->Asp/Asp61-->Gly mutant of ATP synthase subunit
AB  - A mutant of ATP synthase subunit c was isolated in which the essential aspartate was exchanged from position 61 on transmembrane helix-2 to position 24 on transmembrane helix-1 (Miller, M. J., Oldenburg, M., and Fillingame, R. H. (1990) Proc. Natl. Acad. Sci. U. S. A. 87, 4900- 4904). The H+ transporting ATP synthase function of the Ala24-- >Asp/Asp61-->Gly mutant is not optimal, and cells grow more slowly than wild type. Twenty-three third-site suppressor mutants with optimized function were isolated in this study. Ten of the optimizing mutations were located to helix-2 of subunit c, and seven of these fell in residues Phe53, Met57, and Met65. The side chains of these three residues are proposed to form a hydrophobic surface on transmembrane helix-2, which participates in the presentation or occlusion of the essential aspartate carboxyl group during proton translocation. The other 13 optimizing mutations were located to subunit a, and 10 of these fell in residues Ala217, Ile221, and Leu224. These three residues are proposed to lie on one face of a transmembrane alpha-helix that includes the essential Arg210 residue. This helix is proposed to interact with the transmembrane bihelical unit of subunit c during protonation and deprotonation of the essential Asp24 in the mutant or Asp61 in wild type
RP  - NOT IN FILE
NT  - UI - 94132018LA - engRN - 0 (DNA, Bacterial)RN - 0 (Macromolecular Systems)RN - 56-40-6 (Glycine)RN - 56-84-8 (Aspartic Acid)RN - 6898-94-8 (Alanine)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - 5-T32-GM07133/GM/NIGMSID - GM23105/GM/NIGMSDA - 19940304IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:8300584
SO  - J Biol Chem 1994 Jan 28 ;269(4):2562-2567

1211
UI  - 113
AU  - Fraga D
AU  - Hermolin J
AU  - Oldenburg M
AU  - Miller MJ
AU  - Fillingame RH
AD  - Department of Biomolecular Chemistry, University of Wisconsin Medical School, Madison 53706
TI  - Arginine 41 of subunit c of Escherichia coli H(+)-ATP synthase is essential in binding and coupling of F1 to F0
AB  - Two substitutions were made for Arg41 in the polar loop of subunit c of the Escherichia coli F1F0 H(+)-transporting ATP synthase. The R41K and R41H mutants were initially studied by use of a plasmid carrying the complete c R41K or c R41H unc (F1F0) operon in a chromosomal strain deleted for the unc operon. The extent of F0 incorporation into membranes of these cells was quite variable, and the system was concluded to be unsuitable for biochemical characterization. Ultimately, the mutant genes were recombined into the chromosome using a novel method for the unc system. The biochemical phenotype of the chromosomally expressed mutants proved to be reproducible. The c R41H mutation causes a specific defect in assembly of F0, i.e. subunit a was not incorporated into the membrane despite near normal incorporation of subunits b and c. On the other hand, c R41K mutant F0 assembled normally in one of two background strains studied. (In the second genetic background, subunit a was inefficiently incorporated into the c R41K membrane.) In membranes prepared from a c R41K strain assembling a complete F0, R41K F0 was found to bind F1 with near normal affinity and to transport H+ at near normal rates. Although R41K F0 binds F1, F1- ATPase activity and H+ transport remained uncoupled. The uncoupling was indicated by a lack of ATP-driven H+ translocation and by the high proton permeability of membranes with F1 bound to F0. The uncoupled phenotype of the R41K mutant closely resembles that previously reported for the c Q42E mutant
RP  - NOT IN FILE
NT  - UI - 94171781LA - engRN - 0 (Oligodeoxyribonucleotides)RN - 0 (Protons)RN - 56-65-5 (Adenosine Triphosphate)RN - 7004-12-8 (Arginine)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM23105/GM/NIGMSDA - 19940412IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:8125974
SO  - J Biol Chem 1994 Mar 11 ;269(10):7532-7537

1212
UI  - 20855
AU  - Futai M
AU  - Park M
AU  - Iwamoto A
AU  - Omote H
AU  - Maeda M
AD  - Department of Organic Chemistry and Biochemistry, Osaka University, Japan
TI  - Catalysis and energy coupling of H(+)-ATPase (ATP synthase): molecular biological approaches
AB  - The molecular biological approach has provided important information for understanding the F0F1 H(+)-ATPase. This article focuses on our recent results on the catalytic site in the beta subunit, and the roles of alpha/beta subunit interaction and amino/carboxyl terminal interaction of the gamma subunit in energy coupling. Extensive mutagenesis of the beta subunit revealed that beta Lys-155, beta Thr- 156, beta Glu-181 and beta Arg-182 are essential catalytic residues. beta Glu-185 is not absolutely essential, but a carboxyl residue may be necessary at this position. A pseudo-revertant analysis positioned beta Gly-172, beta Ser-174, beta Glu-192 and beta Val-198 in the proximity of beta Gly-149. The finding of the roles of beta Gly-149, beta Lys- 155, and beta Thr-156 emphasized the importance of the glycine-rich sequence (Gly-X-X-X-X-Gly-Lys-Thr/Ser, E. coli beta residues between beta Gly-149 and beta Thr-156) conserved in many nucleotide binding proteins. The A subunits of vacuolar type ATPases may have a similar catalytic mechanism because they have conserved glycine-rich and Gly- Glu-Arg (corresponding to beta Gly-180-beta Arg-182) sequences. The results of these mutational studies are consistent with the labeling of beta Lys-155 and beta Lys-201 with AP3-PL, and of beta Glu-192 with DCCD [15]. The DCCD-binding residue of a thermophilic Bacillus corresponds to beta Glu-181, an essential catalytic residue discussed above. The defective coupling of the beta Ser-174-->Phe mutant was suppressed by the second mutation alpha Arg-296-->Cys, indicating the importance of alpha/beta interaction in energy coupling. The gamma subunit, especially its amino/carboxyl interaction, seems to be essential for energy coupling between catalysis and transport judging from studies on gamma Met-23-->Lys or Arg mutation and second-site mutations which suppressed the gamma Lys-23 mutation. Thus the conserved gamma Met-23 is not absolutely essential but is located in the important region for amino/carboxyl interaction for energy coupling
MH  - A
MH  - analysis
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Bacillus
MH  - BETA-SUBUNIT
MH  - BINDING
MH  - Biochemistry
MH  - Catalysis
MH  - Chemistry
MH  - coupling
MH  - F0F1
MH  - H(+)ATPase
MH  - mechanism
MH  - mutagenesis
MH  - mutant
MH  - nucleotide binding
MH  - protein
MH  - Proteins
MH  - RESIDUE
MH  - Site
MH  - SUBUNIT
MH  - SYNTHASE
MH  - transport
RP  - NOT IN FILE
NT  - UI - 94355334LA - engRN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19941003IS - 0006-3002SB - IMCY - NETHERLANDS
UR  - PM:8075111
SO  - Biochim Biophys Acta 1994 Aug 30 ;1187(2):165-170

1213
UI  - 60
AU  - Gerike U
AU  - Dimroth P
AD  - Mikrobiologisches Institut, Eidgenossische Technische Hochschule, ETH Zentrum, Zurich, Switzerland
TI  - Expression of subunits a and c of the sodium-dependent ATPase of Propionigenium modestum in Escherichia coli
AB  - The aim of the present study was to construct functional hybrid ATPases consisting of all Escherichia coli ATPase subunits excepts the F0 subunits a or c which were replaced by the respective subunits of the Propionigenium modestum ATPase. This would give valuable information on the subunit(s) conferring the coupling ion specificity. Plasmids were constructed that carried the gene for subunit c (uncE) or subunit a (uncB) behind a tac promoter. These plasmids were transformed into E. coli strains which differed with respect to the unc operon and the expression of the P. modestum genes was verified biochemically. Enhanced expression of the P. modestum genes led to strong growth inhibition of all E. coli strains tested. However, the expressed P. modestum proteins could not functionally complement E. coli strains that lacked the homologous subunit
RP  - NOT IN FILE
NT  - UI - 94324847LA - engRN - 0 (Bacterial Proteins)RN - 0 (Chimeric Proteins)RN - 0 (Protons)RN - 0 (uncB protein, Escherichia coli)RN - 0 (uncE protein, Escherichia coli)RN - 7440-23-5 (Sodium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19940901IS - 0302-8933SB - IMCY - GERMANYJC - 7YN
UR  - PM:8048841
SO  - Arch Microbiol 1994  ;161(6):495-500

1214
UI  - 114
AU  - Girvin ME
AU  - Fillingame RH
AD  - Department of Biomolecular Chemistry, University of Wisconsin, Madison 53706
TI  - Hairpin folding of subunit c of F1Fo ATP synthase: 1H distance measurements to nitroxide-derivatized aspartyl-61
AB  - Subunit c from the F1Fo ATP synthase of Escherichia coli folds in a hairpinlike structure of two alpha-helices in a solution of chloroform- methanol-H2O, and thus resembles the structure predicted for the folded protein in the membrane. The relevance of the structure in solution to the native structure was demonstrated. Asp61 in the second helical arm was shown to retain its unique reactivity with dicyclohexylcarbodiimide (DCCD) in chloroform-methanol-H2O solution. Further, the protein purified from the Ile28-->Thr DCCD-resistant mutant proved to be less reactive with DCCD in solution. This suggested that the protein folded with Ile28 of the first helical arm close to Asp61 in the second helical arm. Subunit c in wild-type E. coli membranes was specifically labeled with a nitroxide analog of DCCD (NCCD), and the derivative protein was purified. DQF COSY spectra were recorded, and the distances between the paramagnetic nitroxide and resolved protons in the spectra were calculated based upon paramagnetic broadening of the 1H resonances. The paramagnetic contribution to T2 relaxation in the NCCD- labeled sample was calculated by an iterative computer-fitting method, where a control spectrum of a phenylhydrazine-reduced sample was broadened until the line shape of one-dimensional slices through each COSY cross-peak maximally mimicked the line shape of the paramagnetic sample. The distances calculated from paramagnetic broadening indicate that Ala24 and Ala25 in helix-1 lie close (ca. 12 A) to the derivatized Asp61 in helix-2. A model for the interaction of helices in the NCCD- modified protein was generated by restrained molecular mechanics and molecular dynamics using 25 distances of < 10-20 A derived from paramagnetic broadening in combination with 15 long-range nuclear Overhauser enhancement (NOE) restraints (2-5 A) for distances between helices and the 89 intrahelical NOEs that defined helical structure in the DCCD-modified protein
RP  - NOT IN FILE
NT  - UI - 94122177LA - engRN - 0 (Cyclic N-Oxides)RN - 42249-40-1 (N-(2,2,6,6-tetramethylpiperidyl-1-oxyl) N'- (cyclohexyl)carbodiimide)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-84-8 (Aspartic Acid)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM-23105/GM/NIGMSID - RR02301/RR/NCRRID - RR02781/RR/NCRRDA - 19940228IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:8292594
SO  - Biochemistry 1994 Jan 25 ;33(3):665-674

1215
UI  - 297
AU  - Gruber G
AU  - Engelbrecht S
AU  - Junge W
AU  - Dose K
AU  - Nawroth T
AD  - Institut fur Biochemie, Joh Gutenberg-Universitat, Mainz, Germany
TI  - Purification and characterization of the inhibitory subunit (delta) of the ATP-synthase from Micrococcus luteus
AB  - Subunit delta was isolated from the ATP-synthase from Micrococcus luteus strain (ATCC 4698). delta, in the case of M. luteus F0F1-ATPase, acts as an inhibitor of ATP hydrolysis and thus resembles subunits in E. coli and chloroplast ATP-synthase. After treatment with 1.5 M LiCl the ATP-synthase dissociated, and subsequently subunit delta (27 kDa) was purified by hydrophobic interaction chromatography. Inhibition of ATP-synthase lacking delta by addition of delta showed non-competitive kinetics with a Ki of approximately 5.9 nM. Subunit epsilon from chloroplast F1, which corresponds functionally to the M. luteus F0F1- delta, and chloroplast delta were tested for ATPase inhibitory activity by addition to the partially delta-depleted ATP-synthase from M. luteus. CF1-epsilon inhibited M. luteus ATP-synthase up to 80%, whereas CF1-delta did not show any influence
RP  - NOT IN FILE
NT  - UI - 95104420LA - engRN - 0 (Macromolecular Systems)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19950127IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:7805843
SO  - FEBS Lett 1994 Dec 19 ;356(2-3):226-228

1216
UI  - 21079
AU  - Guffanti AA
AU  - Krulwich TA
AD  - Department of Biochemistry, Mount Sinai School of Medicine, City University of New York, New York 10029
TI  - Oxidative phosphorylation by ADP + P(i)-loaded membrane vesicles of alkaliphilic Bacillus firmus OF4
AB  - ATP synthesis in ADP + P(i)-loaded membrane vesicles of the facultative alkaliphile Bacillus firmus OF4 at an external pH of 10.5 did not depend upon the presence of cell wall polymers, e.g. as a proton barrier or sequestration device. Upon energization with ascorbate plus phenazine methosulfate, vesicles at pH(out) = 7.5 generated an electrochemical proton gradient (delta p) of -160 mV, acid and positive out, whereas at pH(out) = 10.5, the delta p was -40 mV, alkaline and positive out. Nonetheless, ATP synthesis was more rapid at the more alkaline pH value, especially in the presence of 200 mM K2SO4, which markedly lowered the surface potential. No synthesis was observed upon abolition of the delta p. Respiration-derived transmembrane potentials (delta psi) energized ATP synthesis much better than an equally large diffusion potential. The diffusion potential failed to energize ATP synthesis above pH 9.5. When delta p, all in the form of a delta psi, was titrated downward at either pH 7.8 or 9.5, ATP synthesis by the latter vesicles was much less adversely affected in the delta p range of -150 to -50 mV, supporting the existence of a sparing, non- chemiosmotic energy component at high pH
MH  - A
MH  - ACID
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - ADP
MH  - Alkalies
MH  - atp
MH  - ATP synthesis
MH  - Bacillus
MH  - Biochemistry
MH  - buffer
MH  - Buffers
MH  - delta
MH  - Diffusion
MH  - diffusion potential
MH  - membrane
MH  - membrane vesicles
MH  - Oxidative Phosphorylation
MH  - P
MH  - pH
MH  - Phosphates
MH  - Phosphorylation
MH  - Potassium
MH  - proton
MH  - Protons
MH  - PSI
MH  - SURFACE
MH  - synthesis
MH  - vesicles
RP  - NOT IN FILE
NT  - UI - 94342345LA - engRN - 0 (Alkalies)RN - 0 (Aminoisobutyric Acids)RN - 0 (Buffers)RN - 0 (Phosphates)RN - 0 (Protons)RN - 0 (Salts)RN - 50-81-7 (Ascorbic Acid)RN - 58-64-0 (Adenosine Diphosphate)RN - 62-57-7 (2-aminoisobutyric acid)RN - 7440-09-7 (Potassium)PT - Journal ArticleID - NIGMS GM28454/GM/NIGMSDA - 19940922IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:8063796
SO  - J Biol Chem 1994 Aug 26 ;269(34):21576-21582

1217
UI  - 9934
AU  - Harrison MA
AU  - Jones PC
AU  - Kim YI
AU  - Finbow ME
AU  - Findlay JB
AD  - Department of Biochemistry and Molecular Biology, University of Leeds, England
TI  - Functional properties of a hybrid vacuolar H(+)-ATPase in Saccharomyces cells expressing the Nephrops 16-kDa proteolipid
AB  - The hydrophobic 16-kDa polypeptide which forms gap-junction-like structures in the crustacean Nephrops norvegicus is a member of a highly conserved family of proteolipids involved in a variety of membrane transport functions in eukaryotic cells. This family also includes the product of the Saccharomyces cerevisiae VMA3 gene which encodes an integral membrane component of the vacuolar membrane H(+)- ATPase. The cDNA for the Nephrops proteolipid complements a mutation in the yeast VMA3 gene, resulting in assembly of a hybrid H(+)-ATPase comprising yeast catalytic subunits and Nephrops integral membrane components. The hybrid vacuolar ATPase was capable of ATP hydrolysis which was coupled to proton translocation and showed inhibitor binding and enzymological properties similar to those of wild-type V-ATPases (Km for ATP, 0.4 mM), suggesting that both yeast and crustacean proteolipids share conserved structure at regions of protein interaction. To facilitate isolation of the Nephrops proteolipid by affinity chromatography on a Ni(2+)-binding support, six C-terminal histidine residues were added to the proteolipid. This modification did not prohibit assembly into the hybrid H(+)-ATPase, although the resultant enzyme did have a markedly elevated Km (1.8 mM). The membrane- bound Vo sector of the ATPase was isolated by the affinity- chromatography procedure and reconstituted into synthetic vesicles. This complex was found to be impermeable to small cations in the absence of catalytic ATPase subunits either in situ in the vacuolar membrane or in the reconstituted system. The functional significance of this impermeability and the structure/function relationships between proteolipids from different sources are discussed
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BINDING
MH  - Biochemistry
MH  - Cations
MH  - Cells
MH  - COMPLEX
MH  - England
MH  - H(+)-Transporting ATP Synthase
MH  - H(+)ATPase
MH  - Histidine
MH  - Hydrolysis
MH  - Nickel
MH  - Proteins
MH  - Proteolipids
MH  - proton
MH  - Protons
MH  - Saccharomyces cerevisiae
MH  - SUBUNIT
MH  - SYNTHASE
MH  - transport
RP  - NOT IN FILE
NT  - UI - 94222051LA - engRN - 0 (Proteolipids)RN - 0 (Protons)RN - 0 (Recombinant Proteins)RN - 0 (Vanadates)RN - 0 (proteolipid, Nephrops)RN - 139-13-9 (Nitrilotriacetic Acid)RN - 56-65-5 (Adenosine Triphosphate)RN - 7440-02-0 (Nickel)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19940602IS - 0014-2956SB - IMCY - GERMANYJC - EMZ
UR  - PM:8168500
SO  - Eur J Biochem 1994 Apr 1 ;221(1):111-120

1218
UI  - 20814
AU  - Hartzog PE
AU  - Cain BD
AD  - Department of Biochemistry and Molecular Biology, University of Florida, Gainesville 32610
TI  - Second-site suppressor mutations at glycine 218 and histidine 245 in the alpha subunit of F1F0 ATP synthase in Escherichia coli
AB  - The alpha-like subunits of F1F0 ATP synthases share primary structural homology in two segments near their carboxyl termini. However, the amino acids at the functionally important positions occupied by alpha Gly-218 and alpha His-245 in Escherichia coli vary depending upon organism and organelle. The alpha G218-->D,H245-G and alpha G218-- >K,H245-->G double mutations were constructed in the E. coli uncB(alpha) gene to model the chloroplast ATPase IV subunit and alkaliphilic bacterial alpha subunit, respectively. Strains carrying each of the single mutations, alpha G218-->D, alpha G218-->K, and alpha H245-->G, had marked reductions in F1F0 ATP synthase function. The alpha G218-->K mutation was alone sufficient to virtually eliminate enzyme function. Membranes prepared from the alpha G218-->D mutant exhibited increased levels of ATP hydrolysis activity without a corresponding increase in active proton transport, suggesting a mechanistic uncoupling of catalytic activity and proton translocation. However, much of the lost F1F0 ATP synthase activity was restored in the alpha G218-->D,H245-->G and alpha G218-->K,H245-->G double mutant strains demonstrating that these mutations act as mutual intragenic second-site suppressors. The evidence is consistent with a close spatial interaction between alpha Gly-218 and alpha His-245
MH  - A
MH  - ACID
MH  - ACTIVE
MH  - ALPHA-SUBUNIT
MH  - Amino Acids
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Biochemistry
MH  - chloroplast
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - function
MH  - Histidine
MH  - Hydrolysis
MH  - membrane
MH  - Membranes
MH  - model
MH  - mutant
MH  - proton
MH  - Protons
MH  - SUBUNIT
MH  - SYNTHASE
MH  - translocation
MH  - transport
RP  - NOT IN FILE
NT  - UI - 95096079LA - engRN - 0 (DNA, Bacterial)RN - 0 (Protons)RN - 56-40-6 (Glycine)RN - 71-00-1 (Histidine)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleID - GM43495/GM/NIGMSDA - 19950124IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:7798232
SO  - J Biol Chem 1994 Dec 23 ;269(51):32313-32317

1219
UI  - 21270
AU  - Haumann M
AU  - Junge W
AD  - Biophysik, FB Biologie/Chemie, Universitat Osnabruck, Germany
TI  - The rates of proton uptake and electron transfer at the reducing side of photosystem II in thylakoids
AB  - Proton and electron transfer at the reducing side of photosystem II of green plants was studied under flashing light, the former at improved time resolution by using Neutral red. The rates of electron transfer within QAFeQB were determined by pump-probe flashes through electrochromic transients. The extent of proton binding was about 1 H+/e-. The rates of proton transfer were proportional to the concentration of Neutral red (collisional transfer), whereas the rates of electron transfer out of QA- and from QAFeQB- to the cytochrome b6f complex were constant. The half-rise times of electron transfer (tau e) and the apparent times of proton binding (tau h) at 30 microM Neutral red were: QA- --> FeIIIQB (tau c < or = 100 microseconds, tau h = 230 microseconds); QA- --> FeIIQB (tau c = 150 microseconds, tau h = 760 microseconds); and QA- --> FeIIQB (tau c = 150 microseconds, tau h = 760 microseconds); and QA- --> FeIIQB (tau c = 620 microseconds, tau h = 310 microseconds)
MH  - b6f
MH  - BINDING
MH  - COLLISIONAL TRANSFER
MH  - COMPLEX
MH  - CONSTANT
MH  - cytochrome
MH  - electron
MH  - electron transfer
MH  - Electron-transfer
MH  - Light
MH  - microsecond
MH  - neutral red
MH  - photosystem II
MH  - plant
MH  - Plants
MH  - proton
MH  - Proton transfer
MH  - Proton-transfer
MH  - Protons
MH  - pump-probe
MH  - reaction center
MH  - resolution
MH  - thylakoid
MH  - thylakoids
MH  - Time
MH  - TRANSFER
RP  - NOT IN FILE
NT  - UI - 94283626LA - engRN - 0 (Photosynthetic Reaction Center, Plant)RN - 0 (Protons)RN - 553-24-2 (Neutral Red)PT - Journal ArticleDA - 19940728IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:8013659
SO  - FEBS Lett 1994 Jun 20 ;347(1):45-50

1220
UI  - 429
AU  - Hazard AL
AU  - Senior AE
AD  - Department of Biochemistry, University of Rochester Medical Center, New York 14642
TI  - Defective energy coupling in delta-subunit mutants of Escherichia coli F1F0-ATP synthase
AB  - Membrane vesicles from 13 strains carrying mutations in the C-terminal region of the delta-subunit of Escherichia coli F1F0-ATP synthase were characterized in respect to ATPase activity, ATP-driven proton-pumping, dicyclohexylcarbodiimide sensitivity of ATPase, and oxidative phosphorylation. The salient finding was that energy-coupling between F1 and F0 sectors of the enzyme is impaired by several of the mutations. The delta G150N mutant appeared completely uncoupled in vitro. The data emphasize the role of the C-terminal region of delta- subunit in integration of the proton conduction machinery in F0 with the three F1 catalytic sites. It is suggested that the C-terminal region of delta-subunit, speculatively located in the central region of the alpha 3 beta 3 hexagon, acts functionally at the interface between the helical domain of the stalk and the F1 subunits to relay conformational signals which alter the affinities of the catalytic sites for substrates and products
RP  - NOT IN FILE
NT  - UI - 94103249LA - engRN - 0 (Aurovertins)RN - 0 (Proton Pump)RN - 53-84-9 (NAD)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM 25349/GM/NIGMSDA - 19940208IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:8276831
SO  - J Biol Chem 1994 Jan 7 ;269(1):427-432

1221
UI  - 430
AU  - Hazard AL
AU  - Senior AE
AD  - Department of Biochemistry, University of Rochester Medical Center, New York 14642
TI  - Mutagenesis of subunit delta from Escherichia coli F1F0-ATP synthase
AB  - In Escherichia coli, the F1 sector of the F1F0-ATP synthase is connected to the membrane-embedded F0 sector by a narrow stalk, thought to be formed by subunits delta and b. Mutagenic analysis was used here to study the structure and function of subunit delta. First, random mutations in the protein were generated by bisulfite mutagenesis. Two single missense mutations causing impaired growth by oxidative phosphorylation were found, namely delta A149T and delta G150D. Both occur at the conserved C-terminal region, which has been suggested previously to be functionally important. Two techniques were applied to study the C-terminal region in greater detail. Cassette mutagenesis was used to randomly mutate the sequence from delta 145 to delta 167, and residues delta A149 and delta G150 were specifically mutated by site- directed mutagenesis to obtain multiple substitutions at each position. Fifteen of the residues between delta 145 and delta 167 were mutated. None was found to be absolutely essential for function. However, the properties of the mutants obtained, which included partial impairment of growth by oxidative phosphorylation, temperature sensitivity, and specific structural requirements at residues delta A149 and delta G150, confirmed that this region is important for enzyme function. Based on these studies, and on secondary and tertiary structure predictions, a model for subunit delta and its orientation in F1F0-ATP synthase is proposed
RP  - NOT IN FILE
NT  - UI - 94103248LA - engRN - 0 (Culture Media)RN - 0 (Oligodeoxyribonucleotides)RN - 0 (Succinates)RN - 0 (Sulfites)RN - 110-15-6 (Succinic Acid)RN - 7631-90-5 (sodium bisulfite)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSDA - 19940208IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:8276830
SO  - J Biol Chem 1994 Jan 7 ;269(1):418-426

1222
UI  - 6030
AU  - Heberle J
AU  - Riesle J
AU  - Thiedemann G
AU  - Oesterhelt D
AU  - Dencher NA
TI  - Proton migration along the membrane surface and retarded surface to bulk transfer
MH  - A
MH  - ATPase
MH  - buffer
MH  - Diffusion
MH  - equilibration
MH  - membrane
MH  - Membranes
MH  - pH
MH  - protein
MH  - proton
MH  - proton release
MH  - Proton transfer
MH  - Proton-transfer
MH  - Protons
MH  - SURFACE
MH  - SYNTHASE
MH  - theory
MH  - TRANSFER
MH  - Water
RP  - NOT IN FILE
NT  - Since the proposal of the chemiosmotic theory(1) there has been a continuing debate about how protons that have been pumped across membranes reach another membrane protein that utilizes the established pH gradient. Evidence has been gathered in favour of a 'delocalized' theory, in which the pumped protons equilibrate with the aqueous bulk phase before being consumed, and a 'localized' one, in which protons move exclusively along the membrane surface(2,3). We report here that after proton release by an integral membrane protein, long-range proton transfer along the membrane surface is faster than proton exchange with the bulk water phase. The rate of lateral proton diffusion can be calculated by considering the buffer capacity of the membrane surface. Our results suggest that protons can efficiently diffuse along the membrane surface between a source and a sink (for example H+-ATP synthase) without dissipation losses into the aqueous bulk
SO  - Nature 1994  ;370():379-382

1223
UI  - 21081
AU  - Hicks DB
AU  - Cohen DM
AU  - Krulwich TA
AD  - Department of Biochemistry, Mount Sinai School of Medicine of the City University of New York, New York 10029
TI  - Reconstitution of energy-linked activities of the solubilized F1F0 ATP synthase from Bacillus subtilis
AB  - The F1F0 ATP synthases from wild-type Bacillus subtilis and an uncoupler-resistant mutant have comparable subunit structures. In accord with an earlier hypothesis, ATP hydrolysis and ATP-Pi exchange by the two synthases were equally stimulated and inhibited by protonophores, respectively, when reconstituted alone in either wild- type or mutant lipids
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Bacillus
MH  - Biochemistry
MH  - Carbonyl Cyanide m-Chlorophenyl Hydrazone
MH  - Hydrolysis
MH  - Lipids
MH  - mutant
MH  - Phosphates
MH  - Proteolipids
MH  - proteoliposome
MH  - proton
MH  - Proton Pump
MH  - Proton-Translocating ATPases
MH  - protonophore
MH  - reconstitution
MH  - structure
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 94292444LA - engRN - 0 (Peptides)RN - 0 (Phosphates)RN - 0 (Proteolipids)RN - 0 (Proton Pumps)RN - 0 (proteoliposomes)RN - 555-60-2 (Carbonyl Cyanide m-Chlorophenyl Hydrazone)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleID - GM28454/GM/NIGMSDA - 19940729IS - 0021-9193SB - IMCY - UNITED STATES
UR  - PM:8021203
SO  - J Bacteriol 1994 Jul ;176(13):4192-4195

1224
UI  - 739
AU  - Hisabori T
AU  - Kobayashi H
AU  - Kaibara C
AU  - Yoshida M
AD  - Department of Biology, Yokohama City University, Kanagawa
TI  - Asymmetry of the three catalytic sites on beta subunits of TF1 from a thermophilic Bacillus strain PS3
AB  - F1-ATPase isolated from plasma membrane of a thermophilic Bacillus strain PS3 (TF1) has very little or no endogenously bound adenine nucleotides. However, it can bind one ADP per mol of the enzyme on one of three beta subunits to form a stable TF1.ADP complex when incubated with a high concentration of ADP [Yoshida, M. & Allison, W.S. (1986) J. Biol. Chem. 261, 5714-5721]. The same TF1.ADP complex was recovered after filling all ADP binding sites with [3H]ADP and repeated gel filtration. Direct binding assay revealed that the TF1.ADP complex had lost the highest affinity site for TNP-ADP. When a substoichiometric amount of TNP-ATP was added, the complex hydrolyzed TNP-ATP slowly (single site hydrolysis), like native TF1. However, this hydrolysis was not promoted by chase-addition of excess ATP. The optimal pH of the ATPase activity of TF1 or the TF1.ADP complex measured with a short reaction period, 6.5, was lower than the reported value, 9.0, under the steady-state condition. Although the bound ADP was released from the complex only when the enzyme underwent multiple catalytic turnover, the rate of this release was much slower than the turnover. These results suggest that when one ADP binds to a site on one of the beta subunits and stays there for a long time, the enzyme will change form and the bound ADP will become a special species which is not able to be directly involved in the enzyme catalysis. This binding site for ADP appears to be the first site responsible for the single-site catalysis reaction observed for native TF1
RP  - NOT IN FILE
NT  - UI - 94334292LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 61-19-8 (Adenosine Monophosphate)RN - 61368-63-6 (2',3'-O-(2,4,6-trinitro-cyclohexadienylidine)adenosine 5'- triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19940914IS - 0021-924XSB - IMCY - JAPANJC - HIF
UR  - PM:8056763
SO  - J Biochem (Tokyo ) 1994 Mar ;115(3):497-501

1225
UI  - 19862
AU  - Hyndman DJ
AU  - Milgrom YM
AU  - Bramhall EA
AU  - Cross RL
AD  - Department of Biochemistry and Molecular Biology, State University of New York, Health Science Center, Syracuse 13210
TI  - Nucleotide-binding sites on Escherichia coli F1-ATPase. Specificity of noncatalytic sites and inhibition at catalytic sites by MgADP
AB  - Nucleotide-depleted EcF1 binds a maximum of two GTP, ATP, or ADP at noncatalytic sites, whereas all three sites can only be filled by a combination of nucleoside di- and triphosphates. MgPPi prevents binding of GTP and significantly slows ATP binding, suggesting that non- catalytic sites also bind PPi. No binding of GDP at non-catalytic sites could be detected. The slow rate of GTP dissociation from noncatalytic sites (t1/2 = 175 min) is increased 2-8-fold by EDTA, MgPPi, MgADP, or EDTA/ATP, but 23-fold by conditions for ATP hydrolysis. ATP hydrolysis by EcF1, depleted of both its inhibitory epsilon-subunit and endogenous nucleotides, shows a burst of activity. However, it shows a lag if preincubated with MgADP but not when preincubated with Mg2+ alone. For epsilon-depleted EcF1 containing endogenous inhibitory ADP, preincubation with an ATP-regenerating system results in a burst of activity, whereas the control shows a lag. This same enzyme form shows significant inhibition with increasing concentrations of Mg2+ during ATP hydrolysis but lesser levels of inhibition when other NTP substrates are used. With the five-subunit enzyme, increasing amounts of azide cause an increase in the level of inhibition with a corresponding increase in amount of bound nucleotide resistant to rapid chase. Azide-trappable nucleotide is bound at catalytic sites as shown by covalent incorporation of 2-azido-ADP. The results suggest that ligand specificity may not be a reliable means of distinguishing between catalytic and noncatalytic sites and that MgADP inhibition should be taken into account in the kinetic analysis of EcF1 mutants
MH  - A
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - ADP
MH  - analysis
MH  - atp
MH  - ATP synthase
MH  - BINDING
MH  - Biochemistry
MH  - COLI F1 ATPASE
MH  - EPSILON-SUBUNIT
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - Guanosine Triphosphate
MH  - H(+)-Transporting ATP Synthase
MH  - Hydrolysis
MH  - non-catalytic site
MH  - NONCATALYTIC SITES
MH  - nucleotide binding
MH  - nucleotide binding sites
MH  - NUCLEOTIDE-BINDING-SITES
MH  - Nucleotides
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 95050700LA - engRN - 0 (Nucleotides)RN - 58-64-0 (Adenosine Diphosphate)RN - 86-01-1 (Guanosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM 23152/GM/NIGMSDA - 19941219IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:7961847
SO  - J Biol Chem 1994 Nov 18 ;269(46):28871-28877

1226
UI  - 21080
AU  - Ivey DM
AU  - Sturr MG
AU  - Krulwich TA
AU  - Hicks DB
AD  - Department of Biochemistry, Mount Sinai School of Medicine of City University of New York, New York 10029
TI  - The abundance of atp gene transcript and of the membrane F1F0-ATPase as a function of the growth pH of alkaliphilic Bacillus firmus OF4
MH  - A
MH  - atp
MH  - ATPase
MH  - Bacillus
MH  - Biochemistry
MH  - function
MH  - membrane
MH  - pH
MH  - Proton-Translocating ATPases
RP  - NOT IN FILE
NT  - UI - 94327493LA - engRN - 0 (RNA, Bacterial)RN - 0 (RNA, Messenger)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleID - GM28454/GM/NIGMSDA - 19940907IS - 0021-9193SB - IMCY - UNITED STATES
UR  - PM:7519597
SO  - J Bacteriol 1994 Aug ;176(16):5167-5170

1227
UI  - 20854
AU  - Iwamoto A
AU  - Orita-Saita Y
AU  - Maeda M
AU  - Futai M
AD  - Department of Organic Chemistry and Biochemistry, Osaka University, Japan
TI  - N-ethylmaleimide-sensitive mutant (beta Val-153-->Cys) Escherichia coli F1-ATPase: cross-linking of the mutant beta subunit with the alpha subunit
AB  - A beta subunit mutation, beta Val-153-->Cys, in the glycine-rich sequence (phosphate-binding loop) of Escherichia coli F1 was constructed. Like vacuolar-type ATPase, the mutant enzyme was inhibited by N-ethylmaleimide (NEM) and labeled with [14C]NEM. The inhibition and labeling were prevented by ATP. m-Maleimidobenzoyl-N-hydroxysuccinimide (MBS) (3 microM) almost completely inhibited the mutant enzyme, and cross-linked one pair of alpha and beta subunits. These results suggest that the interaction of the domain near beta Val-153 with the alpha subunit is essential for catalytic cooperativity of the enzyme and that beta Val-153 is within 10 A of the alpha subunit
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - ALPHA-SUBUNIT
MH  - atp
MH  - ATPase
MH  - BETA-SUBUNIT
MH  - Biochemistry
MH  - Chemistry
MH  - COLI F1 ATPASE
MH  - CROSS-LINKING
MH  - Cysteine
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - Ethylmaleimide
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - mutant
MH  - SUBUNIT
RP  - NOT IN FILE
NT  - UI - 95010717LA - engRN - 0 (Cross-Linking Reagents)RN - 0 (Succinimides)RN - 128-53-0 (Ethylmaleimide)RN - 52-90-4 (Cysteine)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19941031IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:7925981
SO  - FEBS Lett 1994 Sep 26 ;352(2):243-246

1228
UI  - 737
AU  - Jault JM
AU  - Kaibara C
AU  - Yoshida M
AU  - Garrod S
AU  - Allison WS
AD  - Department of Chemistry, University of California at San Diego, La Jolla 92093-0601
TI  - Probing the specificity of nucleotide binding to the F1-ATPase from thermophilic Bacillus PS3 and its isolated alpha and beta subunits with 2-N3-[beta, gamma-32P]ATP
AB  - Irradiation of the F1-ATPase from Bacillus PS3 (TF1) in the presence of 134 microM 2-N3-[beta, gamma-32P]ATP plus Mg2+ for 90 min led to 95% inactivation of the ATPase activity which was accompanied by exclusive labeling of the beta subunit. The isolated alpha and beta subunits were also treated separately with 2-N3-[beta, gamma-32P]ATP under similar conditions. Fractionation of a tryptic digest of photolabeled TF1 by reversed-phase HPLC resolved a major and a minor radioactive peptide. Sequence analyses showed that the major peptide contained labeled Tyr- beta 364, whereas the minor one contained labeled Tyr-beta 341, residues known to be part of noncatalytic and catalytic sites, respectively. Two closely eluting radioactive peptides were obtained when a tryptic digest of the photolabeled, isolated beta subunit was fractionated by HPLC. Sequence analyses revealed that both contained labeled Tyr-beta 341. Fractionation of a tryptic digest of the photolabeled, isolated alpha subunit by HPLC resolved two peptides which contained the majority of the radioactivity incorporated. When subjected to eight cycles of automatic Edman degradation, one gave the sequence APGVXDR, corresponding to residues 133-139, in which X is a gap and corresponds to Met-alpha 137, which presumably is the derivatized residue. Only the first five cycles yielded phenylthiohydantoin derivatives when the other radioactive peptide derived from the alpha subunit was submitted to automatic Edman degradation which revealed the sequence APGVM, suggesting that Asp- alpha 138 is derivatized. The overall results suggest that the isolated beta subunit is a useful model for studying binding of nucleotides to catalytic sites, whereas the isolated alpha subunit may be of limited value in modeling interactions of nucleotides with noncatalytic sites
RP  - NOT IN FILE
NT  - UI - 94213500LA - engRN - 0 (Affinity Labels)RN - 0 (Azides)RN - 0 (Nucleotides)RN - 0 (Peptide Fragments)RN - 56-65-5 (Adenosine Triphosphate)RN - 72884-75-4 (2-azidoadenosine 5'-triphosphate)RN - EC 3.4.21.4 (Trypsin)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - CM-16,974/CM/NCIDA - 19940518IS - 0003-9861SB - IMCY - UNITED STATESJC - 6SK
UR  - PM:8161217
SO  - Arch Biochem Biophys 1994 Apr ;310(1):282-288

1229
UI  - 9933
AU  - Jones PC
AU  - Harrison MA
AU  - Kim YI
AU  - Finbow ME
AU  - Findlay JB
AD  - Department of Biochemistry and Molecular Biology, University of Leeds, UK
TI  - Structure and function of the proton-conducting sector of the vacuolar H(+)-ATPase
MH  - atp
MH  - ATP synthase
MH  - Biochemistry
MH  - England
MH  - H(+)-Transporting ATP Synthase
MH  - H(+)ATPase
MH  - Proteolipids
MH  - review
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 95121732LA - engRN - 0 (Proteolipids)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 19950216IS - 0300-5127SB - IMCY - ENGLANDJC - E48
UR  - PM:7821690
SO  - Biochem Soc Trans 1994 Aug ;22(3):805-809

1230
UI  - 21037
AU  - Kakinuma Y
AU  - Igarashi K
AD  - Faculty of Pharmaceutical Sciences, Chiba University
TI  - Purification and characterization of the catalytic moiety of vacuolar- type Na(+)-ATPase from Enterococcus hirae
AB  - We have previously reported the molecular cloning and sequences of the ntp genes for Enterococcus hirae Na(+)-translocating ATPase [Takase, K., Kakinuma, S., Yamato, I., Konishi, K., Igarashi, K., and Kakinuma, Y. (1994) J. Biol. Chem. 269, 11037-11044]; the expected structure of this enzyme complex resembles those of the vacuolar H(+)-ATPase complexes in eukaryotes. In this paper we report purification and characterization of the catalytic moiety of Na(+)-ATPase, whose molecular size was about 400 kDa, consisting of polypeptides of 69 kDa (NtpA), 52 kDa (NtpB), and 29 kDa (NtpD) with a probable stoichiometry of 3:3:1. Purified enzyme hydrolyzed GTP as the best substrate (GTP > CTP > UTP > ATP), and the activity was maximal at around pH 6.0. The activity was not stimulated by sodium ions, and was selectively inhibited by nitrate. These properties were different from those of membrane-bound Na(+)-ATPase, suggesting that a significant conformational change of the catalytic moiety may take place upon dissociation from the membrane-embedded moiety and probably also loss of other hydrophilic subunits. Antiserum against purified enzyme inhibited the Na(+)-stimulated ATPase activity of the membranes. Immunoblotting analysis revealed that the change in the amounts of A and B subunits of the membranes paralleled that of the Na(+)-ATPase activity. Furthermore, the A subunit was missing in the membranes of a Na(+)-ATPase mutant, and recovered in those of its revertant. These immunochemical data are consistent with the notion that this enzyme is the hydrophilic catalytic moiety of the V-type Na(+)-ATPase in E. hirae
MH  - A
MH  - Adenosinetriphosphatase
MH  - analysis
MH  - atp
MH  - ATPase
MH  - Bacterial Proteins
MH  - COMPLEX
MH  - conformational change
MH  - H(+)ATPase
MH  - ion
MH  - Ions
MH  - membrane
MH  - Membranes
MH  - mutant
MH  - pH
MH  - protein
MH  - Proteins
MH  - purification
MH  - Sodium
MH  - structure
MH  - SUBUNIT
RP  - NOT IN FILE
NT  - UI - 95221328LA - engRN - 0 (Bacterial Proteins)RN - EC 3.6.1.- (sodium-translocating ATPase)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19950511IS - 0021-924XSB - IMCY - JAPAN
UR  - PM:7706221
SO  - J Biochem (Tokyo ) 1994 Dec ;116(6):1302-1308

1231
UI  - 59
AU  - Kluge C
AU  - Dimroth P
AD  - Mikrobiologisches Institut, Eidgenossische Technische Hochschule, ETH- Zentrum, Zurich, Switzerland
TI  - Modification of isolated subunit c of the F1Fo-ATPase from Propionigenium modestum by dicyclohexylcarbodiimide
AB  - Subunit c of the F1Fo-ATPase from Propionigenium modestum was extracted from the particulate cell fraction with chloroform/methanol. The protein was further purified by carboxymethyl cellulose chromatography and anion exchange HPLC in the organic solvent. SDS-PAGE of the purified protein indicated a single stained protein band migrating as expected for the c-subunit. Incubation of isolated subunit c in chloroform/methanol or aqueous buffer containing dodecyl-beta-D- maltoside with [14C]dicyclohexylcarbodiimide (DCCD) resulted in the incorporation of radioactivity into the protein. The rate of this reaction depended on the external pH; it was significantly faster in the more acidic than in the alkaline pH range. In the presence of Na+ subunit c was partially protected from labeling with [14C]DCCD at pH 6.1 and at pH 7.5, whereas no protection was evident at pH 5.5. At pH 7.5, the rate of subunit c labeling by [14C]DCCD in the presence of 20 mM NaCl was about 50% lower than in the absence of Na+ ions. The isolated c-subunit therefore apparently retains in part the Na+ binding site which, when occupied, diminishes the reactivity of the protein towards DCCD
RP  - NOT IN FILE
NT  - UI - 94178374LA - engRN - 0 (Cations)RN - 0 (Glutamates)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-86-0 (Glutamic Acid)RN - 7440-23-5 (Sodium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19940415IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:7907551
SO  - FEBS Lett 1994 Mar 7 ;340(3):245-248

1232
UI  - 21190
AU  - Komrakov AY
AU  - Kaulen AD
AD  - AN Belozersky Institute of Physico-Chemical Biology, Moscow State University, Russian Federation
TI  - On the two forms of bacteriorhodopsin
AB  - In our previous work [(1993) FEBS Lett. 313, 248-250; (1993) Biochem. Int. 30, 461-469] M-intermediate formation of wild-type bacteriorhodopsin was shown to involve two components differing in time constants (tau 1 = 60-70 microseconds and tau 2 = 220-250 microseconds), which were suggested to reflect two independent pathways of M-intermediate formation. The contribution of the fast M was 4-times higher than the slow one. Our present research on M-intermediate formation in the D115N bacteriorhodopsin mutant revealed the same components but at a contribution ratio of 1:1. Upon lowering the pH, the slow phase of M-formation vanished at a pK of 6.2, and in the pH region 3.0-5.5 only the M-intermediate with a rise time of 60 microseconds was present. A 5-6 h incubation of D115N bacteriorhodopsin at pH 10.6 resulted in the irreversible transformation of 50% of the protein into a form with a difference absorbance maximum at 460 nm. This form was stable at pH 7.5 and had no photocycle, including M- intermediate formation. The remaining bacteriorhodopsin contained 100% fast M-intermediate. The disappearance of the 250-microseconds phase concomitant with bR460 formation indicates that at neutral pH bacteriorhodopsin exists as two spectroscopically indistinguishable forms
MH  - A
MH  - Bacteriorhodopsin
MH  - CONSTANT
MH  - intermediate
MH  - M
MH  - M-intermediate
MH  - microsecond
MH  - mutant
MH  - pH
MH  - protein
MH  - RISE TIME
MH  - Time
RP  - NOT IN FILE
NT  - UI - 94178366LA - engRN - 53026-44-1 (Bacteriorhodopsins)PT - Journal ArticleDA - 19940415IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:8131847
SO  - FEBS Lett 1994 Mar 7 ;340(3):207-210

1233
UI  - 21238
AU  - Leikin S
AU  - Rau DC
AU  - Parsegian VA
AD  - Laboratory of Structural Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland 20892
TI  - Direct measurement of forces between self-assembled proteins: temperature-dependent exponential forces between collagen triple helices
AB  - We report direct measurements of force vs. separation between self- assembled proteins. These forces are observed between collagen triple helices in native and reconstituted fibers. They are a combination of a short-range repulsion, which varies exponentially over at least five decay lengths, and an inferred, longer-ranged attraction responsible for spontaneous assembly. From 5 degrees C to 35 degrees C the relative contribution of the attraction to the net force increases with temperature. These forces are strikingly similar to the "hydration" forces measured between several other linear macromolecules (DNA, polysaccharides) and between lipid bilayer membranes. The decay length of the repulsive force agrees well with a theoretical estimate based on axial periodicity of the triple helix, suggesting another connection between molecular architecture and protein-protein interaction
MH  - A
MH  - Kidney
MH  - Macromolecular Systems
MH  - membrane
MH  - Membranes
MH  - protein
MH  - Proteins
MH  - SYSTEM
MH  - SYSTEMS
MH  - Temperature
MH  - Water
RP  - NOT IN FILE
NT  - UI - 94105150LA - engRN - 0 (Macromolecular Systems)RN - 7732-18-5 (Water)RN - 9007-34-5 (Collagen)RN - 9007-49-2 (DNA)PT - Journal ArticleDA - 19940204IS - 0027-8424SB - IMCY - UNITED STATES
UR  - PM:8278378
SO  - Proc Natl Acad Sci U S A 1994 Jan 4 ;91(1):276-280

1234
UI  - 21256
AU  - Lengeler JW
AU  - Jahreis K
AU  - Wehmeier UF
AD  - Arbeitsgruppe Genetik, Fachbereich Biologie/Chemie, Universitat Osnabruck, Germany
TI  - Enzymes II of the phospho enol pyruvate-dependent phosphotransferase systems: their structure and function in carbohydrate transport
MH  - Carbohydrates
MH  - Enzymes
MH  - function
MH  - review
MH  - structure
MH  - SYSTEM
MH  - SYSTEMS
MH  - transport
RP  - NOT IN FILE
NT  - UI - 95034965LA - engRN - 0 (Carbohydrates)RN - EC 2.7.1.- (Phosphoenolpyruvate Sugar Phosphotransferase System)PT - Journal ArticlePT - ReviewPT - Review, AcademicDA - 19941201IS - 0006-3002SB - IMCY - NETHERLANDS
UR  - PM:7947897
SO  - Biochim Biophys Acta 1994 Nov 1 ;1188(1-2):1-28

1235
UI  - 632
AU  - Martins IS
AU  - Penefsky HS
AD  - Department of Biochemistry and Molecular Biology, State University of New York, Health Science Center at Syracuse 13210
TI  - Covalent modification of catalytic sites on membrane-bound beef heart mitochondrial ATPase by 2-azido-adenine nucleotides
AB  - Incubation in the dark of 32P-labeled 2-azido-adenine nucleotides with submitochondrial particles from beef heart led to tight binding of the label by membrane-bound F1. That is, the label remained with the particles following two passages through centrifuge columns. After removal of free nucleotides and ultraviolet irradiation, the radioactive label was covalently bound exclusively to the beta subunit of the ATPase. Extraction of the modified enzyme from the membrane with chloroform followed by tryptic digestion and separation of peptides by reverse-phase high-pressure liquid chromatography indicated that the radioactive label had been inserted into a peptide fragment that included part of the catalytic site. Covalent modification of catalytic sites by 2-azido-ADP was accompanied by parallel inhibition of both ATP synthesis and ATP hydrolysis by submitochondrial particles. Estimation of the likely amount of F1 participating in the reaction and extrapolation to complete inhibition suggested that modification of no more than a single site was sufficient to block both reactions. The results support suggestions of cooperative interactions between catalytic sites as well as a single catalytic pathway for both enzymic reactions
RP  - NOT IN FILE
NT  - UI - 95010066LA - engRN - 0 (Azides)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 64020-53-7 (2-azidoadenosine 3',5'-diphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleID - GM21737/GM/NIGMSDA - 19941027IS - 0014-2956SB - IMCY - GERMANYJC - EMZ
UR  - PM:7925403
SO  - Eur J Biochem 1994 Sep 15 ;224(3):1057-1065

1236
UI  - 735
AU  - Matsuda C
AU  - Muneyuki E
AU  - Endo H
AU  - Yoshida M
AU  - Kagawa Y
AD  - Department of Biochemistry, Jichi Medical School, Tochigi, Japan
TI  - Comparison of the ATPase activities of bovine heart and liver mitochondrial ATP synthases with different tissue-specific gamma subunit isoforms
AB  - The kinetics of heart and liver mitochondrial ATPase (FoF1) were examined using submitochondrial particles (SMPs) purified from the two tissues to obtain information on the role of gamma subunit isoforms. The F1 portion is mainly composed of the catalytic, common alpha beta subunits and tissue-specific gamma subunits. In contrast to the previous reports on the kinetics and crystallography of various F1's, the Vmax and Km of the two isoforms of FoF1 were identical although the SMPs were prepared from different tissues. Moreover sodium azide inhibited the two equally. The ATPase activity of liver SMP showed slightly steeper pH-dependency than that of heart SMP but the pH optima of the two were the same (pH 8)
RP  - NOT IN FILE
NT  - UI - 94235011LA - engRN - 0 (Azides)RN - 26628-22-8 (Sodium Azide)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19940603IS - 0006-291XSB - IMCY - UNITED STATESJC - 9Y8
UR  - PM:8179599
SO  - Biochem Biophys Res Commun 1994 Apr 29 ;200(2):671-678

1237
UI  - 21161
AU  - Monticello RA
AU  - Brusilow WS
AD  - Department of Biochemistry, Wayne State University School of Medicine, Detroit, Michigan 48201
TI  - Role of the delta subunit in enhancing proton conduction through the F0 of the Escherichia coli F1F0 ATPase
AB  - We studied the effect of the delta subunit of the Escherichia coli F1 ATPase on the proton permeability of the F0 proton channel synthesized and assembled in vivo. Membranes isolated from an unc deletion strain carrying a plasmid containing the genes for the F0 subunits and the delta subunit were significantly more permeable to protons than membranes isolated from the same strain carrying a plasmid containing the genes for the F0 subunits alone. This increased proton permeability could be blocked by treatment with either dicyclohexyl-carbodiimide or purified F1, both of which block proton conduction through the F0. After reconstitution with purified F1 in vitro, both membrane preparations could couple proton pumping to ATP hydrolysis. These results demonstrate that an interaction between the delta subunit and the F0 during synthesis and assembly produces a significant change in the proton permeability of the F0 proton channel
MH  - A
MH  - atp
MH  - ATPase
MH  - Biochemistry
MH  - COLI F1 ATPASE
MH  - delta
MH  - DELTA-SUBUNIT
MH  - Dicyclohexylcarbodiimide
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - Hydrolysis
MH  - In Vitro
MH  - Macromolecular Systems
MH  - membrane
MH  - Membranes
MH  - Permeability
MH  - proton
MH  - Proton-Translocating ATPases
MH  - Protons
MH  - reconstitution
MH  - SUBUNIT
MH  - synthesis
MH  - SYSTEM
MH  - SYSTEMS
RP  - NOT IN FILE
NT  - UI - 94156843LA - engRN - 0 (Macromolecular Systems)RN - 0 (Oligodeoxyribonucleotides)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleID - KO4-AI00882/AI/NIAIDDA - 19940330IS - 0021-9193SB - IMCY - UNITED STATES
UR  - PM:8113178
SO  - J Bacteriol 1994 Mar ;176(5):1383-1389

1238
UI  - 298
AU  - Mulkidjanian AY
AU  - Junge W
AD  - Biophysik, Fachbereich Biologie/Chemie, Universitat Osnabruck, Germany
TI  - Calibration and time resolution of lumenal pH-transients in chromatophores of Rhodobacter capsulatus following a single turnover flash of light: proton release by the cytochrome bc1-complex is strongly electrogenic
AB  - The flash-induced proton release into the lumen of chromatophores from Rhodobacter capsulatus was studied with Neutral red as pH-indicator. Calibration of the acidification jump after a single flash yielded a much larger figure, at least 0.8 units, than previously thought. A slow kinetic phase of proton release (85-90% of total) was sensitive to inhibitors of the cytochrome bc1-complex. Its half-rise time, about 10 ms, was the same as the rise time of the electrogenic reaction in the cytochrome bc1-complex that was recorded by electrochromism of carotenoids. The oxidoreduction of the two b-hemes was significantly faster (t1/2 approximately equal to 3 ms). Thus the major electrogenic event in the cytochome bc1-complex is proton and not electron transfer
RP  - NOT IN FILE
NT  - UI - 95010792LA - engRN - 0 (Protons)RN - 553-24-2 (Neutral Red)RN - EC 1.10.2.2 (Ubiquinol-Cytochrome-c Reductase)PT - Journal ArticleDA - 19941123IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:7926049
SO  - FEBS Lett 1994 Oct 17 ;353(2):189-193

1239
UI  - 731
AU  - Muneyuki E
AU  - Hisabori T
AU  - Allison WS
AU  - Jault JM
AU  - Sasayama T
AU  - Yoshida M
AD  - Research Laboratory of Resources Utilization, Tokyo Institute of Technology, Yokohama, Japan
TI  - Catalytic cooperativity of beef heart mitochondrial F1-ATPase revealed by using 2',3'-O-(2,4,6-trinitrophenyl)-ATP as a substrate; an indication of mutually activating catalytic sites
AB  - The interaction of 2',3'-O-(2,4,6-trinitrophenyl)ATP (TNP-ATP) with bovine mitochondrial F1-ATPase (MF1) was examined under substoichiometric and stoichiometric conditions to investigate the relationship between the amount of bound TNP-AT(D)P and extent of inhibition on steady state ATP hydrolysis. The hydrolysis of bound TNP- ATP under substoichiometric condition proceeded slowly, with a first order rate constant of 0.014 s-1. However, hydrolysis was greatly accelerated by addition of excess ATP. The hydrolyzed product, TNP-ADP, did not dissociate from the enzyme even after the addition of excess ATP. These properties were the same for both native and nucleotide depleted enzyme. The difference spectrum induced by binding TNP-ATP to MF1 had a distinct peak at 410 nm and a deep trough at 395 nm, which were similar to those induced when TNP-ATP bound to the isolated beta subunit of the thermophilic F1-ATPase. The magnitude of difference spectra as a function of TNP-ATP concentration suggested the presence of at least two types of binding sites on the MF1 molecule. The first site, where substoichiometric TNP-ATP was hydrolyzed, had a very high affinity for TNP-ATP. TNP-AT(D)P bound to this site did not dissociate even in the presence of excess ATP. TNP-AT(D)P bound to the second site dissociated slowly when excess ATP was added. The steady state ATPase activity at 100 microM ATP was linearly suppressed as pre-loaded TNP- ATP increased. The binding of 2 mol of TNP-ATP per mol of MF1 was required to abolish ATPase activity. A model which assumes mutually- activating two catalytic sites is presented to explain these results
RP  - NOT IN FILE
NT  - UI - 95034967LA - engRN - 0 (2',3'-O-(2,4,6-trinitrophenyl)adenosine 5'-triphosphate)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19941201IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:7947899
SO  - Biochim Biophys Acta 1994 Nov 1 ;1188(1-2):108-116

1240
UI  - 19774
AU  - Murataliev MB
AU  - Boyer PD
AD  - Scripps Research Institute, La Jolla, California 92037
TI  - Interaction of mitochondrial F1-ATPase with trinitrophenyl derivatives of ATP and ADP. Participation of third catalytic site and role of Mg2+ in enzyme inactivation
AB  - Relatively high ATP concentrations show an unexpected lack of inhibition of the hydrolysis of low concentrations of trinitrophenyl ATP (TNP-ATP) by mitochondrial F1-ATPase. In striking contrast low TNP- ATP concentrations markedly inhibit the hydrolysis of much higher ATP concentrations. The three catalytic sites undergoing sequential conformational changes have different conformations at any instant of catalysis, and only two need to be filled for rapid, steady-state ATP hydrolysis. The remaining site has low affinity for ATP (Kd 2 mM) but about 10(4) greater affinity for TNP-ATP (Km and Kd about 0.2 microM). Thus 500 microM ATP does not prevent binding of less than 1 microM TNP- ATP. As the site binding the TNP-ATP undergoes sequential conformational changes the TNP-ATP undergoes sequential conformational changes the TNP-ATP is hydrolyzed and products are released. The results give strong support to the view that all three catalytic sites proceed equivalently in ATP as well as TNP-ATP hydrolysis. The conformation that has the lowest affinity for ATP has over a 10-fold greater affinity for ADP (Kd 150 microM) and may be akin to the conformation to which ADP binds during net ATP synthesis by the ATP synthase. The recognition of these features was made possible by new information obtained from detailed studies of the interactions of Mg2+, TNP-ADP, TNP-ATP, ATP, and noncatalytic sites on initial and steady- state hydrolysis rates
MH  - A
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - ADP
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - BINDING
MH  - Catalysis
MH  - conformation
MH  - conformational change
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - H(+)-Transporting ATP Synthase
MH  - Hydrolysis
MH  - Magnesium
MH  - MITOCHONDRIAL F1-ATPASE
MH  - NONCATALYTIC SITES
MH  - Phosphorus
MH  - Phosphorus Radioisotopes
MH  - SYNTHASE
MH  - synthesis
RP  - NOT IN FILE
NT  - UI - 94253118LA - engRN - 0 (Phosphorus Radioisotopes)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 61368-63-6 (2',3'-O-(2,4,6-trinitro-cyclohexadienylidine)adenosine 5'- triphosphate)RN - 7439-95-4 (Magnesium)RN - 84430-17-1 (2',3'-(O-(2,4,6-trinitrocyclohexadienylidine))adenosine 5'- diphosphate)RN - EC 3.6.1.- (Ca(2+) Mg(2+)-ATPase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - DK08126/DK/NIDDKID - GM11094/GM/NIGMSDA - 19940630IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:8195184
SO  - J Biol Chem 1994 Jun 3 ;269(22):15431-15439

1241
UI  - 738
AU  - Odaka M
AU  - Kaibara C
AU  - Amano T
AU  - Matsui T
AU  - Muneyuki E
AU  - Ogasahara K
AU  - Yutani K
AU  - Yoshida M
AD  - Chemical Engineering Laboratory, Institute of Physical and Chemical Research, Saitama
TI  - Tyr-341 of the beta subunit is a major Km-determining residue of TF1- ATPase: parallel effect of its mutations on Kd(ATP) of the beta subunit and on Km(ATP) of the alpha 3 beta 3 gamma complex
AB  - Residue Tyr-341 of the F1-ATPase beta subunit from a thermophilic Bacillus strain, PS3, was mutagenized to leucine, cysteine or alanine. Each of the mutated beta subunits was isolated and its affinity for ATP- Mg was examined by means of difference circular dichroism and differential titration calorimetry. The Kd values for ATP-Mg obtained were: beta Y341 (wild type), 0.015 mM; beta Y341L, 0.7 mM; beta Y341C and beta Y341A, > 3 mM. All the mutant beta subunits could be reconstituted into the alpha 3 beta 3 gamma complex with alpha and gamma subunits. The alpha 3 beta (mutant)3 gamma complexes hydrolyzed ATP with apparent Vmax values larger than that of the alpha 3 beta (WILD)3 gamma complex. The apparent Km values of the alpha 3 beta (mutant)3 gamma complexes increased in parallel with the Kd values for ATP-Mg of the isolated mutant beta subunits. These results indicate that residue beta Y341 is directly involved in the catalytic ATP-Mg binding and is a major Km-determining residue of F1-ATPase
RP  - NOT IN FILE
NT  - UI - 94375427LA - engRN - 0 (Peptide Fragments)RN - 52-90-4 (Cysteine)RN - 55520-40-6 (Tyrosine)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19941018IS - 0021-924XSB - IMCY - JAPANJC - HIF
UR  - PM:8089097
SO  - J Biochem (Tokyo ) 1994 Apr ;115(4):789-796

1242
UI  - 20856
AU  - Omote H
AU  - Park MY
AU  - Maeda M
AU  - Futai M
AD  - Department of Organic Chemistry and Biochemistry, Osaka University, Japan
TI  - The alpha/beta subunit interaction in H(+)-ATPase (ATP synthase). An Escherichia coli alpha subunit mutation (Arg-alpha 296-->Cys) restores coupling efficiency to the deleterious beta subunit mutant (Ser-beta 174-->Phe)
AB  - The Ser-beta 174 residue of the Escherichia coli H(+)-ATPase beta subunit has been shown to be near the catalytic site together with Gly- beta 149, Gly-beta 172, Glu-beta 192, and Val-beta 198 (Iwamoto, A., Park, M.-Y., Maeda, M., and Futai, M. (1993) J. Biol. Chem. 268, 3156- 3160). In this study, we introduced various residues at position 174 and found that the larger the side chain volume of the residue introduced, the lower the enzyme activity became. The Phe-beta 174 mutant was defective in energy coupling between catalysis and transport, whereas the Leu-beta 174 mutant could couple efficiently, although both mutants had essentially the same ATPase activities (approximately 10% of the wild type). The defective energy coupling of the Phe-beta 174 mutant was suppressed by the second mutation (Arg- alpha 296-->Cys) in the alpha subunit. The Cys-alpha 296/Phe-beta 174 mutant had essentially the same membrane ATPase activity as the Phe- beta 174 single mutant when assayed under the conditions that stabilize the double mutant enzyme. These results indicate the importance of the alpha/beta interaction, especially that between the regions near Arg- alpha 296 and Ser-beta 174, for energy coupling in the H(+)-ATPase. The 2 residues (Ser-beta 174 and Arg-alpha 296) may be located nearby at the interface of the two subunits. About 1 mol of N-[14C]ethylmaleimide could bind to 1 mol of the alpha subunit of Cys-alpha 296/Phe-beta 174 or Cys-alpha 296 mutant ATPase, but could not inhibit the enzyme activity. This is the first intersubunit mutation/suppression approach to ATPase catalysis and its energy coupling
MH  - A
MH  - ALPHA-SUBUNIT
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BETA-SUBUNIT
MH  - Biochemistry
MH  - Catalysis
MH  - Chemistry
MH  - coupling
MH  - Cysteine
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - Ethylmaleimide
MH  - H(+)ATPase
MH  - Hydrogen
MH  - M
MH  - membrane
MH  - mutant
MH  - RESIDUE
MH  - Site
MH  - SUBUNIT
MH  - SYNTHASE
MH  - transport
RP  - NOT IN FILE
NT  - UI - 94193712LA - engRN - 128-53-0 (Ethylmaleimide)RN - 1333-74-0 (Hydrogen)RN - 52-90-4 (Cysteine)RN - 56-45-1 (Serine)RN - 63-91-2 (Phenylalanine)RN - 74-79-3 (Arginine)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19940505IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:8144607
SO  - J Biol Chem 1994 Apr 8 ;269(14):10265-10269

1243
UI  - 20853
AU  - Park MY
AU  - Omote H
AU  - Maeda M
AU  - Futai M
AD  - Department of Organic Chemistry and Biochemistry, Osaka University, Ibaraki
TI  - Conserved Glu-181 and Arg-182 residues of Escherichia coli H(+)-ATPase (ATP synthase) beta subunit are essential for catalysis: properties of 33 mutants between beta Glu-161 and beta Lys-201 residues
AB  - Twenty-two mutants between beta Glu-161 and beta Lys-201 of Escherichia coli H(+)-ATPase beta subunit could grow by oxidative phosphorylation, but 11 other such mutants, beta Glu-181-->Gln, Asp, Asn, Thr, Ser, Ala, or Lys and beta Arg-182-->Lys, Ala, Glu, or Gln, could not. The beta Asp-181, beta Lys-182, and other defective mutants had 1.4, 1, and < 0.1%, respectively, of the wild-type membrane ATPase activity. Partially purified F1-ATPases from all mutants at positions 181 and 182, except for the beta Asp-181 and beta Lys-182 mutants, showed very low unisite catalysis. Purified F1-ATPases of the beta Gln-181 and beta Ala-181 mutants showed no multisite (or steady state) catalysis and slow unisite catalysis (< or = 1% of that of the wild type): their defects could be attributed to decreased catalytic rates (low k+2 and k- 2). Changes of the k+2 and k-2 values in the beta Asp-181 enzyme, which showed detectable multi- and unisite catalysis, were less marked (27 and 21%, respectively, of wild-type rates). The beta Gln-182 enzyme showed defective catalysis (< or = 0.1% of the multi- and approximately 1% of the unisite catalyses of the wild type), whereas the beta Lys-182 enzyme showed 1 and 85% of the wild-type multisite and unisite catalytic rates, respectively. beta Lys-182 had wild-type values of k+2 and k-2, but beta Gln-182 had k+2 about 10-fold lower than that of wild type.(ABSTRACT TRUNCATED AT 250 WORDS)
MH  - ACID
MH  - Adenine Nucleotides
MH  - ADENINE-NUCLEOTIDES
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BETA-SUBUNIT
MH  - Biochemistry
MH  - Catalysis
MH  - Chemistry
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - glutamic acid
MH  - H(+)ATPase
MH  - membrane
MH  - mutant
MH  - Nucleotides
MH  - Oxidative Phosphorylation
MH  - Phosphorylation
MH  - RESIDUE
MH  - SUBUNIT
MH  - SYNTHASE
MH  - UNISITE CATALYSIS
RP  - NOT IN FILE
NT  - UI - 95204389LA - engRN - 0 (Adenine Nucleotides)RN - 56-86-0 (Glutamic Acid)RN - 74-79-3 (Arginine)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19950426IS - 0021-924XSB - IMCY - JAPAN
UR  - PM:7896744
SO  - J Biochem (Tokyo ) 1994 Nov ;116(5):1139-1145

1244
UI  - 510
AU  - Pedersen PL
AD  - Department of Biological Chemistry, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205-2185, USA
TI  - ATP synthase. The machine that makes ATP
AB  - The recently determined crystal structure of the F1 part of mitochondrial ATP synthase provides new insights into the workings of one of the most remarkable and complex biochemical machines
RP  - NOT IN FILE
NT  - UI - 95219448LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 19950505IS - 0960-9822SB - IMCY - ENGLANDJC - B44
UR  - PM:7704582
SO  - Curr Biol 1994 Dec 1 ;4(12):1138-1141

1245
UI  - 167
AU  - Phadke AS
AU  - Aggeler R
AU  - Keana JF
AU  - Capaldi RA
AD  - Department of Chemistry, University of Oregon, Eugene 97403-1229
TI  - TFPACD, a novel bifunctional reagent for reacting with DCCD sites in proteins: studies using Escherichia coli ATP synthase
AB  - A novel cross-linker, 1-[6-(4-azido-2,3,5,6-tetrafluorobenzamido)hexyl]- 3-cyclohexylc arbodiimide (TFPACD), has been synthesized and tested by reaction with the Escherichia coli ATP Synthase (ECF1F0). The reagent has a carbodiimide as one reactive group, which is shown to react with ECF1F0 in a similar way to 1,3-dicyclohexylcarbodiimide (DCCD) and modify the beta subunit of the ECF1 part and the c subunit of the F0 part. Reaction with both the ECF1 and F0 parts of the complex inhibited ATPase activity. The second reactive group in the reagent is the photoactivatable tetrafluorophenylazide moiety. Subsequent UV photolysis of TFPACD--modified ECF1 and ECF1F0 led to generation of cross-linked products in significant yields, one between beta and alpha subunits; the second, dimers of the c subunit of the F0 part
RP  - NOT IN FILE
NT  - UI - 94271212LA - engRN - 0 (Azides)RN - 0 (Carbodiimides)RN - 0 (Cross-Linking Reagents)RN - 0 (Indicators and Reagents)RN - 157918-73-5 (1-(6-(4-azido-2,3,5,6-tetrafluorobenzamido)hexyl)-3- cyclohexylcarbodiimide)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM 27137/GM/NIGMSID - HL 24526/HL/NHLBIDA - 19940714IS - 0006-291XSB - IMCY - UNITED STATESJC - 9Y8
UR  - PM:8002996
SO  - Biochem Biophys Res Commun 1994 Jun 15 ;201(2):635-641

1246
UI  - 8833
AU  - Possmayer FE
AU  - Grber P
TI  - The pHin and pHout dependence of the rate of ATP synthesis catalyzed by the chloroplast H+-ATPase, CF0F1, in proteoliposomes
MH  - atp
MH  - ATP synthesis
MH  - CF0F1
MH  - chloroplast
MH  - chloroplast H+-ATPase
MH  - DEPENDENCE
MH  - H+-ATPase
MH  - mechanism
MH  - proteoliposome
MH  - proton
MH  - synthesis
RP  - IN FILE
NT  - ju ATP3
SO  - Journal of Biological Chemistry 1994  ;269():1896-1904

1247
UI  - 20995
AU  - Possmayer FE
AU  - Graber P
AD  - Biologisches Institut, Universitat Stuttgart, Germany
TI  - The pHin and pHout dependence of the rate of ATP synthesis catalyzed by the chloroplast H(+)-ATPase, CF0F1, in proteoliposomes
AB  - The H(+)-ATPase from chloroplasts was isolated, purified, and reconstituted into liposomes from phosphatidylcholine/phosphatidic acid. The rate of ATP synthesis was measured after energization of the proteoliposomes by an acid-base transition as a function pHout and pHin. At any given pHout, the rate increased sigmoidally with increasing internal H+ concentration with half-maximal rates at about pHin 5.3 for all pHout values between 9.3 and 7.2. At any given pHin, the rate decreased sigmoidally with increasing external H+ concentration with half-maximal rates at about pHout 7.8. The dependence on internal H+ concentration was attributed to the protonation of three monovalent groups with the same pKin, and the dependence on external proton concentration to the deprotonation of two groups with the same pKout. The following scheme is proposed. The first step of the reaction is the binding of three (or four) protons from the inside of the F0 part, followed by ADP and Pi binding at the F1 part. Then, the enzyme conformation is changed, and the proton binding sites are exposed to the outside, followed by proton release from F0 and ATP release from F1. The reaction cycle is closed by changing the enzyme conformation to the form where protons can bind from the inside
MH  - A
MH  - ACID
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - ADP
MH  - atp
MH  - ATP synthesis
MH  - ATPase
MH  - BINDING
MH  - Binding Sites
MH  - CF0F1
MH  - chloroplast
MH  - Chloroplasts
MH  - conformation
MH  - DEPENDENCE
MH  - F0
MH  - F1
MH  - function
MH  - H(+)ATPase
MH  - H+
MH  - Liposomes
MH  - Phosphatidic Acids
MH  - Phosphatidylcholines
MH  - Proteolipids
MH  - proteoliposome
MH  - proton
MH  - proton release
MH  - protonation
MH  - Protons
MH  - Site
MH  - synthesis
RP  - NOT IN FILE
NT  - UI - 94124535LA - engRN - 0 (Liposomes)RN - 0 (Phosphatidic Acids)RN - 0 (Phosphatidylcholines)RN - 0 (Proteolipids)RN - 0 (Protons)RN - 0 (proteoliposomes)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19940225IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:8294439
SO  - J Biol Chem 1994 Jan 21 ;269(3):1896-1904

1248
UI  - 10665
AU  - Richard P
AU  - Graber P
TI  - Simulation of the kinetics of the H+-ATPase of the plasma membrane from yeast
MH  - atp
MH  - H+-ATPase
MH  - Kinetics
MH  - membrane
MH  - proton
MH  - SIMULATION
T2  - Biothermokinetics
Y2  - -32676  
PB  - Andover, UK: Intercept
RP  - NOT IN FILE
SO  -  1994  ;():145-152

1249
UI  - 732
AU  - Saika K
AU  - Inaka K
AU  - Matsui T
AU  - Yoshida M
AU  - Miki K
AD  - Research Laboratory of Resources Utilization, Tokyo Institute of Technology, Yokohama, Japan
TI  - Crystallization of mutant beta subunit of F1-ATPase from thermophilic Bacillus PS3
AB  - The mutant beta subunit of F1-ATPase from a thermophilic Bacillus strain, PS3, in which tyrosine at position 341 is replaced by leucine (beta Y341L) was expressed in Escherichia coli and crystallized by the vapor-diffusion procedure. Small needle-like crystals were obtained using ammonium sulfate as a precipitant and grown by the stepwise seeding method. The crystals obtained by this procedure diffracted X- rays to about 3 A resolution. The diffraction patterns indicated that the crystals belong to the orthorhombic system and the space group I222 or I2(1)2(1)2(1) with unit-cell dimensions of a = 232 A, b = 66 A, and c = 80 A. It is thought that the asymmetric unit comprises one beta Y341L molecule
RP  - NOT IN FILE
NT  - UI - 95018238LA - engRN - 0 (Recombinant Proteins)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19941027IS - 0022-2836SB - IMCY - ENGLANDJC - J6V
UR  - PM:7932728
SO  - J Mol Biol 1994 Oct 7 ;242(5):709-711

1250
UI  - 21148
AU  - Scherrer P
AU  - Alexiev U
AU  - Marti T
AU  - Khorana HG
AU  - Heyn MP
AD  - Department of Physics, Freie Universitat Berlin, Germany
TI  - Covalently bound pH-indicator dyes at selected extracellular or cytoplasmic sites in bacteriorhodopsin. 1. Proton migration along the surface of bacteriorhodopsin micelles and its delayed transfer from surface to bulk
AB  - The kinetics of the light-induced release and uptake of protons was monitored with the optical pH-indicator fluorescein covalently bound to various sites on the extracellular and cytoplasmic surfaces of bacteriorhodopsin. Selective labeling was achieved by reacting (iodoacetamido)fluorescein with the single cysteine residues in bacteriorhodopsin introduced at the desired positions by site-directed mutagenesis. All measurements were performed with bacteriorhodopsin micelles in phospholipid/detergent mixtures in 150 mM KCl at 22 degrees C, pH 7.3. Neither the replacements by cysteine nor the subsequent labeling affected the absorption spectrum of bacteriorhodopsin and the rise times of the M intermediate. Only the decay of M was altered for some bacteriorhodopsin mutants with cysteine residues on the cytoplasmic side. The proton release time detected with fluorescein attached to the extracellular surface (the proton release side) at position 72 (in the loop connecting helices B and C) or 130 (DE loop) was 22 +/- 4 microseconds, clearly faster than that measured with pyranine in the aqueous bulk phase (125 +/- 10 microseconds for wild- type and all mutants studied). For bacteriorhodopsin mutants labeled at positions 35, 101, 160, 229, and 231 in the cytoplasmic loop region (the proton uptake side), the released proton was observed with a time of 61 +/- 4 microseconds. This was about 3-fold slower than the release time on the extracellular side, but still significantly faster than that measured with pyranine in the bulk phase. These results suggest that the released protons are retained on the micellar surface and move more rapidly along this surface to the cytoplasmic side than from the surface to the bulk medium.(ABSTRACT TRUNCATED AT 250 WORDS)
MH  - A
MH  - absorption
MH  - Arylsulfonates
MH  - Bacteriorhodopsin
MH  - Cysteine
MH  - DYE
MH  - dyes
MH  - fluorescein
MH  - Fluoresceins
MH  - intermediate
MH  - Kinetics
MH  - M
MH  - M-intermediate
MH  - Micelles
MH  - microsecond
MH  - mutagenesis
MH  - mutant
MH  - pH
MH  - pH-indicator
MH  - protein
MH  - Proteins
MH  - proton
MH  - proton release
MH  - Protons
MH  - RESIDUE
MH  - RISE TIME
MH  - Site
MH  - site-directed
MH  - spectra
MH  - SURFACE
MH  - Time
MH  - TRANSFER
RP  - NOT IN FILE
NT  - UI - 95034803LA - engRN - 0 (Arylsulfonates)RN - 0 (Fluoresceins)RN - 0 (Protons)RN - 0 (Recombinant Proteins)RN - 2321-07-5 (Fluorescein)RN - 52-90-4 (Cysteine)RN - 53026-44-1 (Bacteriorhodopsins)RN - 6358-69-6 (pyranine)PT - Journal ArticleID - GM 28289/GM/NIGMSDA - 19941221IS - 0006-2960SB - IMCY - UNITED STATES
UR  - PM:7947777
SO  - Biochemistry 1994 Nov 22 ;33(46):13684-13692

1251
UI  - 631
AU  - Souid AK
AU  - Penefsky HS
AD  - Department of Biochemistry and Molecular Biology, State University of New York, Health Science Center at Syracuse 13210
TI  - Mechanism of ATP synthesis by mitochondrial ATP synthase from beef heart
AB  - Previous studies of the rate constants for the elementary steps of ATP hydrolysis by the soluble and membrane-bound forms of beef heart mitochondrial F1 supported the proposal that ATP is formed in high- affinity catalytic sites of the enzyme with little or no change in free energy and that the major requirement for energy in oxidative phosphorylation is for the release of product ATP. The affinity of the membrane-bound enzyme for ATP during NADH oxidation was calculated from the ratio of the rate constants for the forward binding step (k+1) and the reverse dissociation step (k-1). k-1 was accelerated several orders of magnitude by NADH oxidation. In the presence of NADH and ADP an additional enhancement of k-1 was observed. These energy-dependent dissociations of ATP were sensitive to the uncoupler FCCP. k+1 was affected little by NADH oxidation. The dissociation constant (KdATP) increased many orders of magnitude during the transition from nonenergized to energized states
RP  - NOT IN FILE
NT  - UI - 95238308LA - engRN - 370-86-5 (Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialID - GM21737/GM/NIGMSDA - 19950523IS - 0145-479XSB - IMCY - UNITED STATESJC - HIO
UR  - PM:7721724
SO  - J Bioenerg Biomembr 1994 Dec ;26(6):627-630

1252
UI  - 20815
AU  - Stack AE
AU  - Cain BD
AD  - Department of Biochemistry and Molecular Biology, University of Florida, Gainesville 32610
TI  - Mutations in the delta subunit influence the assembly of F1F0 ATP synthase in Escherichia coli
AB  - Missense mutations affecting Asp-161 and Ser-163 in the delta subunit of F1F0 ATP synthase have been generated. Although most substitutions allowed substantial enzyme function, the delta Asp-161-->Pro substitution resulted in a loss of enzyme activity. The loss of activity was attributable to a structural failure altering assembly of the enzyme complex
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Bacterial Proteins
MH  - Biochemistry
MH  - COMPLEX
MH  - DELTA-SUBUNIT
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - function
MH  - protein
MH  - Proteins
MH  - proton
MH  - Protons
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 94117395LA - engRN - 0 (Bacterial Proteins)RN - 0 (Protons)RN - 0 (uncH protein, Escherichia coli)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleID - GM43495/GM/NIGMSDA - 19940224IS - 0021-9193SB - IMCY - UNITED STATES
UR  - PM:8288552
SO  - J Bacteriol 1994 Jan ;176(2):540-542

1253
UI  - 299
AU  - Steinemann D
AU  - Lill H
AU  - Junge W
AU  - Engelbrecht S
AD  - Fachbereich Biologie/Chemie, Universitat Osnabruck, Germany
TI  - Over-production, renaturation and reconstitution of delta and epsilon subunits from chloroplast and cyanobacterial F1
AB  - We studied the functioning of chimeric F0F1-ATPases by replacing subunits delta and epsilon of spinach CF1 with their counterparts from Synechocystis sp. PCC 6803. The sequence identities between these subunits are 26 and 41%, respectively. For a systematic approach to such studies and later extension to genetically modified subunits recombinant proteins are required. The genes coding for spinach and Synechocystis delta and epsilon were cloned into pET3 expression vectors and expressed in Escherichia coli. Upon expression at 37 degrees C the recombinant subunits formed inclusion bodies within the host cells except for spinach delta, which was soluble. Synechocystis delta and epsilon could be obtained in soluble form upon expression at 20 degrees C. After purification (and refolding of spinach epsilon) both epsilon subunits inhibited the Ca(2+)-ATPase activity of soluble CF1(- epsilon). Subunits delta and epsilon from both species raised the rate of ATP synthesis in partially CF1-depleted spinach thylakoids when added together with CF1(- delta) or CF1(- delta, epsilon). This showed the functionality of recombinant Synechocystis and spinach delta and epsilon together with spinach alpha 3 beta 3 gamma. The molar excess of epsilon necessary for saturation was higher for Ca(2+)-ATPase inhibition than for reconstitution of photophosphorylation thus pointing to a direct interaction between epsilon and both CF1 and CF0
RP  - NOT IN FILE
NT  - UI - 95002040LA - engRN - 0 (Recombinant Proteins)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19941025IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:7918533
SO  - Biochim Biophys Acta 1994 Sep 27 ;1187(3):354-359

1254
UI  - 21082
AU  - Sturr MG
AU  - Guffanti AA
AU  - Krulwich TA
AD  - Department of Biochemistry, Mount Sinai School of Medicine, City University of New York, New York 10029
TI  - Growth and bioenergetics of alkaliphilic Bacillus firmus OF4 in continuous culture at high pH
AB  - The effect of external pH on growth of alkaliphilic Bacillus firmus OF4 was studied in steady-state, pH-controlled cultures at various pH values. Generation times of 54 and 38 min were observed at external pH values of 7.5 and 10.6, respectively. At more alkaline pH values, generation times increased, reaching 690 min at pH 11.4; this was approximately the upper limit of pH for growth with doubling times below 12 h. Decreasing growth rates above pH 11 correlated with an apparent decrease in the ability to tightly regulate cytoplasmic pH and with the appearance of chains of cells. Whereas the cytoplasmic pH was maintained at pH 8.3 or below up to external pH values of 10.8, there was an increase up to pH 8.9 and 9.6 as the growth pH was increased to 11.2 and 11.4, respectively. Both the transmembrane electrical potential and the phosphorylation potential (delta Gp) generally increased over the total pH range, except for a modest fall-off in the delta Gp at pH 11.4. The capacity for pH homeostasis rather than that for oxidative phosphorylation first appeared to become limiting for growth at the high edge of the pH range. No cytoplasmic or membrane- associated organelles were observed at any growth pH, confirming earlier conclusions that structural sequestration of oxidative phosphorylation was not used to resolve the discordance between the total electrochemical proton gradient (delta p) and the delta Gp as the external pH is raised.(ABSTRACT TRUNCATED AT 250 WORDS)
MH  - A
MH  - ACID
MH  - Alkalies
MH  - Bacillus
MH  - Biochemistry
MH  - bioenergetics
MH  - Cells
MH  - delta
MH  - Homeostasis
MH  - membrane
MH  - Organelles
MH  - Oxidative Phosphorylation
MH  - P
MH  - pH
MH  - Phosphorylation
MH  - proton
MH  - Time
RP  - NOT IN FILE
NT  - UI - 94252976LA - engRN - 0 (Alkalies)RN - 0 (Malates)RN - 6915-15-7 (malic acid)PT - Journal ArticleID - GM28454/GM/NIGMSDA - 19940630IS - 0021-9193SB - IMCY - UNITED STATES
UR  - PM:8195065
SO  - J Bacteriol 1994 Jun ;176(11):3111-3116

1255
UI  - 169
AU  - Tang C
AU  - Wilkens S
AU  - Capaldi RA
AD  - Institute of Molecular Biology, University of Oregon, Eugene 97403
TI  - Structure of the gamma subunit of Escherichia coli F1 ATPase probed in trypsin digestion and biotin-avidin binding studies
AB  - The arrangement and functional role of the gamma subunit of the Escherichia coli F1ATPase (ECF1) has been probed by protease digestion and avidin-biotin labeling experiments using wild-type enzyme and four mutants, gamma S8C, gamma T106C, gamma S179C, and gamma V286C, respectively. Trypsin was found to cleave the gamma subunit at four sites, Arg70, Lys199, Lys201, and Lys212. Cleavage at these four sites did not greatly reduce the high ATPase activity of the enzyme that is obtained when the epsilon subunit is removed by the protease treatment. However, prolonged trypsin cleavage led to loss of inhibition by epsilon subunit added back to the trypsin-treated enzyme. Endoproteinase-Lys-C cleaves the gamma subunit of ECF1 at three of the four sites, i.e. Lys199, Lys201, and Lys212, but not at Arg70. The enzyme was activated by treatment with this protease because of degradation and release of the epsilon subunit, but added pure epsilon subunit still caused inhibition of ATPase activity. Therefore, cleavage at Arg70 by trypsin is responsible for the loss of response to the epsilon subunit inhibition. Biotin was reacted with Cys residues at positions 8, 106, 179, and 286 in different gamma subunit mutants and the accessibility of the biotin to avidin monitored in the intact ECF1 from the different mutants. Avidin was able to react with biotin when incorporated at position 106, not at 8, 179, or 286. The four trypsin cleavage sites, Arg70, Lys199, Lys201, and Lys212, as well as Thr106 are in regions of the gamma subunit predicted to be mainly beta-sheet and beta-turn structures
RP  - NOT IN FILE
NT  - UI - 94140880LA - engRN - 0 (Antibodies, Monoclonal)RN - 0 (Epitopes)RN - 1405-69-2 (Avidin)RN - 52-90-4 (Cysteine)RN - 58-85-5 (Biotin)RN - EC 3.4.21.4 (Trypsin)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - HL 24526/HL/NHLBIDA - 19940317IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:7508444
SO  - J Biol Chem 1994 Feb 11 ;269(6):4467-4472

1256
UI  - 21191
AU  - Tikhonova IM
AU  - Andreyev AY
AU  - Antonenko Y
AU  - Kaulen AD
AU  - Komrakov AY
AU  - Skulachev VP
AD  - AN Belozersky Institute of Physico-Chemical Biology, Moscow State University, Russian Federation
TI  - Ion permeability induced in artificial membranes by the ATP/ADP antiporter
AB  - The hypothesis on the additional function of the ATP/ADP antiporter (ANT) as uncoupling protein has been tested in proteoliposomes and planar bilayer phospholipid membranes (BLM). It is found that dissipation of the light-induced delta pH in the dark is very much faster in ANT-bacteriorhodopsin proteoliposomes than in proteoliposomes containing bacteriorhodopsin as the only protein. Mersalyl treatment of ANT-bacteriorhodopsin proteoliposomes causes further increase in the delta pH dissipation rate due to formation of a high conductance pore. The properties of this pore are studied on ANT incorporated to BLM. They proved to be similar to those of so-called multiple conductance channel or permeability transition pore of inner mitochondrial membrane. The conductance of the single channel is as high as 2.2 nS. The channel fails to discriminate between K+, Na+, H+ and Cl-. Thus the obtained data are consistent with the assumption that native and modified ANT might function as an H(+)-specific conductor and as a permeability transition pore, respectively
MH  - A
MH  - Adenine Nucleotide Translocase
MH  - Anions
MH  - Bacteriorhodopsin
MH  - Cations
MH  - conductance
MH  - delta
MH  - DELTA-PH
MH  - function
MH  - H+
MH  - ion
MH  - Lipid Bilayers
MH  - liposome
MH  - Liposomes
MH  - membrane
MH  - Membranes
MH  - Permeability
MH  - pH
MH  - protein
MH  - proteoliposome
RP  - NOT IN FILE
NT  - UI - 94123758LA - engRN - 0 (Anions)RN - 0 (Cations)RN - 0 (Lipid Bilayers)RN - 0 (Liposomes)RN - 1405-97-6 (Gramicidin)RN - 27061-78-5 (Alamethicin)RN - 492-18-2 (Mersalyl)RN - 53026-44-1 (Bacteriorhodopsins)RN - EC 2.7.7.- (Adenine Nucleotide Translocase)PT - Journal ArticleDA - 19940225IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:7507443
SO  - FEBS Lett 1994 Jan 17 ;337(3):231-234

1257
UI  - 168
AU  - Turina P
AU  - Capaldi RA
AD  - Institute of Molecular Biology, University of Oregon, Eugene 97403
TI  - ATP hydrolysis-driven structural changes in the gamma-subunit of Escherichia coli ATPase monitored by fluorescence from probes bound at introduced cysteine residues
AB  - Four mutants of the Escherichia coli F1ATPase, gamma S8-C, gamma T106- C, gamma S179-C, and gamma V286-C, which have a cysteine introduced at different sites in the gamma-subunit by site-directed mutagenesis, were reacted with the fluorescent reagent N-(4-7-(diethylamino)4- methylcoumarin-3-yl)-maleimide (CM) under conditions that selectively label the introduced Cys residue. With each mutant the effect of nucleotide binding on the fluorescence of the probe has been monitored. The results obtained with the mutants gamma S8-C and gamma T106-C are similar. In both cases, there was a spectral shift and change in fluorescence intensity on adding AMP.PNP or ATP to enzyme emptied of nucleotide from catalytic sites, while no change in the fluorescence spectrum was observed upon adding ADP. The fluorescence spectral changes obtained with ATP were transient and involved an initial rapid fluorescence enhancement followed by a subsequent fluorescence quenching. The kinetics of these ATP-induced fluorescence changes and the kinetics of ATP hydrolysis as monitored by the rates of ATP binding and of Pi formation were the same under conditions of unisite catalysis, indicating that the conformational changes in the gamma- subunit being measured by the fluorescent probe are driven by ATP hydrolysis in catalytic sites. No nucleotide-dependent fluorescence changes were observed with CM bound at a Cys at position 179. Nucleotide-dependent changes in fluorescence were seen with CM bound at position 286, but these appear to reflect structural changes due to binding of ADP or ATP in noncatalytic sites. The fluorescence changes observed in mutants gamma S8-C and gamma T106-C were not seen in subunit epsilon-free E. coli F1ATPase, although such enzyme preparations are highly active ATPases. We conclude that the structural changes monitored by the fluorescent probe are a part of the conformational coupling, whereby catalytic site events are linked to proton channeling
RP  - NOT IN FILE
NT  - UI - 94230455LA - engRN - 0 (Coumarins)RN - 0 (Fluorescent Dyes)RN - 0 (Maleimides)RN - 0 (Nucleotides)RN - 52-90-4 (Cysteine)RN - 56-65-5 (Adenosine Triphosphate)RN - 93111-28-5 (N-(4-(7-(diethylamino)-4-methylcoumarin-3-yl))maleimide)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - LH 24236/PHSDA - 19940609IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:8175779
SO  - J Biol Chem 1994 May 6 ;269(18):13465-13471

1258
UI  - 97
AU  - Turina P
AU  - Capaldi RA
AD  - Institute of Molecular Biology, University of Oregon, Eugene 97403
TI  - ATP binding causes a conformational change in the gamma subunit of the Escherichia coli F1ATPase which is reversed on bond cleavage
AB  - ATP hydrolysis by the Escherichia coli F1 ATPase (ECF1) induces a conformational change in the gamma subunit. This change can be monitored by fluorescence changes in N-[4-[7-(diethylamino)-4- methyl]coumarin-3-yl)]maleimide (CM) bound at a cysteine introduced by site-directed mutagenesis into the gamma subunit at position 106 [Turina, P., & Capaldi, R. A. (1994) J. Biol. Chem. 269, 13465-13471]. In studies reported here, the magnitude of the fluorescence change has been determined with the noncleavable nucleotide analogue AMP-PNP and by rapid measurements using the slowly cleavable ATP gamma S. The data indicate that maximal fluorescence change occurs with binding of 1 mol of nucleotide triphosphate per mole of ECF1. During unisite catalysis, ATP binding causes a fluorescence enhancement from CM bound at position 106, which is then followed by fluorescence quenching. The kinetics of these fluorescence changes have been measured using both ATP and ATP gamma S as substrate. With ATP gamma S, these kinetics can be simulated using rate constants similar to those for ATP except for an approximately 30-fold slower rate of the bond cleavage and resynthesis steps, i.e., k+2 and k-2. The observed rates and amplitudes of the fluorescence changes on hydrolysis of ATP and ATP gamma S were analyzed by simulations in which the bond cleavage or the Pi release step was responsible for fluorescence quenching. The results indicate that ATP or ATP gamma S binding causes the fluorescence enhancement of CM bound to the gamma subunit and that this conformational change is reversed upon bond cleavage to yield ADP.Pi or ADP.PiS in catalytic sites
RP  - NOT IN FILE
NT  - UI - 95034875LA - engRN - 0 (Coumarins)RN - 0 (Fluorescent Dyes)RN - 0 (Maleimides)RN - 25612-73-1 (Adenylyl Imidodiphosphate)RN - 35094-46-3 (adenosine 5'-O-(3-thiotriphosphate))RN - 52-90-4 (Cysteine)RN - 56-65-5 (Adenosine Triphosphate)RN - 93111-28-5 (N-(4-(7-(diethylamino)-4-methylcoumarin-3-yl))maleimide)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - HL 24526/HL/NHLBIDA - 19941228IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:7947838
SO  - Biochemistry 1994 Nov 29 ;33(47):14275-14280

1259
UI  - 19291
AU  - Valerio M
AU  - Diolez P
AU  - Haraux F
TI  - Deactivation of F0F1 ATPase in intact plant mitochondria. Effect of pH and inhibitors.
AB  - By using a method especially adapted to intact (pea leaf) mitochondria, we studied the regulation of the F0F1 ATPase by the electrochemical proton gradient (delta mu H+) and by the matricial pH. The kinetics of decay of the ATP hydrolase activity was studied immediately after the collapse of the electrochemical proton gradient by an uncoupler. At pH 7.5, three inhibitors of the ATPase (venturicidin, tri-n-butyl tin and aurovertin), used at non-saturating concentrations, inhibited ATP hydrolysis to the same extent throughout the decay. This showed that the activity was totally controlled by the ATPase during all the decay and rules out any involvement of the phosphate or nucleotide carriers. This interpretation was confirmed by the fact that carboxyatractyloside, an inhibitor of the ATP/ADP antiporter, had a strong effect only on the initial rate of ATP hydrolysis, but not on the rate measured after some tens of seconds of decay. Oligomycin, at variance with the other ATPase inhibitors, interfered with the deactivation process, suggesting that its effect depends on the conformational state of the enzyme. Between pH 6.5 and 7.5, the hydrolase activity rose continuously and was still kinetically controlled by the ATPase. At higher pH value, the activity slightly decreased and appeared limited by at least one of the carriers. The activity of the ATPase itself, free of any transport process, seemed to increase monotonously with pH from 6.5 to 8. The electrochemical proton gradient is required to maintain the ATPase active, whereas no effect can be observed on transport processes. Matricial pH, while modulating the apparent catalytic turnover, has no marked effect on the rate of deactivation. These results, obtained with intact mitochondria, extend previous observations on the isolated enzyme and question the binding of IF1 as a rate-limiting step for ATPase deactivation.
MH  - A
MH  - ACTIVE
MH  - Adenosine Triphosphate
MH  - antagonists & inhibitors
MH  - atp
MH  - ATPase
MH  - aurovertin
MH  - Aurovertins
MH  - BINDING
MH  - Binding Sites
MH  - carrier
MH  - catalytic
MH  - delta
MH  - drug effects
MH  - Electrochemistry
MH  - enzyme
MH  - Enzyme Stability
MH  - enzymology
MH  - F0F1
MH  - H(+)-Transporting ATP Synthase
MH  - H+
MH  - Hydrogen-Ion Concentration
MH  - Hydrolysis
MH  - inhibitor
MH  - inhibitors
MH  - Kinetics
MH  - metabolism
MH  - method
MH  - Mitochondria
MH  - nucleotide
MH  - oligomycin
MH  - Oxidation-Reduction
MH  - pea
MH  - pea leaves
MH  - pH
MH  - pharmacology
MH  - phosphate
MH  - plant
MH  - Plants
MH  - proton
MH  - PROTON GRADIENT
MH  - regulation
MH  - transport
MH  - trialkyltin compounds
MH  - turnover
MH  - uncoupler
MH  - venturicidin
MH  - venturicidins
RP  - NOT IN FILE
NT  - Biosystemes Membranaires, ERS 30, Centre National de la Recherche Scientifique, Gif-sur-Yvette, FrancePMID- 0008181464
SO  - Eur J Biochem 1994 May 1 ;221(3):1071-1078

1260
UI  - 957
AU  - Van Dam K
TI  - Regulation and control of energy coupling at the cellular level
MH  - Energy Metabolism
MH  - Enterobacteriaceae
MH  - Glucose
MH  - metabolism
MH  - Phosphorylation
MH  - Saccharomyces cerevisiae
MH  - Signal Transduction
MH  - Support,Non-U.S.Gov't
RP  - NOT IN FILE
NT  - EC Slater Institute, BioCentrum, University of Amsterdam, The Netherlands
SO  - Biochim Biophys Acta 1994 Aug 30 ;1187(2):129-131

1261
UI  - 343
AU  - Walker JE
AU  - Collinson IR
AD  - Medical Research Council, Laboratory of Molecular Biology, Cambridge, UK
TI  - The role of the stalk in the coupling mechanism of F1F0-ATPases
AB  - The extrinsic and intrinsic membrane sectors of F1F0-ATPases are linked by a slender stalk 40-50 A in length. The stalk transmits the energy produced by oxidative or photosynthetic phosphorylation from the intrinsic sector, F0, to the catalytic sites in the extrinsic F1 sector. How this is achieved is unknown, but long-range conformational changes linked to transmembrane proton transport may be involved. In bacterial and chloroplast F1F0-ATPases, the stalk is probably a composite of subunits delta and epsilon, part of the gamma-subunit, and the extrinsic membrane domains of 2 subunits (identical or non- identical according to the species) that are bound to the membrane by their N-terminal regions. The stalk in the bovine mitochondrial enzyme appears to be more complex, and the gamma, delta, epsilon, OSCP, F6, b and d subunits all contribute to it. A bovine stalk complex has been assembled in vitro from bacterially expressed OSCP, F6, b and d, both in the presence and in the absence of F1-ATPase. One molecule of each of these subunits is present in the assembled complex, as there is also in each native F1F0-ATPase assembly. Providing that suitable crystals can be obtained, the stalk complex and the F1.stalk complex may permit the high resolution structure of bovine F1-ATPase to be extended into the stalk domain
RP  - NOT IN FILE
NT  - UI - 94265914LA - engRN - 0 (Macromolecular Systems)RN - 0 (Membrane Proteins)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 19940708IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:8206156
SO  - FEBS Lett 1994 Jun 6 ;346(1):39-43

1262
UI  - 338
AU  - Walker JE
AD  - Medical Research Council Laboratory of Molecular Biology, Cambridge, UK
TI  - The regulation of catalysis in ATP synthase
AB  - ATP synthase is regulated so as to prevent futile hydrolysis of ATP when the transmembrane proton electrochemical gradient, delta mu H+, falls. Mitochondria and chloroplasts have different mechanisms for inhibition of ATP synthase: by binding an inhibitor protein, and by stabilization of the ADP-inhibited state by making an intramolecular disulphide bond, respectively. The recently determined structure of bovine F1-ATPase is locked in a conformation that probably represents the ADP-inhibited state of the enzyme
RP  - NOT IN FILE
NT  - UI - 95227825LA - engRN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 19950517IS - 0959-440XSB - IMCY - ENGLANDJC - B9V
UR  - PM:7712295
SO  - Curr Opin Struct Biol 1994 Dec ;4(6):912-918

1263
UI  - 426
AU  - Weber J
AU  - Wilke-Mounts S
AU  - Senior AE
AD  - Department of Biochemistry, University of Rochester Medical Center, New York 14642
TI  - Cooperativity and stoichiometry of substrate binding to the catalytic sites of Escherichia coli F1-ATPase. Effects of magnesium, inhibitors, and mutation
AB  - The fluorescence of residue Trp beta 331 in beta Y331W mutant Escherichia coli F1-ATPase was used as reporter probe to investigate the effects of magnesium ions, inhibitors, and mutation on substrate (ATP) binding stoichiometry and cooperativity. It was found that Mg2+ is required for catalytic site binding cooperativity. In the absence of magnesium, ATP bound to three independent catalytic sites, each with Kd = 76 microM. In contrast, MgATP bound to three catalytic sites with Kd1 < 50 nM, Kd2 = 0.5 microM, and Kd3 = 25 microM. There was no significant ATPase activity in the absence of Mg2+. Catalysis is therefore correlated with substrate binding cooperativity and the formation of the high-affinity catalytic site 1. Catalytic site 3 had properties similar to those of the isolated beta-subunit nucleotide- binding site. The inhibitors dicyclohexylcarbodiimide and N- ethylmaleimide (in alpha S373C/beta Y331W mutant F1) gave potent inhibition of multisite ATPase activity without significantly affecting MgATP binding stoichiometry or cooperativity. Therefore each seems to selectively attenuate positive catalytic cooperativity. The same conclusions held for the alpha S373F mutation (in alpha S373F/beta Y331W mutant F1). 7-Chloro-4-nitrobenzo-2-oxa-1,3-diazole, however, reduced the catalytic site MgATP binding stoichiometry from three to two, and appears to inhibit catalysis by sterically blocking catalytic site 3
RP  - NOT IN FILE
NT  - UI - 94327619LA - engRN - 128-53-0 (Ethylmaleimide)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - 7439-95-4 (Magnesium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSDA - 19940906IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:8051144
SO  - J Biol Chem 1994 Aug 12 ;269(32):20462-20467

1264
UI  - 427
AU  - Weber J
AU  - Wilke-Mounts S
AU  - Grell E
AU  - Senior AE
AD  - Department of Biochemistry, University of Rochester Medical Center, New York 14642
TI  - Tryptophan fluorescence provides a direct probe of nucleotide binding in the noncatalytic sites of Escherichia coli F1-ATPase
AB  - Tryptophan fluorescence was investigated as a tool to study the noncatalytic nucleotide-binding sites of Escherichia coli F1-ATPase. Site-directed mutagenesis, affinity labeling, and lin-benzo-ATP binding studies had shown that residues alpha R365 and beta Y354 are located close to the base moiety of bound nucleotide; here, we mutagenized each to tryptophan. The new tryptophans gave a fluorescence signal indicating an environment of high (alpha W365) or intermediate (beta W354) polarity in unoccupied sites. alpha W365 fluorescence was completely quenched by binding of ATP or ADP, providing a direct, specific probe of noncatalytic site nucleotide occupancy. Using this signal, we measured binding parameters for ATP and ADP, showed that nucleotide binding was magnesium-dependent, and showed that GTP and ITP did bind to some extent, but AMP, GDP, and IDP did not. It was possible to follow initial rates of MgATP hydrolysis and noncatalytic site binding under identical conditions; the results indicated that occupancy of noncatalytic sites was not required for catalysis. Fluorescence from beta W354 was quenched completely by lin-benzo-ATP, but only slightly by ATP or ADP. Probably, residue beta 354 is not as closely juxtaposed to the adenine ring of bound ATP and ADP as is residue alpha 365. With either alpha W365 or beta W354 as donor and catalytic site-bound lin-benzo-ADP as acceptor, no fluorescence resonance energy transfer was detected, indicating that the distance between non-catalytic and catalytic sites is > or = 27 A
RP  - NOT IN FILE
NT  - UI - 94209300LA - engRN - 0 (Oligodeoxyribonucleotides)RN - 0 (Recombinant Proteins)RN - 0 (Ribonucleotides)RN - 73-22-3 (Tryptophan)RN - 7439-95-4 (Magnesium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSDA - 19940519IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:8157656
SO  - J Biol Chem 1994 Apr 15 ;269(15):11261-11268

1265
UI  - 820
AU  - Werner-Grune S
AU  - Gunkel D
AU  - Schumann J
AU  - Strotmann H
AD  - Institut fur Biochemie der Pflanzen, Heinrich-Heine-Universitat Dusseldorf, Germany
TI  - Insertion of a "chloroplast-like" regulatory segment responsible for thiol modulation into gamma-subunit of F0F1-ATPase of the cyanobacterium Synechocystis 6803 by mutagenesis of atpC
AB  - A regulatory sequence in the gamma subunit of the F0F1-ATPase complex of higher plant chloroplasts, responsible for so-called thiol modulation, is absent in the corresponding polypeptides of the cyanobacterial complexes analysed so far. We have modified the atpC gene encoding this gamma subunit in Synechocystis 6803 by site-directed mutagenesis. A segment was introduced coding for nine additional amino acids, including the two functional cysteines, which constitutes the sequence of the respective element in the chloroplast gamma subunit. The growth rate as well as the rate of photosynthesis of the transformant was comparable to that of the wild-type, but the transitory increase in respiration observed immediately after a period of illumination was significantly lower in the mutant than in the wild- type. The F1 subcomplex solubilized from thylakoid membranes of both the wild-type and the transformant can be activated by trypsin to yield Ca(2+)-dependent ATPase activity, but only the F1 from the transformant can be activated by the thiol reagent dithiothreitol
RP  - NOT IN FILE
NT  - UI - 94329066LA - engRN - 0 (DNA Primers)RN - 0 (DNA, Bacterial)RN - 0 (Macromolecular Systems)RN - 50-99-7 (Glucose)PT - Journal ArticleDA - 19940902IS - 0026-8925SB - IMCY - GERMANYJC - NGP
UR  - PM:8052233
SO  - Mol Gen Genet 1994 Jul 25 ;244(2):144-150

1266
UI  - 428
AU  - Wilke-Mounts S
AU  - Weber J
AU  - Grell E
AU  - Senior AE
AD  - Department of Biochemistry, University of Rochester Medical Center, New York 14642
TI  - Tryptophan-free Escherichia coli F1-ATPase
AB  - We have engineered a mutant form of Escherichia coli F1-ATPase which is tryptophan-free and contains five mutations, namely delta W28L/alpha W513F/gamma W108Y/gamma W206Y/beta W107F. A strain carrying all five mutations grew normally by oxidative phosphorylation. Purified mutant F1-ATPase showed Vmax and Km both 65% higher than wild-type, resulting in kcat/Km the same as wild-type. The pH dependence of ATPase activity in mutant enzyme was very similar to that in wild-type. Catalytic-site nucleotide-binding characteristics were measured using the analog lin- benzo-ADP and sensitivity to inhibitors was tested using dicyclohexylcarbodiimide, azide and aurovertin. The mutant enzyme was very similar to wild-type in each of these characteristics. The fluorescence spectrum of mutant enzyme confirmed the absence of tryptophan. We have therefore established that it is possible to generate a tryptophan-free enzyme which retains normal catalytic function, oligomeric stability and in vivo assembly
RP  - NOT IN FILE
NT  - UI - 94182952LA - engRN - 0 (Aurovertins)RN - 0 (Azides)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 61925-59-5 (linear-benzoadenosine diphosphate)RN - 73-22-3 (Tryptophan)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSDA - 19940419IS - 0003-9861SB - IMCY - UNITED STATESJC - 6SK
UR  - PM:8135549
SO  - Arch Biochem Biophys 1994 Mar ;309(2):363-368

1267
UI  - 170
AU  - Wilkens S
AU  - Capaldi RA
AD  - Institute of Molecular Biology, University of Oregon, Eugene 97403
TI  - Asymmetry and structural changes in ECF1 examined by cryoelectronmicroscopy
AB  - The Escherichia coli ATPase (ECF1) has been studied by cryoelectronmicroscopy and an intrinsic asymmetry of the molecule in the hexagonal projection identified. The three beta subunits could be distinguished. One, which we have called beta 1, has a greater density in projection than the other two; the second, beta 2, is of intermediate density in projection, while the third, beta 3, is smeared out in density. These different features of the beta subunits were used to orient images, and the positions of the gamma and epsilon subunits then established. The location of the gamma subunit, as monitored by the central mass, was not fixed. This subunit could be found in positions that followed an arc from close to beta 2 to close to beta 3, a shift of around 10A, with respect to the center of the mass. The location of the epsilon subunit was monitored after reconstituting a complex of epsilon subunit-depleted ECF1 with a mutant epsilon subunit in which His at residue 38 had been replaced by Cys, and this Cys labeled with an approximately 14A gold particle. The epsilon subunit was found in positions described by an arc between an alpha subunit (alpha 1) and the neighboring beta subunit (beta 1), a shift of around 20A, with respect to the center of the gold particle. A nucleotide dependence of the position of the gamma subunit has been established by Gogol, E.P., Johnston, E., Aggeler, R. and Capaldi, R.A. (1990) Proc. Natl. Acad. Sci. USA 87, 9585-9589. A nucleotide dependence of the position of the epsilon subunit is shown here.(ABSTRACT TRUNCATED AT 250 WORDS)
RP  - NOT IN FILE
NT  - UI - 94271495LA - engRN - 0 (Antibodies, Monoclonal)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - HL 24526/HL/NHLBIDA - 19940720IS - 0177-3593SB - IMCY - GERMANYJC - AHC
UR  - PM:8003256
SO  - Biol Chem Hoppe Seyler 1994 Jan ;375(1):43-51

1268
UI  - 163
AU  - Wilkens S
AU  - Dunn SD
AU  - Capaldi RA
AD  - Institute of Molecular Biology, University of Oregon, Eugene 97403
TI  - A cryoelectron microscopy study of the interaction of the Escherichia coli F1-ATPase with subunit b dimer
AB  - A complex between the Escherichia coli F1-ATPase and a truncated form of the ECF0-b subunit was formed and examined by cryoelectron microscopy in amorphous ice. Image analysis of single particles in the hexagonal projection revealed that the polar domain of the b subunit interacts with a beta subunit different from the one which interacts with the epsilon subunit. The cavity in the enzyme, visible in the hexagonal projection, is not filled by the b polypeptide, therefore leaving enough room for extensive conformational changes of the gamma and epsilon subunits within the native F1F0 complex
RP  - NOT IN FILE
NT  - UI - 95046301LA - engRN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - HL 22456/HL/NHLBIDA - 19941202IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:7957897
SO  - FEBS Lett 1994 Oct 31 ;354(1):37-40

1269
UI  - 882
AU  - Williams RJP
TI  - Energy coupling: an introduction
MH  - Adenosine Triphosphate
MH  - biosynthesis
MH  - Electrons
MH  - Energy Metabolism
MH  - Oxidation-Reduction
MH  - Protons
RP  - NOT IN FILE
NT  - Inorganic Chemistry Laboratory, University of Oxford, UK
SO  - Biochim Biophys Acta 1994 Aug 30 ;1187(2):125-128

1270
UI  - 633
AU  - Xiao R
AU  - Penefsky HS
AD  - Department of Biochemistry and Molecular Biology, State University of New York, Syracuse 13210
TI  - Unisite catalysis and the delta subunit of F1-ATPase in Escherichia coli
AB  - The 5-subunit form of the Escherichia coli F1-ATPase, characterized by the subunit composition alpha 3 beta 3 gamma delta epsilon, failed to exhibit a rate acceleration when samples of the enzyme hydrolyzing substoichiometric concentrations of [gamma-32P]ATP were switched from unisite to multisite hydrolysis by the addition of a cold chase. A 4- subunit form of the enzyme lacking in the delta subunit (alpha 3 beta 3 gamma epsilon) did exhibit cold chase-promoted accelerations in the hydrolysis of ATP. Reconstitution of a 5-subunit enzyme by incubating the 4-subunit form of the enzyme with a purified preparation of subunit delta was accompanied by a disappearance in the response to a cold chase. The rate constants and equilibrium constants for unisite catalysis by the 4-subunit enzyme did not differ significantly from previously reported values that may have been based on a mixture of 4- and 5-subunit forms of the enzyme. The vesicular form of Escherichia coli F0F1-ATPase exhibited a response to a cold chase only if the vesicles were first extracted with KCl. [gamma-32P]ATP bound in the high affinity catalytic sites of KCl-extracted membranes partially dissociated in an energy-dependent manner when the vesicles oxidized NADH
RP  - NOT IN FILE
NT  - UI - 94308197LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - 7447-40-7 (Potassium Chloride)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM21737/GM/NIGMSDA - 19940818IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:8034684
SO  - J Biol Chem 1994 Jul 29 ;269(30):19232-19237

1271
UI  - 736
AU  - Yokoyama K
AU  - Akabane Y
AU  - Ishii N
AU  - Yoshida M
AD  - Research Laboratory of Resources Utilization, Tokyo Institute of Technology, Yokohama, Japan
TI  - Isolation of prokaryotic V0V1-ATPase from a thermophilic eubacterium Thermus thermophilus
AB  - The soluble ATPase purified from an aerobic thermophilic eubacterium, Thermus thermophilus, was not a usual F1-ATPase but a V1-ATPase, a peripheral section of plasma membrane V-type ATPase (Yokoyama, K., Oshima, T., and Yoshida, M. (1990) J. Biol. Chem. 265, 21946-21950). Here, we report the purification of V-type ATPase from the same bacterium (V0V1-ATPase) which consists of V1-ATPase and a membrane- integrated section, V0. The V0V1-ATPase, either in the Triton X-100- solubilized membrane fraction or in the purified state, migrates as a single band in a non-denaturing polyacrylamide gel electrophoresis for membrane protein complexes, and eight kinds of polypeptides are found when this band is developed in a second dimension denaturing gel electrophoresis in the presence of sodium dodecyl sulfate. The 66-, 56- , 30-, and 12-kDa polypeptides are the subunits of V1-ATPase and the 100-, 38-, 24-, and 13-kDa polypeptides are candidates for V0 (or V0- associated) subunits. The amino-terminal amino acid sequences of the 38- and 24-kDa subunits do not show obvious similarity to any subunits of eukaryotic V0V1-ATPases. The kinetic properties of the purified V0V1- ATPase are very similar to those of V1-ATPase: a very low ATPase activity, stimulation by bisulfite, inhibition by nitrate, and resistance against inhibitors of eukaryotic V-type ATPases, Bafilomycin A1 and N-ethylmaleimide. The V0 vesicles prepared from reconstituted V0V1-ATPase vesicles by 6 M urea treatment show the H+ channel activity. This H+ channel activity is abolished either by treatment of vesicles with dicyclohexylcarbodiimide or by the back addition of V1- ATPase. These results indicate that the coupling ion of this V0V1- ATPase is H+
RP  - NOT IN FILE
NT  - UI - 94216345LA - engRN - 0 (Macromolecular Systems)RN - 0 (Peptide Fragments)RN - 2001-95-8 (Valinomycin)RN - 555-60-2 (Carbonyl Cyanide m-Chlorophenyl Hydrazone)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19940526IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:8163530
SO  - J Biol Chem 1994 Apr 22 ;269(16):12248-12253

1272
UI  - 20945
AU  - Zanotti F
AU  - Guerrieri F
AU  - Deckers-Hebestreit G
AU  - Fiermonte M
AU  - Altendorf K
AU  - Papa S
AD  - Institute of Medical Biochemistry and Chemistry, University of Bari, Italy
TI  - Cross-reconstitution studies with polypeptides of Escherichia coli and bovine heart mitochondrial F0F1 ATP synthase
AB  - To characterize the role of supernumerary subunits of the mammalian F0F1 ATP synthase, cross-reconstitution of mitochondrial and bacterial F0F1 complexes has been carried out. Escherichia coli F1 (EcF1) can be reconstituted with F1-stripped everted membranes of E. coli (UPEc) and of bovine heart mitochondria (USMP). Bovine heart mitochondrial F1 (BHF1) can also be reconstituted with both membranes. Both EcF1 and BHF1, when reconstituted with UPEc, exhibited oligomycin-insensitive ATP-hydrolase activity. Subunits of the mammalian F0, in particular F0I- PVP protein, F6 and oligomycin-sensitivity-conferring protein (OSCP) conferred oligomycin sensitivity to the catalytic activity of EcF1 or BHF1 reconstituted with UPEc. Reaction of N,N'-dicyclohexylcarbodiimide and development of inhibition of passive H+ conduction was, in UPEc, considerably slower and exhibited a lower apparent affinity than in USMP. The ATP hydrolase activity of UPEc+EcF1 or UPEc+BHF1 was, also, less sensitive to inhibition by N,N'-dicyclohexylcarbodiimide than USMP+EcF1 or USMP+BHF1. Addition of mitochondrial F0I-PVP to UPEc enhanced the sensitivity of H+ conduction to oligomycin. F0I-PVP and OSCP added to UPEc, promoted inhibition by N,N'- dicyclohexylcarbodiimide of passive H+ conduction and increased its binding affinity to subunit c of E. coli F0. The presence of F0I-PVP and OSCP also promoted inhibition by N,N'-dicyclohexylcarbodiimide of the ATP-hydrolase activity of EcF1 or BHF1 reconstituted with UPEc
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BINDING
MH  - Biochemistry
MH  - Chemistry
MH  - COMPLEX
MH  - development
MH  - Dicyclohexylcarbodiimide
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - F0F1
MH  - F1
MH  - H+
MH  - membrane
MH  - Membrane Proteins
MH  - Membranes
MH  - Mitochondria
MH  - Oligomycins
MH  - protein
MH  - Proteins
MH  - proton
MH  - Protons
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 94298812LA - engRN - 0 (Membrane Proteins)RN - 0 (Oligomycins)RN - 0 (Protons)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19940811IS - 0014-2956SB - IMCY - GERMANY
UR  - PM:8026487
SO  - Eur J Biochem 1994 Jun 15 ;222(3):733-741

1273
UI  - 34
AU  - Zhang Y
AU  - Fillingame RH
AD  - Department of Biomolecular Chemistry, University of Wisconsin Medical School, Madison 53706
TI  - Essential aspartate in subunit c of F1F0 ATP synthase. Effect of position 61 substitutions in helix-2 on function of Asp24 in helix-1
AB  - Subunit c of the F1F0 type, H(+)-transporting ATP synthase contains an essential Asp that is thought to function in H+ transport. Subunit c folds as a helical hairpin of two transmembrane helices with the essential Asp centered at residue 61 in transmembrane helix-2. Miller et al. (Miller, M. J., Olderburg, M., and Fillingame, R. H. (1990) Proc. Natl. Acad. Sci. U. S. A. 87, 4900-4904) have described a functional subunit c variant in which the essential Asp was moved from helix-2 to residue 24 on helix-1 with replacement of Asp61 by Gly. The function of the A24D/D61G subunit c variant is not optimal. In this study, 11 position 61 variants of an A24D subunit c were generated by site-directed mutagenesis in order to test the importance of the position 61 residue. Three functional combinations were found with activities in the order:A24D/D61N > A24D/D61G > or = A24D/D61S. Other substitutions at position 61, including Ala and Cys, did not support function in the A24D protein. Although the A24D/D61N variant showed the highest rates of ATPase-coupled H+ transport, its F0 was inactive in passive H+ transport when F1 was stripped from the membrane. On the other hand, passive H+ transport by A24D/D61G and A24D/D61S stripped membranes approached that of wild type. The defect in function in these two mutants must be ascribed to events related to coupling ATPase and H+ transport. An A24D subunit c (with Asp at both position 24 and 61) was also generated. Its function proved to be pH-dependent. Activity approaching that of wild type was observed at pH 7.0, but function was almost completely lost at pH 7.8. The pH-dependent loss of ATP synthase function led to a slowing of growth on succinate as carbon source on raising the pH from 7.0 to 7.8. In the A24D mutant, with a second Asp at position 61, we postulate that 1 Asp must be protonated before the other can function in H+ transport
RP  - NOT IN FILE
NT  - UI - 94149019LA - engRN - 0 (DNA Primers)RN - 0 (Macromolecular Systems)RN - 56-84-8 (Aspartic Acid)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM23105/GM/NIGMSDA - 19940322IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:8106529
SO  - J Biol Chem 1994 Feb 18 ;269(7):5473-5479

1274
UI  - 112
AU  - Zhang Y
AU  - Oldenburg M
AU  - Fillingame RH
AD  - Department of Biomolecular Chemistry, University of Wisconsin Medical School, Madison 53706
TI  - Suppressor mutations in F1 subunit epsilon recouple ATP-driven H+ translocation in uncoupled Q42E subunit c mutant of Escherichia coli F1F0 ATP synthase
AB  - The Q42E mutation in the polar loop of subunit c of the Escherichia coli F1F0 ATP synthase leads to an uncoupling of H+ translocation through F0 and ATP synthesis/hydrolysis in F1. We have isolated four second-site suppressor mutants in which the coupling defect is corrected. Substitutions for Glu31 in F1 subunit epsilon were found in each suppressor mutant, where the substitutions were E31G, E31V, and E31K (the last being found twice). The different substitutions vary in effectiveness in restoring wild type growth properties in the order epsilon E31G > epsilon E31V > epsilon E31K. Biochemical properties of epsilon E31G/cQ42E and epsilon E31K/cQ42E membranes were compared. In epsilon E31G/cQ42E mutant membranes, ATP-driven H+ translocation by F1F0 and the binding and coupling of F1 to F0 showed a striking pH dependence. Near normal function was observed at pH 7.0, but function was lost at pH 7.8. The function of epsilon E31K/cQ42E membranes was much less affected by changes in pH. Relative to epsilon E31G/cQ42E membranes, the ATP-driven H+ transport function of epsilon E31K/cQ42E membranes was approximately the same at pH 7.5, greater at pH 7.8, and less at pH 7.0. The differences between mutants could be explained if cGlu42 ionized at pH 7.8 with loss of function in epsilon E31G/cQ42E membrane and a similar ionization were compensated for by the positively charged Lys in the epsilon E31K/cQ42E membrane
RP  - NOT IN FILE
NT  - UI - 94193705LA - engRN - 0 (Glutamates)RN - 1333-74-0 (Hydrogen)RN - 56-65-5 (Adenosine Triphosphate)RN - 56-86-0 (Glutamic Acid)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM23105/GM/NIGMSDA - 19940505IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:7908291
SO  - J Biol Chem 1994 Apr 8 ;269(14):10221-10224

1275
UI  - 634
AU  - Ziegler M
AU  - Xiao R
AU  - Penefsky HS
AD  - Department of Biochemistry and Molecular Biology, State University of New York, Health Science Center at Syracuse 13210
TI  - Close proximity of Cys64 and Cys140 in the delta subunit of Escherichia coli F1-ATPase
AB  - The delta subunit of the F1-ATPase from Escherichia coli contains 2 cysteine residues, one at position 64 and the second at position 140 of the amino acid sequence. These residues were specifically labeled with sulfhydryl reagents in this study without labeling other -SH groups in the enzyme. Modification of Cys140 by maleimides such as N- ethylmaleimide or fluorescein maleimide resulted in a reconstitutively active enzyme that was indistinguishable from the native protein. Labeling of Cys64 with or without concomitant labeling of Cys140 resulted in a reconstitutively inactive enzyme. The ATPase activity of either form of the labeled enzyme was unaffected. However, labeling of Cys64 was accompanied by dissociation of the delta subunit from the enzyme. These observations suggest a role for the microenvironment of Cys64 in interactions of the delta subunit with other subunits in the enzyme. Two types of evidence support the conclusion that the 2 cysteine residues of the delta subunit are in close proximity. First, incorporation of pyrene maleimide into both delta cysteines led to excimer formation. Second, incubation of F1 with 5,5'-dithiobis(2- nitrobenzoic acid) resulted in quantitative formation of a disulfide bond between Cys64 and Cys140, presumably via disulfide interchange. The enzyme containing the internally cross-linked delta subunit exhibited an undiminished ability to support proton pumping when reconstituted into F1-depleted membrane vesicles. The presence of 2 closely apposed cysteinyl residues in the delta subunit of the native enzyme places constraints on the type of structure that may be proposed for the subunit
RP  - NOT IN FILE
NT  - UI - 94140846LA - engRN - 0 (Membrane Proteins)RN - 0 (Sulfhydryl Reagents)RN - 52-90-4 (Cysteine)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM21737/GM/NIGMSDA - 19940317IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:8307987
SO  - J Biol Chem 1994 Feb 11 ;269(6):4233-4239

1276
UI  - 92
AU  - Aggeler R
AU  - Weinreich F
AU  - Capaldi RA
AD  - Institute of Molecular Biology, University of Oregon, Eugene 97403- 1229, USA
TI  - Arrangement of the epsilon subunit in the Escherichia coli ATP synthase from the reactivity of cysteine residues introduced at different positions in this subunit
AB  - ECF1F0 has been purified from three mutants in which a Cys has been incorporated by site-directed mutagenesis in the epsilon subunit: these mutants are epsilon S10C, epsilon H38C and epsilon S108C, respectively. ECF1F0 from the mutant epsilon S10C had a 2-fold higher activity than wild-type enzyme, due to altered association of the epsilon subunit with the rest of the complex, and yet showed normal proton pumping function. The other two mutants had ATPase activities similar to wild- type enzyme. The introduced Cys was exposed for reaction with maleimides in epsilon S10C and epsilon S108C. In epsilon H38C, the introduced Cys reacted readily with N-ethylmaleimide in isolated ECF1, but was unavailable for reaction with this or other maleimides in ECF1F0. When this Cys at position 38 in the epsilon subunit was reacted with various maleimides in isolated ECF1 and then the ECF1 bound back to F0, the interaction between the two parts was perturbed. While ECF1F0 reconstituted with unmodified ECF1 functioned normally, enzyme with maleimide-reacted Cys-38 showed much reduced proton pumping, had only around 50% of the DCCD inhibition of unmodified or wild-type enzyme, and had a much higher LDAO activation (as much as 8.3-fold, c.f. 4-fold for wild type). Nucleotide-dependent conformational changes have been observed previously, in studies of ECF1 from the mutants epsilon S10C and epsilon S108C. Identical nucleotide-dependent structural changes were observed in cross-linking experiments with tetrafluorophenylazide maleimides when the intact ECF1F0 from these mutants was examined. Taken together, the Cys reactivity data and cross- linking results provide the orientation of the epsilon subunit in the enzyme complex
RP  - NOT IN FILE
NT  - UI - 95337123LA - engRN - 52-90-4 (Cysteine)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - HL24526/HL/NHLBIDA - 19950824IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:7612642
SO  - Biochim Biophys Acta 1995 Jun 1 ;1230(1-2):62-68

1277
UI  - 93
AU  - Aggeler R
AU  - Haughton MA
AU  - Capaldi RA
AD  - Institute of Molecular Biology, University of Oregon, Eugene 97403, USA
TI  - Disulfide bond formation between the COOH-terminal domain of the beta subunits and the gamma and epsilon subunits of the Escherichia coli F1- ATPase. Structural implications and functional consequences
AB  - A set of mutants of the Escherichia coli F1F0-type ATPase has been generated by site-directed mutagenesis as follows: beta E381C, beta S383C, beta E381C/epsilon S108C, and beta S383C/epsilon S108C. Treatment of ECF1 isolated from any of these mutants with CuCl2 induces disulfide bond formation. For the single mutants, beta E381C and beta S383C, a disulfide bond is formed in essentially 100% yield between a beta subunit and the gamma subunit, probably at Cys87 based on the recent structure determination of F1 (Abrahams, J. P., Leslie, A. G. W., Lutter, R., and Walker, J. E. (1994) Nature 370, 621-628). In the double mutants, two disulfide bonds are formed, again in essentially full yield, one between beta and gamma, the other between a beta and the epsilon subunit via Cys108. The same two cross-links are produced with CuCl2 treatment of ECF1F0 isolated from either of the double mutants. These results show that the parts of gamma around residue 87 (a short alpha-helix) and the epsilon subunit interact with different beta subunits. The yield of covalent linkage of beta to gamma is nucleotide dependent and highest in ATP and much lower with ADP in catalytic sites. The yield of covalent linkage of beta to epsilon is also nucleotide dependent but in this case is highest in ADP and much lower in ATP. Disulfide bond formation between either beta and gamma, or beta and epsilon inhibits the ATPase activity of the enzyme in proportion to the yield of the cross-linked product. Chemical modification of the Cys at either position 381 or 383 of the beta subunit inhibits ATPase activity in a manner that appears to be dependent on the size of the modifying reagent. These results are as expected if movements of the catalytic site-containing beta subunits relative to the gamma and epsilon subunits are an essential part of the cooperativity of the enzyme
RP  - NOT IN FILE
NT  - UI - 95238425LA - engRN - 0 (Disulfides)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - HL 24526/HL/NHLBIDA - 19950523IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:7721834
SO  - J Biol Chem 1995 Apr 21 ;270(16):9185-9191

1278
UI  - 21146
AU  - Alexiev U
AU  - Scherrer P
AU  - Marti T
AU  - Khorana HG
AU  - Heyn MP
AD  - Biophysics Group, Freie Universitat Berlin, Germany
TI  - Time-resolved surface charge change on the cytoplasmic side of bacteriorhodopsin
AB  - The pH-sensitive dye 5-iodoacetamidofluorescein was covalently bound to a single cysteine residue introduced by site-directed mutagenesis in position 101 on the cytoplasmic surface or in position 130 on the extracellular surface of the proton pump bacteriorhodopsin. Using time- resolved absorption spectroscopy at 495 nm a transient increase was observed in the apparent pK of the dye attached at residue 101. At pH 7.3 the rise and decay times of this pK-change (approximately 2 ms and approximately 60 ms) correlate well with decay times observed for the M and O intermediates and with the proton uptake time. Interpreting the pK-increase of +0.18 pH-unit in terms of a transiently more negative surface charge density, we calculate a change of -0.80 elementary charge per bacteriorhodopsin at the cytoplasmic surface. It is likely that this charge change is due to the transient deprotonation of aspartate-96. With the label in position 130 on the extracellular surface no transient pK-shift was detected
MH  - A
MH  - absorption
MH  - Bacteriorhodopsin
MH  - Biophysics
MH  - Cysteine
MH  - DYE
MH  - intermediate
MH  - M
MH  - mutagenesis
MH  - pH
MH  - protein
MH  - Proteins
MH  - proton
MH  - Proton Pump
MH  - RESIDUE
MH  - site-directed
MH  - spectroscopy
MH  - SURFACE
MH  - Time
RP  - NOT IN FILE
NT  - UI - 96013891LA - engRN - 0 (Recombinant Proteins)RN - 13699-48-4 (Dimyristoylphosphatidylcholine)RN - 53026-44-1 (Bacteriorhodopsins)PT - Journal ArticleID - AI 11479-18/AI/NIAIDDA - 19951124IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:7589439
SO  - FEBS Lett 1995 Oct 2 ;373(1):81-84

1279
UI  - 18562
AU  - Alexiev U
AU  - Mollaaghababa R
AU  - Scherrer P
AU  - Khorana HG
AU  - Heyn MP
TI  - Rapid long-range proton diffusion along the surface of the purple membrane and delayed proton transfer into the bulk.
AB  - The pH-indicator dye fluorescein was covalently bound to the surface of the purple membrane at position 72 on the extracellular side of bacteriorhopsin and at positions 101, 105, 160, or 231 on the cytoplasmic side by reacting bromomethylfluorescein with the sulfhydryl groups of cysteines introduced by site-directed mutagenesis. At position 72, on the extracellular surface, the light-induced proton release was detected 71 +/- 4 microseconds after the flash (conditions: pH 7.3, 22 degrees C, and 150 mM KCl). On the cytoplasmic side with the dye at positions 101, 105, and 160, the corresponding values were 77, 76, and 74 +/- 5 microseconds, respectively. Under the same conditions, the proton release time in the bulk medium as detected by pyranine was around 880 microseconds--i.e., slower by a factor of more than 10. The fact that the proton that is released on the extracellular side is detected much faster on the cytoplasmic surface than in the aqueous bulk phase demonstrates that it is retained on the surface and migrates along the purple membrane to the other side. These findings have interesting implications for bioenergetics and support models of local proton coupling. From the small difference between the proton detection times by labels on opposite sides of the membrane, we estimate that at 22 degrees C the proton surface diffusion constant is greater than 3 x 10(-5) cm2/s. At 5 degrees C, the proton release detection time at position 72 equals the faster of the two main rise times of the M intermediate (deprotonation of the Schiff base). At higher temperatures this correlation is gradually lost, but the curved Arrhenius plot for the proton release time is tangential to the linear Arrhenius plot for the rise of M at low temperatures. These observations are compatible with kinetic coupling between Schiff base deprotonation and proton release.
MH  - A
MH  - Affinity Labels
MH  - Amino Acid Sequence
MH  - Bacteriorhodopsin
MH  - BASE
MH  - bioenergetics
MH  - Cell Polarity
MH  - CONSTANT
MH  - Cysteine
MH  - Diffusion
MH  - DYE
MH  - flash
MH  - fluorescein
MH  - Fluoresceins
MH  - genetics
MH  - Halobacterium
MH  - Heat
MH  - intermediate
MH  - Kinetics
MH  - Light
MH  - low temperature
MH  - M
MH  - M-intermediate
MH  - membrane
MH  - metabolism
MH  - microsecond
MH  - model
MH  - Models,Biological
MH  - Molecular Sequence Data
MH  - mutagenesis
MH  - pH
MH  - pH-indicator
MH  - Protein Engineering
MH  - proton
MH  - proton release
MH  - Proton transfer
MH  - Proton-transfer
MH  - Protons
MH  - purple membrane
MH  - radiation effects
MH  - RISE TIME
MH  - Schiff base
MH  - Schiff-base
MH  - site-directed
MH  - Spectrophotometry
MH  - Support,Non-U.S.Gov't
MH  - Support,U.S.Gov't,P.H.S.
MH  - SURFACE
MH  - Temperature
MH  - Time
MH  - TRANSFER
RP  - NOT IN FILE
NT  - Department of Physics, Freie Universitat, Berlin, GermanyPMID- 0007831293
SO  - Proc Natl Acad Sci U S A 1995 Jan 17 ;92(2):372-376

1280
UI  - 21149
AU  - Alexiev U
AU  - Mollaaghababa R
AU  - Scherrer P
AU  - Khorana HG
AU  - Heyn MP
AD  - Department of Physics, Freie Universitat, Berlin, Germany
TI  - Rapid long-range proton diffusion along the surface of the purple membrane and delayed proton transfer into the bulk
AB  - The pH-indicator dye fluorescein was covalently bound to the surface of the purple membrane at position 72 on the extracellular side of bacteriorhopsin and at positions 101, 105, 160, or 231 on the cytoplasmic side by reacting bromomethylfluorescein with the sulfhydryl groups of cysteines introduced by site-directed mutagenesis. At position 72, on the extracellular surface, the light-induced proton release was detected 71 +/- 4 microseconds after the flash (conditions: pH 7.3, 22 degrees C, and 150 mM KCl). On the cytoplasmic side with the dye at positions 101, 105, and 160, the corresponding values were 77, 76, and 74 +/- 5 microseconds, respectively. Under the same conditions, the proton release time in the bulk medium as detected by pyranine was around 880 microseconds--i.e., slower by a factor of more than 10. The fact that the proton that is released on the extracellular side is detected much faster on the cytoplasmic surface than in the aqueous bulk phase demonstrates that it is retained on the surface and migrates along the purple membrane to the other side. These findings have interesting implications for bioenergetics and support models of local proton coupling. From the small difference between the proton detection times by labels on opposite sides of the membrane, we estimate that at 22 degrees C the proton surface diffusion constant is greater than 3 x 10(-5) cm2/s. At 5 degrees C, the proton release detection time at position 72 equals the faster of the two main rise times of the M intermediate (deprotonation of the Schiff base). At higher temperatures this correlation is gradually lost, but the curved Arrhenius plot for the proton release time is tangential to the linear Arrhenius plot for the rise of M at low temperatures. These observations are compatible with kinetic coupling between Schiff base deprotonation and proton release
MH  - A
MH  - Affinity Labels
MH  - Bacteriorhodopsin
MH  - BASE
MH  - bioenergetics
MH  - CONSTANT
MH  - coupling
MH  - Cysteine
MH  - Diffusion
MH  - DYE
MH  - flash
MH  - fluorescein
MH  - Fluoresceins
MH  - intermediate
MH  - low temperature
MH  - M
MH  - M-intermediate
MH  - membrane
MH  - microsecond
MH  - model
MH  - mutagenesis
MH  - pH
MH  - pH-indicator
MH  - proton
MH  - proton release
MH  - Proton transfer
MH  - Proton-transfer
MH  - Protons
MH  - purple membrane
MH  - RISE TIME
MH  - Schiff base
MH  - Schiff-base
MH  - site-directed
MH  - SURFACE
MH  - Temperature
MH  - Time
MH  - TRANSFER
RP  - NOT IN FILE
NT  - UI - 95132601LA - engRN - 0 (Affinity Labels)RN - 0 (Fluoresceins)RN - 0 (Protons)RN - 148942-72-7 (5-(bromomethyl)fluorescein)RN - 52-90-4 (Cysteine)RN - 53026-44-1 (Bacteriorhodopsins)PT - Journal ArticleID - GM28289/GM/NIGMSDA - 19950222IS - 0027-8424SB - IMCY - UNITED STATES
UR  - PM:7831293
SO  - Proc Natl Acad Sci U S A 1995 Jan 17 ;92(2):372-376

1281
UI  - 24
AU  - Assadi-Porter FM
AU  - Fillingame RH
AD  - Department of Biomolecular Chemistry, University of Wisconsin Medical School, Madison 53706, USA
TI  - Proton-translocating carboxyl of subunit c of F1Fo H(+)-ATP synthase: the unique environment suggested by the pKa determined by 1H NMR
AB  - Subunit c of the H(+)-transporting F1Fo ATP synthase (EC 3.6.1.34) is thought to fold across the membrane as a hairpin of two alpha helices with a conserved Asp/Glu residue, centered in the second membrane- spanning helix, which is thought to function in H+ translocation. NMR studies indicate that the purified subunit c from Escherichia coli is also folded as a hairpin in a chloroform/methanol/H2O (4:4:1) solvent mixture [Girvin, M. E., & Fillingame, R. H. (1993) Biochemistry 32, 12167-12177] and that the conserved Asp remains uniquely reactive in this solvent mixture [Girvin, M. E., & Fillingame, R. H. (1994) Biochemistry 33, 665-674]. The pKa of Asp61 is of interest because of its unique reactivity and because it is thought to protonate and deprotonate during each proton translocation cycle. We have determined the pKa value of the carboxyl group of the functional Asp in wild type and two functional, mutant subunit c proteins, i.e. the Ala24-->Asp (D24D61) and the Ala24-->Asp/Asp61-->Asn (D24N61) mutant proteins. The pKa values were determined by 1H NMR spectroscopy by measuring changes in the alpha and beta proton chemical shifts by constant time two- dimensional (2D) correlated spectroscopy. The pKa of Asp61 in the purified wild type protein was 7.1. This pKa was significantly higher than the pKa of the other two Asp residues, i.e. Asp7 and Asp44 which were 5.4 and 5.6, respectively. The pKa of the two Glu residues in the protein were determined by 2D total correlation spectroscopy and found to be approximately 5.5.(ABSTRACT TRUNCATED AT 250 WORDS)
RP  - NOT IN FILE
NT  - UI - 96101450LA - engRN - 0 (Macromolecular Systems)RN - 1333-74-0 (Hydrogen)RN - 56-84-8 (Aspartic Acid)RN - 56-86-0 (Glutamic Acid)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM23105/GM/NIGMSID - R02301/PHSID - RR02301/RR/NCRRID - etcDA - 19960125IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:8519776
SO  - Biochemistry 1995 Dec 12 ;34(49):16186-16193

1282
UI  - 20942
AU  - Bartl F
AU  - Deckers-Hebestreit G
AU  - Altendorf K
AU  - Zundel G
AD  - Institute of Physical Chemistry, University of Munich, Germany
TI  - The F0 complex of the ATP synthase of Escherichia coli contains a proton pathway with large proton polarizability caused by collective proton fluctuation
AB  - The F0 complex of the Escherichia coli ATP synthase embedded into cardiolipin liposomes was studied by FT-IR spectroscopy. For comparison, respective studies were performed with dried F0 liposomes and with F0 liposomes treated with N,N'-dicyclohexyl-carbodiimide (DCCD), which binds to Asp-61 of subunit c. Furthermore, the effect of H2O-->D2O exchange on the infrared spectrum was investigated. With F0 liposomes an infrared continuum is observed beginning at about 3000 cm- 1 and extending toward smaller wavenumbers. In the DCCD-treated sample, this continuum is no longer observed. It vanishes also with drying of the liposomes. After H2O-->D2O exchange, this infrared continuum begins at about 2350 cm-1 and is less intense. All of these results demonstrate that a proton pathway in native F0 is present, in which the protons are shifted in a hydrogen-bonded chain with large proton polarizability due to collective proton tunneling. With the D2O- hydrated system, deuteron polarizability due to collective deuteron motion is observed, but the polarizability due to collective deuteron motion is smaller. Such pathways are very efficient, because they conduct protons or deuterons within picoseconds. These pathways lose their polarizability if the F0 complex is blocked by DCCD or if the liposomes are dried. On the basis of our results on the proton polarizability of hydrogen bonds and hydrogen-bonded systems and on the basis of structural data from the literature, the nature of the proton pathway of the F0 complex of E. coli is discussed
MH  - A
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Chemistry
MH  - COLI ATP SYNTHASE
MH  - COMPLEX
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - Hydrogen
MH  - Liposomes
MH  - proton
MH  - Protons
MH  - spectra
MH  - spectroscopy
MH  - SUBUNIT
MH  - SYNTHASE
MH  - SYSTEM
MH  - SYSTEMS
RP  - NOT IN FILE
NT  - UI - 95226635LA - engRN - 0 (Cardiolipins)RN - 0 (Liposomes)RN - 0 (Protons)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19950518IS - 0006-3495SB - IMCY - UNITED STATES
UR  - PM:7711231
SO  - Biophys J 1995 Jan ;68(1):104-110

1283
UI  - 566
AU  - Belogrudov GI
AU  - Tomich JM
AU  - Hatefi Y
AD  - Department of Molecular and Experimental Medicine, Scripps Research Institute, La Jolla, California 92037
TI  - ATP synthase complex. Proximities of subunits in bovine submitochondrial particles
AB  - The catalytic sector, F1, and the membrane sector, F0, of the mitochondrial ATP synthase complex are joined together by a 45-A-long stalk. Knowledge of the composition and structure of the stalk is crucial to investigating the mechanism of conformational energy transfer between F0 and F1. This paper reports on the near neighbor relationships of the stalk subunits with one another and with the subunits of F1 and F0, as revealed by cross-linking experiments. The preparations subjected to cross-linking were bovine heart submitochondrial particles (SMP) and F1-deficient SMP. The cross- linkers were three reagents of different chemical specificities and different lengths of cross-linking from zero to 10 A. Cross-linked products were identified after gel electrophoresis of the particles and immunoblotting with subunit-specific antibodies to the individual subunits alpha, beta, gamma, delta, OSCP, F6, A6L, a (subunit 6), b, c, and d. The results suggested that the two b subunits form the principal stem of the stalk to which OSCP, d, and F6 are bound independent of one another. Subunits b, OSCP, d, and F6 cross-linked to alpha and/or beta, but not to gamma or delta. The COOH-terminal half of A6L, which is extramembranous, cross-linked to d but not to any other stalk or F1 subunit. No cross-links of subunits a and c with any stalk or F1 subunits were detected. In F1-deficient SMP, cross-linked b+b and d+F6 dimers appeared, and the extent of cross-linking between b and OSCP diminished greatly. The addition of F1 to F1-deficient particles appeared to reverse these changes. Treatment of F1-deficient particles with trypsin rapidly hydrolyzed away OSCP and F6, fragmented b to membrane-bound 18-, 12-, and 8-9-kDa antigenic fragments, which cross- linked to d and/or with one another. Trypsin also removed the COOH- terminal part of A6L, but the remainder still cross-linked to subunit d. Models showing the near neighbor relationships of the stalk subunits with one another and with the alpha and beta subunits at a level near the proximal end (bottom) of F1 and at the membrane-matrix interface are presented
RP  - NOT IN FILE
NT  - UI - 95138166LA - engRN - 0 (Cross-Linking Reagents)RN - 0 (Cytochromes)RN - 0 (Membrane Proteins)RN - 0 (Multienzyme Complexes)RN - 0 (Sulfhydryl Compounds)RN - 0 (oligomycin sensitivity-conferring protein)RN - EC 3.4.21.4 (Trypsin)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - DK08126/DK/NIDDKDA - 19950302IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:7836433
SO  - J Biol Chem 1995 Feb 3 ;270(5):2053-2060

1284
UI  - 9949
AU  - Berendsen HJC
AU  - van der Spoel D
AU  - van Drunen D
TI  - GROMACS: A message-passing parallel molecular dynamics implementation.
RP  - NOT IN FILE
SO  - Comp Phys Comm 1995  ;91():43-56

1285
UI  - 20940
AU  - Birkenhager R
AU  - Hoppert M
AU  - Deckers-Hebestreit G
AU  - Mayer F
AU  - Altendorf K
AD  - Universitat Osnabruck, Fachbereich Biologie/Chemie, Arbeitsgruppe Mikrobiologie, Germany
TI  - The F0 complex of the Escherichia coli ATP synthase. Investigation by electron spectroscopic imaging and immunoelectron microscopy
AB  - Cholate-solubilized F0 complexes of the ATP synthase (F0F1) from Escherichia coli were studied by application of conventional transmission electron microscopy and electron spectroscopic imaging (ESI) of negatively stained samples. Using the ESI mode, the structural organization of the F0 complex (diameter of 7.5 +/- 0.5 nm) could be observed in more detail and defined projections could be distinguished. Projection A appears as a deltoid-like structure with bilateral symmetry. Projection B has an overall trapezoidal shape with some similarity in shape to the letter W. Applying the ESI mode to the ac complex dissolved in cholate-containing buffer, an elongated structure consisting of two intensity maxima could be observed. Simulations with models of the F0 and the ac complex revealed that the projections observed can be obtained by tilting and rotating a model in which subunit a and the two copies of subunit b are located outside the subunit c oligomer. This view of structural organization was supported by results obtained with F0 complexes decorated with monoclonal antibodies against subunits a, b or c
MH  - A
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - buffer
MH  - COLI ATP SYNTHASE
MH  - COMPLEX
MH  - electron
MH  - electron microscopy
MH  - ELECTRON-MICROSCOPY
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - F0F1
MH  - IMMUNOELECTRON MICROSCOPY
MH  - Microscopy
MH  - model
MH  - MONOCLONAL-ANTIBODIES
MH  - SIMULATION
MH  - SIMULATIONS
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 95324551LA - engRN - 0 (Antibodies, Monoclonal)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19950809IS - 0014-2956SB - IMCY - GERMANY
UR  - PM:7601125
SO  - Eur J Biochem 1995 May 15 ;230(1):58-67

1286
UI  - 9891
AU  - Bottcher B
AU  - Lucken U
AU  - Graber P
TI  - The structure of the H+-ATPase from chloroplasts by electron cryomicroscopy
MH  - chloroplast
MH  - Chloroplasts
MH  - COMPLEX
MH  - electron
MH  - England
MH  - F0
MH  - H+-ATPase
MH  - Microscopy
MH  - Mitochondria
MH  - SPECIMENS
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - JournalArticlePORTLAND PRESSNOVTH865LONDONFRITZ HABER INST,D-14195 BERLIN,GERMANYBIOCHEM SOC TRANS59 PORTLAND PLACE, LONDON, ENGLAND W1N 3AJ
AV  - FRITZ HABER INST,D-14195 BERLIN,GERMANY UNIV FREIBURG,INST PHYS CHEM,D-79104 FREIBURG,GERMANY
UR  - ISI:A1995TH86500017
SO  - Biochemical Society Transactions 1995  ;23(4):780-785

1287
UI  - 9892
AU  - Bottcher B
AU  - Graber P
AU  - BOEKEMA EJ
AU  - Lucken U
TI  - ELECTRON CRYOMICROSCOPY OF 2-DIMENSIONAL CRYSTALS OF THE H+- ATPASE FROM CHLOROPLASTS
AB  - The H+-ATPase from spinach chloroplasts was isolated and purified, Two-dimensional crystals were obtained from the protein/lipid/detergent micelles by treatment with phospholipase and simultaneous removal of detergent and fatty acids by Biobeads. The resulting two-dimensionally ordered arrays were investigated by electron cryomicroscopy, The ordered arrays showed top view projections of CF0F1. The images were analysed by correlation averaging, In this view CF0F1 has dimensions of 11.4 x 9 am. The average view shows a strongly asymmetric molecule, in contrast to the rather hexagonal features of CF1, previously analyzed from two-dimensional arrays. It is concluded that this is due either to an asymmetric structure and positioning of CP0 relative to CF1 or to a rearrangement of CF1 subunits induced by binding of CF0 to CP1
MH  - 2-DIMENSIONAL CRYSTALS
MH  - ATPase
MH  - BINDING
MH  - CF0F1
MH  - chloroplast
MH  - Chloroplasts
MH  - COMPLEX
MH  - electron
MH  - ELECTRON CRYOMICROSCOPY
MH  - H+-ATPase
MH  - Micelles
MH  - Microscopy
MH  - SPECIMENS
MH  - Spinach
MH  - SPINACH-CHLOROPLASTS
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - JournalArticleELSEVIER SCIENCE BVOCT 16TA662AMSTERDAMUNIV FREIBURG,INST PHYS CHEM,D-79104 FREIBURG,GERMANYFEBS LETTPO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS
AV  - UNIV FREIBURG,INST PHYS CHEM,D-79104 FREIBURG,GERMANY FRITZ HABER INST,D-14195 BERLIN,GERMANY UNIV GRONINGEN,GRONINGEN,NETHERLANDS
UR  - ISI:A1995TA66200017
SO  - Febs Letters 1995  ;373(3):262-264

1288
UI  - 19773
AU  - Boyer PD
TI  - From human serum albumin to rotational catalysis by ATP synthase
MH  - atp
MH  - ATP synthase
MH  - Catalysis
MH  - H(+)-Transporting ATP Synthase
MH  - Human
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 95255612LA - engRN - 0 (Serum Albumin)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - BiographyPT - Historical ArticlePT - Journal ArticleDA - 19950606IS - 0892-6638SB - IMCY - UNITED STATESJC - FAS
UR  - PM:7737466
SO  - FASEB J 1995 Apr ;9(7):559-561

1289
UI  - 88
AU  - Capaldi RA
AU  - Aggeler R
AU  - Wilkens S
AD  - Institute of Molecular Biology, University of Oregon, Eugene 97403, USA
TI  - Conformational changes in the gamma and epsilon subunits are integral to the functioning of the Escherichia coli H(+)-pumping ATPase (ECF1F0)
AB  - ATP synthesis and ATP hydrolysis by F1F0-type ATPases involve conformational changes transmitted from the catalytic site regions to the proton channel, a distance of more than 100 A. Our studies focus attention on the gamma and epsilon subunits that provide a part of the stalk region in the energy-coupling process within the complex. There are conformational changes in the gamma subunit, and translocations of the epsilon unit, linked to nucleotide-binding changes in catalytic sites, which might be expected to alter the interaction of this subunits with c subunits and, hence, be linked to proton translocation
RP  - NOT IN FILE
NT  - UI - 96227125LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - HL24526/HL/NHLBIDA - 19960730IS - 0300-5127SB - IMCY - ENGLANDJC - E48
UR  - PM:8654834
SO  - Biochem Soc Trans 1995 Nov ;23(4):767-770

1290
UI  - 508
AU  - Chuang WJ
AU  - Abeygunawardana C
AU  - Gittis AG
AU  - Pedersen PL
AU  - Mildvan AS
AD  - Department of Biological Chemistry, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205-2185, USA
TI  - Solution structure and function in trifluoroethanol of PP-50, an ATP- binding peptide from F1ATPase
AB  - PP-50, a synthetic peptide, based on residues 141-190 of the beta- subunit of mitochondrial F1ATPase, containing the GX4GKT consensus sequence for nucleoside triphosphate binding, binds ATP tightly (Kd = 17.5 microM) as found by fluorescence titration at pH 4.0. CD and 2D proton NMR studies at pH 4.0 revealed two beta-turns, regions of extended secondary structure, transient tertiary structure, and flexibility in the GX4GKT region (W.J. Chuang, C. Abeygunawardana, P. L. Pedersen, and A. S. Mildvan, 1992, Biochemistry 31, 7915-7921). CD titration of PP-50 with trifluoroethanol (TFE) reveals a decrease in ellipticity at 208 and 222 nm, saturating at 25% TFE. Computer analysis indicates that 25% TFE increases the helix content from 5.8 to 28.6%, decreases the beta-structure from 30.2 to 20.2% and decreases the coil content from 64 to 51.2%. Fluorescence titrations of H2
RP  - NOT IN FILE
NT  - UI - 95289712LA - engRN - 0 (Peptide Fragments)RN - 0 (Solutions)RN - 56-65-5 (Adenosine Triphosphate)RN - 75-89-8 (Trifluoroethanol)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - CA10951/CA/NCIID - DK28616/DK/NIDDKDA - 19950703IS - 0003-9861SB - IMCY - UNITED STATESJC - 6SK
UR  - PM:7771774
SO  - Arch Biochem Biophys 1995 May 10 ;319(1):110-122

1291
UI  - 993
AU  - Cladera J
AU  - Villaverde J
AU  - Hartog AF
AU  - Padros E
AU  - Berden JA
AU  - Rigaud JL
AU  - Dunach M
TI  - Influence of nucleotides on the secondary structure and on the thermal stability of mitochondrial F1 visualized by infrared spectroscopy
AB  - We have studied the secondary structure of mitochondrial F1 using infrared spectroscopy. Our results show that in the absence of added nucleotides this complex contains similar percentages of alpha-helices, beta-structures and reverse turns (30%, 28% and 31%, respectively). The influence of ADP and ATP on the different types of secondary structure was determined; when all the nucleotide-binding sites were occupied, small but reproducible changes were observed, corresponding to a decrease in beta-structure and an increase in alpha-helix and reverse turns. The effect of nucleotide binding on the thermal stability of F1 was also studied; the thermal denaturation temperature, 55 degrees C, was increased by 11 degrees C and 7 degrees C by ATP and ADP, respectively. These results indicate that nucleotide binding affects the secondary structure of F1, stabilizing the complex
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - Chemistry
MH  - Enzyme Stability
MH  - enzymology
MH  - H(+)-Transporting ATP Synthase
MH  - Heat
MH  - metabolism
MH  - Mitochondria
MH  - Nucleotides
MH  - Protein Structure,Secondary
MH  - Spectroscopy,Fourier Transform Infrared
MH  - Support,Non-U.S.Gov't
MH  - Temperature
RP  - NOT IN FILE
NT  - Dept Bioquimica i Biologia Molecular, Fac Medicina, Universitat Autonoma de Barcelona, Bellaterra, Spain
SO  - FEBS Lett 1995 Sep 4 ;371(2):115-118

1292
UI  - 609
AU  - Crimi M
AU  - Fregni V
AU  - Altimari A
AU  - Melandri BA
AD  - Department of Biology, University of Bologna, Italy
TI  - Unreliability of carotenoid electrochromism for the measure of electrical potential differences induced by ATP hydrolysis in bacterial chromatophores
AB  - ATP hydrolysis induces the activation of the proton ATPase in chromatophores of Rhodobacter capsulatus supplemented with nigericine and 50 mM K+ (i.e. when delta pH < 0.2 units). The value of transmembrane electric potential (delta phi) driving this activation was measured using three different approaches: carotenoid electrochromism, uptake of SCN- and responses of the dye oxonol VI. The value of delta phi calculated from the SCN- uptake, on the basis of an internal volume determined experimentally, was about 140 mV, while that indicated by the electrochromic signal ranged between 35 and 70 mV. Only the value indicated by SCN- distribution is consistent with the energetic requirement for the activation of H(+)-ATPase
RP  - NOT IN FILE
NT  - UI - 95317419LA - engRN - 0 (Carotenoids)RN - 0 (Thiocyanates)RN - 302-04-5 (thiocyanate)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19950803IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:7796913
SO  - FEBS Lett 1995 Jun 26 ;367(2):167-172

1293
UI  - 21252
AU  - Deisenhofer J
AU  - Epp O
AU  - Sinning I
AU  - Michel H
AD  - Max-Planck-Institut fur Biochemie, Martinsried, FRG
TI  - Crystallographic refinement at 2.3 A resolution and refined model of the photosynthetic reaction centre from Rhodopseudomonas viridis
AB  - The atomic model of the photosynthetic reaction centre from the purple bacterium Rhodopseudomonas viridis has been refined to an R-value of 0.193 at 2.3 A resolution. The refined model contains 10,288 non- hydrogen atoms; 10,045 of these have well defined electron density. A Luzzati-plot indicates an average co-ordinate error of 0.26 A. During refinement, the positions of a partially ordered carotenoid, a unibiquinone in the partially occupied QB site, a detergent molecule, seven putative sulphate ions, and 201 water molecules were found. More than half of these waters are bound at interfaces between protein subunits and therefore contribute significantly to subunit interactions. Water molecules also play important structural and probably functional roles in the environment of some of the cofactors. Two water molecules form hydrogen bonds to the accessory bacteriochlorophylls and to the protein in the vicinity of the special pair of bacteriophylls, the primary electron donor. A group of about 10 water molecules is bound near the binding site of the secondary quinone QB. These waters are likely to participate in the transfer of protons to the doubly reduced QB
MH  - A
MH  - Bacteria
MH  - BINDING
MH  - carotenoid
MH  - cytochrome
MH  - Cytochromes
MH  - electron
MH  - England
MH  - Hydrogen
MH  - interfaces
MH  - ion
MH  - Ions
MH  - model
MH  - protein
MH  - Protein Subunits
MH  - proton
MH  - Protons
MH  - quinone
MH  - reaction center
MH  - resolution
MH  - rhodopseudomonas
MH  - secondary
MH  - Site
MH  - SOLVENT
MH  - SUBUNIT
MH  - TRANSFER
MH  - Water
RP  - NOT IN FILE
NT  - UI - 95182453LA - engRN - 0 (Cytochromes)RN - 0 (Photosynthetic Reaction Center, Bacterial)RN - 0 (Solvents)RN - 147-85-3 (Proline)PT - Journal ArticleDA - 19950404IS - 0022-2836SB - IMCY - ENGLAND
UR  - PM:7877166
SO  - J Mol Biol 1995 Feb 24 ;246(3):429-457

1294
UI  - 51
AU  - Dimroth P
AD  - Mikrobiologisches Institut, Eidgenosissche Technische Hochschule, ETH- Zentrum, Zurich, Switzerland
TI  - On the way towards the Na(+)-binding site within the F1F0 ATPase of Propionigenium modestum
RP  - NOT IN FILE
NT  - UI - 96227126LA - engRN - 0 (Enzyme Inhibitors)RN - 0 (Recombinant Proteins)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 7440-23-5 (Sodium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19960730IS - 0300-5127SB - IMCY - ENGLANDJC - E48
UR  - PM:8654835
SO  - Biochem Soc Trans 1995 Nov ;23(4):770-775

1295
UI  - 109
AU  - Dmitriev OY
AU  - Altendorf K
AU  - Fillingame RH
AD  - Department of Biomolecular Chemistry, University of Wisconsin Medical School, Madison, USA
TI  - Reconstitution of the Fo complex of Escherichia coli ATP synthase from isolated subunits. Varying the number of essential carboxylates by co- incorporation of wild-type and mutant subunit c after purification in organic solvent
AB  - Subunit c of the Escherichia coli F1F0-ATPase, purified in chloroform/methanol (2:1), was reconstituted with detergent-solubilized F0 subunits a and b to form a functionally active H+ channel. The rates of H+ uptake by the proteoliposomes containing the reconstituted F0 complex were comparable to those observed with native F0 reconstituted without subunit dissociation. The F0 reconstituted from purified subunits was also shown to form an active ATP-driven H+ pump upon binding of the F1-ATPase sector of the complex. Reconstitution of D61N and D61G mutant c subunits with wild-type subunits a and b produced an inactive F0. Hybrid F0 complexes, formed with mixtures of wild-type and D61N or D61G mutant c subunits, were also prepared. Formation of an active F0 was prevented by addition of relatively small proportions of D61N or D61G mutant c subunits, i.e. active F0 formation was gradually disrupted as the mutant/wild-type ratio was increased from 0.05 to 0.2. The hybrid reconstitution studies support a model where inactivation of one of the 9-12 c subunits found in F0 is sufficient to abolish activity
RP  - NOT IN FILE
NT  - UI - 96067688LA - engRN - 0 (Protons)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM23105/GM/NIGMSDA - 19951218IS - 0014-2956SB - IMCY - GERMANYJC - EMZ
UR  - PM:7588791
SO  - Eur J Biochem 1995 Oct 15 ;233(2):478-483

1296
UI  - 19861
AU  - Duncan TM
AU  - Zhou Y
AU  - Bulygin VV
AU  - Hutcheon ML
AU  - Cross RL
AD  - Department of Biochemistry & Molecular Biology, SUNY Health Science Center, Syracuse, USA
TI  - Probing interactions of the Escherichia coli F0F1 ATP synthase beta and gamma subunits with disulphide cross-links
MH  - atp
MH  - ATP synthase
MH  - Biochemistry
MH  - CROSS-LINKING
MH  - England
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0F1
MH  - H(+)-Transporting ATP Synthase
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 96292006LA - engRN - 0 (Cross-Linking Reagents)RN - 0 (DNA Primers)RN - 0 (DNA, Bacterial)RN - 0 (Disulfides)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM 23152/GM/NIGMSDA - 19960730IS - 0300-5127SB - IMCY - ENGLANDJC - E48
UR  - PM:8654828
SO  - Biochem Soc Trans 1995 Nov ;23(4):736-741

1297
UI  - 19860
AU  - Duncan TM
AU  - Bulygin VV
AU  - Zhou Y
AU  - Hutcheon ML
AU  - Cross RL
AD  - Department of Biochemistry and Molecular Biology, State University of New York Health Science Center, Syracuse 13210, USA
TI  - Rotation of subunits during catalysis by Escherichia coli F1-ATPase
AB  - During oxidative and photo-phosphorylation, F0F1-ATP synthases couple the movement of protons down an electrochemical gradient to the synthesis of ATP. One proposed mechanistic feature that has remained speculative is that this coupling process requires the rotation of subunits within F0F1. Guided by a recent, high-resolution structure for bovine F1 [Abrahams, J. P., Leslie, A. G., Lutter, R. & Walker, J. E. (1994) Nature (London) 370, 621-628], we have developed a critical test for rotation of the central gamma subunit relative to the three catalytic beta subunits in soluble F1 from Escherichia coli. In the bovine F1 structure, a specific point of contact between the gamma subunit and one of the three catalytic beta subunits includes positioning of the homolog of E. coli gamma-subunit C87 (gamma C87) close to the beta-subunit 380DELSEED386 sequence. A beta D380C mutation allowed us to induce formation of a specific disulfide bond between beta and gamma C87 in soluble E. coli F1. Formation of the crosslink inactivated beta D380C-F1, and reduction restored full activity. Using a dissociation/reassembly approach with crosslinked beta D380C-F1, we incorporated radiolabeled beta subunits into the two noncrosslinked beta-subunit positions of F1. After reduction of the initial nonradioactive beta-gamma crosslink, only exposure to conditions for catalytic turnover results in similar reactivities of unlabeled and radiolabeled beta subunits with gamma C87 upon reoxidation. The results demonstrate that gamma subunit rotates relative to the beta subunits during catalysis
MH  - A
MH  - atp
MH  - ATP synthase
MH  - BETA-SUBUNIT
MH  - Biochemistry
MH  - Catalysis
MH  - COLI F1 ATPASE
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0F1
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - H(+)-Transporting ATP Synthase
MH  - Macromolecular Systems
MH  - Movement
MH  - P
MH  - Photophosphorylation
MH  - proton
MH  - Protons
MH  - rotation
MH  - SUBUNIT
MH  - SYNTHASE
MH  - synthesis
RP  - NOT IN FILE
NT  - UI - 96074627LA - engRN - 0 (Disulfides)RN - 0 (Macromolecular Systems)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM 23152/GM/NIGMSDA - 19951228IS - 0027-8424SB - IMCY - UNITED STATESJC - PV3
UR  - PM:7479919
SO  - Proc Natl Acad Sci U S A 1995 Nov 21 ;92(24):10964-10968

1298
UI  - 9932
AU  - Dunlop J
AU  - Jones PC
AU  - Finbow ME
AD  - CRC Beatson Laboratories, Beatson Institute for Cancer Research, Bearsden, Glasgow, UK
TI  - Membrane insertion and assembly of ductin: a polytopic channel with dual orientations
AB  - Ductin is a highly conserved and polytopic transmembrane protein which is the subunit c component of the vacuolar H(+)-ATPase (V-ATPase) and a component of a connexon channel of gap junctions. Previous studies have suggested that ductin in the V-ATPase has the opposite orientation of ductin in a connexon. Using an in vitro translation system coupled to microsomes derived from the endoplasmic reticulum, we show that ductin is co-translationally inserted into the membrane bilayer, suggesting a dependency on the signal recognition particle for synthesis. By attaching a C-terminal polypeptide derived from beta-lactamase and by using cysteine replacement coupled to chemical labelling, we show that ductin is inserted into the microsomal membrane in both orientations in similar proportions. In contrast, squid rhodopsin appears to be inserted in a single orientation. Changing conserved charged residues at the N-terminus of ductin does not affect the ratio of the two orientations. Once in the microsomal membrane, ductin assembles into an oligomeric complex which contains a pore accessible to a water-soluble probe, reminiscent of the ductin complex found in the V-ATPase and a connexon
MH  - atp
MH  - ATP synthase
MH  - COMPLEX
MH  - England
MH  - Fluoresceins
MH  - H(+)-Transporting ATP Synthase
MH  - H(+)ATPase
MH  - In Vitro
MH  - Proteins
MH  - Proteolipids
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 95369232LA - engRN - 0 (Fluoresceins)RN - 0 (Proteolipids)RN - 0 (Recombinant Fusion Proteins)RN - 0 (ductin)RN - 52-90-4 (Cysteine)RN - 75350-46-8 (fluorescein 5-maleimide)RN - 9009-81-8 (Rhodopsin)RN - EC 3.5.2.6 (beta-Lactamases)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19950915IS - 0261-4189SB - IMCY - ENGLANDJC - EMB
UR  - PM:7641680
SO  - EMBO J 1995 Aug 1 ;14(15):3609-3616

1299
UI  - 21107
AU  - Ferguson SJ
AD  - Department of Biochemistry, University of Oxford, UK
TI  - Chemiosmotic coupling. Protons fast and slow
AB  - Recent measurements show that the proton movements of chemiosmotic energy coupling may occur preferentially by localized movements along membrane surfaces
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - ATPase
MH  - Biochemistry
MH  - coupling
MH  - England
MH  - membrane
MH  - Movement
MH  - proton
MH  - Proton-Translocating ATPases
MH  - Protons
MH  - review
MH  - Sodium
MH  - SURFACE
RP  - NOT IN FILE
NT  - UI - 95211432LA - engRN - 0 (Protons)RN - 56-65-5 (Adenosine Triphosphate)RN - 7440-23-5 (Sodium)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 19950504IS - 0960-9822SB - IMCY - ENGLAND
UR  - PM:7697341
SO  - Curr Biol 1995 Jan 1 ;5(1):25-27

1300
UI  - 818
AU  - Fiedler HR
AU  - Schmid R
AU  - Leu S
AU  - Shavit N
AU  - Strotmann H
AD  - Institut fur Biochemie der Pflanzen, Heinrich Heine Universitat Dusseldorf, Germany
TI  - Isolation of CF0CF1 from Chlamydomonas reinhardtii cw15 and the N- terminal amino acid sequences of the CF0CF1 subunits
AB  - CF0CF1 was isolated from chloroplasts of the cell wall-deficient Chlamydomonas reinhardtii strain cw15. The subunit pattern was analyzed by SDS-gel electrophoresis and the N-terminal amino acid sequences of all nine subunits were determined by microsequencing. The amino acid sequences of subunits alpha, beta, gamma and epsilon match with those derived from the corresponding Chlamydomonas DNA sequences. In variance with the previously assumed N-terminus of beta; however, it was found that the first 11 amino acids are lacking. The subunits delta, I, II, III and IV were identified by comparison with known sequences of homologous polypeptides of higher plant chloroplasts and cyanobacteria, respectively
RP  - NOT IN FILE
NT  - UI - 96128220LA - engRN - 9007-49-2 (DNA)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19960213IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:8543042
SO  - FEBS Lett 1995 Dec 18 ;377(2):163-166

1301
UI  - 108
AU  - Fillingame RH
AU  - Girvin ME
AU  - Zhang Y
AD  - Department of Biomolecular Chemistry, University of Wisconsin Medical School, Madison 53706, USA
TI  - Correlations of structure and function in subunit c of Escherichia coli F0F1 ATP synthase
RP  - NOT IN FILE
NT  - UI - 96227124LA - engRN - 0 (Protons)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM23105/GM/NIGMSDA - 19960730IS - 0300-5127SB - IMCY - ENGLANDJC - E48
UR  - PM:8654833
SO  - Biochem Soc Trans 1995 Nov ;23(4):760-766

1302
UI  - 20847
AU  - Futai M
AU  - Omote H
AU  - Maeda M
AD  - Department of Biological Science, Osaka University, Japan
TI  - Escherichia coli H(+)-ATPase (ATP synthase): catalytic site and roles of subunit interactions in energy coupling
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - coupling
MH  - England
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - H(+)ATPase
MH  - Site
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 96227129LA - engRN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19960730IS - 0300-5127SB - IMCY - ENGLAND
UR  - PM:8654838
SO  - Biochem Soc Trans 1995 Nov ;23(4):785-789

1303
UI  - 52
AU  - Gerike U
AU  - Kaim G
AU  - Dimroth P
AD  - Mikrobiologisches Institut, Eidgenossische Technische Hochschule, Zurich, Switzerland
TI  - In vivo synthesis of ATPase complexes of Propionigenium modestum and Escherichia coli and analysis of their function
AB  - Expression studies of Propionigenium modestum ATPase genes in various combinations with Escherichia coli ATPase genes were performed in the unc deletion mutant strain E. coli DK8. Plasmids containing the whole unc operon from P. modestum were unable to complement the E. coli unc deletion mutant. Although all ATPase subunits were expressed from the plasmids, there was no detectable ATP hydrolysing activity, indicating that the F1 part was not functional. Transformants expressing an E. coli F1-P. modestum F0 hybrid exhibited considerable ATPase activities. Binding of the F1 part to the membrane was very weak, however, and the coupling between ATP hydrolysis and Na+ transport was impaired. After combining the genes for E. coli ATPase subunits alpha, beta, gamma, delta and epsilon and the hydrophilic part of subunit b with P. modestum ATPase subunits a and c and the hydrophobic part of subunit b on a plasmid, a non-functional hybrid ATPase was expressed in E. coli. The ATPase was only loosely bound to the membrane, from which it was solubilized with Triton X-100 and purified. Subunit b and a proteolytic degradation product were the only F0 subunits detectable in the purified enzyme. A stable F0 complex is thus not formed with the hybrid b subunit. The absence of a functional F0 complex was in accord with proton-conduction measurements with bacterial vesicles. The only functional Na(+)-translocating ATPase expressed in E. coli thus far consists of E. coli subunits alpha, beta, gamma and epsilon, and P. modestum subunits delta, a, b and c [Kaim, G. & Dimroth, P. (1993) Eur. J. Biochem. 218, 937-944]. During the cloning conducted in our present study, errors in the sequence entry into the EMBL data bank (accession no. X58461) for the P. modestum ATPase alpha and beta subunits became evident, which are corrected in this paper
RP  - NOT IN FILE
NT  - UI - 96035897LA - engRN - 0 (DNA, Bacterial)RN - 0 (Plasmids)RN - 0 (Recombinant Proteins)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19951114IS - 0014-2956SB - IMCY - GERMANYJC - EMZ
UR  - PM:7556212
SO  - Eur J Biochem 1995 Sep 1 ;232(2):596-602

1304
UI  - 111
AU  - Girvin ME
AU  - Fillingame RH
AD  - Department of Biomolecular Chemistry, University of Wisconsin Medical School, Madison 53706
TI  - Determination of local protein structure by spin label difference 2D NMR: the region neighboring Asp61 of subunit c of the F1F0 ATP synthase
AB  - Purified subunit c from the H(+)-transporting F1F0 ATP synthase of Escherichia coli folds as an antiparallel pair of extended helices in a solution of chloroform-methanol-water. A similar hairpin-like folding is predicted for the native protein in the multisubunit transmembrane Fo sector of the ATP synthase. A single Cys variant (A67C) of subunit c was created and modified with a maleimido-PROXYL [[3-(maleimidomethyl)- 2,2,5,5-tetramethyl-1-pyrrolidinyl]oxy] spin label. Pairs of 1H 2D correlation and NOE spectra were collected with the nitroxide oxidized (paramagnetic) and reduced (diamagnetic). The pairs of spectra were subtracted, yielding difference spectra containing only cross-peaks from 1H within 15 A of the spin label. These greatly simplified spectra were easily analyzed to provide complete assignments for residues 10-25 and 52-79 of the protein, 150 NOE distance restraints, and 27 hydrogen- bonding restraints. The chemical shifts and NOE patterns observed in the derivatized mutant were virtually identical to those which were resolved in the unmodified wild-type protein, strongly suggesting that the spin label was not perturbing the protein structure. The restaints enabled us to calculate a detailed structure for this region of subunit c. The structure consisted of two gently curved helices, crossing at a slight (30 degrees) angle. The C-terminal helix was disrupted from Val60 to Ala62 near the essential Pro64. Asp61, the residue thought to undergo protonation--deprotonation with each H+ transported across the membrane, was in ver der Waals contact with Ala24. The proximity of these residues had been predicted from mutant analyses, where H+ translocation was retained on moving the Asp from position 61 to 24
RP  - NOT IN FILE
NT  - UI - 95151746LA - engRN - 0 (Spin Labels)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM23105/GM/NIGMSID - R02301/PHSDA - 19950313IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:7849023
SO  - Biochemistry 1995 Feb 7 ;34(5):1635-1645

1305
UI  - 295
AU  - Groth G
AU  - Junge W
AD  - Universitat Osnabruck, Germany
TI  - ATP synthase of chloroplasts: selective action of agents binding to F1 on partial reactions of proton transfer in F0
AB  - We studied the basic steps of proton transfer through ATP synthase of chloroplasts, CF0CF1, under conditions of proton slip, a conducting stage in the absence of added nucleotides. On the background of a steady transmembrane pH difference, voltage steps were induced by flashing light. Proton intake, transfer, and release by CF0CF1 were kinetically resolved by spectrophotometric probes. Kinetic disparities between these three steps were observed. Rapid but limited proton intake from the thylakoid lumen and proton release into the medium (tau 1/2 approximately 5 ms) preceded charge transfer across the dielectric barrier in the enzyme (tau 1/2 approximately 30 ms). The saturation behavior under multiple flashes suggested a sequential reaction mechanism. ADP and dequalinium, when bound to subunit beta of the catalytic portion, CF1, blocked different partial reactions involving protons. ADP in the catalytic cleft blocked the electrogenic transfer step, and dequalinium at the adjacent DELSEED sequence on the same subunit blocked the release of protons. Both effects prove a long-range conformational transmission between remote (approximately 10 nm) domains on F1 and on F0. For the normal sequence of events from protons to ATP they suggest a specific action of certain proton-transfer steps on different domains of the catalytic portion
RP  - NOT IN FILE
NT  - UI - 95337077LA - engRN - 0 (Protons)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19950824IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:7612600
SO  - Biochemistry 1995 Jul 11 ;34(27):8589-8596

1306
UI  - 296
AU  - Groth G
AU  - Junge W
AD  - Abt Biophysik, Universitat Osnabruck, Germany
TI  - ATP synthase: activating versus catalytic proton transfer
AB  - ATP synthase (F-ATPase) of chloroplasts, CF0CF1, is both activated and driven by transmembrane protonmotive force. We dichotomized between activating and driving proton transfer by specific inhibitors, tentoxin and venturicidin. Thylakoids membranes were submitted to voltage steps (by flashing light) superimposed to a steady pH-difference. Transient proton intake, transfer and release by CF0CF1 was monitored by spectroscopic probes. Both activities, activation and catalysis, required all three partial reactions of the proton, however, activating proton transfer rose first (monophasically, tau 1/2 approximately 15 ms) followed by another phase of equal magnitude with a time lag of about 15 ms. Both types of consecutive proton transfer reactions contribute free energy for ATP synthesis
RP  - NOT IN FILE
NT  - UI - 95129698LA - engRN - 0 (Peptides, Cyclic)RN - 0 (Protons)RN - 0 (Venturicidins)RN - 28540-82-1 (tentoxin)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19950223IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:7828724
SO  - FEBS Lett 1995 Jan 23 ;358(2):142-144

1307
UI  - 162
AU  - Haughton MA
AU  - Capaldi RA
AD  - Institute of Molecular Biology, University of Oregon, Eugene 97403- 1229, USA
TI  - Asymmetry of Escherichia coli F1-ATPase as a function of the interaction of alpha-beta subunit pairs with the gamma and epsilon subunits
AB  - The asymmetry of Escherichia coli F1-ATPase (ECF1) has been explored in chemical modification experiments involving two mutant enzyme preparations. One mutant contains a cysteine (Cys) at position 149 of the beta subunit, along with conversion of a Val to Ala at residue 198 to suppress the deleterious effect of the Cys for Gly at 149 mutation (mutant beta G149C:V198A). The second mutant has these mutations and also Cys residues at positions 381 of beta and 108 of the epsilon subunit (mutant beta G149C:V198A:E381C/epsilon S108C). On CuCl2 treatment of this second mutant, there is cross-linking of one copy of the beta subunit to gamma via the Cys at 381, a second to the epsilon subunit (between beta Cys381 and epsilon Cys108), while the third beta subunit in the ECF1 complex is mostly free (some cross-linking to delta); thereby distinguishing the three beta subunits as beta gamma, beta epsilon, and beta free, respectively. Both mutants have ATPase activities similar to wild-type enzyme. Under all nucleotide conditions, including with essentially nucleotide-free enzyme, the three different beta subunits were found to react differently with N- ethylmaleimide (NEM) which reacts with Cys149, dicyclohexyl carbodiimide (DCCD) which reacts with Glu192, and 7-chloro-4- nitrobenzofurazan (NbfCl) which reacts with Tyr297. Thus, beta gamma reacted with DCCD but not NEM or NbfCl; beta free was reactive with all three reagents; beta epsilon reacted with NEM, but was poorly reactive to DCCD or NbfCl. There was a strong nucleotide dependence of the reaction of Cys149 in beta epsilon (but not in beta free) with NEM, indicative of the important role that the epsilon subunit plays in functioning of the enzyme
RP  - NOT IN FILE
NT  - UI - 95386505LA - engRN - 0 (Benzofurans)RN - 0 (Cross-Linking Reagents)RN - 0 (Macromolecular Systems)RN - 0 (Recombinant Proteins)RN - 10199-89-0 (4-Chloro-7-nitrobenzofurazan)RN - 128-53-0 (Ethylmaleimide)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 7440-50-8 (Copper)RN - 7447-39-4 (cupric chloride)RN - 76054-80-3 (4-chloro-7-nitrobenzofuran)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - HL 24526/HL/NHLBIDA - 19951004IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:7657634
SO  - J Biol Chem 1995 Sep 1 ;270(35):20568-20574

1308
UI  - 20941
AU  - Hensel M
AU  - Lill H
AU  - Schmid R
AU  - Deckers-Hebestreit G
AU  - Altendorf K
AD  - Arbeitsgruppe Mikrobiologie, Universitat Osnabruck, Germany
TI  - The ATP synthase (F1F0) of Streptomyces lividans: sequencing of the atp operon and phylogenetic considerations with subunit beta
AB  - The DNA encoding the subunits of the ATP synthase (F1F0) of Streptomyces lividans 66 strain 1326 was identified using oligodeoxyribonucleotide probes derived from the N-terminal sequence of subunit gamma of the F1 complex. The complete nucleotide sequence of the operon was determined. The atp operon contains nine genes, atpIBEFHAGDC, encoding the eight structural components of the ATP synthase complex and the i protein, a polypeptide of unknown function. The gene order found is identical to that in other non-photosynthetic eubacteria. The determination of the N-terminal amino acid (aa) sequences of the F1 subunits alpha, beta, gamma, delta and epsilon allowed us to identify the translational start points and to define the primary structures of the proteins. The aa sequence deduced for subunit delta revealed an N-terminal extension of about 90 aa, which is not present in any delta subunit or OSCP (oligomycin sensitivity-conferral protein) of other species studied so far. The phylogenetic relationship of eu- and archaebacteria was investigated using sequencing data of the highly conserved beta subunit of different ATP synthases including that of S. lividans. The calculations revealed that S. lividans beta does not form a phylogenetic group together with the Gram+ taxa of low G+C contents, but is more closely related to the beta subunit of Rhodobacteria
MH  - A
MH  - ACID
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BETA-SUBUNIT
MH  - COMPLEX
MH  - DELTA-SUBUNIT
MH  - F1
MH  - function
MH  - Oligomycins
MH  - protein
MH  - Proteins
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 95129902LA - engRN - 0 (Oligomycins)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19950223IS - 0378-1119SB - IMCY - NETHERLANDS
UR  - PM:7828915
SO  - Gene 1995 Jan 11 ;152(1):11-17

1309
UI  - 29
AU  - Hermolin J
AU  - Fillingame RH
AD  - Department of Biomolecular Chemistry, University of Wisconsin Medical School, Madison 53706
TI  - Assembly of F0 sector of Escherichia coli H+ ATP synthase. Interdependence of subunit insertion into the membrane
AB  - The F0 sector of the Escherichia coli H+ transporting ATP synthase is composed of a complex of three subunits, each of which traverses the inner membrane. We have studied the interdependence of subunit insertion into the membrane in a series of chromosomal mutants in which the primary mutation prevented insertion of one of the F0 subunits. Subunit insertion was assessed using Western blots of mutant membrane preparations. Subunit b and subunit c were found to insert into the membrane independently of the other two F0 subunits. On the other hand, subunit a was not inserted into membranes that lacked either subunit b or subunit c. The conclusion that subunit a insertion is dependent upon the co-insertion of subunits b and c differs from the conclusion of several studies, where subunits were expressed from multicopy plasmids
RP  - NOT IN FILE
NT  - UI - 95155353LA - engRN - 0 (DNA, Bacterial)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM23105/GM/NIGMSDA - 19950316IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:7852354
SO  - J Biol Chem 1995 Feb 10 ;270(6):2815-2817

1310
UI  - 21158
AU  - Hsu DK
AU  - Brusilow WS
AD  - Department of Biochemistry, Wayne State University School of Medicine, Detroit, MI 48201, USA
TI  - Effects of the uncI gene on expression of uncB, the gene coding for the a subunit of the F1F0 ATPase of Escherichia coli
AB  - The eight genes coding for the subunits of the E. coli F1F0 ATPase are preceded by a gene, designated uncI. A homologous gene, or a gene coding for an analagous protein, is found preceding the ATPase genes of several microorganisms. No function for the 1 gene has been described. Using lac fusions to measure gene expression in vivo, we tested the effects of deleting uncI on the expression of the adjacent gene uncB, which codes for the a subunit of the F0 sector of the ATPase. Deleting uncI reduced the expression of three uncB'-'lacZ fusion genes in vivo, but had no effect on the expression of two uncB'-'lacZ fusion genes containing a relatively smaller amount of the uncB coding region. The uncI deletion also reduced the relative synthesis of the a subunit in vitro. The I gene therefore appears to specifically affect the expression of uncB or the synthesis of the a subunit at some step after translational initiation of uncB
MH  - A
MH  - ATPase
MH  - Biochemistry
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - function
MH  - In Vitro
MH  - protein
MH  - Proteins
MH  - Proton-Translocating ATPases
MH  - SUBUNIT
MH  - synthesis
RP  - NOT IN FILE
NT  - UI - 95402193LA - engRN - 0 (Recombinant Fusion Proteins)RN - EC 3.2.1.23 (beta-Galactosidase)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19951016IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:7672111
SO  - FEBS Lett 1995 Sep 4 ;371(2):127-131

1311
UI  - 725
AU  - Jault JM
AU  - Matsui T
AU  - Jault FM
AU  - Kaibara C
AU  - Muneyuki E
AU  - Yoshida M
AU  - Kagawa Y
AU  - Allison WS
AD  - Department of Chemistry and Biochemistry 0601, University of California at San Diego, La Jolla 92093-0601, USA
TI  - The alpha 3 beta 3 gamma complex of the F1-ATPase from thermophilic Bacillus PS3 containing the alpha D261N substitution fails to dissociate inhibitory MgADP from a catalytic site when ATP binds to noncatalytic sites
AB  - ATP hydrolyses by the wild-type alpha 3 beta 3 gamma and mutant (alpha D261N)3 beta 3 gamma subcomplexes of the F1-ATPase from the thermophilic Bacillus PS3 have been compared. The wild-type complex hydrolyzes 50 microM ATP in three kinetic phases: a burst decelerates to an intermediate phase, which then gradually accelerates to a final rate. In contrast, the mutant complex hydrolyzes 50 microM or 2 mM ATP in two kinetic phases. The mutation abolishes acceleration from the intermediate phase to a faster final rate. Both the wild-type and mutant complexes hydrolyze ATP with a lag after loading a catalytic site with MgADP. The rate of the MgADP-loaded wild-type complex rapidly accelerates and approaches that observed for the wild-type apo-complex. The MgADP-loaded mutant complex hydrolyzes ATP with a more pronounced lag, and the gradually accelerating rate approaches the slow, final rate observed with the mutant apo-complex. Lauryl dimethylamide oxide (LDAO) stimulates hydrolysis of 2 mM ATP catalyzed by wild-type and mutant complexes 4- and 7.5-fold, respectively. The rate of release of [3H]ADP from the Mg[3H]ADP-loaded mutant complex during hydrolysis of 40 microM ATP is slower than observed with the wild-type complex. LDAO increases the rate of release of [3H]ADP from the preloaded wild-type and mutant complexes during hydrolysis of 40 microM ATP. Again, release is slower with the mutant complex. When the wild-type and mutant complexes are irradiated in the presence of 2-N3-[3H]ADP plus Mg2+ or 2- N3-[3H]ATP plus Mg2+ and azide, the same extent of labeling of noncatalytic sites is observed. Whereas ADP and ATP protect noncatalytic sites of the wild-type and mutant complexes about equally from labeling by 2-N3-[3H]ADP or 2-N3-[3H[ATP, respectively, AMP-PNP provides little protection of noncatalytic sites of the mutant complex. The results suggest that the substitution does not prevent binding of ADP or ATP to noncatalytic sites, but rather that it affects cross-talk between liganded noncatalytic sites and catalytic sites which is necessary to promote dissociation of inhibitory MgADP
RP  - NOT IN FILE
NT  - UI - 96110720LA - engRN - 0 (Dimethylamines)RN - 0 (Rhodamines)RN - 1643-20-5 (dodecyldimethylamine oxide)RN - 25612-73-1 (Adenylyl Imidodiphosphate)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 989-38-8 (rhodamine 6G)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM-16974/GM/NIGMSDA - 19961022IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:8845368
SO  - Biochemistry 1995 Dec 19 ;34(50):16412-16418

1312
UI  - 20850
AU  - Jeanteur-De Beukelaer C
AU  - Omote H
AU  - Iwamoto-Kihara A
AU  - Maeda M
AU  - Futai M
AD  - Division of Biological Science, Osaka University, Japan
TI  - Beta-gamma subunit interaction is required for catalysis by H(+)-ATPase (ATP synthase). Beta subunit amino acid replacements suppress a gamma subunit mutation having a long unrelated carboxyl terminus
AB  - The mechanisms of energy coupling and catalytic co-operativity are not yet understood for H(+)-ATPase (ATP synthase). An Escherichia coli gamma subunit frameshift mutant (downstream of Thr-gamma 277) could not grow by oxidative phosphorylation because both mechanisms were defective (Iwamoto, A., Miki, J., Maeda, M., and Futai, M. (1990) J. Biol. Chem. 265, 5043-5048). The defect(s) of the gamma frameshift was obvious, because the mutant subunit had a carboxyl terminus comprising 16 residues different from those in the wild type. However, in this study, we surprisingly found that an Arg-beta 52-->Cys or Gly-beta 150-- >Asp replacement could suppress the deleterious effects of the gamma frameshift. The membranes of the two mutants (gamma frameshift/Cys-beta 52 with or without a third mutation, Val-beta 77-->Ala) exhibited increased oxidative phosphorylation, together with 70-100% of the wild type ATPase activity. Similarly, the gamma frameshift/Asp-beta 150 mutant could grow by oxidative phosphorylation, although this mutant had low membrane ATPase activity. These results suggest that the beta subunit mutation suppressed the defects of catalytic cooperativity and/or energy coupling in the gamma mutant, consistent with the notion that conformational transmission between the two subunits is pertinent for this enzyme
MH  - A
MH  - ACID
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BETA-SUBUNIT
MH  - Catalysis
MH  - coupling
MH  - Cysteine
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - H(+)ATPase
MH  - M
MH  - Macromolecular Systems
MH  - mechanism
MH  - MECHANISMS
MH  - membrane
MH  - Membranes
MH  - mutant
MH  - Oxidative Phosphorylation
MH  - Phosphorylation
MH  - protein
MH  - Proteins
MH  - RESIDUE
MH  - SUBUNIT
MH  - SYNTHASE
MH  - SYSTEM
MH  - SYSTEMS
RP  - NOT IN FILE
NT  - UI - 96032713LA - engRN - 0 (DNA Primers)RN - 0 (Macromolecular Systems)RN - 0 (Plasmids)RN - 0 (Recombinant Proteins)RN - 52-90-4 (Cysteine)RN - 56-40-6 (Glycine)RN - 56-84-8 (Aspartic Acid)RN - 74-79-3 (Arginine)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19951106IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:7559418
SO  - J Biol Chem 1995 Sep 29 ;270(39):22850-22854

1313
UI  - 9931
AU  - Jones PC
AU  - Harrison MA
AU  - Kim YI
AU  - Finbow ME
AU  - Findlay JB
AD  - Department of Biochemistry and Molecular Biology, University of Leeds, UK
TI  - The first putative transmembrane helix of the 16 kDa proteolipid lines a pore in the Vo sector of the vacuolar H(+)-ATPase
AB  - The 16 kDa proteolipid is the major component of the vacuolar H(+)- ATPase membrane sector, responsible for proton translocation. Expression of a related proteolipid from the arythropod Nephrops norvegicus in a Saccharomyces strain in which the VMA3 gene for the endogenous proteolipid has been disrupted results in restored vacuolar H(+)-ATPase function. We have used this complementation system, coupled to cysteine substitution mutagenesis and protein chemistry, to investigate structural features of the proteolipid. Consecutive cysteines were introduced individually into putative transmembrane segment 1 of the proteolipid, and at selected sites in extramembranous regions and in segment 3 and 4. Analysis of restored vacuolar H(+)- ATPase function showed that segment 1 residues sensitive to mutation to cysteine were clustered on a single face, but only if the segment was helical. Only residues insensitive to mutation could be covalently modified by the cysteine-specific reagent fluorescein 5-maleimide. A cysteine introduced into segment 3 was the only residue accessible to a relatively hydrophobic reagent, suggesting accessibility to the lipid phase. Analysis of disulphide bond formation between introduced cysteines indicates that the first transmembrane alpha-helices of each monomer are adjacent to each other at the centre of the proteolipid multimeric complex. The data are consistent with a model in which the fluorescein maleimide-accessible face of helix I lines a pore at the centre of a hexameric complex formed by the proteolipid, with the mutationally sensitive face oriented into the protein core. The implications for ion-transport function in this family of proteins are discussed in the context of this structural model
MH  - analysis
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Biochemistry
MH  - Chemistry
MH  - COMPLEX
MH  - England
MH  - Fluoresceins
MH  - H(+)-Transporting ATP Synthase
MH  - H(+)ATPase
MH  - Indicators and Reagents
MH  - Ion Transport
MH  - Macromolecular Systems
MH  - model
MH  - Proteins
MH  - Proteolipids
MH  - proton
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 96128055LA - engRN - 0 (Disulfides)RN - 0 (Fluoresceins)RN - 0 (Indicators and Reagents)RN - 0 (Macromolecular Systems)RN - 0 (Proteolipids)RN - 52-90-4 (Cysteine)RN - 75350-46-8 (fluorescein 5-maleimide)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19960220IS - 0264-6021SB - IMCY - ENGLANDJC - 9YO
UR  - PM:8554514
SO  - Biochem J 1995 Dec 15 ;312 ( Pt 3)():739-747

1314
UI  - 49
AU  - Kaim G
AU  - Dimroth P
AD  - Mikrobiologisches Institut Eidgenossische Technische Hochschule, ETH- Zentrum, Zurich, Switzerland
TI  - A double mutation in subunit c of the Na(+)-specific F1F0-ATPase of Propionigenium modestum results in a switch from Na+ to H(+)-coupled ATP synthesis in the Escherichia coli host cells
AB  - The in vivo synthesis of an F1F0-ATPase hybrid in Escherichia coli strain PEF42 which harbours the genes for the Propionigenium modestum subunits a, b, c, and delta, a gene for hybrid alpha subunit with the N- terminal portion (amino acids 1 to 173) of P. modestum and the C- terminal region (amino acids 176 to 513) from E. coli, and the genes for the E. coli subunits beta, gamma and epsilon, yielded a functional enzyme complex. This hybrid ATPase coupled ATP synthesis to Na+ transport and required Na+ for growth on succinate. After random mutagenesis of the P. modestum genes of strain PEF42, clones were selected that grew on succinate in the absence of Na+. A double- mutation cPhe84Leu, cLeu87Val that was found in several of these clones, was introduced by site specific mutagenesis into the parent strain PEF42. The resulting strain E. coli MPC8487 also exhibited Na(+)- independent growth on succinate, showing that the double mutation is the only reason for the new phenotype. The mutation causes a change of the coupling ions of the hybrid ATPase from Na+ in strain PEF42 to H+ in strain MPC8487. This conclusion was supported by the biochemical properties of the ATPase from strain MPC8487. Unlike the parent enzyme, the mutated ATPase was not activated by Na+, but retained activation by Li+. The pH optimum of the mutated ATPase (in the absence of Na+ or Li+) was shifted from pH 6.5 to pH 7.5, and the specific ATPase activity of the cell membranes increased about fourfold over that found in membranes of the parent cells. The mutated ATPase pumped protons or Li+ after reconstitution into proteoliposomes, and the transport of both cations was not affected by Na+. The double mutation in the c subunit thus results in the loss of Na+ binding, retention of Li+ binding and an improvement of H+ binding
RP  - NOT IN FILE
NT  - UI - 96074781LA - engRN - 0 (Bacterial Proteins)RN - 0 (Recombinant Fusion Proteins)RN - 1333-74-0 (Hydrogen)RN - 56-65-5 (Adenosine Triphosphate)RN - 7439-93-2 (Lithium)RN - 7440-23-5 (Sodium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19951226IS - 0022-2836SB - IMCY - ENGLANDJC - J6V
UR  - PM:7473747
SO  - J Mol Biol 1995 Nov 10 ;253(5):726-738

1315
UI  - 727
AU  - Kato Y
AU  - Sasayama T
AU  - Muneyuki E
AU  - Yoshida M
AD  - Research Laboratory of Resources Utilization, Tokyo Institute of Technology, Kanagawa, Japan
TI  - Analysis of time-dependent change of Escherichia coli F1-ATPase activity and its relationship with apparent negative cooperativity
AB  - Except for the case of gradual activation of EF1 (F1-ATPase from Escherichia coli) caused by the dissociation of the epsilon subunit [Laget, P. P. and Smith, J. B. (1979) Arch. Biochem. Biophys. 197, 83- 89], EF1 has long been thought not to show a time-dependent change in activity [Senior, A.E. et al. (1992) Arch. Biochem. Biophys. 297, 340- 344]. Here, we report the time-dependent inactivation and activation of EF1, which are apparently similar to those of mitochondrial F1-ATPases [Vasilyeva, E.A. et al. (1982) Biochem. J. 202, 15-23]. Analysis of these changes as a function of ATP concentrations in relation to negative cooperativity revealed that the initial inactivation phase was attributable to the decrease in the Vmax associated with the low Km (around 10 microM), and the following activation, probably due to the dissociation of the epsilon subunit, corresponded to the increase in the Vmax associated with the high Km (in the order of 100 microM). Thus, the time-dependent change in EF1 activity is closely related to the apparent negative cooperativity (multiple Km values) of ATP hydrolysis
RP  - NOT IN FILE
NT  - UI - 96026449LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19951124IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:7578215
SO  - Biochim Biophys Acta 1995 Oct 10 ;1231(3):275-281

1316
UI  - 819
AU  - Krenn BE
AU  - Aardewijn P
AU  - van Walraven HS
AU  - Werner-Grune S
AU  - Strotmann H
AU  - Kraayenhof R
AD  - Institute for Molecular Biological Sciences, BioCentrum Amsterdam, Vrije Universiteit, The Netherlands
TI  - ATP synthase from a cyanobacterial Synechocystis 6803 mutant containing the regulatory segment of the chloroplast gamma subunit shows thiol modulation
RP  - NOT IN FILE
NT  - UI - 96227123LA - engRN - 0 (Sulfhydryl Compounds)RN - 52-90-4 (Cysteine)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19960730IS - 0300-5127SB - IMCY - ENGLANDJC - E48
UR  - PM:8654832
SO  - Biochem Soc Trans 1995 Nov ;23(4):757-760

1317
UI  - 21078
AU  - Krulwich TA
AD  - Department of Biochemistry, Mount Sinai School of Medicine of CUNY, New York 10029, USA
TI  - Alkaliphiles: 'basic' molecular problems of pH tolerance and bioenergetics
AB  - Alkaliphilic Bacillus species provide experimental opportunities for examination of physiological processes under conditions in which the stress of the extreme environment brings issues of general biological importance into special focus. The alkaliphile, like many other cells, uses Na+/H+ antiporters in pH regulation, but its array of these porters, and other ion-flux pathways that energize and support their activity, result in an extraordinary capacity for pH homeostasis; this process nonetheless becomes the factor that limits growth at the upper edge of the pH range. Above pH 9.5, aerobic alkaliphiles maintain a cytoplasmic pH that is two or more units below the external pH. This chemiosmotically adverse delta pH is bypassed by use of an electrochemical gradient of Na+ rather than of protons to energize solute uptake and motility. By contrast, ATP synthesis occurs via completely proton-coupled oxidative phosphorylation that proceeds just as well, or better, at pH 10 and above as it does in the same bacteria growing at lower pH, without the adverse pH gradient. Various mechanisms that might explain this conundrum are described, and the current state of the evidence supporting them is summarized
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthesis
MH  - ATPase
MH  - Bacillus
MH  - Bacteria
MH  - Biochemistry
MH  - bioenergetics
MH  - Cells
MH  - delta
MH  - DELTA-PH
MH  - England
MH  - Homeostasis
MH  - mechanism
MH  - MECHANISMS
MH  - Oxidative Phosphorylation
MH  - pH
MH  - Phosphorylation
MH  - proton
MH  - Proton-Translocating ATPases
MH  - Protons
MH  - regulation
MH  - review
MH  - Sodium
MH  - synthesis
RP  - NOT IN FILE
NT  - UI - 95302951LA - engRN - 0 (Protons)RN - 56-65-5 (Adenosine Triphosphate)RN - 7440-23-5 (Sodium)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticlePT - ReviewPT - Review, TutorialID - GM28454/GM/NIGMSDA - 19950719IS - 0950-382XSB - IMCY - ENGLAND
UR  - PM:7783613
SO  - Mol Microbiol 1995 Feb ;15(3):403-410

1318
UI  - 10671
AU  - Labahn A
AU  - Graber P
TI  - Numerical simulation of uni-site and bi-site ATP-hydrolysis catalyzed by the membrane-bound hydrogen ion-ATPase from chloroplasts
MH  - atp
MH  - bioenergetics
MH  - chloroplast
MH  - Chloroplasts
MH  - Hydrogen
MH  - SIMULATION
MH  - theory
MH  - unisite
RP  - NOT IN FILE
SO  - Acta Physiol Scand 1995  ;146(607):241-244

1319
UI  - 21159
AU  - Matten SR
AU  - Schemidt RA
AU  - Brusilow WS
AD  - Department of Biochemistry, Wayne State University School of Medicine, Detroit, Michigan 48201
TI  - Construction and function of chimeric beta subunits containing regions from the beta subunits of the F1F0 ATPases of Escherichia coli and Bacillus megaterium
AB  - The highly conserved beta subunit of the Escherichia coli F1F0 ATPase was divided into three sections, each of which was exchanged with the homologous section of the beta subunit of the obligate aerobe Bacillus megaterium. Plasmids coding for the resultant six chimeric beta subunits varied in their abilities to complement two E. coli beta mutants as measured by testing transformed cells for aerobic growth on a nonfermentable carbon source or anaerobic growth on rich medium containing glucose. Two chimeras were able to restore both growth on succinate and anaerobic growth on rich medium. The genetic results corresponded to increased levels of membrane-bound ATPase and ATP synthase activities. These chimeric subunits were therefore capable of being assembled into functional E. coli ATPase complexes. The results indicate that chimeric beta subunits can be used to analyze assembly of the beta subunit and that the final 181 amino acids of the beta subunit might contain a region involved in functional energy coupling
MH  - A
MH  - ACID
MH  - Amino Acids
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Bacillus
MH  - BETA
MH  - BETA-SUBUNIT
MH  - Biochemistry
MH  - Cells
MH  - COMPLEX
MH  - coupling
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - function
MH  - Glucose
MH  - Macromolecular Systems
MH  - mutant
MH  - protein
MH  - Proteins
MH  - Proton-Translocating ATPases
MH  - SUBUNIT
MH  - succinate
MH  - SYNTHASE
MH  - SYSTEM
MH  - SYSTEMS
RP  - NOT IN FILE
NT  - UI - 95130514LA - engRN - 0 (Chimeric Proteins)RN - 0 (DNA Primers)RN - 0 (Macromolecular Systems)RN - 0 (Plasmids)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19950222IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:7829474
SO  - J Biol Chem 1995 Jan 27 ;270(4):1489-1492

1320
UI  - 21248
AU  - McDermott G
AU  - Prince S
AU  - Freer A
AU  - Lawlwss A
AU  - Papiz M
AU  - Isaacs N
AU  - Cogdell R
TI  - Crystal structure of an integral membrane light-harvesting complex from photosynthetic bacteria
MH  - structure
MH  - membrane
MH  - COMPLEX
RP  - NOT IN FILE
SO  - Nature 1995  ;374():517-521

1321
UI  - 729
AU  - Miyauchi M
AU  - Tozawa K
AU  - Yoshida M
AD  - Research Laboratory of Resources Utilization, Tokyo Institute of Technology, Yokohama, Japan
TI  - F1-ATPase alpha-subunit made up from two fragments (1-395, 396-503) is stabilized by ATP and complexes containing it obey altered kinetics
AB  - Inferred from the crystal structure of mitochondrial F1-ATPase (Abrahams, J.P. et al. (1994) Nature 370, 621-628), the proteinase- sensitive region around Phe-395 of thermophilic F1-ATPase alpha-subunit corresponds to the loop which connects main part of the carboxyl- terminal helical bundle domain with the ATP binding domain. This loop is in contact with the gamma- and adjacent beta-subunits. Two polypeptides corresponding to the sequence 1-395 and 396-503 of the alpha-subunit were expressed in Escherichia coli cells and they were copurified as an apparently functional alpha-subunit (alpha(395/396)) made up of two polypeptides. The isolated alpha(395/396) was stabilized by ATP-Mg, but not by ADP-Mg, although it bound both ATP-Mg and ADP-Mg with similar affinities (Kd, 11 microM and 14 microM, respectively). The alpha(395/396) was reconstitutable into alpha(395/396)3 beta 3 and alpha(395/396)3 beta 3 gamma complexes. Different from the intact the ATP-Mg-induced dissociation into alpha 1 beta 1 heterodimers. ATP hydrolysis by the alpha(395/396)3 beta 3 gamma complex underwent a slow initial phase, whereas the intact alpha 3 beta 3 gamma complex exhibited an accelerated initial phase. Steady-state ATPase activity at various ATP concentrations showed negative cooperativity for the intact alpha 3 beta 3 gamma complex but apparently positive cooperativity for the alpha(395/396)3 beta 3 gamma complex. The ATPase activities at a saturating ATP concentration of the complexes containing the alpha(395/396) were 180% of those containing intact alpha-subunits. These results indicate that a loop around Phe-395 is involved in intersubunit interaction in F1-ATPase
RP  - NOT IN FILE
NT  - UI - 95244573LA - engRN - 0 (Oligodeoxyribonucleotides)RN - 0 (Peptide Fragments)RN - 0 (Recombinant Proteins)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19950601IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:7727499
SO  - Biochim Biophys Acta 1995 Apr 26 ;1229(2):225-232

1322
UI  - 20852
AU  - Moriyama Y
AU  - Patel V
AU  - Futai M
AD  - Department of Biochemistry and Organic Chemistry, Osaka University, Japan
TI  - Quinacrine mustard and lipophilic cations inhibitory to both vacuolar H(+)-ATPase and F0F1-ATP synthase
AB  - Various lipophilic cations, such as quinacrine mustard and dequalinium, which are known to inhibit mitochondrial F1-ATPase, strongly inhibited vacuolar H(+)-ATPase purified from bovine adrenal chromaffin granules. Quinacrine mustard bound irreversibly to vacuolar H(+)-ATPase subunit A, and the 115 kDa accessory polypeptide and dithiothreitol had no effect. The binding was competitively inhibited by chlorpromazine and quinacrine, and these compounds specifically reduced the amount of labeling of subunit A. Quinacrine mustard also prevented the binding of [alpha-32P]ATP to subunit A but had no effect on the binding of [3H]N- ethylmaleimide to either subunit A or the 115 kDa accessory polypeptide. These results suggest that the binding site of quinacrine mustard in subunit A is not related to the N-ethylmaleimide-binding site(s), which is important for activity
MH  - A
MH  - ATPase
MH  - BINDING
MH  - Biochemistry
MH  - Cations
MH  - Chemistry
MH  - Chromaffin Granules
MH  - Dithiothreitol
MH  - Ethylmaleimide
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - H(+)ATPase
MH  - MITOCHONDRIAL F1-ATPASE
MH  - Site
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 95154449LA - engRN - 0 (Cations)RN - 3483-12-3 (Dithiothreitol)RN - 4213-45-0 (Quinacrine Mustard)RN - 50-53-3 (Chlorpromazine)RN - 6707-58-0 (Dequalinium)RN - 83-89-6 (Quinacrine)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19950313IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:7851533
SO  - FEBS Lett 1995 Feb 6 ;359(1):69-72

1323
UI  - 20851
AU  - Nakamoto RK
AU  - al Shawi MK
AU  - Futai M
AD  - Department of Molecular Physiology and Biological Physics, University of Virginia, Charlottesville 22908, USA
TI  - The ATP synthase gamma subunit. Suppressor mutagenesis reveals three helical regions involved in energy coupling
AB  - A role in coupling proton transport to catalysis of ATP synthesis has been demonstrated for the Escherichia coli F0F1 ATP synthase gamma subunit. Previously, functional interactions between the terminal regions that were important for coupling were shown by finding several mutations in the carboxyl-terminal region of the gamma subunit (involving residues at positions 242 and 269-280) that restored efficient coupling to the mutation, gamma Met-23-->Lys (Nakamoto, R. K., Maeda, M., and Futai, M. (1993) J. Biol. Chem. 268, 867-872). In this study, we used suppressor mutagenesis to establish that the terminal regions can be separated into three interacting segments. Second-site mutations that cause pseudo reversion of the primary mutations, gamma Gln-269-->Glu or gamma Thr-273-->Val, map to an amino- terminal segment with changes at residues 18, 34, and 35, and to a segment near the carboxyl terminus with changes at residues 236, 238, 242, and 246. Each second-site mutation suppressed the effects of both gamma Gln-269-->Glu and gamma Thr-273-->Val, and restored efficient coupling to enzyme complexes containing either of the primary mutations. Mapping of these residues in the recently reported x-ray crystallographic structure of the F1 complex (Abrahams, J. P., Leslie, A. G., Lutter, R., and Walker, J. E. (1994) Nature 370, 621-628), reveals that the second-site mutations do not directly interact with gamma Gln-269 and gamma Thr-273 and that the effect of suppression occurs at a distance. We propose that the three gamma subunit segments defined by suppressor mutagenesis, residues gamma 18-35, gamma 236-246, and gamma 269-280, constitute a domain that is critical for both catalytic function and energy coupling
MH  - A
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - ATPase
MH  - Catalysis
MH  - COMPLEX
MH  - coupling
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0F1
MH  - F1
MH  - function
MH  - M
MH  - mutagenesis
MH  - P
MH  - physiology
MH  - proton
MH  - RESIDUE
MH  - SUBUNIT
MH  - SYNTHASE
MH  - synthesis
MH  - transport
RP  - NOT IN FILE
NT  - UI - 95294008LA - engRN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleID - GM50957/GM/NIGMSDA - 19950710IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:7775464
SO  - J Biol Chem 1995 Jun 9 ;270(23):14042-14046

1324
UI  - 20848
AU  - Omote H
AU  - Le NP
AU  - Park MY
AU  - Maeda M
AU  - Futai M
AD  - Department of Biological Science, Osaka University, Japan
TI  - Beta subunit Glu-185 of Escherichia coli H(+)-ATPase (ATP synthase) is an essential residue for cooperative catalysis
AB  - Glu-beta 185 of the Escherichia coli H(+)-ATPase (ATP synthase) beta subunit was replaced by 19 different amino acid residues. The rates of multisite (steady state) catalysis of all the mutant membrane ATPases except Asp- beta 185 were less than 0.2% of the wild type one; the Asp- beta 185 enzyme exhibited 15% (purified) and 16% (membrane-bound) ATPase activity. The purified inactive Cys- beta 185 F1-ATPase recovered substantial activity after treatment with iodoacetate in the presence of MgCl2; maximal activity was obtained upon the introduction of about 3 mol of carboxymethyl residues/mol of F1. The divalent cation dependences of the S-carboxymethyl- beta 185 and Asp- beta 185 ATPase activities were altered from that of the wild type. The Asp- beta 185, Cys- beta 185, S-carboxymethyl-beta 185, and Gln- beta 185 enzymes showed about 130, 60, 20, and 50% of the wild type unisite catalysis rates, respectively. The S-carboxymethyl- beta 185 and Asp- beta 185 enzymes showed altered divalent cation sensitivities, and the S- carboxymethyl- beta 185 enzyme showed no Mg2+ inhibition. Unlike the wild type, the two mutant enzymes showed low sensitivities to azide, which stabilizes the enzyme Mg-ADP complex. These results suggest that Glu- beta 185 may form a Mg2+ binding site, and its carboxyl moiety is essential for catalytic cooperativity. Consistent with this model, the bovine glutamate residue corresponding to Glu- beta 185 is located close to the catalytic site in the higher order structure (Abrahams, J.P., Leslie, A.G.W., Lutter, R ., and Walker, J.E. (1994) Nature 370, 621-628)
MH  - A
MH  - ACID
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Azides
MH  - BETA-SUBUNIT
MH  - BINDING
MH  - Catalysis
MH  - COMPLEX
MH  - DEPENDENCE
MH  - Enzymes
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - glutamic acid
MH  - H(+)ATPase
MH  - Iodoacetates
MH  - Magnesium
MH  - membrane
MH  - model
MH  - mutant
MH  - RESIDUE
MH  - Site
MH  - SUBUNIT
MH  - SYNTHASE
MH  - UNISITE CATALYSIS
RP  - NOT IN FILE
NT  - UI - 96029656LA - engRN - 0 (Azides)RN - 0 (Iodoacetates)RN - 56-86-0 (Glutamic Acid)RN - 64-69-7 (Iodoacetic Acid)RN - 7447-41-8 (Lithium Chloride)RN - 7786-30-3 (Magnesium Chloride)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19951214IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:7592742
SO  - J Biol Chem 1995 Oct 27 ;270(43):25656-25660

1325
UI  - 509
AU  - Pedersen PL
AU  - Hullihen J
AU  - Bianchet M
AU  - Amzel LM
AU  - Lebowitz MS
AD  - Department of Biological Chemistry, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205-2185
TI  - Rat liver ATP synthase. Relationship of the unique substructure of the F1 moiety to its nucleotide binding properties, enzymatic states, and crystalline form
AB  - The F1 moiety of rat liver ATP synthase has a molecular mass of 370,000, exhibits the unique substructure alpha 3 beta 3 gamma delta epsilon, and fully restores ATP synthesis to F1-depleted membranes. Here we provide new information about rat liver F1 as it relates to the relationship of its unique substructure to its nucleotide binding properties, enzymatic states, and crystalline form. Seven types of experiments were performed in a comprehensive study. First, the capacity of F1 to bind [3H]ADP, the substrate for ATP synthesis and [32P]AMP-PNP (5'-adenylyl-beta,gamma-imidodiphosphate), a nonhydrolyzable ATP analog, was quantified. Second, double-label experiments were performed to establish whether ADP and AMP-PNP bind to the same or different sites. Third, total nucleotide binding was assessed by the luciferin-luciferase assay. Fourth, F1 was subfractionated into an alpha gamma and a beta delta epsilon fraction, both of which were subjected to nucleotide binding assays. Fifth, the nucleotide binding capacity of F1 was quantified after undergoing ATP hydrolysis. Sixth, the intensity of the fluorescence probe pyrene maleimide bound at alpha subunits was monitored before and after F1 experienced ATP hydrolysis. Finally, the catalytic activity and nucleotide content of F1 obtained from crystals being used in x-ray crystallographic studies was determined. The picture of rat liver F1 that emerges is one of an enzyme molecule that 1) loads nucleotide readily at five sites; 2) requires for catalysis both the alpha gamma and the beta delta epsilon fractions; 3) directs the reversible binding of ATP and ADP to different regions of the enzyme's substructure; 4) induces inhibition of ATP hydrolysis only after ADP fills at least five sites; and 5) exists in several distinct forms, one an active, symmetrical form, obtained in the presence of ATP and high P(i) and on which an x-ray map at 3.6 A has been reported (Bianchet, M., Ysern, X., Hullihen, J., Pedersen, P. L., and Amzel, L. M. (1991) J. Biol. Chem. 266, 21197-21201). These results are discussed within the context of a multistate model for rat liver F1 and also discussed relative to those reported for bovine heart F1, which has been crystallized with inhibitors in an asymmetrical form and has a propensity for binding nucleotides more tightly
RP  - NOT IN FILE
NT  - UI - 95130558LA - engRN - 0 (Macromolecular Systems)RN - 25612-73-1 (Adenylyl Imidodiphosphate)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - CA 10951/CA/NCIID - GM 25432/GM/NIGMSDA - 19950222IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:7829514
SO  - J Biol Chem 1995 Jan 27 ;270(4):1775-1784

1326
UI  - 21188
AU  - Radionov AN
AU  - Kaulen AD
AD  - Department of Photobiochemistry, A N Belozersky Institute of Physico- Chemical Biology, Moscow State University, Russian Federation
TI  - Cooperative phenomena in the photocycle of D96N mutant bacteriorhodopsin
AB  - The M intermediate decay in the photocycle of D96N mutant bacteriorhodopsin does not depend on the light intensity of the exciting flash. Cooperative phenomena in the photocycle are revealed after addition of azide causing acceleration of the M decay and making it kinetically well separated from the N decay. Increase in the light intensity induces slight deceleration of the M decay and significant acceleration of the N decay. The data obtained directly confirm our recent model [Komrakov and Kaulen (1995) Biophys. Chem. 56, 113-119], according to which appearance of the Mslow intermediate in the photocycle of the wild type bR at high light intensity is due to destabilization of the N intermediate leading to the acceleration of the N-->M and N-->bR reactions
MH  - A
MH  - Azides
MH  - Bacteriorhodopsin
MH  - flash
MH  - intermediate
MH  - Light
MH  - M
MH  - M-intermediate
MH  - model
MH  - mutant
MH  - proton
MH  - Protons
MH  - Sodium
RP  - NOT IN FILE
NT  - UI - 96140541LA - engRN - 0 (Azides)RN - 0 (Protons)RN - 26628-22-8 (Sodium Azide)RN - 53026-44-1 (Bacteriorhodopsins)PT - Journal ArticleDA - 19960221IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:8549749
SO  - FEBS Lett 1995 Dec 27 ;377(3):330-332

1327
UI  - 992
AU  - Richard P
AU  - Pitard B
AU  - Rigaud JL
TI  - ATP synthesis by the F0F1-ATPase from the thermophilic Bacillus PS3 co-reconstituted with bacteriorhodopsin into liposomes. Evidence for stimulation of ATP synthesis by ATP bound to a noncatalytic binding site
AB  - F-type ATPase from the thermophilic Bacillus PS3, TF0F1, which was essentially free of bound nucleotides after isolation and purification, was co-reconstituted into liposomes with the light-driven proton pump bacteriorhodopsin. The time course of the light-induced ATP synthesis was biphasic; an initial slow phase accelerated to a final steady-state rate two to three times faster. Adding ATP before initiating the reaction suppressed the slow phase, suggesting that the state of occupancy of specific sites by ATP regulated the synthetic activity of TF0F1. Incubating the purified TF0F1 with ADP and ATP revealed one ADP and two ATP binding sites that were stable to gel filtration. We analyzed the time courses of light-induced ATP synthesis for the enzyme with different nucleotide content, after co-reconstitution into liposomes with bacteriorhodopsin. The two ATP sites were identified to have regulatory function. A complex containing TF0F1.ADP, 1:1, was co-reconstituted with various quantities of ATP to obtain a range of molar ratios of TF0F1.ADP:ATP of between 1:0 and 1:1.7. It was found that the initial rate of ATP synthesis increased with the level of ATP bound to the enzyme. After binding one ATP, a stimulation of ATP synthesis by a factor of 2 was observed. The second ATP site also exhibited regulatory properties. It stimulated ATP synthesis but to a much smaller extent; the stimulation did not exceed 20%. Binding of the photoreactive analogues 2-azido-[alpha-32P]ADP and 2-azido-[alpha-32P]ATP to the TF0F1 and their effects on the rate of ATP synthesis are described further.(ABSTRACT TRUNCATED AT 250 WORDS)
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - Amino Acid Sequence
MH  - Bacillus
MH  - Bacteriorhodopsin
MH  - Binding Sites
MH  - Chemistry
MH  - Comparative Study
MH  - enzymology
MH  - H(+)-Transporting ATP Synthase
MH  - Heat
MH  - isolation & purification
MH  - Kinetics
MH  - Light
MH  - Liposomes
MH  - metabolism
MH  - Molecular Sequence Data
MH  - Peptide Fragments
MH  - Peptide Mapping
MH  - Proteolipids
MH  - Proton Pump
MH  - purification
MH  - Support,Non-U.S.Gov't
MH  - Time
MH  - Trypsin
RP  - NOT IN FILE
NT  - Departement Biologie Cellulaire et Moleculaire, CE Saclay, Gif-sur-Yvette, France
SO  - J Biol Chem 1995 Sep 15 ;270(37):21571-21578

1328
UI  - 991
AU  - Rigaud JL
AU  - Pitard B
AU  - Levy D
TI  - Reconstitution of membrane proteins into liposomes: application to energy-transducing membrane proteins
MH  - Chemistry
MH  - Detergents
MH  - Energy Metabolism
MH  - Liposomes
MH  - Membrane Proteins
MH  - metabolism
MH  - Micelles
MH  - Proteins
MH  - Proteolipids
MH  - Solubility
MH  - Support,Non-U.S.Gov't
RP  - NOT IN FILE
NT  - Section de Bioenergetique, DBCM, CEA-Saclay, Gif sur Yvette, France
SO  - Biochim Biophys Acta 1995 Oct 10 ;1231(3):223-246

1329
UI  - 21331
AU  - Roberts G
AU  - Berberian G
AU  - Beauge L
TI  - Evidence for Two Catalytic Sites in the Functional Unit of H+-ATPase from Higher Plants
AB  - We investigated the nature of the complex ATP activation kinetics of plant H+-ATPases. To this aim we analyzed that activation in three isolated isoforms (AHA1, AHA2, and AHA3) of H+-ATPase from Arabidopsis thaliana. The isoforms were obtained by heterologous expression in endoplasmic reticulum of yeast. ATP stimulation was always with low affinity (K0.5 between 500 and 1800 [mu]M). In addition, the curves were not Michaelian and displayed positive cooperativity. Detailed studies with AHA2 showed that (a) enzyme solubilized with lysophosphatidylcholine exhibited Michaelian behavior even in the presence of soybean lecithin liposomes free of enzyme, (b) solubilized enzyme incorporated into the same liposomes displayed two-site kinetics with negative cooperativity, and (c) enzyme partially digested with trypsin lost the C-terminal portion of the molecule. Under this condition the ATP activation kinetics was Michaelian or had a slight negative cooperativity and the K0.5ATP was reduced 3-fold. These data suggest that the functional unit of the H+-ATPase has two catalytic ATP sites with variable cooperativity and kinetics competence of the E(ATP) and E(ATP)2 complexes. Such variability is likely modulated by the association of the enzyme with membrane structures and by a regulatory domain in the C terminus of the enzyme molecule
MH  - A
MH  - ACTIVATION
MH  - affinity
MH  - atp
MH  - catalytic
MH  - COMPLEX
MH  - data
MH  - enzyme
MH  - H+-ATPase
MH  - Kinetics
MH  - liposome
MH  - Liposomes
MH  - membrane
MH  - plant
MH  - Plants
MH  - Site
MH  - structure
MH  - Trypsin
MH  - YEAST
RP  - NOT IN FILE
NT  - Division de Biofisica, Instituto de Investigacion Medica "Mercedes y Martin Ferreyra," Casilla de Correo 389, 5000 Cordoba, Argentina
SO  - Plant Physiol 1995 Jun ;108(2):813-819

1330
UI  - 728
AU  - Saika K
AU  - Yoshida M
AD  - Research Laboratory of Resources Utilization, Tokyo Institute of Technology, Yokohama, Japan
TI  - A minimum catalytic unit of F1-ATPase shows non-cooperative ATPase activity inherent in a single catalytic site with a Km 70 microM
AB  - F1-ATPase has three interacting catalytic sites and shows complicated kinetics. Here, we report reconstitution of a complex, most likely composed of one alpha subunit and one beta subunit, with a single catalytic site from thermophilic Bacillus PS3 F1-ATPase on the solid surface. The complex has an ATPase activity which obeys a simple non- cooperative kinetics with a Km(ATP) of 70 microM and a Vmax of 0.1 unit/mg. Different from F1-ATPase, the complex is not inactivated by 7- chrolo-4-nitrobenzofrazan. Thus, the inherent activity attributable to a single catalytic site unaffected by other catalytic sites of F1- ATPase is characterized
RP  - NOT IN FILE
NT  - UI - 95354837LA - engRN - 10199-89-0 (4-Chloro-7-nitrobenzofurazan)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19950907IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:7628606
SO  - FEBS Lett 1995 Jul 17 ;368(2):207-210

1331
UI  - 20939
AU  - Schemidt RA
AU  - Hsu DK
AU  - Deckers-Hebestreit G
AU  - Altendorf K
AU  - Brusilow WS
AD  - Department of Biochemistry, Wayne State University School of Medicine, Detroit, Michigan 48201, USA
TI  - The effects of an atpE ribosome-binding site mutation on the stoichiometry of the c subunit in the F1F0 ATPase of Escherichia coli
AB  - We tested the hypothesis that the stoichiometry of the c subunit in the F0 sector of the Escherichia coli F1F0 ATPase is dependent upon the level of atpE gene expression. F0 was purified from cells carrying plasmids encoding the F0 subunits with and without a ribosome-binding site mutation preceding atpE, the gene which codes for the c subunit. Subunit-specific antibodies were used to quantitate the relative amounts of the b and c subunits. The decreased expression of atpE resulted in a significantly decreased amount of the c subunit in the purified F0. Immunoblot quantitation of the amounts of b and c subunits in F1F0 precipitated by anti-F1 antiserum also showed that the mutation produced significant differences in the stoichiometry of subunit c. The amount of c subunit assembled into the F1F0 synthesized from a plasmid carrying the atpE ribosome binding site mutation was 2-5 times less than the amount found in the F1F0 synthesized from a wild-type plasmid. Therefore, the stoichiometry of the c subunit assembled into the F1F0 complex appears to be variable, depending on the expression of atpE
MH  - A
MH  - ATPase
MH  - BINDING
MH  - Biochemistry
MH  - Cells
MH  - COMPLEX
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - Macromolecular Systems
MH  - Site
MH  - SUBUNIT
MH  - SYSTEM
MH  - SYSTEMS
MH  - Time
RP  - NOT IN FILE
NT  - UI - 96063619LA - engRN - 0 (Macromolecular Systems)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleID - KO4-AI00882/AI/NIAIDDA - 19951214IS - 0003-9861SB - IMCY - UNITED STATES
UR  - PM:7487107
SO  - Arch Biochem Biophys 1995 Nov 10 ;323(2):423-428

1332
UI  - 424
AU  - Schmidt G
AU  - Senior AE
AD  - Department of Biochemistry, University of Rochester Medical Center, New York 14642, USA
TI  - ATP-dependent inactivation of the beta-Ser339Cys mutant F1-ATPase from Escherichia coli by N-ethylmaleimide
AB  - We introduced mutations at the highly-conserved residue Ser-339 in subunit beta of Escherichia coli F1-ATPase. The mutations beta S339Y and beta S339F abolished ATPase activity and impaired enzyme assembly. In contrast beta S339C F1 retained function to a substantial degree. N- Ethylmaleimide (NEM) at 0.2-0.3 mM inactivated beta S339C F1-ATPase by 80-95% in the presence of MgATP or MgADP but did not inactivate appreciably in absence of nucleotide or presence of EDTA. In absence of nucleotide, 0.7 mol of [14C-NEM] was incorporated into beta-subunits of 1.0 mol F1: in presence of MgATP the amount was 1.7 mol/mol, i.e. the introduced Cys residue became more accessible to reaction in the presence of MgATP. In the X-ray structure of F1 (Abrahams et al. (1994) Nature 370, 621-628) one of the catalytic nucleotide-binding domains is empty (on the "beta E subunit") and contains a cleft. Residue beta-339 lies within this cleft; the cleft does not occur in the other two beta- subunits. Our data are consistent with the conclusion that in wild-type enzyme under physiological conditions, MgATP or MgADP induce an enzyme conformation in which residue beta-Ser-339 becomes more exposed, possibly similar to the situation seen in the "beta E-subunit" in the X- ray structure
RP  - NOT IN FILE
NT  - UI - 95352655LA - engRN - 128-53-0 (Ethylmaleimide)RN - 52-90-4 (Cysteine)RN - 56-45-1 (Serine)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 60-00-4 (Edetic Acid)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSDA - 19950905IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:7626639
SO  - Biochemistry 1995 Aug 1 ;34(30):9694-9699

1333
UI  - 19895
AU  - Schweighofer KJ
AU  - Benjamin I
TI  - Transfer of small ions across the water/1,2-dichloroethane interface
MH  - ion
MH  - Ions
MH  - TRANSFER
RP  - IN FILE
SO  - J Chem Phys 1995  ;99():9974-9985

1334
UI  - 422
AU  - Senior AE
AU  - Weber J
AU  - al Shawi MK
AD  - Department of Biochemistry, University of Rochester Medical Center, NY 14642, USA
TI  - Catalytic mechanism of Escherichia coli F1-ATPase
RP  - NOT IN FILE
NT  - UI - 96227121LA - engRN - 0 (Nucleotides)RN - 56-65-5 (Adenosine Triphosphate)RN - 7439-95-4 (Magnesium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSDA - 19960730IS - 0300-5127SB - IMCY - ENGLANDJC - E48
UR  - PM:8654830
SO  - Biochem Soc Trans 1995 Nov ;23(4):747-752

1335
UI  - 21287
AU  - Skaf MS
AU  - Ladanyi BM
TI  - Molecular dynamics simulation of the wave vector-dependent static dielectric properties of methanol-water mixtures
MH  - MOLECULAR-DYNAMICS
MH  - SIMULATION
RP  - IN FILE
SO  - J Chem Phys 1995  ;102():6542-6551

1336
UI  - 20849
AU  - Someya Y
AU  - Moriyama Y
AU  - Futai M
AU  - Sawai T
AU  - Yamaguchi A
AD  - Division of Microbial Chemistry, Faculty of Pharmaceutical Sciences, Chiba University, Japan
TI  - Reconstitution of the metal-tetracycline/H+ antiporter of Escherichia coli in proteoliposomes including F0F1-ATPase
AB  - The tetracycline resistance gene (tetA) was cloned downstream of the lac promoter. When expression of the tetA gene in E. coli cells carrying the lac Iq gene was induced with isopropyl beta-D- thiogalactopyranoside, the tetracycline resistance protein (TetA) was overproduced, amounting to about 30% of the integral cytoplasmic membrane protein. Essentially pure TetA protein could be obtained by solubilization with 1.25% n-octyl-beta-D-glucopyranoside and one-step purification by DEAE Sepharose CL-6B column chromatography. The TetA protein was incorporated into proteoliposomes with F0F1-ATPase. The proteoliposomes exhibited [3H]tetracycline transport dependent on ATP hydrolysis. The specific activity was about 2 nmol/mg protein/min. The proteoliposomes also showed H+ efflux coupled with tetracycline influx. Tetracycline/H+ antiport by proteoliposomes reconstituted with the Ser- 65-->Cys mutant TetA protein was inhibited by N-ethylmaleimide. These results proved for the first time that the tetracycline/H+ antiport is only mediated by the TetA protein
MH  - A
MH  - atp
MH  - ATPase
MH  - Bacterial Proteins
MH  - Cells
MH  - Chemistry
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - H+
MH  - Hydrogen
MH  - Hydrolysis
MH  - membrane
MH  - mutant
MH  - protein
MH  - Proteins
MH  - Proteolipids
MH  - proteoliposome
MH  - purification
MH  - reconstitution
MH  - Time
MH  - transport
RP  - NOT IN FILE
NT  - UI - 96049558LA - engRN - 0 (Antiporters)RN - 0 (Bacterial Proteins)RN - 0 (DNA, Bacterial)RN - 0 (Metals)RN - 0 (Proteolipids)RN - 0 (Sulfhydryl Reagents)RN - 0 (proteoliposomes)RN - 0 (tet A protein)RN - 1333-74-0 (Hydrogen)RN - 60-54-8 (Tetracycline)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19951205IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:7589516
SO  - FEBS Lett 1995 Oct 23 ;374(1):72-76

1337
UI  - 630
AU  - Souid AK
AU  - Penefsky HS
AD  - Department of Biochemistry and Molecular Biology, State University of New York, Syracuse 13210, USA
TI  - Energetics of ATP dissociation from the mitochondrial ATPase during oxidative phosphorylation
AB  - The dissociation constant (KdATP) for ATP bound in the high affinity catalytic site of membrane-bound beef heart mitochondrial ATPase (F1) was calculated from the ratio of the rate constants for the reverse dissociation step (k-1) and the forward binding step (k+1). k-1 for ATP bound to submitochondrial particles or to submitochondrial particles washed with KCl so as to activate ATPase activity was accelerated by about five orders of magnitude during respiratory chain-linked oxidations of NADH. In the presence of NADH and 0.1 mM ADP, k-1 increased more than six orders of magnitude. These energy-dependent dissociations of ATP were sensitive to the uncoupler carbonyl cyanide p- trifluoromethyloxyphenylhydrazone. Only small changes in k+1 were observed in the presence of NADH or NADH and ADP. KdATP at 23 degrees C in the absence of NADH and ADP was 10(-12) M, in the presence of NADH, 3 microM, and in the presence of NADH and 0.1 mM ADP, 60 microM. Thus, the dissociation of ATP during the transition from non-energized to energized states was, under these conditions, accompanied by observed free energy changes of 8 and 9.7 kcal/mol, respectively
RP  - NOT IN FILE
NT  - UI - 95238412LA - engRN - 53-84-9 (NAD)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleID - GM21737/GM/NIGMSDA - 19950523IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:7721821
SO  - J Biol Chem 1995 Apr 21 ;270(16):9074-9082

1338
UI  - 386
AU  - Syroeshkin AV
AU  - Vasilyeva EA
AU  - Vinogradov AD
AD  - Department of Biochemistry, School of Biology, Moscow State University, Russian Federation
TI  - ATP synthesis catalyzed by the mitochondrial F1-F0 ATP synthase is not a reversal of its ATPase activity
AB  - The ADP(Mg2+)-deactivated oligomycin-sensitive F1-F0 ATPase of coupled submitochondrial particles treated with the substoichiometric amount of oligomycin was studied to test whether ATP synthesis and hydrolysis proceed in either direction through the same intermediates. The initial rates of ATP hydrolysis, oxidative phosphorylation, ATP-dependent, succinate-supported NAD+ reduction, and ATP-induced delta microH+ generation were measured using deactivated ATPase trapped by azide [Biochem. J. (1982) 202, 15-23]. Three ATP consuming reactions were strongly inhibited when azide was present in the assay mixtures, whereas ATP synthesis was not altered by azide. The unidirectional effect of azide is not consistent with three alternating binding sites mechanism operating in ATP synthesis and support our hypothesis on the existence of nucleotide(Mg2+)-controlled 'synthase' and 'hydrolase' states of the mitochondrial F1-F0 ATPase
RP  - NOT IN FILE
NT  - UI - 95309397LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19950726IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:7789510
SO  - FEBS Lett 1995 Jun 5 ;366(1):29-32

1339
UI  - 726
AU  - Tozawa K
AU  - Sekino N
AU  - Soga M
AU  - Yagi H
AU  - Yoshida M
AU  - Akutsu H
AD  - Department of Bioengineering, Faculty of Engineering, Yokohama National University, Japan
TI  - Conformational dynamics monitored by His-179 and His-200 of isolated thermophilic F1-ATPase beta subunit which reside at the entrance of the 'conical tunnel' in holoenzyme
AB  - When monitored by 1H NMR at various pH values, most of the C-2 proton signals from 12 His residues of the isolated beta subunit of thermophilic F1-ATPase (TF1) could be separately observed. Two of them were assigned to His-179 and His-200 which reside at the entrance of a 'conical tunnel' to reach catalytic site in the crystal structure of F1- ATPase. His-200 gave doublet, suggesting that this region is not a rigid alpha-helix in the isolated beta subunit. The binding of Mg.AMP- PNP changed the chemical shifts of His-179 and His-200 significantly. Although His-119 located at the opposite side of the conical tunnel was not affected by the nucleotide-binding, it contributed to the stability of beta subunit and the efficiency of the catalysis of the holoenzyme
RP  - NOT IN FILE
NT  - UI - 96105378LA - engRN - 7006-35-1 (Histidine)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19960118IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:7498539
SO  - FEBS Lett 1995 Dec 4 ;376(3):190-194

1340
UI  - 337
AU  - Walker JE
AU  - Collinson IR
AU  - van Raaij MJ
AU  - Runswick MJ
AD  - Medical Research Council Laboratory of Molecular Biology, Cambridge, United Kingdom
TI  - Structural analysis of ATP synthase from bovine heart mitochondria
RP  - NOT IN FILE
NT  - UI - 96130706LA - engRN - 0 (DNA Primers)RN - 0 (Indicators and Reagents)RN - 0 (Macromolecular Systems)RN - 0 (Plasmids)RN - 0 (Recombinant Proteins)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19960401IS - 0076-6879SB - IMCY - UNITED STATESJC - MVA
UR  - PM:8592443
SO  - Methods Enzymol 1995  ;260():163-190

1341
UI  - 110
AU  - Watts SD
AU  - Zhang Y
AU  - Fillingame RH
AU  - Capaldi RA
AD  - Institute of Molecular Biology, University of Oregon, Eugene 97403, USA
TI  - The gamma subunit in the Escherichia coli ATP synthase complex (ECF1F0) extends through the stalk and contacts the c subunits of the F0 part
AB  - A mutant, in which a cysteine has been site-directed into the polar loop region of the c subunit at residue 44, has been studied. Cross- linking of the c subunit to both the gamma and epsilon subunits was observed with cupric 1,10-phenanthrolinate treatment. The linkage between the c and gamma subunits was localized to that part of the gamma subunit between residues 202-286, based on peptide analysis. Reference to the high resolution structure of F1 [Abrahams et al. (1994) Nature 370, 621-628] appears to limit this contact site to the region including residues 202-230. This segment contains 4 tyrosines and 1 tryptophan as possible reactive residues for cross-linking with the c subunit cysteine
RP  - NOT IN FILE
NT  - UI - 95354843LA - engRN - 0 (Cross-Linking Reagents)RN - 0 (Disulfides)RN - 0 (Phenanthrolines)RN - 15823-71-9 (bis(1,10-phenanthroline)copper(2+) ion)RN - 52-90-4 (Cysteine)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM 07759/GM/NIGMSID - GM23105/GM/NIGMSID - HL24526/HL/NHLBIDA - 19950907IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:7628612
SO  - FEBS Lett 1995 Jul 17 ;368(2):235-238

1342
UI  - 425
AU  - Weber J
AU  - Senior AE
AD  - Department of Biochemistry, University of Rochester Medical Center, New York 14642, USA
TI  - Location and properties of pyrophosphate-binding sites in Escherichia coli F1-ATPase
AB  - Binding of pyrophosphate (PPi) to the three catalytic ("C") and three noncatalytic ("NC") nucleotide sites of Escherichia coli F1-ATPase was determined by fluorescence spectroscopy using mutant enzymes with tryptophan inserted specifically in either C sites (beta Y331W) or NC sites (alpha R365W). Fluorescence of the tryptophan is quenched on binding of nucleotide; PPi binding parameters were determined by competition with ATP or adenyl-5'-yl imidodiphosphate. It was found that MgPPi binds to each NC site with Kd = 20 microM. In contrast, even at millimolar concentration, neither MgPPi nor free PPi showed significant binding to C sites. We confirmed that free PPi displaces nucleotide from C sites, but this was shown to be due to complexation of Mg2+ ions rather than to occupancy of the sites. MgPPi bound at NC sites was found not to affect ATP hydrolysis rates. From the data we propose a two-phase model for nucleotide binding at NC sites. In phase one, NC sites recognize the pyrophosphate "end" of the nucleotide, which binds initially with Kd similar to MgPPi; in phase two, a slow conformational change occurs which tightly sequesters adenine nucleotide. Phase two does not occur with guanine nucleotide. This model explains the preference of NC sites for adenine nucleotides. Pi (5 mM) did not bind to either C or NC sites
RP  - NOT IN FILE
NT  - UI - 95279401LA - engRN - 0 (Adenine Nucleotides)RN - 0 (Diphosphates)RN - 0 (Ligands)RN - 0 (Magnesium Compounds)RN - 0 (Phosphates)RN - 13446-24-7 (magnesium pyrophosphate)RN - 25612-73-1 (Adenylyl Imidodiphosphate)RN - 56-65-5 (Adenosine Triphosphate)RN - 61-19-8 (Adenosine Monophosphate)RN - 73-22-3 (Tryptophan)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSDA - 19950628IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:7759515
SO  - J Biol Chem 1995 May 26 ;270(21):12653-12658

1343
UI  - 423
AU  - Weber J
AU  - Bowman C
AU  - Wilke-Mounts S
AU  - Senior AE
AD  - Department of Biochemistry, University of Rochester Medical Center, New York 14642, USA
TI  - alpha-Aspartate 261 is a key residue in noncatalytic sites of Escherichia coli F1-ATPase
AB  - X-ray structure analysis of the noncatalytic sites of F1-ATPase revealed that residue alpha-Asp261 lies close to the Mg of bound Mg-5'- adenylyl-beta,gamma-imidodiphosphate. Here, the mutation alpha D261N was generated in Escherichia coli and combined with the alpha R365W mutation, allowing nucleotide binding at F1 noncatalytic sites to be specifically monitored by tryptophan fluorescence spectroscopy. Purified alpha D261N/alpha R365W F1-ATPase showed catalytic activity similar to wild-type. An important feature was that, without any resort to nucleotide-depletion procedures, the noncatalytic sites in purified native enzyme were already empty. Binding studies with MgATP, MgADP, and the corresponding free nucleotides led to the following conclusions. Residue alpha-Asp261 interacts with the Mg of Mg- nucleotide in noncatalytic sites and provides a large component of the binding energy (approximately 3 kcal/mol). It is the primary determinant of the preference of noncatalytic sites for Mg-nucleotide. The natural ligands at these sites in wild-type enzyme are the Mg- nucleotides and free nucleotides bind poorly. Under conditions where noncatalytic sites were empty, alpha D261N/alpha R365W F1 showed significant hydrolysis of MgATP. This established unequivocally that occupancy of noncatalytic sites by nucleotide is not required for catalysis
RP  - NOT IN FILE
NT  - UI - 95403331LA - engRN - 0 (Oligodeoxyribonucleotides)RN - 56-65-5 (Adenosine Triphosphate)RN - 56-84-8 (Aspartic Acid)RN - EC 3.6.1.- (Ca(2+) Mg(2+)-ATPase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSDA - 19951018IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:7673131
SO  - J Biol Chem 1995 Sep 8 ;270(36):21045-21049

1344
UI  - 421
AU  - Wilke-Mounts S
AU  - Pagan J
AU  - Senior AE
AD  - Department of Biochemistry, University of Rochester Medical Center, New York 14642, USA
TI  - Mutagenesis and reversion analysis of residue Met-209 of the beta- subunit of Escherichia coli ATP synthase
AB  - Residue beta-Met-209 is conserved in all known F1-ATPase sequences, and the mutation beta M209I in Escherichia coli causes profound inhibition of ATP synthesis and hydrolysis. Based on the properties of this mutant it had previously been proposed that residue beta-209 lies close to the site of catalysis. Two approaches were used to study this residue further. First, revertants were sought. Only wild-type and beta-Ser-209 were found; the Ser revertants involved a two-base change. Significantly, Ser is found at the equivalent position in the homologous vacuolar and archaebacterial ATPases. Second, all 20 natural amino acids were placed at position beta-209 by mutagenesis, and catalytic properties of the mutants were analyzed. The results showed that only a limited set of residues supported significant growth or ATPase activity, and that many of the mutations impacted severely on catalysis. X-ray structure analysis of the bovine enzyme has revealed that residue beta-Met-209 lies only 3.1. A from residue beta-Glu-181, which has been proposed to act as catalytic base. The results reported here emphasize that, in this discrete region of the catalytic site, specific stereo-chemical constraints on structure are critical for catalysis
RP  - NOT IN FILE
NT  - UI - 96095704LA - engRN - 0 (Proton Pump)RN - 56-65-5 (Adenosine Triphosphate)RN - 7005-18-7 (Methionine)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSDA - 19960116IS - 0003-9861SB - IMCY - UNITED STATESJC - 6SK
UR  - PM:7503551
SO  - Arch Biochem Biophys 1995 Dec 1 ;324(1):153-158

1345
UI  - 161
AU  - Wilkens S
AU  - Dahlquist FW
AU  - McIntosh LP
AU  - Donaldson LW
AU  - Capaldi RA
AD  - Institute of Molecular Biology, University of Oregon, Eugene 97401, USA
TI  - Structural features of the epsilon subunit of the Escherichia coli ATP synthase determined by NMR spectroscopy
AB  - The tertiary fold of the epsilon subunit of the Escherichia coli F1F0 ATPsynthase (ECF1F0) has been determined by two- and three-dimensional heteronuclear (13C, 15N) NMR spectroscopy. The epsilon subunit exhibits a distinct two domain structure, with the N-terminal 84 residues of the protein forming a 10-stranded beta-structure, and with the C-terminal 48 amino acids arranged as two alpha-helices running antiparallel to one another (two helix hairpin). The beta-domain folds as a beta- sandwich with a hydrophobic interior between the two layers of the sandwich. The C-terminal two-helix hairpin folds back to the N-terminal domain and interacts with one side of the beta-domain. The arrangement of the epsilon subunit in the intact F1F0 ATP synthase involves interaction of the two helix hairpin with the F1 part, and binding of the open side of the beta-sandwich to the c subunits of the membrane- embedded F0 part
RP  - NOT IN FILE
NT  - UI - 96069782LA - engRN - 0 (Carbon Isotopes)RN - 0 (Nitrogen Isotopes)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19951219IS - 1072-8368SB - IMCY - UNITED STATESJC - B98
UR  - PM:7583669
SO  - Nat Struct Biol 1995 Nov ;2(11):961-967

1346
UI  - 730
AU  - Yoshida M
AU  - Amano T
AD  - Research Laboratory of Resources Utilization, Tokyo Institute of Technology, Yokohama, Japan
TI  - A common topology of proteins catalyzing ATP-triggered reactions
AB  - A protein fold, six parallel beta strands surrounding the central alpha helix, is likely to be a common structure in protein families known to have a typical set of nucleotide binding consensus sequence motifs A and B and to catalyze ATP-triggered reactions. According to this ATP- triggered protein fold, the conserved Glu (or Asp), which acts as a general base to activate a water molecule for an in-line attack of the gamma-phosphate, is at the exit of the second beta strand. The fifth beta strand may be involved in propagation of conformational change triggered by ATP hydrolysis
RP  - NOT IN FILE
NT  - UI - 95154435LA - engRN - 0 (Proteins)RN - 56-65-5 (Adenosine Triphosphate)RN - 56-86-0 (Glutamic Acid)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 19950313IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:7851521
SO  - FEBS Lett 1995 Feb 6 ;359(1):1-5

1347
UI  - 21189
AU  - Zaslavsky DL
AU  - Smirnova IA
AU  - Siletsky SA
AU  - Kaulen AD
AU  - Millett F
AU  - Konstantinov AA
AD  - AN Belozersky Institute of Physico-Chemical Biology, Moscow State University, Russian Federation
TI  - Rapid kinetics of membrane potential generation by cytochrome c oxidase with the photoactive Ru(II)-tris-bipyridyl derivative of cytochrome c as electron donor
AB  - Yeast iso-1-cytochrome c covalently modified at cysteine-102 with (4- bromomethyl-4'-methylbipyridine)[bis(bipyridine)]Ru2+ (Ru-102-Cyt c) has been used as a photoactive electron donor to mitochondrial cytochrome c oxidase (COX) reconstituted into phospholipid vesicles. Rapid kinetics of membrane potential generation by the enzyme following flash-induced photoreduction of Ru-102-Cyt c heme has been measured and compared to photovoltaic responses observed with Ru(II)(bipyridyl)3 (RuBpy) as the photoreductant [D.L. Zaslavsky et al. (1993) FEBS Lett. 336, 389-393]. At low ionic strength, when Ru-102-Cyt c forms a tight electrostatic complex with COX, flash-activation results in a polyphasic electrogenic response corresponding to transfer of a negative charge to the interior of the vesicles. The initial rapid phase is virtually identical to the 50 microsecond transient observed in the presence of RuBpy as the photoactive electron donor which originates from electrogenic reduction of heme a by CuA. CuA reduction by Ru-102-Cyt c turns out to be not electrogenic in agreement with the peripheral location of visible copper in the enzyme. A millisecond phase (tau ca. 4 ms) following the 50 microsecond initial part of the response and associated with vectorial translocation of protons linked to oxygen intermediate interconversion in the binuclear centre, can be resolved both with RuBpy and Ru-102-Cyt c as electron donors; however, this phase is small in the absence of added H2O2. In addition to these two transients, the flash-induced electrogenic response in the presence of Ru-102-Cyt c reveals a large slow phase of delta psi generation not observed with RuBpy. This phase is completely quenched upon inclusion of 100 microM ferricyanide in the medium and originates from a second order reaction of COX with the excess Ru-102-Cyt c2+ generated by the flash in a solution
MH  - A
MH  - COMPLEX
MH  - Copper
MH  - Cysteine
MH  - cytochrome
MH  - Cytochrome c
MH  - Cytochrome-c Oxidase
MH  - delta
MH  - electrogenic
MH  - electron
MH  - flash
MH  - intermediate
MH  - Kinetics
MH  - liposome
MH  - Liposomes
MH  - membrane
MH  - Membrane Potential
MH  - microsecond
MH  - Oxygen
MH  - proton
MH  - Protons
MH  - PSI
MH  - TRANSFER
MH  - translocation
MH  - vesicles
RP  - NOT IN FILE
NT  - UI - 95154439LA - engRN - 0 ((4-bromomethyl-4'-methylbipyridine)(bis(bipyridine))ruthenium(II))RN - 0 (Liposomes)RN - 0 (Organometallic Compounds)RN - 52-90-4 (Cysteine)RN - 7440-18-8 (Ruthenium)RN - 9007-43-6 (Cytochrome c)RN - EC 1.9.3.1 (Cytochrome-c Oxidase)PT - Journal ArticleID - GM 20488/GM/NIGMSDA - 19950313IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:7851525
SO  - FEBS Lett 1995 Feb 6 ;359(1):27-30

1348
UI  - 27
AU  - Zhang Y
AU  - Fillingame RH
AD  - Department of Biomolecular Chemistry, University of Wisconsin Medical School, Madison 53706, USA
TI  - Subunits coupling H+ transport and ATP synthesis in the Escherichia coli ATP synthase. Cys-Cys cross-linking of F1 subunit epsilon to the polar loop of F0 subunit c
AB  - Second site suppressor mutations at position 31 of F1 subunit epsilon recouple ATP-driven H+ translocation in the uncoupled Q42E mutant of subunit c of the Escherichia coli F1F0 ATP synthase (Zhang, Y., Oldenburg, M., and Fillingame, R. H. (1994) J. Biol. Chem. 269, 10221- 10224). This finding suggests a functional interaction between subunit c and subunit epsilon during the coupling of H+ transport through F0 to ATP synthesis of F1. However, the physical proximity of the two subunits remained to be defined. In this study, Cys residues were introduced into residues in the polar loop region of subunit c surrounding Gln42 and at position 31 of subunit epsilon to see whether the subunits could be cross-linked. Disulfide bridge formation between subunit c and subunit epsilon was observed in membranes of three double mutants, i.e. cA40C/epsilon E31C, cQ42C/epsilon E31C, and cP43C/epsilon E31C, but not in wild type membranes or in membranes of the cA39C/epsilon E31C double mutant. These results indicate that the polar loop of subunit c and the region around residue 31 of subunit epsilon are physically close to each other in the F1F0 complex and support the hypothesis that these two subunits interact directly in the coupling of H+ transport to ATP synthesis. Disulfide cross-linking of the Q42C subunit c and E31C subunit epsilon leads to inhibition of ATPase coupled H+ transport, as might be expected in a model where the catalytic sites of the F1 ATPase alternate during H+ transport-coupled ATP hydrolysis/synthesis. However, a quantitative relationship between the extent of inhibition of transport and the extent of cross-linking could not be established by the methods used here, and the possibility remains that the epsilon-c cross-linked F1F0 complex retains residual H+ transporting activity
RP  - NOT IN FILE
NT  - UI - 96025863LA - engRN - 0 (Macromolecular Systems)RN - 0 (Oligodeoxyribonucleotides)RN - 0 (Plasmids)RN - 0 (Recombinant Proteins)RN - 52-90-4 (Cysteine)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM23105/GM/NIGMSDA - 19951204IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:7592682
SO  - J Biol Chem 1995 Oct 13 ;270(41):24609-24614

1349
UI  - 31
AU  - Zhang Y
AU  - Fillingame RH
AD  - Department of Biomolecular Chemistry, University of Wisconsin Medical School, Madison 53706
TI  - Changing the ion binding specificity of the Escherichia coli H(+)- transporting ATP synthase by directed mutagenesis of subunit c
AB  - Most F1F0 type ATP synthases, including that in Escherichia coli, use H+ as the coupling ion for ATP synthesis. However, the structurally related F1F0 ATP synthase in Propionigenium modestum uses Na+ instead. The binding site for Na+ residues in the F0 sector of the P. modestum enzyme. We postulated that Na+ might interact with subunit c of F0. Subunit c of P. modestum and E. coli are reasonably homologous (19% identity) but show striking variations around the H(+)-translocating, dicyclohexylcarbodiimide-reactive carboxyl (Asp61 in E. coli). Several hydrophobic residues around Asp61 were replaced with polar residues according to the P. modestum sequence in the hope that the polar replacements might provide liganding groups for Na+. One mutant from 31 different mutation combinations did generate an active enzyme that binds Li+, the combination being V60A, D61E, A62S, and I63T. Li+ binding was detected by Li+ inhibition of ATP-driven H+ transport, Li+ inhibition of F1F0-ATPase activity, and Li+ inhibition of F0-mediated H+ transport. The Li+ effects were observed with membrane vesicles prepared from a delta nhaA, delta nhaB mutant background which lacks Na+/H+ antiporters, and with purified, reconstituted preparations of F0 prepared from this background strain. Li+ inhibition was observed at pH 8.5 but not at pH 7.0. H+ thus appears to compete with Li+ for the binding site. Li+ binding was abolished by replacement of Glu61 by Asp or Ser62 by Ala. The side chains at Ala60 and Thr63 may act in a supporting structural role by providing a more flexible conformation for the Li+ binding cavity. Thr63 does not appear to provide a liganding group since H+ transport in two other mutants, with Gly or Ala in place of Thr63, was also inhibited by Li+. We suggest that a X- Glu-Ser-Y or X-Glu-Thr-Y sequence may provide a general structural motif for monovalent cation binding, and that the flexibility provided by residues X and Y will prove crucial to this structure
RP  - NOT IN FILE
NT  - UI - 95113882LA - engRN - 0 (Ions)RN - 0 (Oligodeoxyribonucleotides)RN - 0 (Protons)RN - 7439-93-2 (Lithium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM23105/GM/NIGMSDA - 19950203IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:7814424
SO  - J Biol Chem 1995 Jan 6 ;270(1):87-93

1350
UI  - 331
AU  - Abrahams JP
AU  - Buchanan SK
AU  - van Raaij MJ
AU  - Fearnley IM
AU  - Leslie AG
AU  - Walker JE
AD  - Medical Research Council Laboratory of Molecular Biology, Cambridge, United Kingdom
TI  - The structure of bovine F1-ATPase complexed with the peptide antibiotic efrapeptin
AB  - In the previously determined structure of mitochondrial F1-ATPase determined with crystals grown in the presence of adenylyl- imidodiphosphate (AMP-PNP) and ADP, the three catalytic beta-subunits have different conformations and nucleotide occupancies. AMP-PNP and ADP are bound to subunits beta TP and beta DP, respectively, and the third beta-subunit (beta E) has no bound nucleotide. The efrapeptins are a closely related family of modified linear peptides containing 15 amino acids that inhibit both ATP synthesis and hydrolysis by binding to the F1 catalytic domain of F1F0-ATP synthase. In crystals of F1- ATPase grown in the presence of both nucleotides and inhibitor, efrapeptin is bound to a unique site in the central cavity of the enzyme. Its binding is associated with small structural changes in side chains of F1-ATPase around the binding pocket. Efrapeptin makes hydrophobic contacts with the alpha-helical structure in the gamma- subunit, which traverses the cavity, and with subunit beta E and the two adjacent alpha-subunits. Two intermolecular hydrogen bonds could also form. Intramolecular hydrogen bonds probably help to stabilize efrapeptin's two domains (residues 1-6 and 9-15, respectively), which are connected by a flexible region (beta Ala-7 and Gly-8). Efrapeptin appears to inhibit F1-ATPase by blocking the conversion of subunit beta E to a nucleotide binding conformation, as would be required by an enzyme mechanism involving cyclic interconversion of catalytic sites
RP  - NOT IN FILE
NT  - UI - 96382481LA - engRN - 0 (Antibiotics, Peptide)RN - 56645-91-1 (efrapeptin)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19961024IS - 0027-8424SB - IMCY - UNITED STATESJC - PV3
UR  - PM:8790345
SO  - Proc Natl Acad Sci U S A 1996 Sep 3 ;93(18):9420-9424

1351
UI  - 83
AU  - Aggeler R
AU  - Capaldi RA
AD  - Institute of Molecular Biology, University of Oregon, Eugene, Oregon 97403-1229, USA
TI  - Nucleotide-dependent movement of the epsilon subunit between alpha and beta subunits in the Escherichia coli F1F0-type ATPase
AB  - Mutants of ECF1-ATPase were generated, containing cysteine residues in one or more of the following positions: alphaSer-411, betaGlu-381, and epsilonSer-108, after which disulfide bridges could be created by CuCl2 induced oxidation in high yield between alpha and epsilon, beta and epsilon, alpha and gamma, beta and gamma (endogenous Cys-87), and alpha and beta. All of these cross-links lead to inhibition of ATP hydrolysis activity. In the two double mutants, containing a cysteine in epsilonSer-108 along with either the DELSEED region of beta (Glu-381) or the homologous region in alpha (Ser-411), there was a clear nucleotide dependence of the cross-link formation with the epsilon subunit. In betaE381C/epsilonS108C the beta-epsilon cross-link was obtained preferentially when Mg2+ and ADP + Pi (addition of MgCl2 + ATP) was present, while the alpha-epsilon cross-link product was strongly favored in the alphaS411C/epsilonS108C mutant in the Mg2+ ATP state (addition of MgCl2 + 5'-adenylyl-beta,gamma-imidodiphosphate). In the triple mutant alphaS411C/betaE381C/epsilonS108C, the epsilon subunit bound to the beta subunit in Mg2+-ADP and to the alpha subunit in Mg2+-ATP, indicating a significant movement of this subunit. The gamma subunit cross-linked to the beta subunit in higher yield in Mg2+- ATP than in Mg2+-ADP, and when possible, i.e. in the triple mutant, always preferred the interaction with the beta over the alpha subunit
RP  - NOT IN FILE
NT  - UI - 96278829LA - engRN - 0 (Cross-Linking Reagents)RN - 0 (Oligodeoxyribonucleotides)RN - 52-90-4 (Cysteine)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - HL24526/HL/NHLBIDA - 19960826IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:8662953
SO  - J Biol Chem 1996 Jun 7 ;271(23):13888-13891

1352
UI  - 722
AU  - Amano T
AU  - Hisabori T
AU  - Muneyuki E
AU  - Yoshida M
AD  - Research Laboratory of Resources Utilization, Tokyo Institute of Technology, 4259 Nagatsuta, Yokohama 226, Japan
TI  - Catalytic activities of alpha3beta3gamma complexes of F1-ATPase with 1, 2, or 3 incompetent catalytic sites
AB  - In order to know how many functional catalytic sites are necessary for ATPase activity of F1-ATPase from a thermophilic Bacillus PS3, a new method of isolating homogeneous preparations of the alpha3beta3gamma complex with 1, 2, or 3 incompetent catalytic sites was developed. Ten glutamic acids (Glu.Tag) were linked to the C terminus of the catalytically incompetent beta(E190Q) subunit. The Glu.Tag itself did not affect ATPase activity of the complexes. Two kinds of alpha3beta3gamma complexes, one containing beta(wild-type) and the other Glu.Tag-linked beta(E190Q), were mixed, urea-denatured, and dialyzed, and alpha3beta3gamma complexes were reconstituted. Each of the complexes containing a different number of Glu.Tag-linked beta(E190Q) was separated by anion-exchange chromatography and analyzed. The results were as follows. 1) Normal steady-state ATPase activity requires three intact catalytic sites. 2) Chase-acceleration, a catalytic cooperativity, requires at least two intact catalytic sites. 3) Single-site catalysis can be mediated by a single intact catalytic site alone. Rescrambling of subunits between complexes could occur when the complex was aged under certain conditions, and this might be one of the reasons for previous contradictory results (Miwa, K., Ohtsubo, M., Denda, K., Hisabori, T., Date, T., and Yoshida, M.(1989) J. Biochem. (Tokyo) 106, 730-734)
RP  - NOT IN FILE
NT  - UI - 96279363LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 61368-63-6 (2',3'-O-(2,4,6-trinitro-cyclohexadienylidine)adenosine 5'- triphosphate)RN - 84430-17-1 (2',3'-(O-(2,4,6-trinitrocyclohexadienylidine))adenosine 5'- diphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19960903IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:8663463
SO  - J Biol Chem 1996 Jul 26 ;271(30):18128-18133

1353
UI  - 717
AU  - Bar-Zvi D
AU  - Yoshida M
AU  - Shavit N
AD  - Doris and Bertie Black Center of Bioenergetics in Life Sciences, Ben Gurion University of the Negev, Beer Sheva, Israel
TI  - Modification of domains of alpha and beta subunits of F1-ATPase from the thermophylic bacterium PS3, in their isolated and associated forms, by 3'-O-(4-benzoyl)benzoyl adenosine 5'-triphosphate (BzATP)
AB  - Photoaffinity labeling by 3'-O-(4-benzoyl)benzoyl adenosine 5'- triphosphate (BzATP) of the adenine nucleotide binding site(s) on isolated and complexed alpha and beta subunits of F1-ATPase from the thermophilic bacterium PS3 (TF1) is described. BzATP binds to both isolated alpha and beta subunits, to complexed beta subunit but not to complexed alpha subunit. Amino acid sequence determination of radiolabeled peptides obtained by proteolytic digestion of [gamma- 32P]BzATP-labeled alpha subunit indicates that residues on both the amino-terminal (residues A41-E67) and carboxy-terminal (residues Q422- Q476) were modified by BzATP. One of the residues in the carboxy- terminal modified by BzATP is most probably alpha Q422. Although the binding stoichiometry of 1 mol of BzATP incorporated by either isolated or complexed beta subunit was maintained, the spatial conformation of the polypeptide determines which amino acid residue(s) is more accessible to the reactive radical. CNBr derived fragments beta G10- M64, beta E75-M233, and beta D390-M469 were labeled with the isolated beta subunit. With complexed beta subunit the label was found only in CNBr fragments: beta E75-M233 and beta G339-M389. The locations where the covalently bound BzATP was found, in the soluble and assembled subunits, indicate that different conformational states exist. In the isolated form of the alpha and beta subunits the amino- and carboxy- termini can fold and reach the central domain of the polypeptide, the domain containing the adenine nucleotide binding site. When alpha combines with beta to form the alpha 3 beta 3 core complex the new conformation of the subunits is such that covalent labeling by BzATP of alpha and of the amino terminal of beta subunit is excluded
RP  - NOT IN FILE
NT  - UI - 97111577LA - engRN - 0 (Affinity Labels)RN - 56-65-5 (Adenosine Triphosphate)RN - 81790-82-1 (3'-O-(4-benzoyl)benzoyladenosine 5'-triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19970318IS - 0145-479XSB - IMCY - UNITED STATESJC - HIO
UR  - PM:8953379
SO  - J Bioenerg Biomembr 1996 Dec ;28(6):471-481

1354
UI  - 565
AU  - Belogrudov GI
AU  - Tomich JM
AU  - Hatefi Y
AD  - Department of Molecular and Experimental Medicine, Scripps Research Institute, La Jolla, California 92037, USA
TI  - Membrane topography and near-neighbor relationships of the mitochondrial ATP synthase subunits e, f, and g
AB  - The well characterized subunits of the bovine ATP synthase complex are the alpha, beta, gamma, delta, and epsilon subunits of the catalytic sector, F1; the ATPase inhibitor protein; and subunits a, b, c, and d, OSCP (oligomycin sensitivity-conferring protein), F6, and A6L, which are present in the membrane sector, F0, and the 45-A-long stalk that connects F1 to F0. It has been shown recently that bovine ATP synthase preparations also contain three small polypeptides, designated e, f, and g, with respective molecular masses of 8.2, 10. 2, and 11.3 kDa. To ascertain their involvement as bona fide subunits of the ATP synthase and to investigate their membrane topography and proximity to the above ATP synthase subunits, polyclonal antipeptide antibodies were raised in the rabbit to the COOH-terminal amino acid residues 57-70 of e, 75-86 of f, and 91-102 of g. It was shown that (i) e, f, and g could be immunoprecipitated with anti-OSCP IgG from a fraction of bovine submitochondrial particles enriched in oligomycin-sensitive ATPase; (ii) the NH2 termini of f and g are exposed on the matrix side of the mitochondrial inner membrane and can be curtailed by proteolysis; (iii) the COOH termini of all three polypeptides are exposed on the cytosolic side of the inner membrane; and (iv) f cross-links to A6L and to g, and e cross-links to g and appears to form an e-e dimer. Thus, the bovine ATP synthase complex appears to have 16 unlike subunits, twice as many as its counterpart in Escherichia coli
RP  - NOT IN FILE
NT  - UI - 96355358LA - engRN - 0 (Cross-Linking Reagents)RN - 0 (Macromolecular Systems)RN - EC 3.4.21.1 (Chymotrypsin)RN - EC 3.4.21.4 (Trypsin)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - DK08126/DK/NIDDKDA - 19961011IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:8702768
SO  - J Biol Chem 1996 Aug 23 ;271(34):20340-20345

1355
UI  - 19909
AU  - Bopp PA
AU  - Kornyshev AA
AU  - Sutmann G
TI  - Static nonlocal dielectric function of liquid water
MH  - function
MH  - LIQUID
MH  - Water
RP  - IN FILE
SO  - Phys Rev Lett 1996  ;76():1280-1283

1356
UI  - 333
AU  - Buchanan SK
AU  - Walker JE
AD  - Medical Research Council Laboratory of Molecular Biology, Cambridge, UK
TI  - Large-scale chromatographic purification of F1F0-ATPase and complex I from bovine heart mitochondria
AB  - A new chromatographic procedure has been developed for the isolation of F1F0-ATPase and NADH:ubiquinone oxidoreductase (complex I) from a single batch of bovine heart mitochondria. The method employed dodecyl beta-delta-maltoside, a monodisperse, homogeneous detergent in which many respiratory complexes exhibit high activity, for solubilization and subsequent purification by ammonium sulphate fractionation and column chromatography. A combination of anion-exchange, gel-filtration, and dye-ligand affinity chromatography was used to purify both complexes to homogeneity. The F1F0-ATPase preparation contains only the 16 known subunits of the enzyme. It has oligomycin-sensitive ATP hydrolysis activity and, as demonstrated elsewhere, when reconstituted into lipid vesicles it is capable of ATP-dependent proton pumping and of ATP synthesis driven by a proton gradient [Groth and Walker (1996) Biochem. J. 318, 351-357]. The complex I preparation contains all of the subunits identified in other preparations of the enzyme, and has rotenone-sensitive NADH:ubiquinone oxidoreductase and NADH:ferricyanide oxidoreductase activities. The procedure is rapid and reproducible, yielding 50-80 mg of purified F1F0-ATPase and 20-40 mg of purified complex I from 1 g of mitochondrial membranes. Both preparations are devoid of phospholipids, and gel filtration and dynamic light scattering experiments indicate that they are monodisperse. Therefore, the preparations fulfil important prerequisites for structural analysis
RP  - NOT IN FILE
NT  - UI - 96358525LA - engRN - 0 (Detergents)RN - 0 (Glucosides)RN - 0 (Phospholipids)RN - 69227-93-6 (dodecyl maltoside)RN - EC 1.10.2.2 (Ubiquinol-Cytochrome-c Reductase)RN - EC 1.6.99.2 (NAD(P)H Dehydrogenase (Quinone))RN - EC 1.9.3.1 (Cytochrome-c Oxidase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19960926IS - 0264-6021SB - IMCY - ENGLANDJC - 9YO
UR  - PM:8761491
SO  - Biochem J 1996 Aug 15 ;318 ( Pt 1)():343-349

1357
UI  - 157
AU  - Capaldi RA
AU  - Aggeler R
AU  - Wilkens S
AU  - Gruber G
AD  - Institute of Molecular Biology, University of Oregon, Eugene 97403- 1229, USA
TI  - Structural changes in the gamma and epsilon subunits of the Escherichia coli F1F0-type ATPase during energy coupling
AB  - Structural changes in the Escherichia coli ATP synthase (ECF1F0) occur as part of catalysis, cooperativity and energy coupling within the complex. The gamma and epsilon subunits, two major components of the stalk that links the F1 and F0 parts, are intimately involved in conformational coupling that links catalytic site events in the F1 part with proton pumping through the membrane embedded F0 section. Movements of the gamma subunit have been observed by electron microscopy, and by cross-linking and fluorescence studies in which reagents are bound to Cys residues introduced at selected sites by mutagenesis. Conformational changes and shifts of the epsilon subunit related to changes in nucleotide occupancy sites have been followed by similar approaches
RP  - NOT IN FILE
NT  - UI - 97108764LA - engRN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 19970226IS - 0145-479XSB - IMCY - UNITED STATESJC - HIO
UR  - PM:8951085
SO  - J Bioenerg Biomembr 1996 Oct ;28(5):397-401

1358
UI  - 19894
AU  - Chang T
AU  - Dang LX
TI  - Molecular dynamics simulations of CCl4-H2O liquid-liquid interface with polarizable potential models
MH  - model
RP  - IN FILE
SO  - J Chem Phys 1996  ;104():6772-6782

1359
UI  - 330
AU  - Collinson IR
AU  - Skehel JM
AU  - Fearnley IM
AU  - Runswick MJ
AU  - Walker JE
AD  - MRC Laboratory of Molecular Biology, Cambridge, UK
TI  - The F1F0-ATPase complex from bovine heart mitochondria: the molar ratio of the subunits in the stalk region linking the F1 and F0 domains
AB  - The F1 globular catalytic domain and the F0 intrinsic membrane domain of the F1F0-ATPases in bacteria, chloroplasts, and mitochondria are connected by a slender stalk. In the F1F0 complex from bovine heart mitochondria, the stalk is thought to contain subunits OSCP, d, and F6, and the globular part of the membrane bound subunit b, referred to as b'. It has been shown previously that the OSCP, b', d, and F6 proteins can be assembled in vitro into a water soluble complex named the "stalk". The stalk and F1-ATPase together form another complex named F1.stalk. In this paper, the molar ratios of the OSCP, b (or b'), d, and F6 in the stalk, F.stalk, and F1F0-ATPase complexes have been investigated by three independent methods. By quantitation of radioactivity incorporated by S-carboxymethylation with iodo-2- [14C]acetic acid into a stalk complex containing a form of F6 with the mutation Glu3-Cys, it was shown that the stalk consists of equimolar quantities of its four constituent proteins. In the stalk complex containing the natural F6 sequence, this conclusion was confirmed both by quantitation of radioactivity incorporated by Nepsilon-acetimidation with ethyl [1-14C]acetimidate, and by quantitative N-terminal sequence analysis of subunits. By similar Nepsilon-acetimidation experiments, it has been demonstrated that the F1.stalk complex contains one copy per assembly of the OSCP, b', d, and F6 proteins and that the F1F0-ATPase contains one copy per enzyme complex of subunits OSCP, b, and d. The presence of one copy per complex of the OSCP, b' (or b), d, and F6 proteins in the F1.stalk and F1F0-ATPase complexes, respectively, was confirmed by quantitative sequencing
RP  - NOT IN FILE
NT  - UI - 96420503LA - engRN - 0 (2-(4-isothiocyanatophenoxy)-1,3,2-dioxaphosphinene 2-oxide)RN - 0 (Imidoesters)RN - 0 (Iodoacetates)RN - 0 (Membrane Proteins)RN - 0 (Organophosphorus Compounds)RN - 0 (oligomycin sensitivity-conferring protein)RN - 1000-84-6 (ethyl acetimidate)RN - 64-69-7 (Iodoacetic Acid)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19961105IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:8823202
SO  - Biochemistry 1996 Sep 24 ;35(38):12640-12646

1360
UI  - 19858
AU  - Cross RL
AU  - Duncan TM
AD  - Department of Biochemistry and Molecular Biology, State University of New York Health Science Center, Syracuse 13210, USA
TI  - Subunit rotation in F0F1-ATP synthases as a means of coupling proton transport through F0 to the binding changes in F1
AB  - The rotation of an asymmetric core of subunits in F0F1-ATP synthases has been proposed as a means of coupling the exergonic transport of protons through F0 to the endergonic conformational changes in F1 required for substrate binding and produce release. Here we review earlier evidence both for and against subunit rotation and then discuss our most recent studies using reversible intersubunit disulfide cross- links to test for rotation. We conclude that the gamma subunit of F1 rotates relative to the surrounding catalytic subunits during catalytic turnover by both soluble F1 and membrane-bound F0F1. Furthermore, the inhibition of this rotation by the modification of F0 with DCCD suggests that rotation in F1 is obligatorily coupled to rotation in F0 as an integral part of the coupling mechanism
MH  - A
MH  - atp
MH  - ATP synthase
MH  - BINDING
MH  - Biochemistry
MH  - conformational change
MH  - F0
MH  - F0F1
MH  - F1
MH  - H(+)-Transporting ATP Synthase
MH  - mechanism
MH  - proton
MH  - Protons
MH  - review
MH  - rotation
MH  - SUBUNIT
MH  - SYNTHASE
MH  - transport
RP  - NOT IN FILE
NT  - UI - 97108765LA - engRN - 0 (Protons)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialID - GM23152/GM/NIGMSDA - 19970226IS - 0145-479XSB - IMCY - UNITED STATESJC - HIO
UR  - PM:8951086
SO  - J Bioenerg Biomembr 1996 Oct ;28(5):403-408

1361
UI  - 20938
AU  - Deckers-Hebestreit G
AU  - Altendorf K
AD  - Universitat Osnabruck, Fachbereich Biologie/Chemie, Arbeitsgruppe Mikrobiologie, Germany
TI  - The F0F1-type ATP synthases of bacteria: structure and function of the F0 complex
AB  - Membrane-bound ATP synthases (F0F1-ATPases) of bacteria serve two important physiological functions. The enzyme catalyzes the synthesis of ATP from ADP and inorganic phosphate utilizing the energy of an electrochemical ion gradient. On the other hand, under conditions of low driving force, ATP synthases function as ATPases, thereby generating a transmembrane ion gradient at the expense of ATP hydrolysis. The enzyme complex consists of two structurally and functionally distinct parts: the membrane-integrated ion-translocating F0 complex and the peripheral F1 complex, which carries the catalytic sites for ATP synthesis and hydrolysis. The ATP synthase of Escherichia coli, which has been the most intensively studied one, is composed of eight different subunits, five of which belong to F1, subunits alpha, beta, gamma, delta, and epsilon (3:3:1:1:1), and three to F0, subunits a, b, and c (1:2:10 +/- 1). The similar overall structure and the high amino acid sequence homology indicate that the mechanism of ion translocation and catalysis and their mode of coupling is the same in all organisms
MH  - A
MH  - ACID
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - ADP
MH  - Amino Acid Sequence
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - ATPase
MH  - Bacteria
MH  - Catalysis
MH  - COMPLEX
MH  - coupling
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - F1
MH  - function
MH  - Hydrolysis
MH  - INORGANIC-PHOSPHATE
MH  - ion
MH  - Macromolecular Systems
MH  - mechanism
MH  - membrane
MH  - Membrane Proteins
MH  - protein
MH  - Proteins
MH  - proton
MH  - Proton Pump
MH  - review
MH  - Site
MH  - SUBUNIT
MH  - SYNTHASE
MH  - synthesis
MH  - SYSTEM
MH  - SYSTEMS
MH  - translocation
RP  - NOT IN FILE
NT  - UI - 97061053LA - engRN - 0 (Macromolecular Systems)RN - 0 (Membrane Proteins)RN - 0 (Proton Pumps)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticlePT - ReviewPT - Review, AcademicDA - 19970218IS - 0066-4227SB - IMCY - UNITED STATES
UR  - PM:8905099
SO  - Annu Rev Microbiol 1996  ;50():791-824

1362
UI  - 158
AU  - Feng Z
AU  - Aggeler R
AU  - Haughton MA
AU  - Capaldi RA
AD  - Institute of Molecular Biology, University of Oregon, Eugene, 97403- 1229, USA
TI  - Conformational changes in the Escherichia coli ATP synthase (ECF1F0) monitored by nucleotide-dependent differences in the reactivity of Cys- 87 of the gamma subunit in the mutant betaGlu-381 --> Ala
AB  - Cys-87, one of two intrinsic cysteines of the gamma subunit of the Escherichia coli ATP synthase (ECF1F0), is in a short segment of this subunit that binds to the bottom domain of a beta subunit close to a glutamate (Glu-381). Cys-87 was unreactive to maleimides under all conditions in wild-type ECF1 and ECF1F0 but became reactive when Glu- 381 of beta was replaced by a cysteine or alanine. The reactivity of Cys-87 with maleimides was nucleotide-dependent, occurring with ATP or ADP + EDTA in catalytic sites, in the presence of AMP.PNP + Mg2+ but not with ADP + Mg2+ bound, whether Pi was present or not, and not when nucleotide binding sites were empty. Binding of N-ethylmaleimide had no effect, whereas 7-diethyl-amino-3-(4'-maleimidylphenyl)-4- methylcoumarin increased the ATPase activity of ECF1 more than 2-fold by reaction with Cys-87. In ECF1F0, these reagents inhibited activity. The nucleotide dependence of the reaction of Cys-87 of the gamma subunit depended on the presence of the epsilon subunit. In epsilon subunit-free ECF1, maleimides reacted with Cys-87 under all nucleotide conditions, including when catalytic sites were empty. These results are discussed in terms of nucleotide-dependent movements of the gamma subunit during functioning of the F1F0-type ATPase
RP  - NOT IN FILE
NT  - UI - 96279344LA - engRN - 0 (Maleimides)RN - 0 (Nucleotides)RN - 52-90-4 (Cysteine)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - HL24526/HL/NHLBIDA - 19960903IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:8663500
SO  - J Biol Chem 1996 Jul 26 ;271(30):17986-17989

1363
UI  - 23
AU  - Fillingame RH
AD  - Department of Biomolecular Chemistry, University of Wisconsin Medical School, Madison, WI 53706 USA
TI  - Membrane sectors of F- and V-type H+-transporting ATPases
AB  - H+ transporting, reversible F-type ATPases (ATP synthases) and V-type ATPases share major structural features and function by similar mechanisms. Recent structural and genetic experiments provide new insights into the organization of the transmembrane and coupling sectors of the enzymes and the molecular mechanics of coupling H+ transport to ATP
RP  - NOT IN FILE
NT  - UI - 96391945LA - engRN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialID - GM23105/GM/NIGMSDA - 19961018IS - 0959-440XSB - IMCY - ENGLANDJC - B9V
UR  - PM:8794160
SO  - Curr Opin Struct Biol 1996 Aug ;6(4):491-498

1364
UI  - 21332
AU  - Forti G
AU  - Elli G
TI  - Stimulation of Photophosphorylation by Ascorbate as a Function of Light Intensity
AB  - When isolated, stroma-free thylakoids are illuminated in the presence of ADP and orthophosphate in the absence of any electron acceptor except O2, the addition of ascorbate stimulates electron transport through the formation of the radical monodehydroascorbate and the coupled synthesis of ATP (G. Forti and G. Elli [1995] Plant Physiol 109: 1207-1211). The stimulation is shown here to be higher at low light intensity. These observations are explained in terms of the increase of the electron transport rate by ascorbate, which established a higher value of the steady-state pH gradient, causing activation of ATP synthase, which is known to be dependent on the level of the H+-electrochemical potential difference, and a higher rate of proton flux across the membranes available for utilization by ATP synthesis
MH  - A
MH  - acceptor
MH  - ACTIVATION
MH  - ADP
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - electron
MH  - electron acceptor
MH  - Electron Transport
MH  - function
MH  - Light
MH  - membrane
MH  - Membranes
MH  - pH
MH  - Photophosphorylation
MH  - plant
MH  - proton
MH  - SYNTHASE
MH  - synthesis
MH  - thylakoid
MH  - thylakoids
MH  - transport
RP  - NOT IN FILE
NT  - Centro di Studio Consiglio Nazionale delle Ricerche sulla Biologia Cellulare e Molecolare delle Piante, Dipartimento di Biologia, Universita di Milano, Via Celoria 26, Milan 20133, Italy
SO  - Plant Physiol 1996 Dec ;112(4):1509-1511

1365
UI  - 9899
AU  - Frank V
AU  - Kadenbach B
TI  - Regulation of the H+/e- stoichiometry of cytochrome c oxidase from bovine heart by intramitochondrial ATP/ADP ratios.
AB  - This paper describes the effect of intramitochondrial ATP/ADP ratios on the H+/e- stoichiometry of reconstituted cytochrome c oxidase (COX) from bovine heart. At 100% intraliposomal ATP the H+/e- stoichiometry of the reconstituted enzyme is decreased to half of the value measured below 98% intraliposomal ATP (above 2% ADP), while it remains constant up to 100% ADP. The decrease is obtained with different COX preparations, independent of the absolute value of the H+/e- stoichiometry. Decrease of H+/e- stoichiometry is prevented by preincubation of the enzyme with a tissue-specific monoclonal antibody to subunit VIa-H (heart type). Tissue-specific regulation of the efficiency of energy transduction in COX of muscle mitochondria could have a physiological function in maintaining the body temperature at rest or sleep, i.e. at low ATP expenditure.
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - ADP
MH  - Animal
MH  - atp
MH  - Carbonyl Cyanide m-Chlorophenyl Hydrazone
MH  - Cattle
MH  - Cytochrome c
MH  - Cytochrome-c Oxidase
MH  - Electrons
MH  - enzymology
MH  - Ion Transport
MH  - Liposomes
MH  - metabolism
MH  - Mitochondria
MH  - Mitochondria,Heart
MH  - Mitochondria,Liver
MH  - MONOCLONAL-ANTIBODIES
MH  - Oxidation-Reduction
MH  - pharmacology
MH  - physiology
MH  - Proteolipids
MH  - Protons
MH  - regulation
MH  - SUBUNIT
MH  - Support,Non-U.S.Gov't
MH  - Temperature
MH  - Uncoupling Agents
RP  - NOT IN FILE
NT  - Fachbereich Chemie, Philipps-Universitat, Marburg, GermanyPMID- 0008612732EDAT- 1996/03/11 00:00MHDA- 1996/03/11 00:00
SO  - FEBS Lett 1996 Mar 11 ;382(1-2):121-124

1366
UI  - 20844
AU  - Futai M
AU  - Omote H
AD  - Division of Biological Sciences, Osaka University, Japan
TI  - Conformational transmission in ATP synthase during catalysis: search for large structural changes
AB  - Escherichia coli ATP synthase has eight subunits and functions through transmission of conformational changes between subunits. Defective mutation at beta Gly-149 was suppressed by the second mutations at the outer surface of the beta subunit, indicating that the defect by the first mutation was suppressed by the second mutation through long range conformation transmission. Extensive mutant/pseudorevertant studies revealed that beta/alpha and beta/gamma subunits interactions are important for the energy coupling between catalysis and H+ translocation. In addition, long range interaction between amino and carboxyl terminal regions of the gamma subunit has a critical role(s) for energy coupling. These results suggest that the dynamic conformation change and its transmission are essential for ATP synthase
MH  - A
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BETA-SUBUNIT
MH  - Catalysis
MH  - COLI ATP SYNTHASE
MH  - conformation
MH  - conformational change
MH  - coupling
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - function
MH  - H+
MH  - review
MH  - SUBUNIT
MH  - SURFACE
MH  - SYNTHASE
MH  - translocation
RP  - NOT IN FILE
NT  - UI - 97108766LA - engRN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 19970226IS - 0145-479XSB - IMCY - UNITED STATES
UR  - PM:8951087
SO  - J Bioenerg Biomembr 1996 Oct ;28(5):409-414

1367
UI  - 21187
AU  - Grishanin RN
AU  - Bibikov SI
AU  - Altschuler IM
AU  - Kaulen AD
AU  - Kazimirchuk SB
AU  - Armitage JP
AU  - Skulachev VP
AD  - AN Belozersky Institute of Physico-Chemical Biology, Moscow State University, Russia
TI  - delta psi-mediated signalling in the bacteriorhodopsin-dependent photoresponse
AB  - It has been shown previously that the proton-pumping activity of bacteriorhodopsin from Halobacterium salinarium can transmit an attractant signal to the bacterial flagella upon an increase in light intensity over a wide range of wavelengths. Here, we studied the effect of blue light on phototactic responses by the mutant strain Pho8l-B4, which lacks both sensory rhodopsins but has the ability to synthesize bacteriorhodopsin. Under conditions in which bacteriorhodopsin was largely accumulated as the M412 bacteriorhodopsin photocycle intermediate, halobacterial cells responded to blue light as a repellent. This response was pronounced when the membrane electric potential level was high in the presence of arginine, active oxygen consumption, or high-background long-wavelength light intensity but was inhibited by an uncoupler of oxidative phosphorylation (carbonyl cyanide 3-chlorophenylhydrazone) and was inverted in a background of low long-wavelength light intensity. The response to changes in the intensity of blue light under high background light was asymmetric, since removal of blue light did not produce an expected suppression of reversals. Addition of ammonium acetate, which is known to reduce the pH gradient changes across the membrane, did not inhibit the repellent effect of blue light, while the discharge of the membrane electric potential by tetraphenylphosphonium ions inhibited this sensory reaction. We conclude that the primary signal from bacteriorhodopsin to the sensory pathway involves changes in membrane potential
MH  - A
MH  - acetate
MH  - Acetates
MH  - ACTIVE
MH  - Bacteriorhodopsin
MH  - Cells
MH  - delta
MH  - Flagella
MH  - Halobacterium
MH  - Halobacterium salinarium
MH  - intermediate
MH  - ion
MH  - Ions
MH  - Light
MH  - membrane
MH  - Membrane Potential
MH  - mutant
MH  - Onium Compounds
MH  - Oxidative Phosphorylation
MH  - Oxygen
MH  - Oxygen Consumption
MH  - pH
MH  - Phosphorylation
RP  - NOT IN FILE
NT  - UI - 96236010LA - engRN - 0 (Acetates)RN - 0 (Onium Compounds)RN - 0 (Organophosphorus Compounds)RN - 18198-39-5 (tetraphenylphosphonium)RN - 53026-44-1 (Bacteriorhodopsins)RN - 631-61-8 (ammonium acetate)PT - Journal ArticleDA - 19960726IS - 0021-9193SB - IMCY - UNITED STATES
UR  - PM:8655473
SO  - J Bacteriol 1996 Jun ;178(11):3008-3014

1368
UI  - 332
AU  - Groth G
AU  - Walker JE
AD  - Medical Research Council, Laboratory of Molecular Biology, Cambridge, UK
TI  - ATP synthase from bovine heart mitochondria: reconstitution into unilamellar phospholipid vesicles of the pure enzyme in a functional state
AB  - A highly purified and monodisperse preparation of proton-translocating F1F0-ATPase from bovine heart mitochondria is an assembly of 16 unlike polypeptides. This preparation has been reconstituted in the presence of various detergents into unilamellar phospholipid vesicles. Incorporation of the enzyme into vesicles increases the ATP hydrolase activity of the enzyme by 10-20-fold, depending on the detergent, and the highest activities of ATP hydrolysis, 70 units/mg, were obtained by reconstitution from dodecylmaltoside or CHAPS. This activity is mostly sensitive to inhibitors that act on the F0 membrane sector of the complex. From the quenching of the pH-sensitive probe, 9-amino-6-chloro- 2-methoxyacridine, it was shown that the reconstituted enzyme was able to form a transmembrane proton gradient in an ATP-dependent manner. By co-reconstitution of the enzyme with bacteriorhodopsin, it was demonstrated that in the presence of a light-induced proton gradient the enzyme can synthesize ATP from ADP and phosphate. Therefore, the characteristic biological functions of the F1F0-ATPase in mitochondria have been demonstrated with the purified enzyme. Thus, in terms of both its physical and biochemical properties, the purified enzyme fulfils important pre-requisites for formation of two- and three-dimensional crystals
RP  - NOT IN FILE
NT  - UI - 96358526LA - engRN - 0 (Aminoacridines)RN - 0 (Detergents)RN - 0 (Enzyme Inhibitors)RN - 0 (Fluorescent Dyes)RN - 0 (Liposomes)RN - 0 (Oligomycins)RN - 0 (Proteolipids)RN - 0 (proteoliposomes)RN - 28380-24-7 (Nigericin)RN - 3548-09-2 (9-amino-6-chloro-2-methoxyacridine)RN - 370-86-5 (Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone)RN - 53026-44-1 (Bacteriorhodopsin)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19960926IS - 0264-6021SB - IMCY - ENGLANDJC - 9YO
UR  - PM:8761492
SO  - Biochem J 1996 Aug 15 ;318 ( Pt 1)():351-357

1369
UI  - 154
AU  - Gruber G
AU  - Capaldi RA
AD  - Institute of Molecular Biology, University of Oregon, Eugene, Oregon 97403-1229, USA
TI  - The trapping of different conformations of the Escherichia coli F1 ATPase by disulfide bond formation. Effect on nucleotide binding affinities of the catalytic sites
AB  - Two mutants of the Escherichia coli F1 ATPase, betaY331W:E381C/epsilonS108C and alphaS411C/betaY331W/epsilonS108C, have been used to relate nucleotide binding in catalytic sites with different interactions of the stalk-forming subunits gamma and epsilon at the alpha3beta3 subunit domain. Essentially full yield cross-linking between beta + gamma and beta + epsilon, or between alpha + gamma and alpha + epsilon, was obtained in these mutants by Cu2+-induced disulfide bond formation, thereby trapping the enzyme in states with the small subunits interacting either with beta or alpha subunits. The presence of the Trp for beta Tyr-331 in both mutants allowed direct measurement of nucleotide occupancy of catalytic sites. Before cross- linking, Mg2+ATP could be bound in all three catalytic sites in both mutants with a Kd of around 0.1 microM for the highest affinity site and Kd values of approximately 2 microM and 30-40 microM for the second and third sites, respectively. In the absence of Mg2+, ATP also bound in all three catalytic sites but with a single low affinity (above 100 microM) in both mutants. Cu2+-induced cross-linking of ECF1 from the mutant betaY331W:E381C/epsilonS108C had very little effect on nucleotide binding. The binding affinities of the three catalytic sites for Mg2+ATP were not significantly altered from those obtained before cross-linking, and the enzyme still switched between cooperative binding and equal binding affinities of the three catalytic sites (when Mg2+ was absent). When the gamma and epsilon subunits were cross-linked to alpha subunits, ATP binding in the highest affinity catalytic site was dramatically altered. This site became closed so that nucleotide (ATP or ADP) that had been bound into it prior to cross-linking was trapped and could not exchange out. Also, ATP or ADP could not enter this site, although empty, once the enzyme had been cross-linked. Finally, cross-linking of the gamma and epsilon to the alpha subunits prevented the switching between cooperative binding and the state where the three catalytic sites are equivalent. We argue that the conformation of the enzyme in which the small subunits are at alpha subunits occurs during functioning of the enzyme in the course of the rotation of gamma and epsilon subunits within the alpha3beta3 hexamer and that this may be the activated state for ATP synthesis
RP  - NOT IN FILE
NT  - UI - 97115789LA - engRN - 0 (Bacterial Proteins)RN - 0 (Cross-Linking Reagents)RN - 0 (Disulfides)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 60-00-4 (Edetic Acid)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - HL24526/HL/NHLBIDA - 19970123IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:8955091
SO  - J Biol Chem 1996 Dec 20 ;271(51):32623-32628

1370
UI  - 159
AU  - Gruber G
AU  - Capaldi RA
AD  - Institute of Molecular Biology, University of Oregon, Eugene, 97403- 1229, USA
TI  - Differentiation of catalytic sites on Escherichia coli F1ATPase by laser photoactivated labeling with [3H]-2-Azido-ATP using the mutant beta Glu381Cys:epsilonSer108Cys to identify different beta subunits by their interactions with gamma and epsilon subunits
AB  - The ATP binding affinities of the catalytic sites in the three beta subunits of the Escherichia coli F1 ATPase (ECF1) have been explored in relation to the interaction of these subunits with the small subunits gamma and epsilon. ECF1 from the mutant beta E381C:epsilonS108C was reacted with different concentrations of [3H]-2-azido-ATP and covalent insertion of the nucleotide analogue induced by photoactivation of the azide group to a nitrene with single-pulse UV laser excitation. The enzyme showed cooperative binding of [3H]-2-azido-ATP in the presence of Mg2+. The highest affinity site was located at betafree, the one of the three beta subunits in the mutant that does not form disulfide bonds with either the gamma or the epsilon subunit. This beta subunit is, therefore, the site of unisite catalysis in the enzyme. The second mole of [3H]-2-azido-ATP to bind was located in the beta subunit that links to epsilon (betaepsilon), while the lowest affinity binding of the substrate analogue was with the beta subunit that links to gamma (betagamma). In the absence of Mg2+, all three beta subunits bound [3H]- 2-azido-ATP with a similar, low affinity. The results show that binding of MgATP is determined by, and/or must determine, the interactions of the different alpha-beta subunit pairs with the single-copy subunits gamma, delta, and epsilon of the enzyme
RP  - NOT IN FILE
NT  - UI - 96264762LA - engRN - 0 (Affinity Labels)RN - 0 (Azides)RN - 0 (Macromolecular Systems)RN - 0 (Recombinant Proteins)RN - 10028-17-8 (Tritium)RN - 52-90-4 (Cysteine)RN - 56-45-1 (Serine)RN - 56-65-5 (Adenosine Triphosphate)RN - 56-86-0 (Glutamic Acid)RN - 60-00-4 (Edetic Acid)RN - 72884-75-4 (2-azidoadenosine 5'-triphosphate)RN - 7439-95-4 (Magnesium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - HL24526/HL/NHLBIDA - 19960813IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:8672416
SO  - Biochemistry 1996 Apr 2 ;35(13):3875-3879

1371
UI  - 958
AU  - Guerrieri F
AU  - Vendemiale G
AU  - Turturro N
AU  - Fratello A
AU  - Furio A
AU  - Muolo L
AU  - Grattagliano I
AU  - Papa S
TI  - Alteration of mitochondrial F0F1 ATP synthase during aging. Possible involvement of oxygen free radicals
MH  - Adenosine Triphosphate
MH  - Aging
MH  - Animal
MH  - Biochemistry
MH  - biosynthesis
MH  - Chemistry
MH  - Electrophoresis
MH  - Energy Metabolism
MH  - Liver
MH  - Male
MH  - metabolism
MH  - Mitochondria
MH  - Myocardium
MH  - Oxygen
MH  - Rats
MH  - Rats,Wistar
MH  - Support,Non-U.S.Gov't
RP  - NOT IN FILE
NT  - Institute of Medical Biochemistry and Chemistry, University of Bari, Italy
SO  - Ann N Y Acad Sci 1996 Jun 15 ;786:62-71.():62-71

1372
UI  - 817
AU  - Hase B
AU  - Werner-Grune S
AU  - Deckers-Hebestreit G
AU  - Strotmann H
AD  - Institut fur Biochemie der Pflanzen, Heinrich Heine Universitat Dusseldorf, Germany
TI  - Site-directed mutagenesis of two conserved charged amino acids in the N- terminal region of alpha subunit of E. coli-F(0)F(1)
AB  - Two conserved charged amino acids of the N-terminal 'crown' region of the alpha subunit of E. coli-F(1), alpha-D36 and alpha-R40 were exchanged for chemically related (alpha-D36-->E, alpha-R40-->K) or unrelated amino acids (alpha D-36-->K, alpha R40-->G), respectively, by employing oligonucleotide-directed mutagenesis. ATP formation and ATP hydrolyzing activity of isolated plasma membrane vesicles was strongly inhibited in mutant HS2 (alpha-D36-->K), but only slightly affected in the other mutants. The inhibition is not due to a lower content of F0F1 in HS2. In this mutant the extent of the proton gradient generated by ATP hydrolysis was more than 80% inhibited; in all other transformants much smaller effects were observed. The proton gradient established by NADH oxidation was 33% decreased in HS2, but was decreased to a lesser extent in all other mutants. After blockage of F0 by DCCD treatment, the same NADH-induced proton gradient was obtained in all transformants including HS2. This and the fact that the activity of NADH oxidation was unchanged indicate increased proton leakiness of F0F1 carrying the alpha-D36-->K mutation. In F1 alpha-D36 is located in a domain contacting the beta subunit in the vicinity of the arginine beta-R52. The effect of alpha-D36-->K replacement on catalysis and coupling thus may be due to an electrostatic repulsive effect in the crown region which alters the alpha and beta interaction
RP  - NOT IN FILE
NT  - UI - 96196588LA - engRN - 0 (Proton Pump)RN - 53-84-9 (NAD)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - 56-84-8 (Aspartic Acid)RN - 7004-12-8 (Arginine)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19960605IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:8612744
SO  - FEBS Lett 1996 Mar 11 ;382(1-2):171-174

1373
UI  - 80
AU  - Haughton MA
AU  - Capaldi RA
AD  - Institute of Molecular Biology, University of Oregon, Eugene 97403- 1229, USA
TI  - The Escherichia coli F1-ATPase mutant beta Tyr-297-->Cys: functional studies and asymmetry of the enzyme under various nucleotide conditions based on reaction of the introduced Cys with N-ethylmaleimide and 7- chloro-4-nitrobenzofurazan
AB  - Conversion of residue beta Tyr-297 of the Escherichia coli F1-ATPase (ECF1) to a Cys in the mutant beta Y297C led to impaired oxidative phosphorylation based on growth curves. The ATPase activity of ECF1 isolated from the mutant beta Y297C was only 1% of wild-type activity, but the residual activity involves cooperative multi-site enzyme turnover based on inhibition by DCCD and azide. ATPase activity could be increased to 8%, and 13% of wild-type by reaction of the introduced Cys with N-ethyl maleimide (NEM), and 7-chloro-4-nitrobenzofurazan (NbfCl), respectively, suggesting that enzymatic function is improved by an increased hydrophobicity of residue beta Cys-297. The mutation beta Tyr-297-->Cys had no effect on nucleotide binding in studies with the fluorescent analog lin-benzo-ADP. The asymmetry of ECF1 was investigated in the mutants beta Y297C and beta Y297C:E381C/epsilon S108C by examining the relative reactivity of Cys-297 in the three copies of the beta subunit under different nucleotide binding conditions. In agreement with a previous study (Haughton, M.A. and Capaldi, R.A. (1995) J. Biol. Chem., 270, 20568-20574), the asymmetry was maintained under all nucleotide conditions. The NbfCl reaction site was found to be beta free, which is also the site most reactive to NEM, beta epsilon is the second site which reacts with NbfCl or NEM, while the third site, beta gamma, is poorly reactive to either reagent
RP  - NOT IN FILE
NT  - UI - 96413812LA - engRN - 0 (Benzofurans)RN - 0 (Cross-Linking Reagents)RN - 0 (Nucleotides)RN - 0 (Sulfhydryl Reagents)RN - 115491-60-6 (7-chloro-4-nitrobenzofuran)RN - 128-53-0 (Ethylmaleimide)RN - 52-90-4 (Cysteine)RN - 7440-50-8 (Copper)RN - 7758-89-6 (cuprous chloride)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - HL24526/HL/NHLBIDA - 19961031IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:8816947
SO  - Biochim Biophys Acta 1996 Sep 12 ;1276(2):154-160

1374
UI  - 21328
AU  - He XD
AU  - Wu SL
AU  - Li YZ
TI  - Effects of CF(0) Proton Flow on the Distribution of Light Energy Between PSI and PSII
AB  - The Influence of energy transfer inhibitors of ATP synthase on the chlorophyll fluorescence quenching of thylakoids was studied by the saturation pulse method. Triphenyltin chloride (TPT) treatment could result in an increase of q(Q) and a decrease of q(E) of the thylakoids, but DCCD could not. This increase could be abolished when the deltapH across the thylakoid membrane was dissipated by uncouplers (1O mM NH(4)Cl plus 1&mgr;M nigericin) or in higher salt medium which could relieve localized protons on the thylakoid membrane. In the electron transport system of H(2)O right curved arrow PD(0X) or H(2)O right curved arrow PBQ coupled with PSII. The stimulatory effect of TPT on q(Q) also diminished. By the methods of modulated fluorescence and low temperature fluorescence measurement we observed that TPT markedly increased the imbalance of the distribution of light energy between PSI and PSII in favor of PSII. These results suggest that the CF(0) proton flow could influence the light energy distribution between the two photosystems. This might be materialized by the regulation of the membrane-localized protons on the photochemical efficiency of photosystem II
MH  - A
MH  - atp
MH  - ATP synthase
MH  - Chlorophyll
MH  - electron
MH  - Electron Transport
MH  - energy
MH  - Energy Transfer
MH  - fluorescence
MH  - inhibitor
MH  - inhibitors
MH  - Light
MH  - low temperature
MH  - membrane
MH  - method
MH  - Methods
MH  - photosystem II
MH  - physiology
MH  - plant
MH  - proton
MH  - Protons
MH  - PSI
MH  - regulation
MH  - SYNTHASE
MH  - SYSTEM
MH  - Temperature
MH  - thylakoid
MH  - thylakoid membrane
MH  - thylakoids
MH  - TRANSFER
MH  - transport
MH  - uncoupler
RP  - NOT IN FILE
NT  - Shanghai Institute of Plant Physiology, Academia Sinica, Shanghai 200032, China
SO  - Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai ) 1996  ;28(6):678-685

1375
UI  - 21241
AU  - Israelachvili JN
AU  - Wennerstrom H
AD  - Department of Chemical Engineering, University of California, Santa Barbara 93106, USA
TI  - Role of hydration and water structure in biological and colloidal interactions
AB  - The conventional explanation of why hydrophilic surfaces and macromolecules remain well separated in water is that they experience a monotonically repulsive hydration force owing to structuring of water molecules at the surfaces. A consideration of recent experimental and theoretical results suggests an alternative interpretation in which hydration forces are either attractive or oscillatory, and where repulsions have a totally different origin. Further experiments are needed to distinguish between these possibilities
MH  - A
MH  - England
MH  - review
MH  - SOLVENT
MH  - structure
MH  - SURFACE
MH  - Water
RP  - NOT IN FILE
NT  - UI - 96138377LA - engRN - 0 (Solvents)RN - 7732-18-5 (Water)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 19960208IS - 0028-0836SB - IMCY - ENGLAND
UR  - PM:8538786
SO  - Nature 1996 Jan 18 ;379(6562):219-225

1376
UI  - 718
AU  - Jault JM
AU  - Dou C
AU  - Grodsky NB
AU  - Matsui T
AU  - Yoshida M
AU  - Allison WS
AD  - Department of Chemistry and Biochemistry, School of Medicine, University of California at San Diego, La Jolla, California 92093-0601, USA wallison@ucsdedu
TI  - The alpha3beta3gamma subcomplex of the F1-ATPase from the thermophilic bacillus PS3 with the betaT165S substitution does not entrap inhibitory MgADP in a catalytic site during turnover
AB  - The hydrolytic properties of the mutant alpha3(betaT165S)3gamma and wild-type alpha3beta3gamma subcomplexes of TF1 have been compared. Whereas the wild-type complex hydrolyzes 50 &mgr;M ATP in three kinetic phases, the mutant complex hydrolyzes 50 &mgr;M ATP with a linear rate. After incubation with a slight excess of ADP in the presence of Mg2+, the wild-type complex hydrolyzes 2 mM ATP with a long lag. In contrast, prior incubation of the mutant complex under these conditions does not affect the kinetics of ATP hydrolysis. The ATPase activity of the wild- type complex is stimulated 4-fold by 0. 1% lauryl dimethylamine oxide, whereas this concentration of lauryl dimethylamine oxide inhibits the mutant complex by 25%. Compared with the wild-type complex, the activity of the mutant complex is much less sensitive to turnover- dependent inhibition by azide. This comparison suggests that the mutant complex does not entrap substantial inhibitory MgADP in a catalytic site during turnover, which is supported by the following observations. ATP hydrolysis catalyzed by the wild-type complex is progressively inhibited by increasing concentrations of Mg2+ in the assay medium, whereas the mutant complex is insensitive to increasing concentrations of Mg2+. A Lineweaver-Burk plot constructed from rates of hydrolysis of 20-2000 &mgr;M ATP by the wild-type complex is biphasic, exhibiting apparent Km values of 30 &mgr;M and 470 &mgr;M with corresponding kcat values of 26 and 77 s-1. In contrast, a Lineweaver-Burk plot for the mutant complex is linear in this range of ATP concentration, displaying a Km of 133 &mgr;M and a kcat of 360 s-1
RP  - NOT IN FILE
NT  - UI - 97067124LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM-16974/GM/NIGMSDA - 19970107IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:8910526
SO  - J Biol Chem 1996 Nov 15 ;271(46):28818-28824

1377
UI  - 11188
AU  - Junge W
AU  - Sabbert D
AU  - Engelbrecht S
TI  - Rotatory Catalysis by F-ATPase: Real-Time Recording of Intersubunit Rotation
MH  - atp
MH  - Catalysis
MH  - F-ATPASE
MH  - rotation
RP  - IN FILE
SO  - Ber Bunsenges Phys Chem 1996  ;12():

1378
UI  - 724
AU  - Kaibara C
AU  - Matsui T
AU  - Hisabori T
AU  - Yoshida M
AD  - Research Laboratory of Resources Utilization, Tokyo Institute of Technology, Yokohama, Japan
TI  - Structural asymmetry of F1-ATPase caused by the gamma subunit generates a high affinity nucleotide binding site
AB  - The alpha 3 beta 3 gamma and alpha 3 beta 3 complexes of F1-ATPase from a thermophilic Bacillus PS3 were compared in terms of interaction with trinitrophenyl analogs of ATP and ADP (TNP-ATP and TNP-ADP) that differed from ATP and ADP and did not destabilize the alpha 3 beta 3 complex. The results of equilibrium dialysis show that the alpha 3 beta 3 gamma complex has a high affinity nucleotide binding site and several low affinity sites, whereas the alpha 3 beta 2 complex has only low affinity sites. This is also supported from analysis of spectral change induced by TNP-ADP, which in addition indicates that this high affinity site is located on the beta subunit. Single-site hydrolysis of substoichiometric amounts of TNP-ATP by the alpha 3 beta 3 gamma complex is accelerated by the chase addition of excess ATP, whereas that by the alpha 3 beta 3 complex is not. We further examined the complexes containing mutant beta subunits (Y341L, Y341A, and Y341C). Surprisingly, in spite of very weak affinity of the isolated mutant beta subunits to nucleotides (Odaka, M., Kaibara, C., Amano, T., Matsui, T., Muneyuki, E., Ogasawara, K, Yutani, K., and Yoshida, M. (1994) J. Biochem. (Tokyo) 115, 789-796), a high affinity TNP-ADP binding site is generated on the beta subunit in the mutant alpha 3 beta 3 gamma complexes where single-site TNP-ATP hydrolysis can occur. ATP concentrations required for the chase acceleration of the mutant complexes are higher than that of the wild-type complex. The mutant alpha 3 beta 3 complexes, on the contrary, catalyze single-site hydrolysis of TNP-ATP rather slowly, and there is no chase acceleration. Thus, the gamma subunit is responsible for the generation of a high affinity nucleotide binding site on the beta subunit in F1- ATPase where cooperative catalysis can proceed
RP  - NOT IN FILE
NT  - UI - 96161973LA - engRN - 0 (Fluorescent Dyes)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 61368-63-6 (2',3'-O-(2,4,6-trinitro-cyclohexadienylidine)adenosine 5'- triphosphate)RN - 84430-17-1 (2',3'-(O-(2,4,6-trinitrocyclohexadienylidine))adenosine 5'- diphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19960312IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:8576203
SO  - J Biol Chem 1996 Feb 2 ;271(5):2433-2438

1379
UI  - 21249
AU  - Koepke J
AU  - Hu X
AU  - Muenke C
AU  - Schulten K
AU  - Michel H
AD  - Max-Planck-Institut fur Biochemie, Abteilung Molekulare Membranbiologie, Frankfurt, Germany
TI  - The crystal structure of the light-harvesting complex II (B800-850) from Rhodospirillum molischianum
AB  - BACKGROUND: The light-harvesting complexes II (LH-2s) are integral membrane proteins that form ring-like structures, oligomers of alpha beta-heterodimers, in the photosynthetic membranes of purple bacteria. They contain a large number of chromophores organized optimally for light absorption and rapid light energy migration. Recently, the structure of the nonameric LH-2 of Rhodopseudomonas acidophila has been determined; we report here the crystal structure of the octameric LH-2 from Rhodospirillum molischianum. The unveiling of similarities and differences in the architecture of these proteins may provide valuable insight into the efficient energy transfer mechanisms of bacterial photosynthesis. RESULTS: The crystal structure of LH-2 from Rs. molischianum has been determined by molecular replacement at 2.4 A resolution using X-ray diffraction. The crystal structure displays two concentric cylinders of sixteen membrane-spanning helical subunits, containing two rings of bacteriochlorophyll-a (BChl-a) molecules. One ring comprises sixteen B850 BChl-as perpendicular to the membrane plane and the other eight B800 BChl-as that are nearly parallel to the membrane plane; eight membrane-spanning lycopenes (the major carotenoid in this complex) stretch out between the B800 and B850 BChl-as. The B800 BChl-as exhibit a different ligation from that of Rps. acidophila (aspartate is the Mg ligand as opposed to formyl-methionine in Rps. acidophila). CONCLUSIONS: The light-harvesting complexes from different bacteria assume various ring sizes. In LH-2 of Rs. molischianum, the Qy transition dipole moments of neighbouring B850 and B800 BChl-as are nearly parallel to each other, that is, they are optimally aligned for Foster exciton transfer. Dexter energy transfer between these chlorophylls is also possible through interactions mediated by lycopenes and B850 BChl-a phytyl tails; the B800 BChl-a and one of the two B850 BChl-as associated with each heterodimeric unit are in van der Waals distance to a lycopene, such that singlet and triplet energy transfer between lycopene and the BChl-as can occur by the Dexter mechanism. The ring structure of the B850 BChl-as is optimal for light energy transfer in that it samples all spatial absorption and emission characteristics and places all oscillator strength into energetically low lying, thermally accessible exciton states
MH  - A
MH  - absorption
MH  - alpha
MH  - ALPHA-SUBUNIT
MH  - Bacteria
MH  - BETA-SUBUNIT
MH  - carotenoid
MH  - Carotenoids
MH  - Chlorophyll
MH  - COMPLEX
MH  - Energy Transfer
MH  - England
MH  - Light
MH  - mechanism
MH  - MECHANISMS
MH  - membrane
MH  - Membrane Proteins
MH  - Membranes
MH  - Photosynthesis
MH  - protein
MH  - Proteins
MH  - reaction center
MH  - resolution
MH  - rhodopseudomonas
MH  - Rhodospirillum
MH  - structure
MH  - SUBUNIT
MH  - TRANSFER
MH  - X-Ray Diffraction
RP  - NOT IN FILE
NT  - UI - 96347357LA - engRN - 0 (Apoproteins)RN - 0 (B800-850 light-harvesting complex, alpha-subunit)RN - 0 (B800-850 light-harvesting complex, beta-subunit)RN - 0 (Bacteriochlorophylls)RN - 0 (Membrane Proteins)RN - 0 (Photosynthetic Reaction Center, Bacterial)RN - 36-88-4 (Carotenoids)PT - Journal ArticleID - P41 RR05969/RR/NCRRDA - 19961230IS - 0969-2126SB - IMCY - ENGLAND
UR  - PM:8736556
SO  - Structure 1996 May 15 ;4(5):581-597

1380
UI  - 21077
AU  - Krulwich TA
AU  - Ito M
AU  - Gilmour R
AU  - Sturr MG
AU  - Guffanti AA
AU  - Hicks DB
AD  - Department of Biochemistry, Mount Sinai School of Medicine of the City University of New York, NY 10029, USA krulwich@msvaxmssmedu
TI  - Energetic problems of extremely alkaliphilic aerobes
AB  - Over a decade of work on extremely alkaliphilic Bacillus species has clarified the extraordinary capacity that these bacteria have for regulating their cytoplasmic pH during growth at pH values well over 10. However, a variety of interesting energetic problems related to their Na(+)-dependent pH homeostatic mechanism are yet to be solved. They include: (1) the clarification of how cell surface layers play a role in a category of alkaliphiles for which this is the case; (2) identification of the putative, electrogenic Na+/H+ antiporter(s) that, in at least some alkaliphiles, may completely account for a cytoplasmic pH that is over 2 pH units lower than the external pH; (3) the determination of whether specific modules or accessory proteins are essential for the efficacy of such antiporters; (4) the mechanistic basis for the increase in the transmembrane electrical potential at the high external pH values at which the potential-consuming antiporter(s) must be most active; and (5) an explanation for the Na(+)-specificity of pH homeostasis in the extremely alkaliphilic bacilli as opposed to the almost equivalent efficacy of K+ for pH homeostasis in at least some non-alkaliphilic aerobes. The current status of such studies and future strategies will be outlined for this central area of alkaliphile energetics. Also considered, will be strategies to elucidate the basis for robust H(+)-coupled oxidative phosphorylation by alkaliphiles at pH values over 10. The maintenance of a cytoplasmic pH over 2 units below the high external pH results in a low bulk electrochemical proton gradient (delta p). To bypass this low delta p, Na(+)-coupling is used for solute uptake even by alkaliphiles that are mesophiles from environments that are not especially Na(+)-rich. This indicates that these bacteria indeed experience a low delta p, to which such coupling is an adaptation. Possible reasons and mechanisms for using a H(+)- coupled rather than a Na(+)-coupled ATP synthase under such circumstances will be discussed
MH  - A
MH  - ACTIVE
MH  - atp
MH  - ATP synthase
MH  - Bacillus
MH  - Bacteria
MH  - Biochemistry
MH  - coupling
MH  - delta
MH  - electrogenic
MH  - Homeostasis
MH  - mechanism
MH  - MECHANISMS
MH  - Oxidative Phosphorylation
MH  - P
MH  - pH
MH  - Phosphorylation
MH  - protein
MH  - Proteins
MH  - proton
MH  - review
MH  - SURFACE
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 96305567LA - engRN - 0 (Sodium-Hydrogen Antiporter)PT - Journal ArticlePT - ReviewPT - Review, TutorialID - GM28454/GM/NIGMSDA - 19960829IS - 0006-3002SB - IMCY - NETHERLANDS
UR  - PM:8688448
SO  - Biochim Biophys Acta 1996 Jul 18 ;1275(1-2):21-26

1381
UI  - 507
AU  - Lebowitz MS
AU  - Pedersen PL
AD  - Laboratory for Molecular and Cellular Bioenergetics, Department of Biological Chemistry, Johns Hopkins School of Medicine, Baltimore, Maryland 21205, USA
TI  - Protein inhibitor of mitochondrial ATP synthase: relationship of inhibitor structure to pH-dependent regulation
AB  - In the absence of an electrochemical proton gradient, the F1 moiety of the mitochondrial ATP synthase catalyzes the hydrolysis of ATP. This reaction is inhibited by a natural protein inhibitor, in a process characterized by an increase in ATPase inhibition as pH is decreased from 8.0 to 6.0. In order to gain greater insight into the molecular and chemical events underlying this regulatory process, the relationships among pH, helicity of the inhibitor protein, and its capacity to inhibit F1-ATPase activity were examined. First, peptides corresponding to four regions of the 82-amino-acid inhibitor protein were chemically synthesized and assessed for both retention of secondary structure, and capacity to inhibit F1-ATPase activity. These studies showed that a region of only 24-amino-acid residues, from Phe 22 through Len 45, accounts for the inhibitory capacity of the inhibitor protein, and that retention of native helical structure in this region is not essential for inhibition. Second, three mutants (33P34, 39P40, and 43P44) of the intact inhibitor protein were prepared in which a proline residue was inserted within the inhibitory region to disrupt native helical structure. The secondary structures and inhibitory capacities of these mutants were analyzed as a function of pH. These studies revealed that, despite the initial loss of helical structure within the inhibitory region due to proline insertion, a further loss of helical structure is required to modulate inhibitory activity. These results suggest that a loss of helical structure outside the inhibitory region correlates with an increase in inhibitory capacity. Finally, two separate mutants (H48A and H55A) were prepared in which a conserved histidine residue in the wild-type inhibitor protein was replaced with an alanine. The secondary structures and inhibitory capacities of these mutants were also investigated as a function of pH. Results indicated that, although histidine residues do not directly affect the inhibitory capacity of the protein, they are important for maintaining the inhibitor protein in an inactive form at high pH. Furthermore, these results show that loss in helical structure, although correlated with an increase in inhibitory capacity, is not essential for this function. These novel experiments are consistent with a model in which the inhibitor protein is envisioned as consisting of two regions, an inhibitory region and a regulatory region. It is suggested that reduction of pH allows for the protonation of a histidine residue blocking the interaction between the two regions, thus activating the inhibitory response. The pH reduction also correlates with a partial unfolding of the protein that may either cause or result from the loss of interaction between the two helices. This unfolding may be necessary for further optimization of inhibitor function
RP  - NOT IN FILE
NT  - UI - 96239150LA - engRN - 0 (Enzyme Inhibitors)RN - 0 (Oligodeoxyribonucleotides)RN - 0 (Proteins)RN - 147-85-3 (Proline)RN - 7006-35-1 (Histidine)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - CA 10951/CA/NCIDA - 19960820IS - 0003-9861SB - IMCY - UNITED STATESJC - 6SK
UR  - PM:8660664
SO  - Arch Biochem Biophys 1996 Jun 15 ;330(2):342-354

1382
UI  - 293
AU  - Lill H
AU  - Hensel F
AU  - Junge W
AU  - Engelbrecht S
AD  - Biophysik, Fachbereich Biologie/Chemie, Universitat Osnabruck, Barbarastrasse 11, D-49069 Osnabruck, Federal Republic of Germany engel@ax3301biologieUni-OsnabrueckDE
TI  - Cross-linking of engineered subunit delta to (alphabeta)3 in chloroplast F-ATPase
AB  - Ser --> Cys mutations were introduced into subunit delta of spinach chloroplast F0F1-ATPase (CF0CF1) by site-directed mutagenesis. The engineered delta subunits were overexpressed in Escherichia coli, purified, and reassembled with spinach chloroplast F1-ATPase (CF1) lacking the delta subunit (CF1(-delta)). By modification with eosin-5- maleimide, it was shown that residues 10, 57, 82, 160, and 166 were solvent-accessible in isolated CF1 and all but residue 166 also in membrane-bound CF0CF1. Modification of the engineered delta subunit with photolabile cross-linkers, binding of delta to CF1(-delta), and photolysis yielded the same SDS gel pattern of cross-link products in the presence or absence of ADP, phosphate, and ATP and both in soluble CF1 and in CF0CF1. By chemical hydrolysis of cross-linked CF1, it was shown that deltaS10C was cross-linked within the N-terminal 62 residues of subunit beta. deltaS57C, deltaS82C, and deltaS166C were cross-linked within the N-terminal 192 residues of subunit alpha. Cross-linking affected neither ATP hydrolysis by soluble CF1 nor its ability to reassemble with CF0 and to structurally reconstitute ATP synthesis. Functional reconstitution, however, seemed to be impaired
RP  - NOT IN FILE
NT  - UI - 97115805LA - engRN - 0 (Cross-Linking Reagents)RN - 0 (Macromolecular Systems)RN - 0 (Recombinant Proteins)RN - 0 (Sulfhydryl Reagents)RN - 52-90-4 (Cysteine)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19970123IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:8955107
SO  - J Biol Chem 1996 Dec 20 ;271(51):32737-32742

1383
UI  - 723
AU  - Matsui T
AU  - Jault JM
AU  - Allison WS
AU  - Yoshida M
AD  - Research Laboratory of Resource Utilization, R-1, Tokyo Institute of Technology, Yokohama 226, Japan
TI  - An attempt to convert noncatalytic nucleotide binding site of F1-ATPase to the catalytic site: hydrolysis of tethered ATP by mutated alpha subunits in the enzyme
AB  - The alpha and beta subunits of F1-ATPase are homologous in primary structure and have similar folding topologies. The position of the essential Glu residue in the catalytic sites which reside in the beta subunits is occupied by a Gln residue in the noncatalytic nucleotide binding sites which reside in the alpha subunits. To test if an exchange of catalytic and noncatalytic binding sites is possible, we have replaced the Gln-Lys sequence in the noncatalytic binding site of the alpha subunit with Glu-Arg and, reciprocally, the Glu in the catalytic site of the beta subunit with Gln. The resultant mutant alpha3beta3gamma complex lost steady-state ATPase activity. However, HPLC analysis of tryptic digests of the mutant alpha3beta3gamma complex which had been photolabeled with 2-N3-[8-3H]ATP revealed that ATP tethered to the noncatalytic binding side was hydrolyzed, indicating that a primitive catalytic ability was generated at the alpha subunit by the introduced Glu
RP  - NOT IN FILE
NT  - UI - 96176961LA - engRN - 0 (DNA, Bacterial)RN - 0 (Oligodeoxyribonucleotides)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19960509IS - 0006-291XSB - IMCY - UNITED STATESJC - 9Y8
UR  - PM:8602864
SO  - Biochem Biophys Res Commun 1996 Mar 7 ;220(1):94-97

1384
UI  - 334
AU  - Miroux B
AU  - Walker JE
AD  - The Medical Research Council Laboratory of Molecular Biology, Cambridge, UK
TI  - Over-production of proteins in Escherichia coli: mutant hosts that allow synthesis of some membrane proteins and globular proteins at high levels
AB  - We have investigated the over-production of seven membrane proteins in an Escherichia coli-bacteriophage T7 RNA polymerase expression system. In all seven cases, when expression of the target membrane protein was induced, most of the BL21(DE3) host cells died. Similar effects were also observed with expression vectors for ten globular proteins. Therefore, protein over-production in this expression system is either limited or prevented by bacterial cell death. From the few survivors of BL21(DE3) expressing the oxoglutarate-malate carrier protein from mitochondrial membranes, a mutant host C41(DE3) was selected that grew to high saturation cell density, and produced the protein as inclusion bodies at an elevated level without toxic effect. Some proteins that were expressed poorly in BL21(DE3), and others where the toxicity of the expression plasmids prevented transformation into this host, were also over-produced successfully in C41(DE3). The examples include globular proteins as well as membrane proteins, and therefore, strain C41(DE3) is generally superior to BL21(DE3) as a host for protein over- expression. However, the toxicity of over-expression of some of the membrane proteins persisted partially in strain C41(DE3). Therefore, a double mutant host C43(DE3) was selected from C41(DE3) cells containing the expression plasmid for subunit b of bacterial F-ATPase. In strain C43(DE3), both subunits b and c of the F-ATPase, an alanine-H(+) symporter, and the ADP/ATP and the phosphate carriers from mitochondria were all over-produced. The transcription of the gene for the OGCP and subunit b was lower in C41(DE3) and C43(DE3), respectively, than in BL21(DE3). In C43(DE3), the onset of transcription of the gene for subunit b was delayed after induction, and the over-produced protein was incorporated into the membrane. The procedure used for selection of C41(DE3) and C43(DE3) could be employed to tailor expression hosts in order to overcome other toxic effects associated with over-expression
RP  - NOT IN FILE
NT  - UI - 96300262LA - engRN - 0 (Carrier Proteins)RN - 0 (Genetic Vectors)RN - 0 (Membrane Proteins)RN - 0 (Recombinant Proteins)RN - 130433-50-0 (2-oxoglutarate-malate carrier protein)RN - 367-93-1 (Isopropyl Thiogalactoside)RN - 69-53-4 (Ampicillin)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 19960916IS - 0022-2836SB - IMCY - ENGLANDJC - J6V
UR  - PM:8757792
SO  - J Mol Biol 1996 Jul 19 ;260(3):289-298

1385
UI  - 720
AU  - Muneyuki E
AU  - Hisabori T
AU  - Sasayama T
AU  - Mochizuki K
AU  - Yoshida M
AD  - Research Laboratory of Resources Utilization, Tokyo Institute of Technology, Yokohama emuneyuk@restitechacjp
TI  - The heterogeneous interaction of substoichiometric TNP-ATP and F1- ATPase from Escherichia coli
AB  - The interactions of substoichiometric TNP-ATP and F1-ATPase from Escherichia coli (EF1) were examined and compared with those in the case of mitochondrial F1-ATPase (MF1) and F1-ATPase from thermophilic Bacillus PS3 (TF1). EF1 hydrolyzed substoichiometric TNP-ATP faster than TF1 or MF1, although some 20% of the TNP-ATP remained unhydrolyzed even in the presence of excess chase ATP. The affinity of the catalytic site of EF1 for the product, TNP-ADP, was weaker than that of TF1 or MF1, and the TNP-ADP was readily released upon addition of excess ATP. The amplitude of the difference absorption spectrum induced by binding of TNP-AT(D)P to EF1 was smaller than that of MF1 or TF1 under similar experimental conditions. When an excess amount of TNP-ATP was added to EF1 and the change of the difference spectrum was measured, the shape of the difference spectrum of the ATP-replaceable fraction was very similar to that in the case of binding of TNP-ATP to the isolated beta subunit of TF1, indicating that the rapidly replaceable fraction of bound TNP-ATP was actually at the catalytic site and most of the non- replaceable portion was bound at noncatalytic sites. Weaker affinity of the catalytic site for TNP-ATP may account for the heterogeneous binding and hydrolysis under the conditions described in this paper
RP  - NOT IN FILE
NT  - UI - 97137523LA - engRN - 0 (Fluorescent Dyes)RN - 56-65-5 (Adenosine Triphosphate)RN - 61368-63-6 (2',3'-O-(2,4,6-trinitro-cyclohexadienylidine)adenosine 5'- triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19970320IS - 0021-924XSB - IMCY - JAPANJC - HIF
UR  - PM:8982860
SO  - J Biochem (Tokyo ) 1996 Nov ;120(5):940-945

1386
UI  - 21040
AU  - Murata T
AU  - Yamato I
AU  - Igarashi K
AU  - Kakinuma Y
AD  - Department of Biological Science and Technology, Science University of Tokyo, 2641 Yamazaki, Noda-shi, Chiba 278, Japan
TI  - Intracellular Na+ regulates transcription of the ntp operon encoding a vacuolar-type Na+-translocating ATPase in Enterococcus hirae
AB  - The Gram-positive bacterium Enterococcus hirae has a vacuolar-type Na+- translocating ATPase that is encoded by the ntp operon (ntpFIKECGABDHJ) (Takase, K., Kakinuma, S., Yamato, I., Konishi, K., Igarashi, K., and Kakinuma, Y. (1994) J. Biol. Chem. 269, 11037-11044). Primer extension experiments identified the start site of transcription of this operon upstream of the ntpF gene. In parallel with the increases of both Na+- pumping activity in whole cells and Na+-stimulated ATPase activity in the membranes, the amounts of the two major subunits (A and B) of this enzyme increased remarkably in cells grown on medium containing high concentrations of NaCl but not on medium containing KCl or sorbitol. Chloramphenicol completely abolished the increases of the enzyme activity and the amounts of A and B subunits, suggesting that the Na+- ATPase level increased by de novo synthesis of the enzyme with the stimulation of high concentrations of the external sodium ions. Finally, Western blot and Northern blot experiments revealed that the increase in the Na+-ATPase level with the external Na+ was further accelerated by addition of an ionophore, such as monensin, which rendered the cell membrane permeable to Na+. These results suggest that the transcription of the Na+-ATPase operon is regulated by the intracellular concentration of sodium ions
MH  - A
MH  - Adenosinetriphosphatase
MH  - ATPase
MH  - Bacteria
MH  - Cell Membrane
MH  - Cells
MH  - ion
MH  - Ions
MH  - membrane
MH  - Membranes
MH  - Site
MH  - Sodium
MH  - SUBUNIT
MH  - synthesis
RP  - NOT IN FILE
NT  - UI - 96394481LA - engRN - 0 (RNA, Bacterial)RN - 0 (RNA, Messenger)RN - 7440-23-5 (Sodium)RN - EC 3.6.1.- (sodium-translocating ATPase)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19961118IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:8798587
SO  - J Biol Chem 1996 Sep 27 ;271(39):23661-23666

1387
UI  - 21308
AU  - Nachliel E
AU  - Gutman M
AD  - Department of Biochemistry, Tel Aviv University, Ramat Aviv, Israel
TI  - Quantitative evaluation of the dynamics of proton transfer from photoactivated bacteriorhodopsin to the bulk
AB  - It has been reported by many research groups that protons released during the photocycle of bacteriorhodopsin are detected by surface bound indicators much faster than by indicators in the bulk. In this study we used numerical simulation of chemical reaction's dynamics for analyzing the delayed appearance of protons in the bulk. The results indicate that the low pK surface groups of the membrane, which form an undilutable concentrated matrix of proton binding sites, retain the protons in this space according to the mass action law. The main sites for proton accumulation are the cluster of carboxylates on the cytoplasmic side of the membrane. The protonation of an indicator in the bulk does not proceed by its reaction with free proton, but rather through self-diffusion of the indicator to the membrane and abstraction of proton from the protonated surface group. The detailed mechanisms which correspond with these reactions are reported
MH  - Bacteriorhodopsin
MH  - BINDING
MH  - BINDING SITE
MH  - Binding Sites
MH  - Biochemistry
MH  - indicator
MH  - mechanism
MH  - MECHANISMS
MH  - membrane
MH  - proton
MH  - Proton transfer
MH  - Proton-transfer
MH  - protonation
MH  - Protons
MH  - SIMULATION
MH  - Site
MH  - SURFACE
MH  - TRANSFER
MH  - universities
RP  - NOT IN FILE
NT  - UI - 96409312DA - 19961107IS - 0014-5793LA - engPT - Journal ArticleCY - NETHERLANDSRN - 0 (Protons)RN - 53026-44-1 (Bacteriorhodopsins)SB - IM
UR  - PM:8814294
SO  - FEBS Lett 1996 Sep 16 ;393(2-3):221-225

1388
UI  - 21307
AU  - Nachliel E
AU  - Gutman M
AU  - Kiryati S
AU  - Dencher NA
AD  - Department of Biochemistry, Tel Aviv University, Ramat Aviv, Israel
TI  - Protonation dynamics of the extracellular and cytoplasmic surface of bacteriorhodopsin in the purple membrane
AB  - The dynamics of proton binding to the extracellular and the cytoplasmic surfaces of the purple membrane were measured by laser-induced proton pulses. Purple membranes, selectively labeled by fluorescein at Lys-129 of bacteriorhodopsin, were pulsed by protons released in the aqueous bulk from excited pyranine (8-hydroxy-1,3,6-pyrenetrisulfonate) and the reaction of protons with the indicators was measured. Kinetic analysis of the data imply that the two faces of the membrane differ in their buffer capacities and in their rates of interaction with bulk protons. The extracellular surface of the purple membrane contains one anionic proton binding site per protein molecule with pK = 5.1. This site is within a Coulomb cage radius (approximately 15 A) from Lys-129. The cytoplasmic surface of the purple membrane bears 4-5 protonable moieties (pK = 5.1) that, due to close proximity, function as a common proton binding site. The reaction of the proton with this cluster is at a very fast rate (3.10(10) M-1.s-1). The proximity between the elements is sufficiently high that even in 100 mM NaCl they still function as a cluster. Extraction of the chromophore retinal from the protein has a marked effect on the carboxylates of the cytoplasmic surface, and two to three of them assume positions that almost bar their reaction with bulk protons. The protonation dynamics determined at the surface of the purple membrane is of relevance both for the vectorial proton transport mechanism of bacteriorhodopsin and for energy coupling, not only in halobacteria, but also in complex chemiosmotic systems such as mitochondrial and thylakoid membranes
MH  - A
MH  - analysis
MH  - Bacteriorhodopsin
MH  - BINDING
MH  - BINDING SITE
MH  - Biochemistry
MH  - buffer
MH  - Buffers
MH  - COMPLEX
MH  - coupling
MH  - data
MH  - energy
MH  - fluorescein
MH  - Fluoresceins
MH  - function
MH  - Halobacterium
MH  - indicator
MH  - INTERACTION
MH  - mechanism
MH  - membrane
MH  - Membranes
MH  - protein
MH  - proton
MH  - protonation
MH  - Protons
MH  - purple membrane
MH  - retinal
MH  - Site
MH  - SURFACE
MH  - SYSTEM
MH  - SYSTEMS
MH  - thylakoid
MH  - thylakoid membrane
MH  - transport
MH  - united states
MH  - universities
RP  - NOT IN FILE
NT  - UI - 97008075DA - 19961125IS - 0027-8424LA - engPT - Journal ArticleCY - UNITED STATESRN - 0 (Buffers)RN - 0 (Fluoresceins)RN - 0 (Protons)RN - 2321-07-5 (Fluorescein)RN - 53026-44-1 (Bacteriorhodopsins)RN - 56-87-1 (Lysine)SB - IM
UR  - PM:8855251
SO  - Proc Natl Acad Sci U S A 1996 Oct 1 ;93(20):10747-10752

1389
UI  - 721
AU  - Noji H
AU  - Amano T
AU  - Yoshida M
AD  - Research laboratory of Resources Utilization, Tokyo Institute of Technology, Yokohama, Japan
TI  - Molecular switch of F0F1-ATP synthase, G-protein, and other ATP-driven enzymes
AB  - Exchange-out of amide tritium from labeled gamma-subunit of alpha 3 beta 3 gamma complex of F0F1-ATP synthase was not accelerated by ATP, suggesting that hemagglutinin-type transition of coiled-coil structure did not occur in gamma-subunit. Local topology of nucleotide binding site and "switch II" region of G-protein alpha resemble those of F1- beta subunit and other proteins which catalyze ATP-triggered reactions. Probably, binding of nucleotide to F0F1-ATP synthase induces conformational change of the switch II-like region with transforming beta subunit structure from "open" to "close" for and this transformation results in loss of hydrogen bonds with gamma subunit, thus enabling the gamma subunit to move
RP  - NOT IN FILE
NT  - UI - 97108772LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.- (GTP-Binding Proteins)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 19970226IS - 0145-479XSB - IMCY - UNITED STATESJC - HIO
UR  - PM:8951093
SO  - J Bioenerg Biomembr 1996 Oct ;28(5):451-457

1390
UI  - 336
AU  - Orriss GL
AU  - Runswick MJ
AU  - Collinson IR
AU  - Miroux B
AU  - Fearnley IM
AU  - Skehel JM
AU  - Walker JE
AD  - The Medical Research Council Laboratory of Molecular Biology, Cambridge, UK
TI  - The delta- and epsilon-subunits of bovine F1-ATPase interact to form a heterodimeric subcomplex
AB  - The delta-subunit of bovine F1-ATPase was expressed from a bacterial vector at fairly high level in Escherichia coli, but the yield of bovine epsilon-subunit was rather low under similar conditions. However, co-expression of the proteins from a dicistronic operon delta- epsilon in the same expression vector, produced both of them in good yield in a soluble form in the bacterial cytoplasm, and by chromatography it was found that the delta- and epsilon-subunits were associated in a stable complex. The amino groups in the complex were labelled exhaustively by chemical reaction under denaturing conditions with ethyl-[1-14C]acetimidate. The alpha-amino groups of the proteins were unmodified, but complete reaction of all epsilon-amino groups in both proteins was demonstrated by determination of the molecular masses of the modified proteins by electrospray MS. The modified subunits were separated by denaturing gel electrophoresis, and from measurements of the ratio of incorporated radioactivities and the lysine contents of the proteins, it was calculated that the subcomplex contains equimolar amounts of the two proteins. As the apparent molecular mass of the complex determined by gel filtration was 29 kDa, it appears that the complex contains one copy of each protein. It is likely that the delta- and epsilon subunits are associated in a similar manner in the bovine F1-ATPase complex, and that, like a bacterial homologue of the delta- subunit, they interact with the gamma- and beta-subunits
RP  - NOT IN FILE
NT  - UI - 96239132LA - engRN - 0 (Biopolymers)RN - 0 (DNA Primers)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19960805IS - 0264-6021SB - IMCY - ENGLANDJC - 9YO
UR  - PM:8670087
SO  - Biochem J 1996 Mar 1 ;314 ( Pt 2)():695-700

1391
UI  - 800
AU  - Panke O
AU  - Rumberg B
AD  - Max-Volmer-Institut fur Biophysikalische und Physikalische Chemie, Technische Universitat, Berlin, Germany
TI  - Kinetic modelling of the proton translocating CF0CF1-ATP synthase from spinach
AB  - The rate of both ATP synthase and hydrolysis catalysed by the thiol- modulated and activated ATP synthase from spinach is measured as a function of all substrates including the protons inside the thylakoid lumen. The most important findings are: (1) sigmoid kinetics with respect to H+in, (2) hyperbolic kinetics with respect to ADP, ATP and phosphate, with Km for phosphate and ADP decreasing upon increasing H+in, (3) binding of ADP and phosphate in random order and competitive to ATP. Simulation of the complete set of experimental data is obtained by a kinetic model featuring Boyer's binding-chain mechanism
RP  - NOT IN FILE
NT  - UI - 96198601LA - engRN - 0 (Phosphates)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19961114IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:8925895
SO  - FEBS Lett 1996 Apr 1 ;383(3):196-200

1392
UI  - 21281
AU  - Paunov VN
AU  - Dimova RI
AU  - Kralchevsky PA
AU  - Broze G
AU  - Mehreteab A
TI  - The hydration repulsion between charged surfaces as an interplay of volume exclusion and dielectric saturation effects
MH  - SURFACE
RP  - IN FILE
SO  - J Colloid Interface Sci 1996  ;182():239-248

1393
UI  - 506
AU  - Pedersen PL
AD  - Department of Biological Chemistry, Johns Hopkins University, School of Medicine, Baltimore, Maryland 21205-2185, USA
TI  - Frontiers in ATP synthase research: understanding the relationship between subunit movements and ATP synthesis
AB  - How biological systems make ATP has intrigued many scientists for well over half the 20th century, and because of the importance and complexity of the problem it seems likely to continue to be a source of fascination to both senior and younger investigators well into the 21st century. Scientific battles fought to unravel the vast secrets by which ATP synthases work have been fierce, and great victories have been short-lived, tempered with the realization that more structures are needed, additional subunits remain to be conquered, and that during ATP synthesis, not one, but several subunits may undergo either significant conformational changes, repositioning, or perhaps even physical "rotation" similar to bacterial flagella (1,2). In this introductory article, the author briefly summarizes our current knowledge about the complex substructure of ATP synthases, what we have learned from X-ray crystallography of the F1 unit, and current evidence for subunit movements
RP  - NOT IN FILE
NT  - UI - 97108763LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialID - CA 10951/CA/NCIDA - 19970226IS - 0145-479XSB - IMCY - UNITED STATESJC - HIO
UR  - PM:8951084
SO  - J Bioenerg Biomembr 1996 Oct ;28(5):389-395

1394
UI  - 990
AU  - Pitard B
AU  - Richard P
AU  - Dunach M
AU  - Girault G
AU  - Rigaud JL
TI  - ATP synthesis by the F0F1 ATP synthase from thermophilic Bacillus PS3 reconstituted into liposomes with bacteriorhodopsin. 1. Factors defining the optimal reconstitution of ATP synthases with bacteriorhodopsin
AB  - Optimal conditions for the reconstitution of bacteriorhodopsin and H+-transporting ATP synthase from thermophilic Bacillus PS3 (TF0F1) were determined. Phosphatidylcholine/phosphatidic acid liposomes prepared by reverse-phase evaporation were treated with various amounts of Triton X-100, octyl glucoside, octaethylene glycol n-dodecylether, sodium cholate or sodium deoxycholate and the incorporation of proteins by these detergents was studied at each step of the solubilization process. After removal of detergent by means of SM-2 Bio-Beads, the light-driven ATP synthase activities of the resulting proteoliposomes were analyzed at 40 degrees C. The nature of the detergent used for reconstitution was important for determining the mechanism of protein insertions. The most efficient reconstitutions were obtained with octyl glucoside or Triton X-100 by insertion of the proteins into detergent-saturated liposomes. The conditions for reconstitutions were further optimized with regard to functional coupling between bacteriorhodopsin and TF0F1. It was demonstrated that one of the main factors limiting the production of efficient reconstituted proteoliposomes was related to activation of the highly stable TFO-F1. Activation was accomplished by total solubilization of phospholipids and proteins in a Triton X-100/octyl glucoside mixture containing 20 mM octyl glucoside, leading to a threefold stimulation of the ATP synthase activity. Final ATP synthase activities depended greatly on the lipid/bacteriorhodopsin and the lipid/TF0F1 ratios as well as on the phospholipid used. In particular, light-driven ATP synthesis depended upon the presence of negatively charged phospholipids. Cholesterol was found to induce a fourfold increase in ATP synthase activity with a concomitant 65% decrease in the Km for ADP, suggesting that sterols can modulate catalytic events mediated by F1. Preparations obtained by this step-by-step reconstitution procedure displayed activities up to 20-fold higher (500-800 nmol ATP x min(-1) x mg TF0F1(-1) in the presence of cholesterol) than the maximal values reported in the literature for light-driven ATP synthesis TF0F1 measured under similar conditions. This study also allowed rationalization of the different parameters involved in reconstitution experiments and the present simple method is shown to be of general use for preparation of efficient proteoliposomes containing bacteriorhodopsin and choloroplast or mitochondrial F0F1-type ATP synthases
MH  - Adenosine Triphosphate
MH  - Animal
MH  - Bacillus
MH  - Bacteriorhodopsin
MH  - biosynthesis
MH  - Chloroplasts
MH  - Detergents
MH  - Enzyme Activation
MH  - enzymology
MH  - H(+)-Transporting ATP Synthase
MH  - Liposomes
MH  - Magnesium
MH  - metabolism
MH  - Mitochondria,Heart
MH  - Phospholipids
MH  - Polyethylene Glycols
MH  - Proteins
MH  - Proteolipids
MH  - Sodium
MH  - Support,Non-U.S.Gov't
MH  - Swine
MH  - Temperature
RP  - NOT IN FILE
NT  - Section de Bioenergetique, DBCM, CEA-Saclay, Gif sur Yvette, France
SO  - Eur J Biochem 1996 Feb 1 ;235(3):769-778

1395
UI  - 989
AU  - Pitard B
AU  - Richard P
AU  - Dunach M
AU  - Rigaud JL
TI  - ATP synthesis by the F0F1 ATP synthase from thermophilic Bacillus PS3 reconstituted into liposomes with bacteriorhodopsin. 2. Relationships between proton motive force and ATP synthesis
AB  - The correlation between the rate of ATP synthesis and light-induced proton flux was investigated in proteoliposomes reconstituted with bacteriorhodopsin and ATP synthase from thermophilic Bacillus PS3. By variation of the actinic light intensity it was found that ATP synthase activity depended in a sigmoidal manner on the amplitude of the transmembrane light-induced pH gradient. Maximal rates of ATP synthesis (up to to 200 nmol ATP x min(-1) x mg protein (-1) were obtained at saturating light intensities under a steady-state pH gradient of about pH 1.25. It was demonstrated that this was the maximal deltapH attainable at 40 degrees C in reconstituted proteoliposomes, due to the feedback inhibition of bacteriorhodopsin by the proton gradient it generates. In the absence of valinomycin, a small but significant transmembrane electrical potential could develop at 40 degrees C, contributing to an increase in the rate of ATP synthesis. The H+/ATP stoichiometry was measured at the static-head (equilibrium) conditions from the ratio of the phosphate potential to the size of the light-induced pH gradient and a value of about four was obtained under the maximal electrochemical proton gradient. Increasing the amount of bacteriorhodopsin in the proteoliposomes at a constant F0F1 concentration led to a large increase in the rate of ATP synthesis whereas the magnitude of delta pH remained the same or, at very high bacteriorhodopsin levels, decreased. Consequently the H+/ATP stoichiometry was found to increase significantly with increasing bacteriorhodopsin content. Reconstitutions with mixtures of native and impaired bacteriorhodopsin (Asp96-->Asn mutated bacteriorhodopsin) further demonstrated that this increase in the coupling efficiency could not be related to protein-protein interactions but rather to bacteriorhodopsin donating H+ to the ATP synthase. Increasing the amount of negatively charged phospholipids in the proteoliposomes also increased the coupling efficiency between bacteriorhodopsin and ATP synthase at a constant transmembrane pH gradient. Similar results were obtained with chloroplast ATP synthase. Furthermore, ATP synthase activities induced by delta pH/delta psi transitions were independent of bacteriorhodopsin or anionic lipid levels. These observations were interpreted as indicating that, in bacteriorhodopsin/ATP synthase, proteoliposomes, a localized pathway for coupling light-driven H+ transport by bacteriorhodopsin to ATP synthesis by F0F1 might exist under specific experimental conditions
MH  - Adenosine Triphosphate
MH  - Bacillus
MH  - Bacteriorhodopsin
MH  - biosynthesis
MH  - Chemistry
MH  - Electrochemistry
MH  - enzymology
MH  - H(+)-Transporting ATP Synthase
MH  - Hydrogen-Ion Concentration
MH  - Light
MH  - Liposomes
MH  - metabolism
MH  - Phospholipids
MH  - Proteolipids
MH  - Proton-Motive Force
MH  - Protons
MH  - Support,Non-U.S.Gov't
MH  - Thermodynamics
MH  - Valinomycin
RP  - NOT IN FILE
NT  - Section de Bioenergetique, DBCM, CEA-Saclay, Gif sur Yvette, France
SO  - Eur J Biochem 1996 Feb 1 ;235(3):779-788

1396
UI  - 21185
AU  - Radionov AN
AU  - Kalaidzidis IV
AU  - Kaulen AD
AD  - Department of Photobiochemistry, AN Belozersky Institute of Physico- Chemical Biology, Moscow State University, Russian Federation
TI  - Complicated character of the M decay pH dependence in the D96N mutant is due to the two pathways of the M conversion
AB  - At high ionic strength, the pH dependence of the M intermediate decay in a photocycle of the D96N mutant bacteriorhodopsin shows a complicated behavior which is found to be due to the coexistence of two pathways of the M conversion. The M decay which dominates at pH < 5 is coupled to the proton uptake from the cytoplasmic surface and proceeds probably through the N intermediate. This pathway is inhibited by glutaraldehyde, the potent inhibitor of M decay in the wild-type bacteriorhodopsin and of the azide-facilitated M decay in the D96N mutant. Another pathway of the M decay is predominant at pH > 5. This pathway is insensitive to glutaraldehyde and some other similar inhibitors (lutetium ions, sucrose and glycerol). On the other hand, it is sensitive to the pK changes of the group X (Glu-204) in the outward proton pathway. Possibly, the M decay through this pathway represents a reverse H+ transport process (the proton uptake from the external surface) and proceeds via the L intermediate
MH  - A
MH  - Bacteriorhodopsin
MH  - DEPENDENCE
MH  - H+
MH  - inhibitor
MH  - intermediate
MH  - ion
MH  - Ions
MH  - M
MH  - M-intermediate
MH  - mutant
MH  - pH
MH  - proton
MH  - Sucrose
MH  - SURFACE
MH  - transport
RP  - NOT IN FILE
NT  - UI - 97138130LA - engRN - 53026-44-1 (Bacteriorhodopsins)PT - Journal ArticleDA - 19970130IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:8985156
SO  - FEBS Lett 1996 Dec 16 ;399(3):251-254

1397
UI  - 21186
AU  - Radionov AN
AU  - Kaulen AD
AD  - Department of Photobiochemistry, AN Belozersky Institute of Physico- Chemical Biology, Moscow State University, Moscow, Russian Federation
TI  - Two bacteriorhodopsin M intermediates differing in accesibility of the Schiff base for azide
AB  - Glutaraldehyde treatment leads to the inhibition (i) of the M intermediate decay in wild-type bacteriorhodopsin (bR) and (ii) of the azide-facilitated M decay in the D96N mutant bR. LuCl3 is shown to be a more potent inhibitor of both processes. Glycerol and sucrose are also inhibitors. None of these agents change the linearity of the azide concentration dependency of the M decay in the D96N mutant but they do shift this dependency to higher azide concentrations. It is concluded that the two M forms are in equilibrium. These M forms differ in the accessibility of the Schiff base for azide and, probably, also for water molecules. The above-mentioned agents shift the equilibrium toward the less accessible M form. The data obtained are in line with the model of azide action as the penetrating proton donor and can hardly be realized within the framework of the model of Le Coutre et al. [(1995) Proc. Natl. Acad. Sci. USA 92, 4962-4966] which assumes that a bound anionic form of azide catalyzes proton transfer to the Schiff base
MH  - A
MH  - Azides
MH  - Bacteriorhodopsin
MH  - BASE
MH  - inhibitor
MH  - intermediate
MH  - M
MH  - M-intermediate
MH  - model
MH  - mutant
MH  - proton
MH  - Proton transfer
MH  - Proton-transfer
MH  - Schiff base
MH  - Schiff Bases
MH  - Schiff-base
MH  - Sucrose
MH  - TRANSFER
MH  - Water
RP  - NOT IN FILE
NT  - UI - 96244507LA - engRN - 0 (Azides)RN - 0 (Schiff Bases)RN - 111-30-8 (Glutaral)RN - 53026-44-1 (Bacteriorhodopsins)RN - 56-81-5 (Glycerol)RN - 57-50-1 (Sucrose)RN - 7439-94-3 (Lutetium)PT - Journal ArticleDA - 19960815IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:8674532
SO  - FEBS Lett 1996 Jun 3 ;387(2-3):122-126

1398
UI  - 873
AU  - Repke KR
TI  - Reinstatement of the ATP high energy paradigm
AB  - The paradigm that the hydrolysis of ATP releases high Gibbs energy able to perform work has increasingly been questioned over the last two decades. Results from theoretical and experimental studies have been interpreted to indicate that the synthesis of ATP from ADP and P(i) does not require energy supply and that binding of ATP per se can transmit utilizable energy to an enzyme. As has recently been concluded, all this has led to a change of the ATP high energy paradigm in bioenergetics. Starting from this challenge, the present review singles out the striking sources of the apparent dichotomy in bioenergetics, and endeavours to eliminate the apparent contradictions by the application of the prior knowledge on both the participation of the enzyme protein in energy exchange processes and the particular reactivities of phosphorus that make it an outstanding element for functionally variable work assignments in enzymatic systems
MH  - Adenosine Triphosphate
MH  - Animal
MH  - biosynthesis
MH  - Energy Metabolism
MH  - Hydrolysis
MH  - Models,Chemical
MH  - physiology
RP  - NOT IN FILE
NT  - Max Delbruck Centre for Molecular Medicine, Berlin-Buch, Germany
SO  - Mol Cell Biochem 1996 Jul ;160-161:95-9.():95-99

1399
UI  - 505
AU  - Reynafarje BD
AU  - Pedersen PL
AD  - Department of Biological Chemistry, The Johns Hopkins University, School of Medicine, Baltimore, Maryland 21205-2185, USA
TI  - ATP synthase. Conditions under which all catalytic sites of the F1 moiety are kinetically equivalent in hydrolyzing ATP
AB  - Conditions have been reported under which the F1 moiety of bovine heart ATP synthase catalyzes the hydrolysis of ATP by an apparently cooperative mechanism in which the slow rate of hydrolysis at a single catalytic site (unisite catalysis) is enhanced more than 10(6)-fold when ATP is added in excess to occupy one or both of the other two catalytic sites (multisite catalysis) (Cross, R. L., Grubmeyer, C., and Penefsky, H. S. (1982) J. Biol. Chem. 257, 12101-12105). In the novel studies reported here, and in contrast to the earlier report, we have (a) monitored the kinetics of ATP hydrolysis of F1 by using nucleotide- depleted preparations and a highly sensitive chemiluminescent assay; (b) followed the reaction immediately upon addition of F1 to ATP, rather than after prior incubation with ATP; and (c) used a reaction medium with Pi as the only buffer. The following observations were noted. First, regardless of the source of enzyme, bovine or rat, and catalytic conditions (unisite or multisite), the rates of hydrolysis depend on ATP concentration to the first power. Second, the first order rate constant for ATP hydrolysis remains relatively constant under both unisite and multisite conditions declining only slightly at high ATP concentration. Third, the initial rates of ATP hydrolysis exhibit Michaelis-Menten kinetic behavior with a single Vmax exceeding 100 micromol of ATP hydrolyzed per min/mg of F1 (turnover number = 635 s-1) and a single Km for ATP of about 57 microM. Finally, the reaction is inhibited markedly by low concentrations of ADP. It is concluded that, under the conditions described here, all catalytic sites that participate in the hydrolysis of ATP within the F1 moiety of mitochondrial ATP synthase function in a kinetically equivalent manner
RP  - NOT IN FILE
NT  - UI - 97115777LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - CA 10951/CA/NCIDA - 19970123IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:8955079
SO  - J Biol Chem 1996 Dec 20 ;271(51):32546-32550

1400
UI  - 294
AU  - Sabbert D
AU  - Engelbrecht S
AU  - Junge W
AD  - Abteilung Biophysik, Fachbereich Biologie/Chemie, Universitat Osnabruck, Germany
TI  - Intersubunit rotation in active F-ATPase
AB  - The enzyme ATP synthase, or F-ATPase, is present in the membranes of bacteria, chloroplasts and mitochondria. Its structure is bipartite, with a proton-conducting, integral membrane portion, F0, and a peripheral portion, F1. Solubilized F1 is composed of five different subunits, (alpha beta)3 gamma delta epsilon, and is active as an ATPase. The function of F-ATPase is to couple proton translocation through F0 with ATP synthesis in F1 (ref.3). Several lines of evidence support the spontaneous formation of ATP on F1 (refs 4,5) and its endergonic release at cooperative and rotating (or at least alternating) sites. The release of ATP at the expense of protonmotive force might involve mechanical energy transduction from F0 into F1 by rotation of the smaller subunits (mainly gamma) within (alpha beta)3, the catalytic hexagon of F1 as suggested by electron microscopy, by X- ray crystal structure analysis and by the use of cleavable crosslinkers. Here we record an intersubunit rotation in real time in the functional enzyme by applying polarized absorption relaxation after photobleaching to immobilized F1 with eosin-labelled gamma. We observe the rotation of gamma relative to immobilized (alpha beta)3 in a timespan of 100 ms, compatible with the rate of ATP hydrolysis by immobilized F1. Its angular range, which is of at least 200 degrees, favours a triple-site mechanism of catalysis, with gamma acting as a crankshaft in (alpha beta)3. The rotation of gamma is blocked when ATP is substituted with its non-hydrolysable analogue AMP-PNP
RP  - NOT IN FILE
NT  - UI - 96238867LA - engRN - 0 (Enzymes, Immobilized)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19960709IS - 0028-0836SB - IMCY - ENGLANDJC - NSC
UR  - PM:8637601
SO  - Nature 1996 Jun 13 ;381(6583):623-625

1401
UI  - 19893
AU  - Samec Z
TI  - Kinetics of Charge Transfer
MH  - Kinetics
MH  - TRANSFER
MH  - interfaces
MH  - theory
MH  - Methods
MH  - method
T2  - Liqid-Liquid Interfaces. Theory and Methods
A2  - Volkov AG
A2  - Deamer DW
PB  - New York: CRC Press
RP  - IN FILE
SO  -  1996  ;(8):155-178

1402
UI  - 21157
AU  - Schemidt RA
AU  - Brauning CK
AU  - Bouvier A
AU  - Brusilow WS
AD  - Department of Biochemistry and Molecular Biology, Wayne State University School of Medicine, Detroit, Michigan 48201, USA wbrusil@cmsccwayneedu
TI  - Localization of a conformational energy-coupling determinant near the C terminus of the beta subunit of the F1F0-ATPase
AB  - Escherichia coli mutants in the beta subunit of the F1F0-ATPase can be complemented with the beta subunit from the obligate aerobe Bacillus megaterium. It has been shown that cells carrying such hybrid ATPases have an unusual energy-coupling phenotype. Although they are able to grow on minimal succinate medium, and therefore carry a functional ATP synthase, they are defective in the ability to grow anaerobically, indicating some defect in ATP-driven proton pumping (Scarpetta, M., Hawthorne, C. A., and Brusilow, W. S. A. (1991) J. Biol. Chem. 266, 18567-18572). In this study, chimeric beta subunits were constructed consisting of the E. coli or the B. megaterium beta subunit carrying the C-terminal 18% of the other's beta subunit. The phenotypes of an E. coli beta mutant complemented with these chimeric subunits showed that the energy-coupling defect was located in this C-terminal region. The E. coli beta subunit carrying the B. megaterium C-terminal region displayed the energy-coupling defect, while the B. megaterium beta subunit carrying the E. coli C-terminal region did not. In ATP- dependent fluorescence quenching assays, membranes isolated from cells displaying the energy-coupling defect also pumped protons less well than membranes isolated from cells that were able to grow anaerobically. These results demonstrate that the C terminus of the beta subunit is involved in the conformational coupling pathway, which, through the polypeptide backbone of the beta subunit, physically links ATP synthesis or hydrolysis to the energy of proton translocation
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - ATPase
MH  - Bacillus
MH  - BETA
MH  - BETA-SUBUNIT
MH  - Biochemistry
MH  - Cells
MH  - coupling
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - fluorescence
MH  - Hydrolysis
MH  - M
MH  - membrane
MH  - Membranes
MH  - mutant
MH  - proton
MH  - Proton-Translocating ATPases
MH  - Protons
MH  - SUBUNIT
MH  - succinate
MH  - SYNTHASE
MH  - synthesis
MH  - translocation
RP  - NOT IN FILE
NT  - UI - 97125978LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19970128IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:8969200
SO  - J Biol Chem 1996 Dec 27 ;271(52):33390-33393

1403
UI  - 155
AU  - Singh S
AU  - Turina P
AU  - Bustamante CJ
AU  - Keller DJ
AU  - Capaldi R
AD  - Department of Chemistry, University of New Mexico, Albuquerque 87131, USA
TI  - Topographical structure of membrane-bound Escherichia coli F1F0 ATP synthase in aqueous buffer
AB  - Scanning force microscope images of membrane-bound Escherichia coli ATP synthase F0 complexes have been obtained in aqueous solution. The images show a consistent set of internal features: a ring structure which surrounds a central dimple and contains an asymmetric lateral mass. Images of trypsin-treated F0 complexes, which have lost part of their b subunits, show a reduced asymmetric mass, while images of c- subunit oligomers, which lack both the a and b subunits, show a ring and dimple but do not have an asymmetric mass. These results support models in which the F0 complex contains a ring of 9-12 c subunits with the b subunits located outside this ring, and show that scanning force microscopy is able to provide structural information on membrane proteins of molecular mass less than 200 000 Da
RP  - NOT IN FILE
NT  - UI - 97096887LA - engRN - 0 (Phosphatidylethanolamines)RN - EC 3.4.21.4 (Trypsin)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM32543/GM/NIGMSID - HL 24526/HL/NHLBIDA - 19970114IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:8941708
SO  - FEBS Lett 1996 Nov 11 ;397(1):30-34

1404
UI  - 961
AU  - Stroud RM
TI  - Balancing ATP in the cell
MH  - Adenosine Triphosphate
MH  - Animal
MH  - Cells
MH  - Chemistry
MH  - Coenzymes
MH  - Creatine Kinase
MH  - Cytoplasm
MH  - Energy Metabolism
MH  - enzymology
MH  - metabolism
MH  - Mitochondria
MH  - Models,Structural
MH  - Oligopeptides
RP  - NOT IN FILE
SO  - Nat Struct Biol 1996 Jul ;3(7):567-569

1405
UI  - 20845
AU  - Takeyasu K
AU  - Omote H
AU  - Nettikadan S
AU  - Tokumasu F
AU  - Iwamoto-Kihara A
AU  - Futai M
AD  - Department of Medical Biochemistry, Ohio State University, Columbus, USA
TI  - Molecular imaging of Escherichia coli F0F1-ATPase in reconstituted membranes using atomic force microscopy
AB  - The structure of Escherichia coli F0F1-ATPase (ATP synthase), and its F0 sector reconstituted in lipid membranes was analyzed using atomic force microscopy (AFM) by tapping-mode operation. The majority of F0F1- ATPases were visualized as spheres with a calculated diameter of approximately 90 angstroms, and a height of approximately 100 angstroms from the membrane surface. F0 sectors were visualized as two different ring-like structures (one with a central mass and the other with a central hollow of greater than or equal to 18 angstroms depth) with a calculated outer diameter of approximately 130 angstroms. The two different images possibly represent the opposite orientations of the complex in the membranes. The ring-like projections of both images suggest inherently asymmetric assemblies of the subunits in the F0 sector. Considering the stoichiometry of F0 subunits, the area of the image observed is large enough to accommodate all three F0 subunits in an asymmetric manner
MH  - A
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Biochemistry
MH  - COMPLEX
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - F0F1
MH  - membrane
MH  - Membrane Proteins
MH  - Membranes
MH  - Microscopy
MH  - protein
MH  - Proteins
MH  - SUBUNIT
MH  - SURFACE
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 96368032LA - engRN - 0 (Membrane Proteins)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleID - GM44373/GM/NIGMSDA - 19961021IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:8772185
SO  - FEBS Lett 1996 Aug 26 ;392(2):110-113

1406
UI  - 160
AU  - Tang C
AU  - Capaldi RA
AD  - Institute of Molecular Biology, University of Oregon, Eugene, Oregon 97403-1229, USA
TI  - Characterization of the interface between gamma and epsilon subunits of Escherichia coli F1-ATPase
AB  - The interaction faces of the gamma and epsilon subunits in the Escherichia coli F1-ATPase have been explored by a combination of cross- linking and chemical modification experiments using several mutant epsilon subunits as follows: epsilonS10C, epsilonH38C, epsilonT43C, epsilonS65C, epsilonS108C, and epsilonM138C, along with a mutant of the gamma subunit, gammaT106C. The replacement of Ser-10 by a Cys or Met- 138 by a Cys reduced the inhibition of ECF1 by the epsilon subunit, while the mutation S65C increased this inhibitory effect. Modification of the Cys at position 10 with N-ethylmaleimide or fluoroscein maleimide further reduced the binding affinity of, and the maximal inhibition by, the epsilon subunit. Similar chemical modification of the Cys at position 43 of the epsilon subunit (in the mutant epsilonT43C) and a Cys at position 106 of the gamma subunit (gammaT106C) also affected the inhibition of ECF1 by the epsilon subunit. The various epsilon subunit mutants were reacted with TFPAM3, and the site(s) of cross-linking within the ECF1 complex was determined. Previous studies have shown cross-linking from the Cys at positions 10 and 38 with the gamma subunit and from a Cys at position 108 to an alpha subunit (Aggeler, R., Chicas-Cruz, K., Cai, S. X., Keana, J. F. W., and Capaldi, R. A. (1992) Biochemistry 31, 2956-2961; Aggeler, R., Weinreich, F., and Capaldi, R. A. (1995) Biochim. Biophys. Acta 1230, 62-68). Here, cross-linking was found from a Cys at position 43 to the gamma subunit and from the Cys at position 138 to a beta subunit. The site of cross-linking from Cys-10 of epsilon to the gamma subunit was localized by peptide mapping to a region of the gamma subunit between residues 222 and 242. Cross-linking from a Cys at position 38 and at position 43 was with the C-terminal part of the gamma subunit, between residues 202 and 286. ECF1 treated with trypsin at pH 7.0 still binds purified epsilon subunit, while enzyme treated with the protease at pH 8.0 does not. This identifies sites around residue 70 and/or between 202 and 212 of the gamma subunit as involved in epsilon subunit binding
RP  - NOT IN FILE
NT  - UI - 96216373LA - engRN - 0 (Cross-Linking Reagents)RN - 0 (Macromolecular Systems)RN - 0 (Recombinant Proteins)RN - 128-53-0 (Ethylmaleimide)RN - 52-90-4 (Cysteine)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - HL24526/HL/NHLBIDA - 19960619IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:8621695
SO  - J Biol Chem 1996 Feb 9 ;271(6):3018-3024

1407
UI  - 719
AU  - Tozawa K
AU  - Ohbuchi H
AU  - Yagi H
AU  - Amano T
AU  - Matsui T
AU  - Yoshida M
AU  - Akutsu H
AD  - Department of Bioengineering, Faculty of Engineering, Yokohama National University, Japan
TI  - Unusual pKa of the carboxylate at the putative catalytic position of the thermophilic F1-ATPase beta subunit determined by 13C-NMR
AB  - Glutamic acid-190 in the beta subunit of F1-ATPase from thermophilic Bacillus PS-3 (TF1) was reported to be essential for the ATPase activity. The mutant TF1beta subunit in which Glu-190 had been substituted by cysteine was carboxymethylated with 13C-labeled monoiodoacetic acid. The pKa value of the carboxymethylene group at the 190 position was determined as 5.6 +/- 0.4 by 13C-NMR. On the basis of this value, the pKa of the carboxylate of Glu-190 of the TF1beta subunit was estimated to be 6.8 +/- 0.5. The unusually high pKa could play a role in the catalytic mechanism of F1-ATPase
RP  - NOT IN FILE
NT  - UI - 97096906LA - engRN - 0 (Carboxylic Acids)RN - 0 (Iodoacetates)RN - 2387-59-9 (Carbocysteine)RN - 56-86-0 (Glutamic Acid)RN - 58-64-0 (Adenosine Diphosphate)RN - 64-69-7 (Iodoacetic Acid)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19970114IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:8941727
SO  - FEBS Lett 1996 Nov 11 ;397(1):122-126

1408
UI  - 329
AU  - van Raaij MJ
AU  - Orriss GL
AU  - Montgomery MG
AU  - Runswick MJ
AU  - Fearnley IM
AU  - Skehel JM
AU  - Walker JE
AD  - MRC Laboratory of Molecular Biology, Cambridge, UK
TI  - The ATPase inhibitor protein from bovine heart mitochondria: the minimal inhibitory sequence
AB  - The mitochondrial ATPase inhibitor subunit is a basic protein of 84 amino acids that helps to regulate the activity of F1F0-ATPase. In order to obtain structural information on the mechanism of inhibition, the bovine inhibitor subunit has been expressed in Escherichia coli and purified in high yield. The recombinant protein has a similar inhibitory activity to the inhibitor subunit isolated from bovine mitochondria. Progressive N-terminal and C-terminal deletion mutants of the inhibitor subunit have been produced either by overexpression and purification, or by chemical synthesis. By assaying the truncated proteins for inhibitory activity, the minimal inhibitory sequence of the inhibitor subunit has been defined as consisting of residues 14-47. The immediately adjacent sequences 10-13 and 48-56 help to stabilize the complex between F1F0-ATPase and the inhibitor protein, and residues 1-9 and 57-84 appear to be dispensable. At physiological pH values, the inhibitor subunit is mainly alpha-helical and forms monodisperse aggregates in solution. Smaller inhibitory fragments of the inhibitor protein, such as residues 10-50, seem to have a mainly random coil structure in solution, but they can adopt the correct inhibitory conformation when they from a complex with the ATPase. However, these latter fragments are mainly monomeric in solution, suggesting that the aggregation of the inhibitor subunit in solution may be due to intermolecular alpha-helical coiled-coil formation via the C-terminal region. The noninhibitory peptides consisting of residues 10-40 and 23- 84 of the inhibitor protein can bind to F1F0-ATPase, and interfere with inhibition by the intact inhibitor subunit. The noninhibitory fragments of the inhibitor protein consisting of residues 22-46 and 44-84 do not compete with the inhibitor subunit for its binding site on F1F0-ATPase
RP  - NOT IN FILE
NT  - UI - 97121257LA - engRN - 0 (ATPase inhibitory protein)RN - 0 (Deoxyribonucleotides)RN - 0 (Enzyme Inhibitors)RN - 0 (Peptide Fragments)RN - 0 (Proteins)RN - 0 (Recombinant Proteins)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19970116IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:8961923
SO  - Biochemistry 1996 Dec 10 ;35(49):15618-15625

1409
UI  - 335
AU  - van Raaij MJ
AU  - Abrahams JP
AU  - Leslie AG
AU  - Walker JE
AD  - Medical Research Council Laboratory of Molecular Biology, Cambridge, United Kingdom
TI  - The structure of bovine F1-ATPase complexed with the antibiotic inhibitor aurovertin B
AB  - In the structure of bovine mitochondrial F1-ATPase that was previously determined with crystals grown in the presence of adenylyl- imidodiphosphate (AMP-PNP) and ADP, the three catalytic beta-subunits have different conformations and nucleotide occupancies. Adenylyl- imidodiphosphate is bound to one beta-subunit (betaTP), ADP is bound to the second (betaDP), and no nucleotide is bound to the third (betaE). Here we show that the uncompetitive inhibitor aurovertin B binds to bovine F1 at two equivalent sites in betaTP and betaE, in a cleft between the nucleotide binding and C-terminal domains. In betaDP, the aurovertin B pocket is incomplete and is inaccessible to the inhibitor. The aurovertin B bound to betaTP interacts with alpha-Glu399 in the adjacent alphaTP subunit, whereas the aurovertin B bound to betaE is too distant from alphaE to make an equivalent interaction. Both sites encompass betaArg-412, which was shown by mutational studies to be involved in binding aurovertin. Except for minor changes around the aurovertin pockets, the structure of bovine F1-ATPase is the same as determined previously. Aurovertin B appears to act by preventing closure of the catalytic interfaces, which is essential for a catalytic mechanism involving cyclic interconversion of catalytic sites
RP  - NOT IN FILE
NT  - UI - 96293448LA - engRN - 0 (Aurovertins)RN - 0 (Enzyme Inhibitors)RN - 0 (Macromolecular Systems)RN - 25612-73-1 (Adenylyl Imidodiphosphate)RN - 55350-03-3 (aurovertin B)RN - 56-86-0 (Glutamic Acid)RN - 7004-12-8 (Arginine)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19960829IS - 0027-8424SB - IMCY - UNITED STATESJC - PV3
UR  - PM:8692918
SO  - Proc Natl Acad Sci U S A 1996 Jul 9 ;93(14):6913-6917

1410
UI  - 801
AU  - van Walraven HS
AU  - Strotmann H
AU  - Schwarz O
AU  - Rumberg B
AD  - Department of Physiology and Biochemistry of Plants, Institute for Molecular Biological Sciences (IMBW), BioCentrum Amsterdam, The Netherlands
TI  - The H+/ATP coupling ratio of the ATP synthase from thiol-modulated chloroplasts and two cyanobacterial strains is four
AB  - In this paper the authors emphasise that the proton translocating ATP synthase from thiol-modulated chloroplasts and two cyanobacterial strains has a coupling ratio of 4 protons per ATP synthesised or hydrolysed. This ratio is determined by several thermodynamic studies at equilibrium between phosphate potential (Delta Gp) and proton gradient (Delta(mu)H+), and is confirmed by measurement of proton flux during ATP hydrolysis. Ratios lower than 4 H+/ATP that have been published in the past have predominantly been determined with the oxidised chloroplast enzyme. Errors in these measurements will be discussed
RP  - NOT IN FILE
NT  - UI - 96184589LA - engRN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19960515IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:8603713
SO  - FEBS Lett 1996 Feb 5 ;379(3):309-313

1411
UI  - 156
AU  - Watts SD
AU  - Tang C
AU  - Capaldi RA
AD  - Institute of Molecular Biology, University of Oregon, Eugene, Oregon 97403-1229, USA
TI  - The stalk region of the Escherichia coli ATP synthase. Tyrosine 205 of the gamma subunit is in the interface between the F1 and F0 parts and can interact with both the epsilon and c oligomer
AB  - The soluble portion of the Escherichia coli F1F0 ATP synthase (ECF1) and E. coli F1F0 ATP synthase (ECF1F0) have been isolated from a novel mutant gammaY205C. ECF1 isolated from this mutant had an ATPase activity 3.5-fold higher than that of wild-type enzyme and could be activated further by maleimide modification of the introduced cysteine. This effect was not seen in ECF1F0. The mutation partly disrupts the F1 to F0 interaction, as indicated by a reduced efficiency of proton pumping. ECF1 containing the mutation gammaY205C was bound to the membrane-bound portion of the E. coli F1F0 ATP synthase (ECF0) isolated from mutants cA39C, cQ42C, cP43C, and cD44C to reconstitute hybrid enzymes. Cu2+ treatment or reaction with 5,5'-dithio-bis(2-nitro- benzoic acid) induced disulfide bond formation between the Cys at gamma position 205 and a Cys residue at positions 42, 43, or 44 in the c subunit but not at position 39. Using Cu2+ treatment, this covalent cross-linking was obtained in yields as high as 95% in the hybrid ECF1 gammaY205C/cQ42C and in ECF1F0 isolated from the double mutant of the same composition. The covalent linkage of the gamma to a c subunit had little effect on ATPase activity. However, ATP hydrolysis-linked proton translocation was lost, by modification of both gamma Cys-205 and c Cys- 42 by bulky reagents such as 5,5'-dithio-bis (2-nitro-benzoic acid) or benzophenone-4-maleimide. In both ECF1 and ECF1F0 containing a Cys at gamma 205 and a Cys in the epsilon subunit (at position 38 or 43), cross-linking of the gamma to the epsilon subunit was induced in high yield by Cu2+. No cross-linking was observed in hybrid enzymes in which the Cys was at position 10, 65, or 108 of the epsilon subunit. Cross- linking of gamma to epsilon had only a minimal effect on ATP hydrolysis. The reactivity of the Cys at gamma 205 showed a nucleotide dependence of reactivity to maleimides in both ECF1 and ECF1F0, which was lost in ECF1 when the epsilon subunit was removed. Our results show that there is close interaction of the gamma and epsilon subunits for the full-length of the stalk region in ECF1F0. We argue that this interaction controls the coupling between nucleotide binding sites and the proton channel in ECF1F0
RP  - NOT IN FILE
NT  - UI - 97067055LA - engRN - 0 (Disulfides)RN - 0 (Maleimides)RN - 25612-73-1 (Adenylyl Imidodiphosphate)RN - 55520-40-6 (Tyrosine)RN - 56-89-3 (Cystine)RN - 69-78-3 (Dithionitrobenzoic Acid)RN - 7439-95-4 (Magnesium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - HL 24526/HL/NHLBIDA - 19961230IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:8910457
SO  - J Biol Chem 1996 Nov 8 ;271(45):28341-28347

1412
UI  - 418
AU  - Weber J
AU  - Bowman C
AU  - Senior AE
AD  - Department of Biochemistry, University of Rochester Medical Center, Rochester, New York 14642, USA
TI  - Specific tryptophan substitution in catalytic sites of Escherichia coli F1-ATPase allows differentiation between bound substrate ATP and product ADP in steady-state catalysis
AB  - Tryptophan was specifically inserted as the residue immediately preceding the P-loop sequence in F1-ATPase catalytic sites. The mutant enzyme (betaF148W) showed normal enzymatic characteristics. The fluorescence responses of beta-tryptophan 148 enabled us to differentiate between nucleoside di- and triphosphate bound in catalytic sites; MgADP quenched at 350 nm, whereas MgAMPPNP and MgADP.BeFx complex enhanced the fluorescence at 325 nm. With MgATP, both effects were seen simultaneously. This allowed analysis of bound catalytic site nucleotides directly under steady-state MgATP hydrolysis conditions. At mM concentration of MgATP (Vmax conditions) one of the three catalytic sites was filled with substrate MgATP and the other two sites were filled with product MgADP. A model for F1-ATPase steady- state turnover is presented that encompasses these findings. Given the structural similarity of the P-loop in nucleotide-binding proteins, this approach may prove widely useful
RP  - NOT IN FILE
NT  - UI - 96324950LA - engRN - 0 (Fluorescent Dyes)RN - 0 (Oligodeoxyribonucleotides)RN - 25612-73-1 (Adenylyl Imidodiphosphate)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 73-22-3 (Tryptophan)RN - 7439-95-4 (Magnesium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSDA - 19960924IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:8702526
SO  - J Biol Chem 1996 Aug 2 ;271(31):18711-18718

1413
UI  - 420
AU  - Weber J
AU  - Senior AE
AD  - Department of Biochemistry, University of Rochester Medical Center, Rochester, New York 14642, USA
TI  - Binding and hydrolysis of TNP-ATP by Escherichia coli F1-ATPase
AB  - It had previously been suggested that Vmax hydrolysis rate of 2', 3'-O- (2,4,6-trinitrophenyl)adenosine 5'-triphosphate (TNP-ATP) by F1-ATPase required filling of only two catalytic sites on the enzyme (Grubmeyer, C., and Penefsky, H. S. (1981) J. Biol. Chem. 256, 3718-3727), whereas recently it was shown that Vmax rate of ATP hydrolysis requires that all three catalytic sites are filled (Weber, J., Wilke-Mounts, S., Lee, R. S. F., Grell, E., and Senior, A. E. (1993) J. Biol. Chem. 268, 20126- 20133). To resolve this apparent discrepancy, we measured equilibrium binding and hydrolysis of MgTNP-ATP under identical conditions, using betaY331W mutant Escherichia coli F1-ATPase, in which the genetically engineered tryptophan provides a direct fluorescent probe of catalytic site occupancy. We found that MgTNP-ATP hydrolysis at Vmax rate did require filling of all three catalytic sites, but in contrast to the situation with MgATP, "bisite hydrolysis" of MgTNP-ATP amounted to a substantial fraction (approximately 40%) of Vmax. Binding of MgTNP-ATP to the three catalytic sites showed strong binding cooperativity (Kd1 < 1 nm, Kd2 = 23 nm, Kd3 = 1.4 microM). Free TNP-ATP (i.e. in presence of EDTA) bound to all three catalytic sites with lower affinity but was not hydrolyzed. These data emphasize that the presence of Mg2+ is critical for cooperativity of substrate binding, formation of the very high affinity first catalytic site, and hydrolytic activity in F1- ATPases and that these three properties are strongly correlated
RP  - NOT IN FILE
NT  - UI - 96216441LA - engRN - 0 (Fluorescent Dyes)RN - 56-65-5 (Adenosine Triphosphate)RN - 61368-63-6 (2',3'-O-(2,4,6-trinitro-cyclohexadienylidine)adenosine 5'- triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSDA - 19960702IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:8631950
SO  - J Biol Chem 1996 Feb 16 ;271(7):3474-3477

1414
UI  - 419
AU  - Weber J
AU  - Senior AE
AD  - University of Rochester Medical Center, Department of Biochemistry, NY 14642, USA weber@medinforochesteredu
TI  - F1F0-ATP synthase: development of direct optical probes of the catalytic mechanism
AB  - Using strategically-placed tryptophan (Trp) residues as optical probes to monitor nucleotide binding and hydrolysis, we demonstrate that all three catalytic nucleotide binding sites in F1-ATPase must be filled to obtain physiological (Vmax) MgATP hydrolysis rates. At Vmax hydrolysis rates, the predominant enzyme species has one of the three catalytic sites filled with unhydrolyzed substrate MgATP, the other two sites are filled with product MgADP. A specifically-inserted Trp probe was also developed to characterize nucleotide binding to the noncatalytic sites, and a model to explain the specificity of these sites is shown. These sites appear to play no role in ATP hydrolysis
RP  - NOT IN FILE
NT  - UI - 96305573LA - engRN - 0 (Nucleotides)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialID - GM25349/GM/NIGMSDA - 19960829IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:8688441
SO  - Biochim Biophys Acta 1996 Jul 18 ;1275(1-2):101-104

1415
UI  - 20846
AU  - Yamada H
AU  - Moriyama Y
AU  - Maeda M
AU  - Futai M
AD  - Department of Biological Science, Institute of Scientific and Industrial Research, Osaka University, Japan
TI  - Transmembrane topology of Escherichia coli H(+)-ATPase (ATP synthase) subunit a
AB  - Escherichia coli H(+)-ATPase subunit a is a hydrophobic F0 subunit. To investigate the topology of the subunit in the membrane, we prepared site-specific polyclonal antibodies against amino-terminal (Ser-3 to Leu-16), middle loop (Lys-167 to Gln-181), and carboxyl-terminal (Thr- 259 to His-271) peptide segments. Enzyme-linked immunosorbent assay revealed that these antibodies specifically reacted with subunit a of inside-out membrane vesicles, but not with that of right-side-out spheroplasts. Full reactivity appeared when spheroplasts were disrupted with Triton X-100 (0.5%) or by sonication. These results suggest that at least parts of the three peptide segments of subunit a face the cytoplasm. Based on these observations, we propose a novel transmembrane topology of subunit a
MH  - A
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Cytoplasm
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - H(+)ATPase
MH  - Macromolecular Systems
MH  - membrane
MH  - membrane vesicles
MH  - Peptide Fragments
MH  - protein
MH  - Proteins
MH  - SUBUNIT
MH  - SYNTHASE
MH  - SYSTEM
MH  - SYSTEMS
RP  - NOT IN FILE
NT  - UI - 96314544LA - engRN - 0 (Antibodies)RN - 0 (Macromolecular Systems)RN - 0 (Peptide Fragments)RN - 0 (Recombinant Proteins)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19960910IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:8706824
SO  - FEBS Lett 1996 Jul 15 ;390(1):34-38

1416
UI  - 19859
AU  - Zhou Y
AU  - Duncan TM
AU  - Bulygin VV
AU  - Hutcheon ML
AU  - Cross RL
AD  - Department of Biochemistry and Molecular Biology, State University of New York Health Science Center, Syracuse 13210, USA
TI  - ATP hydrolysis by membrane-bound Escherichia coli F0F1 causes rotation of the gamma subunit relative to the beta subunits
AB  - We recently demonstrated that the gamma subunit in soluble F1-ATPase from Escherichia coli rotates relative to surrounding beta subunits during catalytic turnover (Duncan et al. (1995) Proc. Natl. Acad. Sci. USA 92, 10964-10968). Here, we extend our studies to the more physiologically relevant membrane-bound F0F1 complex. It is shown that beta D380C-F1, containing a beta-gamma intersubunit disulfide bond, can bind to F1-depleted membranes and can restore coupled membrane activities upon reduction of the disulfide. Using a dissociation/reconstitution approach with crosslinked beta D380C-F1, beta subunits containing an N-terminal Flag epitope (beta flag) were incorporated into the two non-crosslinked beta positions and the hybrid F1 was reconstituted with membrane-bound F0. Following reduction and ATP hydrolysis, reoxidation resulted in a significant amount of crosslinking of beta flag to the gamma subunit. This demonstrates that gamma rotates within F1 during catalytic turnover by membrane-bound F0- F1. Furthermore, the rotation of gamma is functionally coupled to F0, since preincubation with DCCD to modify F0 blocked rotation
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthase
MH  - BETA-SUBUNIT
MH  - Biochemistry
MH  - COMPLEX
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - F0F1
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - H(+)-Transporting ATP Synthase
MH  - Hydrolysis
MH  - membrane
MH  - Membranes
MH  - review
MH  - rotation
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 96305572LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialID - GM 23152/GM/NIGMSDA - 19960829IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:8688454
SO  - Biochim Biophys Acta 1996 Jul 18 ;1275(1-2):96-100

1417
UI  - 151
AU  - Aggeler R
AU  - Ogilvie I
AU  - Capaldi RA
AD  - Institute of Molecular Biology, University of Oregon, Eugene, Oregon 97403-1229, USA
TI  - Rotation of a gamma-epsilon subunit domain in the Escherichia coli F1F0- ATP synthase complex. The gamma-epsilon subunits are essentially randomly distributed relative to the alpha3beta3delta domain in the intact complex
AB  - A triple mutant of Escherichia coli F1F0-ATP synthase, alphaQ2C/alphaS411C/epsilonS108C, has been generated for studying movements of the gamma and epsilon subunits during functioning of the enzyme. It includes mutations that allow disulfide bond formation between the Cys at alpha411 and both Cys-87 of gamma and Cys-108 of epsilon, two covalent cross-links that block enzyme function (Aggeler, R., and Capaldi, R. A. (1996) J. Biol. Chem. 271, 13888-13891). A cross- link is also generated between the Cys at alpha2 and Cys-140 of the delta subunit, which has no effect on functioning (Ogilvie, I., Aggeler, R., and Capaldi, R. A. (1997) J. Biol. Chem. 272, 16652- 16656). CuCl2 treatment of the mutant alphaQ2C/alphaS411C/epsilonS108C generated five major cross-linked products. These are alpha-gamma- delta, alpha-gamma, alpha-delta-epsilon, alpha-delta, and alpha- epsilon. The ratio of alpha-gamma-delta to the alpha-gamma product was close to 1:2, i.e. in one-third of the ECF1F0 molecules the gamma subunit was attached to the alpha subunit at which the delta subunit is bound. Also, 20% of the epsilon subunit was present as a alpha-delta- epsilon product. With regard to the delta subunit, 30% was in the alpha- gamma-delta, 20% in the alpha-delta-epsilon, and 50% in the alpha-delta products when the cross-linking was done after incubation in ATP + MgCl2. The amounts of these three products were 40, 22, and 38%, respectively, in experiments where Cu2+ was added after preincubation in ATP + Mg2+ + azide. The delta subunit is fixed to, and therefore identifies, one specific alpha subunit (alphadelta). A distribution of the gamma and epsilon subunits, which is essentially random with respect to the alpha subunits, can only be explained by rotation of gamma-epsilon relative to the alpha3beta3 domain in ECF1F0
RP  - NOT IN FILE
NT  - UI - 97382300LA - engRN - 3483-12-3 (Dithiothreitol)RN - 56-65-5 (Adenosine Triphosphate)RN - 7440-50-8 (Copper)RN - 7447-39-4 (cupric chloride)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - HL 24526/HL/NHLBIDA - 19970821IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:9235970
SO  - J Biol Chem 1997 Aug 1 ;272(31):19621-19624

1418
UI  - 20926
AU  - al Shawi MK
AU  - Ketchum CJ
AU  - Nakamoto RK
AD  - Department of Molecular Physiology and Biological Physics, University of Virginia, Charlottesville, Virginia 22908, USA
TI  - Energy coupling, turnover, and stability of the F0F1 ATP synthase are dependent on the energy of interaction between gamma and beta subunits
AB  - Replacement of the F0F1 ATP synthase gamma subunit Met-23 with Lys (gammaM23K) perturbs coupling efficiency between transport and catalysis (Shin, K., Nakamoto, R. K., Maeda, M., and Futai, M. (1992) J. Biol. Chem. 267, 20835-20839). We demonstrate here that the gammaM23K mutation causes altered interactions between subunits. Binding of delta or epsilon subunits stabilizes the alpha3beta3gamma complex, which becomes destabilized by the mutation. Significantly, the inhibition of F1 ATP hydrolysis by the epsilon subunit is no longer relieved when the gammaM23K mutant F1 is bound to F0. Steady state Arrhenius analysis reveals that the gammaM23K enzyme has increased activation energies for the catalytic transition state. These results suggest that the mutation causes the formation of additional bonds within the enzyme that must be broken in order to achieve the transition state. Based on the x-ray crystallographic structure of Abrahams et al. (Abrahams, J. P., Leslie, A. G. W., Lutter, R., and Walker, J. E. (1994) Nature 370, 621-628), the additional bond is likely due to gammaM23K forming an ionized hydrogen bond with one of the betaGlu-381 residues. Two second site mutations, gammaQ269R and gammaR242C, suppress the effects of gammaM23K and decrease activation energies for the gammaM23K enzyme. We conclude that gammaM23K is an added function mutation that increases the energy of interaction between gamma and beta subunits. The additional interaction perturbs transmission of conformational information such that epsilon inhibition of ATPase activity is not relieved and coupling efficiency is lowered
MH  - A
MH  - ACTIVATION
MH  - analysis
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BETA-SUBUNIT
MH  - BINDING
MH  - Catalysis
MH  - COMPLEX
MH  - coupling
MH  - EPSILON-SUBUNIT
MH  - F0
MH  - F0F1
MH  - F1
MH  - function
MH  - Hydrogen
MH  - Hydrolysis
MH  - M
MH  - mutant
MH  - P
MH  - physiology
MH  - RESIDUE
MH  - Site
MH  - SUBUNIT
MH  - SYNTHASE
MH  - transport
RP  - NOT IN FILE
NT  - UI - 97153003LA - engRN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleID - HL07284/HL/NHLBIID - R01-GM50957/GM/NIGMSID - R01-GM52502/GM/NIGMSDA - 19970221IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:8999937
SO  - J Biol Chem 1997 Jan 24 ;272(4):2300-2306

1419
UI  - 20924
AU  - al Shawi MK
AU  - Ketchum CJ
AU  - Nakamoto RK
AD  - Department of Molecular Physiology and Biological Physics, University of Virginia, Charlottesville, Virginia 22906-0011, USA
TI  - The Escherichia coli FOF1 gammaM23K uncoupling mutant has a higher K0.5 for Pi. Transition state analysis of this mutant and others reveals that synthesis and hydrolysis utilize the same kinetic pathway
AB  - The Escherichia coli FOF1 ATP synthase uncoupling mutation, gammaM23K, was found to increase the energy of interaction between gamma and beta subunits, prevent the proper utilization of binding energy to drive catalysis, and block the enzyme in a Pi release mode. In this paper, the effects of this mutation on substrate binding in cooperative ATP synthesis are assessed. Activation of ATP synthesis by ADP and Pi was determined for the gammaM23K FOF1. The K0.5 for ADP was not affected, but K0.5 for Pi was approximately 7-fold higher even though the apparent Vmax was close to the wild-type level. Wild-type enzyme had a turnover number of 82 s-1 at pH 7.5 and 30 degrees C. During oxidative phosphorylation, the apparent dissociation constant (KI) for ATP was not affected and was 5-6 mM for both wild-type and gammaM23K enzymes. Thus, the apparent binding affinity for ATP in the presence of DeltamuH+ was lowered by 7 orders of magnitude from the affinity measured at the high-affinity catalytic site. Arrhenius analysis of ATP synthesis for the gammaM23K FOF1 revealed that, like those of ATP hydrolysis, the transition state DeltaH was much more positive and TDeltaS was much less negative, adding up to little change in DeltaG. These results suggested that ATP synthesis is inefficient because of an extra bond between gamma and beta subunits which must be broken to achieve the transition state. Analysis of the transition state structures using isokinetic plots demonstrate that ATP hydrolysis and synthesis utilize the same kinetic pathway. Incorporating this information into a model for rotational catalysis suggests that at saturating substrate concentrations, the rate-limiting step for hydrolysis and synthesis is the rotational power stroke where each of the beta subunits changes conformation and affinity for nucleotide
MH  - A
MH  - ACTIVATION
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - ADP
MH  - analysis
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - ATPase
MH  - BETA-SUBUNIT
MH  - BINDING
MH  - Catalysis
MH  - conformation
MH  - CONSTANT
MH  - Enzymes
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - Hydrolysis
MH  - model
MH  - mutant
MH  - Oxidative Phosphorylation
MH  - pH
MH  - Phosphates
MH  - Phosphorylation
MH  - physiology
MH  - Site
MH  - SUBUNIT
MH  - SYNTHASE
MH  - synthesis
RP  - NOT IN FILE
NT  - UI - 97477346LA - engRN - 0 (Phosphates)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleID - GM50957/GM/NIGMSID - GM52502/GM/NIGMSID - HL07284/HL/NHLBIDA - 19971120IS - 0006-2960SB - IMCY - UNITED STATES
UR  - PM:9335556
SO  - Biochemistry 1997 Oct 21 ;36(42):12961-12969

1420
UI  - 20925
AU  - al Shawi MK
AU  - Nakamoto RK
AD  - Department of Molecular Physiology and Biological Physics, University of Virginia, Charlottesville, Virginia 22906-0011, USA
TI  - Mechanism of energy coupling in the FOF1-ATP synthase: the uncoupling mutation, gammaM23K, disrupts the use of binding energy to drive catalysis
AB  - The Escherichia coli FOF1 ATP synthase uncoupling mutation, gammaM23K, was found to increase the energy of interaction between gamma and beta subunits which caused inefficient transmission of coupling information between transport and catalysis [Al-Shawi, M. K. , Ketchum, C. J., and Nakamoto, R. K. (1997) J. Biol. Chem. 272, 2300-2306]. We hypothesized that the gammaM23K mutation, because of its effect on coupling, should alter the fundamental reactions steps that are normally modulated by DeltamuH+ via the coupling mechanism. In this paper, we address this issue by studying the thermodynamics of individual catalytic steps through the use of energy profiles to gain information regarding enzyme mechanism and the effects of the mutation. Compared to wild-type enzyme, the gammaM23K F1 had significant differences of two partial reactions: the rate constant for Pi release was 49-fold faster and the rate constant for ATP release was 8.4-fold faster than wild-type. These rate constants were considered together with characteristics of a group of F1 ATPase mutant enzymes and were analyzed quantitatively by linear free energy relationships [Al-Shawi, M. K., Parsonage, D., and Senior, A. E., (1990) J. Biol. Chem. 265, 4402-4410]. We found that the gammaM23K mutation prevents the proper utilization of binding energy to drive catalysis and blocks the enzyme in a Pi release mode. This finding is consistent with the use of energy from DeltamuH+ for increasing the affinity for Pi so that the substrate binds in a catalytically competent manner for synthesis of ATP. These results support the notion that the communication of coupling information is transmitted through the gamma-beta interface near gammaMet23 and beta380DELSEED386 segment
MH  - A
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BETA-SUBUNIT
MH  - BINDING
MH  - Catalysis
MH  - CONSTANT
MH  - coupling
MH  - Enzymes
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - M
MH  - mechanism
MH  - mutant
MH  - Phosphates
MH  - physiology
MH  - protein
MH  - Proteins
MH  - rate constant
MH  - SUBUNIT
MH  - SYNTHASE
MH  - synthesis
MH  - Thermodynamics
MH  - transport
RP  - NOT IN FILE
NT  - UI - 97477345LA - engRN - 0 (Phosphates)RN - 0 (Recombinant Proteins)RN - 56-65-5 (Adenosine Triphosphate)RN - 56-87-1 (Lysine)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleID - GM50957/GM/NIGMSID - GM52502/GM/NIGMSDA - 19971120IS - 0006-2960SB - IMCY - UNITED STATES
UR  - PM:9335555
SO  - Biochemistry 1997 Oct 21 ;36(42):12954-12960

1421
UI  - 19772
AU  - Boyer PD
AD  - Molecular Biology Institute, University of California, Los Angeles 90095-1570, USA
TI  - The ATP synthase--a splendid molecular machine
AB  - An X-ray structure of the F1 portion of the mitochondrial ATP synthase shows asymmetry and differences in nucleotide binding of the catalytic beta subunits that support the binding change mechanism with an internal rotation of the gamma subunit. Other structural and mutational probes of the F1 and F0 portions of the ATP synthase are reviewed, together with kinetic and other evaluations of catalytic site occupancy and behavior during hydrolysis or synthesis of ATP. Subunit function as related to proton translocation and rotational catalysis is considered. Physical demonstrations of the gamma subunit rotation have been achieved. The findings have implications for other enzymatic catalyses
MH  - atp
MH  - ATP synthase
MH  - BETA-SUBUNIT
MH  - BINDING
MH  - Catalysis
MH  - CHANGE MECHANISM
MH  - F0
MH  - F1
MH  - H(+)-Transporting ATP Synthase
MH  - Hydrolysis
MH  - mechanism
MH  - nucleotide binding
MH  - proton
MH  - review
MH  - rotation
MH  - SUBUNIT
MH  - SYNTHASE
MH  - synthesis
MH  - translocation
RP  - NOT IN FILE
NT  - UI - 97386832LA - engRN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, AcademicDA - 19971007IS - 0066-4154SB - IMCY - UNITED STATESJC - 6DJ
UR  - PM:9242922
SO  - Annu Rev Biochem 1997  ;66():717-749

1422
UI  - 608
AU  - Cappellini P
AU  - Turina P
AU  - Fregni V
AU  - Melandri BA
AD  - Department of Biology, University of Bologna, Italy
TI  - Sulfite stimulates the ATP hydrolysis activity of but not proton translocation by the ATP synthase of Rhodobacter capsulatus and interferes with its activation by delta muH+
AB  - Sulfite stimulates the rate of ATP hydrolysis by the ATP synthase in chromatophores of Rhodobacter capsulatus. The stimulated activity is inhibited by oligomycin. The activation takes place also in uncoupled chromatophores. The activation consists in an increase of about 12-15- fold of the Vmax for the ATP hydrolysis reaction, while the Km for MgATP is unaffected at 0.16+/-0.03 mM. The dependence of Vmax on the sulfite concentration follows a hyperbolic pattern with half maximum effect at 12 mM. Sulfite affects the ability of the enzyme in translocating protons. Concomitant measurements of the rate of ATP hydrolysis and of ATP-induced protonic flows demonstrate that at sulfite concentrations of greater than 10 mM the hydrolytic reaction becomes progressively uncoupled from the process of proton translocation. This is accompanied by an inhibition of ATP synthesis, either driven by light or by artificially induced ionic gradients. ATP synthesis is totally inhibited at concentrations of at least 80 mM. Sulfite interferes with the mechanism of activation by delta muH+. Low concentrations of this anion (< or = 2 mM) prevent the activation by delta muH+. At higher concentrations a marked stimulation of the activity prevails, regardless of the occurrence of a delta muH+ across the membrane. Phosphate at millimolar concentrations can reverse the inhibition by sulfite. These experimental results can be simulated by a model assuming multiple and competitive equilibria for phosphate or sulfite binding with two binding sites for the two ligands (for sulfite K1S = 0.26 and K2S = 37 mM, and for phosphate K1P = 0.06 and K2P = 4.22 mM), and in which the state bound only to one sulfite molecule is totally inactive in hydrolysis. The competition between phosphate and sulfite is consistent with the molecular structures of the two ligands and of the enzyme
RP  - NOT IN FILE
NT  - UI - 98004294LA - engRN - 0 (Protons)RN - 0 (Sulfites)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19971121IS - 0014-2956SB - IMCY - GERMANYJC - EMZ
UR  - PM:9346308
SO  - Eur J Biochem 1997 Sep 1 ;248(2):496-506

1423
UI  - 21305
AU  - Checover S
AU  - Nachliel E
AU  - Dencher NA
AU  - Gutman M
AD  - Department of Biochemistry, Tel Aviv University, Israel
TI  - Mechanism of proton entry into the cytoplasmic section of the proton-conducting channel of bacteriorhodopsin
AB  - Bacteriorhodopsin is the light-driven proton-pumping protein of Halobacterium salinarum that extracts protons from the well-buffered cytoplasmic space within the time limits set by the photocycle turnover. The specific mechanism of the proton uptake by the cytoplasmic surface of the protein was investigated in this study by the laser-induced proton pulse technique. The purple membrane preparations were labeled by fluorescein at two residues (36 or 38) of the cytoplasmic surface of the protein, sites that are close to the orifice of the proton-conducting channel. The membranes were pulsed by protons discharged from photoexcited pyranine [Nachliel, E., Gutman, M., Kiryati, S., and Dencher, N.A. (1996) Proc. Nat Acad. Sci. U.S.A. 93, 10747-10752). The reaction of the discharged protons with the pyranine anion and the fluorescein was measured with sub-microsecond resolution. The experimental signals were reconstructed through numeric integration of differential rate equations which quantitated the rates of all proton transfer reactions between all reactants present in the system. The interaction of protons with the orifice of the cytoplasmic channel is enhanced by the exposed carboxylates of the protein. A cluster of three carboxylates acts as a strong proton attractor site while one carboxylate, identified as D36, acts as a mediator that delivers the proton to the channel. The combination of these reactions render the surface of the protein with properties of a proton-collecting antenna. The size of the collecting area is less than that of the protein's surface
MH  - A
MH  - Bacteriorhodopsin
MH  - Biochemistry
MH  - DYE
MH  - dyes
MH  - fluorescein
MH  - Fluoresceins
MH  - Fluorescent Dyes
MH  - Halobacterium
MH  - INTERACTION
MH  - ion
MH  - Ion Channels
MH  - M
MH  - mechanism
MH  - membrane
MH  - Membranes
MH  - protein
MH  - proton
MH  - Proton Pump
MH  - Proton transfer
MH  - Proton-transfer
MH  - Protons
MH  - purple membrane
MH  - RESIDUE
MH  - resolution
MH  - Site
MH  - SURFACE
MH  - SYSTEM
MH  - Time
MH  - TRANSFER
MH  - turnover
MH  - united states
MH  - universities
RP  - NOT IN FILE
NT  - UI - 98042284DA - 19971208IS - 0006-2960LA - engPT - Journal ArticleCY - UNITED STATESRN - 0 (Fluoresceins)RN - 0 (Fluorescent Dyes)RN - 0 (Ion Channels)RN - 0 (Proton Pumps)RN - 0 (Protons)RN - 53026-44-1 (Bacteriorhodopsins)RN - 75350-46-8 (fluorescein 5-maleimide)SB - IM
UR  - PM:9374871
SO  - Biochemistry 1997 Nov 11 ;36(45):13919-13928

1424
UI  - 988
AU  - Cladera J
AU  - Rigaud JL
AU  - Villaverde J
AU  - Dunach M
TI  - Liposome solubilization and membrane protein reconstitution using Chaps and Chapso
AB  - The process of liposome solubilization and reconstitution of two transport proteins have been studied using Chaps and Chapso (3-[(3-cholamidopropyl)dimethylammonio]-2- hydroxy-1-propanesulfonate). The solubilization of unilamellar liposomes was followed by absorption experiments and the process was shown to fit well to the three-stage model previously proposed for other detergents. The solubilization parameters have been determined and the detergent to phospholipid ratios at which the lamellar-to-micellar transition initiates and ends were estimated to be 0.21 mol/mol and 0.74 mol/mol, for Chapso and 0.4 mol/mol and 1.04 mol/mol for Chaps, respectively. The best conditions for the incorporation of two membrane proteins, bacteriorhodopsin and the H(+)-ATP synthase from chloroplasts, were analyzed at each step of the solubilization process. After detergent removal, the activities of the resulting proteoliposomes were measured indicating that the most efficient reconstitutions were obtained by addition of the proteins to completely solubilized lipid-detergent micelles. The use of Chapso and Chaps for membrane protein reconstitution studies provides a reproducible method of achieving active proteoliposomes, homogeneous in size, with a low permeability and thus, well suited for transport measurements
MH  - Bacterial Outer Membrane Proteins
MH  - Bacteriorhodopsin
MH  - Chemistry
MH  - Chloroplasts
MH  - Cholic Acids
MH  - Detergents
MH  - Halobacterium
MH  - Liposomes
MH  - Membrane Proteins
MH  - Micelles
MH  - Proteins
MH  - Proteolipids
MH  - Solubility
MH  - Support,Non-U.S.Gov't
RP  - NOT IN FILE
NT  - Department de Bioquimica i Biologia Molecular, Facultat de Medicina, Universitat Autonma de Barcelona, Spain
SO  - Eur J Biochem 1997 Feb 1 ;243(3):798-804

1425
UI  - 9890
AU  - CreczynskiPasa TB
AU  - Graber P
AU  - Alves EW
AU  - Ferreira AT
AU  - Scofano HM
TI  - Phosphatase activity of H+-ATPase from chloroplasts
AB  - The chloroplast H+-ATPase (CF0F1) was purified from spinach chloroplasts and studied both in the soluble state and after reconstitution into asolectin liposomes. Hydrolysis of ATP and of p-nitrophenylphosphate (p-NPP) catalysed by CF0F1 was investigated. (1) Soluble, isolated CF0F1 catalyzes ATP hydrolysis and p-NPP hydrolysis. (2) ATP inhibits the phosphatase activity in the latent state (K-1 = 1.7 mM). (3) Addition of 100 mM sulfite increases the rate of ATP hydrolysis by a factor of 10 while p-NPP hydrolysis is completely abolished. (4) CF0F1 reconstituted into asolectin vesicles catalyzes ATP hydrolysis and p-NPP hydrolysis. When the enzyme is brought into its active state by a Delta pH/Delta phi jump, ATP hydrolysis is increased by a factor of 8, and p-NPP hydrolysis is completely abolished. (5) ATP hydrolysis by the activated enzyme is inhibited by p-NPP (K-1 = 1.6 mM). (6) p- NPP also inhibits ATP synthesis by the activated enzyme, competing with phosphate (K-1 = 0.9 mM). These results show that in the active state of CF0F1, p-NPP is not hydrolyzed but acts as a competitive inhibitor; in the inactive state of CF0F1, p-NPP is hydrolyzed. Hydrolysis of p-NPP might be used as an assay for the inactive forms of CF0F1
MH  - ADENOSINE-TRIPHOSPHATASE
MH  - ADP
MH  - ANALOGS
MH  - atp
MH  - ATP synthesis
MH  - BETA- SUBUNIT
MH  - BINDING
MH  - CF0F1
MH  - chloroplast
MH  - chloroplast H+-ATPase
MH  - Chloroplasts
MH  - COUPLING FACTOR-I
MH  - H+-ATPase
MH  - Hydrolysis
MH  - Liposomes
MH  - p-nitrophenylphosphate
MH  - phosphatase
MH  - RECONSTITUTED CF0F1-LIPOSOMES
MH  - Spinach
MH  - SPINACH-CHLOROPLASTS
MH  - SYNTHASE
MH  - THYLAKOID ATPASE
RP  - NOT IN FILE
NT  - JournalArticleELSEVIER SCIENCE BVMAY 16XA452AMSTERDAMUNIV FREIBURG,INST PHYS CHEM,D-79104 FREIBURG,GERMANYBBA-BIOENERGETICSPO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS
AV  - UNIV FREIBURG,INST PHYS CHEM,D-79104 FREIBURG,GERMANY UNIV FED RIO DE JANEIRO,DEPT BIOQUIM MED,ICB,CCS,BR-21941590 RIO JANEIRO,RJ,BRAZIL
UR  - ISI:A1997XA45200007
SO  - Biochimica et Biophysica Acta-Bioenergetics 1997  ;1320(1):58-64

1426
UI  - 21299
AU  - Daizadeh I
AU  - Medvedev ES
AU  - Stuchebrukhov AA
AD  - Department of Chemistry, University of California, Davis 95616, USA
TI  - Effect of protein dynamics on biological electron transfer
AB  - Computer simulations of the effect of protein dynamics on the long distance tunneling mediated by the protein matrix have been carried out for a Ru-modified (His 126) azurin molecule. We find that the tunneling matrix element is a sensitive function of the atomic configuration of the part of the protein matrix in which tunneling currents (pathways) are localized. Molecular dynamics simulations show that fluctuations of the matrix element can occur on a time scale as short as 10 fs. These short time fluctuations are an indication of a strong dynamic coupling of a tunneling electron to vibrational motions of the protein nuclear coordinates. The latter results in a modification of the conventional Marcus picture of electron transfer in proteins. The new element in the modified theory is that the tunneling electron is capable of emitting or absorbing vibrational energy (phonons) from the medium. As a result, some biological reactions may occur in an activationless fashion. An analytical theoretical model is proposed to account for thermal fluctuations of the medium in long distance electron transfer reactions. The model shows that, at long distances, the phonon-modified inelastic tunneling always dominates over the conventional elastic tunneling
MH  - A
MH  - Chemistry
MH  - computer simulations
MH  - COMPUTER-SIMULATION
MH  - COMPUTER-SIMULATIONS
MH  - coupling
MH  - electron
MH  - electron transfer
MH  - Electron-transfer
MH  - energy
MH  - function
MH  - model
MH  - MOLECULAR-DYNAMICS
MH  - protein
MH  - Proteins
MH  - SIMULATION
MH  - SIMULATIONS
MH  - theory
MH  - Time
MH  - TRANSFER
MH  - united states
MH  - universities
RP  - NOT IN FILE
NT  - UI - 97268633DA - 19970522IS - 0027-8424LA - engID - GM54052-02/GM/NIGMSPT - Journal ArticleCY - UNITED STATESRN - 0 (Proteins)SB - IM
UR  - PM:9108041
SO  - Proc Natl Acad Sci U S A 1997 Apr 15 ;94(8):3703-3708

1427
UI  - 41
AU  - Dimroth P
AD  - Mikrobiologisches Institut, Eidgenossische Technische Hochschule, Zurich, Switzerland
TI  - Primary sodium ion translocating enzymes
RP  - NOT IN FILE
NT  - UI - 97182208LA - engRN - 7440-23-5 (Sodium)RN - 77-92-9 (Citric Acid)RN - EC 1.6. (NADH, NADPH Oxidoreductases)RN - EC 1.6.5.3 (NADH dehydrogenase (ubiquinone))RN - EC 2.1.1. (Methyltransferases)RN - EC 2.1.1.86 (tetrahydromethanopterin S-methyltransferase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)RN - EC 3.6.1.37 (Na(+)-K(+)-Exchanging ATPase)RN - EC 4.1.1. (Carboxy-Lyases)RN - EC 4.1.1.3 (oxaloacetate decarboxylase)RN - EC 4.1.1.41 (propionyl-CoA carboxylase)PT - Journal ArticlePT - ReviewPT - Review, AcademicDA - 19970313IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:9030254
SO  - Biochim Biophys Acta 1997 Jan 16 ;1318(1-2):11-51

1428
UI  - 715
AU  - Dou C
AU  - Grodsky NB
AU  - Matsui T
AU  - Yoshida M
AU  - Allison WS
AD  - Department of Chemistry & Biochemistry, School of Medicine, University of California at San Diego, La Jolla 92093, USA
TI  - ADP-fluoroaluminate complexes are formed cooperatively at two catalytic sites of wild-type and mutant alpha3beta3gamma subcomplexes of the F1- ATPase from the thermophilic Bacillus PS3
AB  - Addition of Al3+ and F- to the alpha3beta3gamma subcomplex of the TF1- ATPase containing MgADP in one catalytic site causes slow, complete inactivation as the ADP-fluoroaluminate complex is formed. This conflicts with the "bisite" stochastic model suggested earlier (Issartel, J. P., Dupuis, A., Lunardi, J. & Vignais, P. V. (1991) Biochemistry 30, 4726-4733] on the finding that complete inactivation of the bovine mitochondrial F1-ATPase by Al3+, F-, Mg2+, and excess ADP occurs as ADP-fluoroaluminate complexes form in two catalytic sites. When Al3+ and F- were added to alpha3beta3gamma containing MgADP in two catalytic sites, inactivation accelerated 8-fold, indicating catalytic to catalytic site cooperativity. When added to alpha3beta3gamma containing MgADP bound to one or two catalytic sites prior to addition of Al3+ and F-, phosphate inhibits formation of the ADP-fluoroaluminate complex. When introduced after adding 200 microM ADP plus Mg2+ to alpha3beta3gamma, but before adding Al3+ and F-, phosphate accelerated formation of the ADP-fluoroaluminate complex 3-fold. Sulfite accelerated formation of the ADP-fluoroaluminate complex 9-fold when 200 microM ADP plus Mg2+ was added to alpha3beta3gamma before adding Al3+ and F-. The accelerations induced by phosphate or sulfite in the presence of excess ADP and Mg2+ suggest noncatalytic to catalytic site cooperativity. When Al3+ and F- were added to the (alphaD261N)3beta3gamma subcomplex containing MgADP in a single catalytic site, the ADP-fluoroaluminate complex formed at least 10-fold more slowly than observed with wild-type under the same conditions. Therefore, the catalytic site containing MgADP recognizes the alphaD261N substitution when noncatalytic sites are empty. Cross- linking alpha to gamma or beta to gamma by oxidizing the (alphaA396C)3beta3(gammaA22C) and alpha3(betaD390C)3(gammaS90C) subcomplexes, respectively, abolishes cooperative formation of ADP- fluoroaluminate complexes in two catalytic sites. ADP-fluoroaluminate complex formation is restricted to a single catalytic site in the oxidized double mutants. The alpha3beta3delta subcomplex does not form an inhibitory ADP-fluoroaluminate complex under any of the conditions examined for the alpha3beta3gamma subcomplexes
RP  - NOT IN FILE
NT  - UI - 97238575LA - engRN - 0 (Macromolecular Systems)RN - 0 (Phosphates)RN - 0 (Sulfites)RN - 21330-18-7 (fluoroaluminum)RN - 58-64-0 (Adenosine Diphosphate)RN - 7429-90-5 (Aluminum)RN - 7782-41-4 (Fluorine)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM-16974/GM/NIGMSDA - 19970429IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:9132025
SO  - Biochemistry 1997 Mar 25 ;36(12):3719-3727

1429
UI  - 19040
AU  - Drose S
AU  - Altendorf K
TI  - Bafilomycins and concanamycins as inhibitors of V-ATPases and P- ATPases
MH  - Adenosinetriphosphatase
MH  - Animal
MH  - ANS
MH  - antagonists & inhibitors
MH  - ANTIBIOTIC
MH  - Antibiotics
MH  - antibiotics,macrolide
MH  - ATPase
MH  - Bacteria
MH  - Biological Transport,Active
MH  - cell
MH  - Cells
MH  - Chemistry
MH  - drug effects
MH  - Enzyme Inhibitors
MH  - function
MH  - fungal proteins
MH  - Fungi
MH  - H(+)-Transporting ATP Synthase
MH  - Human
MH  - Hydrogen-Ion Concentration
MH  - inhibitor
MH  - inhibitors
MH  - P
MH  - pharmacology
MH  - Phosphorylation
MH  - plant
MH  - Proton-Translocating ATPases
MH  - review
MH  - Structure-Activity Relationship
MH  - Support,Non-U.S.Gov't
MH  - YEAST
RP  - NOT IN FILE
NT  - Bafilomycins and concanamycins, two groups of the plecomacrolide- defined class of macrolide antibiotics, have recently been recognized as important tools for studying the physiological role of vacuolar- type, proton-translocating ATPases (V-ATPases) and ATPases with phosphorylated states (P-ATPases) in animal and plant cells as well as in yeast, fungi and bacteria. The following review will give an account of the classification and function of these antibiotics Universitat Osnabruck, Fachbereich Biologie/Chemie, Germany
SO  - J exp Biol 1997  ;200(Pt 1):1-8

1430
UI  - 290
AU  - Engelbrecht S
AU  - Junge W
AD  - Universitat Osnabruck, AG Biophysik, Germany engel@uni-osnabrueckde
TI  - ATP synthase: a tentative structural model
AB  - Adenosine triphosphate (ATP) synthase produces ATP from ADP and inorganic phosphate at the expense of proton- or sodium-motive force across the respective coupling membrane in Archaea, Bacteria and Eucarya. Cation flow through the intrinsic membrane portion of this enzyme (Fo, subunits ab2c9-12) and substrate turnover in the headpiece (F1, subunits alpha3beta3 gammadeltaepsilon) are mechanically coupled by the rotation of subunit gamma in the center of the catalytic hexagon of subunits (alphabeta)3 in F1. ATP synthase is the smallest rotatory engine in nature. With respect to the headpiece alone, it probably operates with three steps. Partial structures of six out of its at least eight different subunits have been published and a 3-dimensional structure is available for the assembly (alphabeta)3gamma. In this article, we review the available structural data and build a tentative topological model of the holoenzyme. The rotor portion is proposed to consist of a wheel of at least nine copies of subunits c, epsilon and a portion of gamma as a spoke, and another portion of gamma as a crankshaft. The stator is made up from a, the transmembrane portion of b2, delta and the catalytic hexagon of (alphabeta)3. As an educated guess, the model may be of heuristic value for ongoing studies on this fascinating electrochemical-to-mechanical-to-chemical transducer
RP  - NOT IN FILE
NT  - UI - 97462794LA - engRN - 0 (Multienzyme Complexes)RN - 0 (Proteolipids)RN - EC 2.7.4 (Phosphotransferases (Phosphate Group Acceptor))RN - EC 2.7.4.- (ATP synthetase complex)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 19971027IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:9323021
SO  - FEBS Lett 1997 Sep 15 ;414(3):485-491

1431
UI  - 20937
AU  - Etzold C
AU  - Deckers-Hebestreit G
AU  - Altendorf K
AD  - Fachbereich Biologie/Chemie, Arbeitsgruppe Mikrobiologie, Universitat Osnabruck, Germany
TI  - Turnover number of Escherichia coli F0F1 ATP synthase for ATP synthesis in membrane vesicles
AB  - The rate of ATP synthesized by the ATP synthase (F0F1-ATPase) is limited by the rate of energy production via the respiratory chain, when measured in everted membrane vesicles of an Escherichia coli atp wild-type strain. After energization of the membranes with NADH, fractional inactivation of F0F1 by the covalent inhibitor N,N'- dicyclohexylcarbodiimide allowed the rate of ATP synthesis/mol remaining active ATP synthase complexes to increase; the active ATP synthase complexes were calculated using ATP hydrolysis rates as the defining parameter. In addition, variation of the assay temperature revealed an increase of the ATP synthesis rate up to a temperature of 37 degrees C, the optimal growth temperature of E. coli. In parallel, the amount of F0F1 complexes present in membrane vesicles was determined by immunoquantitation to be 3.3 +/- 0.3% of the membrane protein for cells grown in rich medium and 6.6 +/- 0.3% for cells grown in minimal medium with glycerol as sole carbon and energy source. Based on these data, a turnover number for ATP synthesis of 270 +/- 40 s(-1) could be determined in the presence of 5% active F0F1 complexes. Therefore, these studies demonstrate that the ATP synthase complex of E. coli has, with respect to maximum rates, the same capacity as the corresponding enzymes of eukaryotic organells
MH  - A
MH  - ACTIVE
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - ATPase
MH  - Cells
MH  - COMPLEX
MH  - Dicyclohexylcarbodiimide
MH  - Enzymes
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0F1
MH  - Hydrolysis
MH  - membrane
MH  - membrane vesicles
MH  - Membranes
MH  - protein
MH  - SYNTHASE
MH  - synthesis
MH  - Temperature
RP  - NOT IN FILE
NT  - UI - 97182618LA - engRN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19970320IS - 0014-2956SB - IMCY - GERMANY
UR  - PM:9030757
SO  - Eur J Biochem 1997 Jan 15 ;243(1-2):336-343

1432
UI  - 22
AU  - Fillingame RH
AD  - Department of Biomolecular Chemistry, University of Wisconsin Medical School, Madison 53706, USA filingam@maccwiscedu
TI  - Coupling H+ transport and ATP synthesis in F1F0-ATP synthases: glimpses of interacting parts in a dynamic molecular machine
AB  - Reversible, F1F0-type ATPases (also termed F-ATP synthases) catalyze the synthesis of ATP during oxidative phosphorylation. In animal cells, the enzyme traverses the inner mitochondrial membrane and uses the energy of an H+ electrochemical gradient, generated by electron transport, in coupling H+ translocation to ATP formation. Closely related enzymes are found in the plasma membrane of bacteria such as Escherichia coli, where the enzymes function reversibly depending upon nutritional circumstance. The F1F0-type enzymes are more distantly related to a second family of H(+)-translocating ATPases, the vacuolar- type or V-ATPases. Recent structural information has provided important hints as to how these enzymes couple H+ transport to the chemical work of ATP synthesis. The simplest F1F0-type enzymes, e.g. as in E. coli, are composed of eight types of subunits in an unusual stoichiometry of alpha 3 beta 3 gamma delta epsilon (F1) and a1b2c12 (F0). F1 extends from the membrane, with the alpha and beta subunits alternating around a central subunit gamma. ATP synthesis occurs alternately in different beta subunits, the cooperative tight binding of ADP + Pi at one catalytic site being coupled to ATP release at a second. The differences in binding affinities appear to be caused by rotation of the gamma subunit in the center of the alpha 3 beta 3 hexamer. The gamma subunit traverses a 4.5 nm stalk connecting the catalytic subunits to the membrane-traversing F0 sector. Subunit c is the H(+)- translocating subunit of F0. Protonation/deprotonation of Asp61 in the center of the membrane is coupled to structural changes in an extramembranous loop of subunit c which interacts with both the gamma and epsilon subunits. Subunits gamma and epsilon appear to move from one subunit c to another as ATP is synthesized. The torque of such movement is proposed to cause the rotation of gamma within the alpha 3 beta 3 complex. Four protons are translocated for each ATP synthesized. The movement of gamma and epsilon therefore probably involves a unit of four c subunits. The organization of subunits in F0 remains a mystery; it will have to be understood if we are to understand the mechanism of torque generation
RP  - NOT IN FILE
NT  - UI - 97202735LA - engRN - 0 (Bacterial Proteins)RN - 0 (Proton Pump)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, AcademicID - GM23105/GM/NIGMSDA - 19970415IS - 0022-0949SB - IMCY - ENGLANDJC - I2F
UR  - PM:9050229
SO  - J Exp Biol 1997 Jan ;200 ( Pt 2)():217-224

1433
UI  - 9930
AU  - Findlay JB
AU  - Finbow ME
AU  - Jones PC
AU  - Kim YI
AU  - Harrison MA
AU  - Hughes G
AD  - Department of Biochemistry and Molecular Biology, University of Leeds, UK
TI  - A structure-based model for the 16 kDa membrane sector of the vacuolar H(+)-ATPase
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Biochemistry
MH  - England
MH  - H(+)-Transporting ATP Synthase
MH  - H(+)ATPase
MH  - Lipid Bilayers
MH  - Macromolecular Systems
MH  - model
MH  - Proteins
MH  - Proteolipids
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 98049962LA - engRN - 0 (Disulfides)RN - 0 (Lipid Bilayers)RN - 0 (Macromolecular Systems)RN - 0 (Proteolipids)RN - 0 (Recombinant Proteins)RN - EC 3.6.1.- (V-type ATPase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19980102IS - 0300-5127SB - IMCY - ENGLANDJC - E48
UR  - PM:9388608
SO  - Biochem Soc Trans 1997 Aug ;25(3):1107-1113

1434
UI  - 327
AU  - Groth G
AU  - Walker JE
AD  - Heinrich-Heine-Universitat Dusseldorf, Biochemie der Pflanzen, Germany georggroth@uni-duesseldorfde
TI  - Model of the c-subunit oligomer in the membrane domain of F-ATPases
AB  - A model is described of a dodecameric complex consisting of the integral membrane component subunit c of the H+-transporting Fo domain of Escherichia coli F-ATPase. A high-resolution partial structure of monomeric subunit c resulting from 1H-NMR studies [1] was used for constructing the model. The validity of the proposed arrangement of protomers in the dodecameric complex was tested by amino acid substitution analysis and chemical, biochemical and genetic data on subunit c
RP  - NOT IN FILE
NT  - UI - 97379290LA - engRN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19970903IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:9237612
SO  - FEBS Lett 1997 Jun 30 ;410(2-3):117-123

1435
UI  - 67
AU  - Gruber G
AU  - Hausrath A
AU  - Sagermann M
AU  - Capaldi RA
AD  - Institute of Molecular Biology, University of Oregon, Eugene 97403- 1229, USA
TI  - An improved purification of ECF1 and ECF1F0 by using a cytochrome bo- deficient strain of Escherichia coli facilitates crystallization of these complexes
AB  - A novel strategy, which employs a cytochrome bo-lacking strain (GO104) and a modified isolation procedure provides an effective approach for obtaining much purer preparations of ECF1F0 than described previously, as well as for isolating homogeneous and protein-chemically pure ECF1. ECF1 obtained in this way could be crystallized by vapor-diffusion using polyethylene glycol (PEG) as a precipitant in a form suitable for X-ray diffraction analysis. The crystals belong to the orthorhombic space group P2(1)2(1)2(1), with lattice parameters a = 110, h = 134, and c = 269 A, and diffract to a resolution of at least 6.4 A
RP  - NOT IN FILE
NT  - UI - 97379300LA - engRN - 9035-37-4 (Cytochrome b)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM20066/GM/NIGMSID - HL24526/HL/NHLBIDA - 19970903IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:9237622
SO  - FEBS Lett 1997 Jun 30 ;410(2-3):165-168

1436
UI  - 21333
AU  - Helguera G
AU  - Beauge L
TI  - Phosphoryl Group Exchange between ATP and ADP Catalyzed by H+-ATPase from Oat Roots
AB  - ATP-ADP exchange was estimated in the presence of plasma membrane H+-ATPase of oat (Avena sativa) roots partially purified with Triton X-100 by measuring [14C]ATP formation from [14C]ADP. Most studies were done at 0[deg]C. At pH 6.0 the exchange showed: (a) Mg2+ requirement with a biphasic response giving maximal activity at 152 [mu]M and (b) insensitivity to ionic strength, [Na+], and [K+]. ATP and ADP dependence were analyzed with a model in which nucleotide-enzyme interactions are at rapid-random equilibrium, whereas E1ATP [left right arrow] E1P-ADP transitions occur in steady state. The results indicated competition between ADP and ATP for the catalytic site, whereas ATP interaction with the ADP site was extremely weak. At 0[deg]C the exchange showed a 3-fold pH increase, from pH 5.5 to 9.0. At an alkaline pH the reaction was not affected by sodium azide and carbonyl cyanide p-trifluometoxyphenyl-hydrazone, had a biphasic response to Mg2+ (maximal at 513 [mu]m), and was insensitive to ionic strength. At 20[deg]C ATP-ADP exchange was pH insensitive. At both temperatures ATP hydrolysis displayed a bell-shaped response, with a maximum around pH 6.0 to 6.5. Because no adenylate kinase activity was detected under any condition, these results demonstrate the existence of an ATP-ADP exchange reaction catalyzed by the plant H+-ATPase
MH  - A
MH  - ADP
MH  - atp
MH  - azide
MH  - catalytic
MH  - DEPENDENCE
MH  - equilibrium
MH  - H+-ATPase
MH  - Hydrolysis
MH  - INTERACTION
MH  - membrane
MH  - model
MH  - pH
MH  - plant
MH  - Site
MH  - Sodium
MH  - sodium azide
MH  - Temperature
RP  - NOT IN FILE
NT  - Division de Biofisica, Instituto de Investigacion Medica "Mercedes y Martin Ferreyra," Casilla de Correo 389, 5000 Cordoba, Argentina
SO  - Plant Physiol 1997 Aug ;114(4):1397-1403

1437
UI  - 813
AU  - Hisabori T
AU  - Kato Y
AU  - Motohashi K
AU  - Kroth-Pancic P
AU  - Strotmann H
AU  - Amano T
AD  - Research Laboratory of Resources Utilization, Tokyo Institute of Technology, Yokohama, Japan thisabor@restitechacjp
TI  - The regulatory functions of the gamma and epsilon subunits from chloroplast CF1 are transferred to the core complex, alpha3beta3, from thermophilic bacterial F1
AB  - The expression plasmids for the subunit gamma (gamma(c)) and the subunit epsilon (epsilon(c)) of chloroplast coupling factor (CF1) from spinach were constructed, and the desired proteins were expressed in Escherichia coli. Both expressed subunits were obtained as inclusion bodies. When recombinant gamma(c) was mixed with recombinant alpha and beta subunits of F1 from thermophilic Bacillus PS3 (TF1), a chimeric subunit complex (alpha3beta3gamma(c)) was reconstituted and it showed significant ATP hydrolysis activity. The ATP hydrolysis activity of this complex was enhanced in the presence of dithiothreitol and suppressed by the addition of CuCl2, which induces formation of a disulfide bond between two cysteine residues in gamma(c). Hence, this complex has similar modulation characteristics as CF1. The effects of recombinant epsilon(c) and epsilon subunit from TF1 (epsilon(t)) on alpha3beta3gamma(c) were also investigated. Epsilon(c) strongly inhibited the ATP hydrolysis activity of chimeric alpha3beta3gamma(c) complex but epsilon(t) did not. The inhibition was abolished and the ATP hydrolysis activity was recovered when methanol was added to the assay medium. The addition of epsilon(c) or epsilon(t) to the alpha3beta3gamma(t) complex, which is the authentic subunit complex from TF1, resulted in weak stimulation of the ATP hydrolysis activity. These results suggest that (a) the specific regulatory function of gamma(c) can be transferred to the bacterial subunit complex; (b) the interaction between the gamma(c) subunit and epsilon(c) strongly affects the enzyme activity, which was catalyzed at the catalytic sites that reside on the alpha3beta3 core
RP  - NOT IN FILE
NT  - UI - 97433303LA - engRN - 0 (Chimeric Proteins)RN - 0 (Recombinant Proteins)RN - 67-56-1 (Methanol)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19971002IS - 0014-2956SB - IMCY - GERMANYJC - EMZ
UR  - PM:9288943
SO  - Eur J Biochem 1997 Aug 1 ;247(3):1158-1165

1438
UI  - 816
AU  - Hu D
AU  - Fiedler HR
AU  - Golan T
AU  - Edelman M
AU  - Strotmann H
AU  - Shavit N
AU  - Leu S
AD  - Doris and Bertie Black Center for Bioenergetics in Life Sciences, Ben Gurion University of the Negev, Beer Sheva 84105, Israel
TI  - Catalytic properties and sensitivity to tentoxin of Chlamydomonas reinhardtii ATP synthases changed in codon 83 of atpB by site-directed mutagenesis
AB  - The participation of the amino acid beta83 in determining the sensitivity of chloroplast ATP synthases to tentoxin was reported previously. We have changed codon 83 of the Chlamydomonas reinhardtii atpB gene by site-directed mutagenesis to further examine the role of this amino acid in the response of the ATP synthase to tentoxin and in the mechanism of ATP synthesis and hydrolysis. Amino acid beta83 was changed from Glu to Asp (betaE83D) and to Lys (betaE83K), and the highly conserved tetrapeptide betaT82-E83-G84-L85 (DeltaTEGL) was deleted. Mutant strains were produced by particle gun transformation of atpB deletion mutants cw15DeltaatpB and FUD50 with the mutated atpB genes. The transformants containing the betaE83D and betaE83K mutant genes grew well photoautotrophically. The DeltaTEGL transformant did not grow photoautotrophically, and no CF1 subunits were detected by immunostaining of Western blots using CF1 specific antibodies. The rates of ATP synthesis at clamped DeltapH with thylakoids isolated from cw15 and the two mutants, betaE83D and betaE83K, were similar. However, only the phosphorylation activity of the mutant betaE83D was inhibited by tentoxin with 50% inhibition attained at 4 microM. These results confirm that amino acid beta83 is critical in determining the response of ATP synthase to tentoxin. The rates of the latent Mg-ATPase activity of the CF1s isolated from cw15, betaE83D, and betaE83K were similar and could be enhanced by heat, alcohols, and octylglucoside. As in the case of the membrane-bound enzyme, only CF1 from the betaE83D mutant was sensitive to tentoxin. A lower alcohol concentration was required for optimal stimulation of the ATPase of the betaE83K-CF1 than that of CF1 from the other two strains. Moreover, the optimal activity of the betaE83K-CF1 was also lower. These results suggest that introduction of an amino acid with a positively charged side chain in position 83 in the "crown" domain affects the active conformation of the CF1-ATPase
RP  - NOT IN FILE
NT  - UI - 97190265LA - engRN - 0 (Codon)RN - 0 (Glucosides)RN - 0 (Multienzyme Complexes)RN - 0 (Mycotoxins)RN - 0 (Peptides, Cyclic)RN - 28540-82-1 (tentoxin)RN - 29836-26-8 (octyl-beta-D-glucoside)RN - 64-17-5 (Ethanol)RN - 67-56-1 (Methanol)RN - EC 2.7.4 (Phosphotransferases (Phosphate Group Acceptor))RN - EC 2.7.4.- (ATP synthetase complex)RN - EC 3.6.1.- (Ca(2+) Mg(2+)-ATPase)PT - Journal ArticleDA - 19970415IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:9038147
SO  - J Biol Chem 1997 Feb 28 ;272(9):5457-5463

1439
UI  - 288
AU  - Junge W
AU  - Lill H
AU  - Engelbrecht S
AD  - Universitat Osnabruck, Abt Biophysik, Germany junge@uni-osnabrueckde
TI  - ATP synthase: an electrochemical transducer with rotatory mechanics
AB  - ATP synthase (F0F1-ATPase) uses proton- or sodium-motive force to produce ATP form ADP and P(i). Three lines of experiment have recently demonstrated large-scale intersubunit rotation during ATP hydrolysis by F1. We discuss how ion flow through the membrane-intrinsic portion, F0, may generate torque and how this might be transmitted between stator and rotor to finally expel spontaneously formed ATP from F1 into water
RP  - NOT IN FILE
NT  - UI - 98060078LA - engRN - 0 (Protons)RN - 56-65-5 (Adenosine Triphosphate)RN - 7732-18-5 (Water)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 19980115IS - 0968-0004SB - IMCY - ENGLANDJC - WEF
UR  - PM:9397682
SO  - Trends Biochem Sci 1997 Nov ;22(11):420-423

1440
UI  - 37
AU  - Kaim G
AU  - Wehrle F
AU  - Gerike U
AU  - Dimroth P
AD  - Mikrobiologisches Institut, Eidgenossische Technische Hochschule, ETH- Zentrum, Schmelzbergstrasse 7, CH-8092 Zurich, Switzerland
TI  - Molecular basis for the coupling ion selectivity of F1F0 ATP synthases: probing the liganding groups for Na+ and Li+ in the c subunit of the ATP synthase from Propionigenium modestum
AB  - The conserved glutamate residue at position 65 of the Propionigenium modestum c subunit is directly involved in binding and translocation of Na+ across the membrane. The site-specific introduction of the cQ32I and cS66A substitutions in the putative vicinity to cE65 inhibited growth of the single-site mutants on succinate minimal agar, indicating that both amino acid residues are important for proper function of the oxidative phosphorylation system. This growth inhibition was abolished, however, if the cF84L/cL87V double mutation was additionally present in the P. modestum c subunit. The newly constructed Escherichia coli strain MPC848732I, harboring the cQ32I/cF84L/cL87V triple mutation, revealed a change in the coupling ion specificity from Na+ to H+. ATP hydrolysis by this enzyme was therefore not activated by NaCl, and ATP- driven H+ transport was not affected by this alkali salt. Both activities were influenced, however, by LiCl. These data demonstrate the loss of the Na+ binding site and retention of Li+ and H+ binding sites within this mutant ATPase. In the E. coli strain MPC848766A (cS66A/cF84L/cL87V), the specificity of the ATPase was further restricted to H+ as the exclusive coupling ion. Therefore, neither Na+ nor Li+ stimulated the ATPase activity, and no ATP-driven Li+ transport was observed. The ATPase of the E. coli mutant MPC32N (cQ32N) was activated by NaCl and LiCl. The mutant ATPase exhibited a 5-fold higher Km for NaCl but no change in the Km for LiCl in comparison to that of the parent strain. These results demonstrate that the binding of Na+ to the c subunit of P. modestum requires liganding groups provided by Q32, E65, and S66. For the coordination of Li+, two liganding partners, E65 and S66, are sufficient, and H+ translocation was mediated by E65 alone
RP  - NOT IN FILE
NT  - UI - 97375643LA - engRN - 0 (Chimeric Proteins)RN - 0 (Ligands)RN - 7439-93-2 (Lithium)RN - 7440-23-5 (Sodium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19970808IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:9230051
SO  - Biochemistry 1997 Jul 29 ;36(30):9185-9194

1441
UI  - 21183
AU  - Kalaidzidis IV
AU  - Kaulen AD
AD  - Department of Photobiochemistry, AN Belozersky Institute of Physico- Chemical Biology, Moscow State University, Russia
TI  - Cl- -dependent photovoltage responses of bacteriorhodopsin: comparison of the D85T and D85S mutants and wild-type acid purple form
AB  - Laser flash-induced photovoltage responses of the D85S and D85T mutants as well as of the wild-type acid blue form are similar and reflect intraprotein charge redistribution caused by retinal isomerization. The Cl- -induced transition of all of these blue forms into purple ones is accompanied by the appearance of electrogenic stages, which is probably associated with Cl- translocation in the cytoplasmic direction. Cl- translocation efficiency of these purple forms is much lower than that of the proton transport by the wild-type bacteriorhodopsin. The values of the efficiency do not exceed 15, 8 and 3% for the D85T, D85S and wild-type acid purple form, respectively. Cl- induces an additional electrogenic phase in the photovoltage responses of the D85S mutant and the wild-type acid purple form. This phase is supposed to be associated with the reversible Cl- movement in the extracellular direction. It is interesting that this component is absent in the photovoltage response of the D85T mutant which has, like halorhodopsin, a threonine residue at position 85
MH  - A
MH  - ACID
MH  - Bacteriorhodopsin
MH  - electrogenic
MH  - Movement
MH  - mutant
MH  - proton
MH  - RESIDUE
MH  - retinal
MH  - translocation
MH  - transport
RP  - NOT IN FILE
NT  - UI - 98088996LA - engRN - 0 (Chlorides)RN - 53026-44-1 (Bacteriorhodopsins)PT - Journal ArticleDA - 19980202IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:9428720
SO  - FEBS Lett 1997 Dec 1 ;418(3):239-242

1442
UI  - 707
AU  - Kato Y
AU  - Matsui T
AU  - Tanaka N
AU  - Muneyuki E
AU  - Hisabori T
AU  - Yoshida M
AD  - Research Laboratory of Resources Utilization, R-1, Tokyo Institute of Technology, 4259 Nagatsuta, Yokohama, 226, Japan
TI  - Thermophilic F1-ATPase is activated without dissociation of an endogenous inhibitor, epsilon subunit
AB  - Subunit complexes (alpha3beta3gamma, alpha3beta3gammadelta, alpha3beta3gammaepsilon, and alpha3beta3gammadeltaepsilon) of thermophilic F1-ATPase were prepared, and their catalytic properties were compared to know the role of delta and epsilon subunits in catalysis. The presence of delta subunit in the complexes had slight inhibitory effect on the ATPase activity. The effect of epsilon subunit was more profound. The (-epsilon) complexes, alpha3beta3gamma and alpha3beta3gammadelta, initiated ATP hydrolysis without a lag. In contrast, the (+epsilon) complexes, alpha3beta3gammaepsilon and alpha3beta3gammadeltaepsilon, started hydrolysis of ATP (<700 microM) with a lag phase that was gradually activated during catalytic turnover. As ATP concentration increased, the lag phase of the (+epsilon) complexes became shorter, and it was not observed above 1 mM ATP. Analysis of binding and hydrolysis of the ATP analog, 2',3'-O- (2,4,6-trinitrophenyl)-ATP, suggested that the (+epsilon) complexes bound substrate only slowly. Differing from Escherichia coli F1-ATPase, the activation of the (+epsilon) complexes from the lag phase was not due to dissociation of epsilon subunit since the re-isolated activated complex retained epsilon subunit. This indicates that there are two alternative forms of the (+epsilon) complex, inhibited form and activated form, and the inhibited one is converted to the activated one during catalytic turnover
RP  - NOT IN FILE
NT  - UI - 97460072LA - engRN - 0 (2',3'-O-(2,4,6-trinitrophenyl)adenosine 5'-triphosphate)RN - 0 (DNA Primers)RN - 0 (Enzyme Inhibitors)RN - 0 (Macromolecular Systems)RN - 0 (Recombinant Proteins)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19971022IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:9312092
SO  - J Biol Chem 1997 Oct 3 ;272(40):24906-24912

1443
UI  - 629
AU  - Knowles AF
AU  - Penefsky HS
AD  - Department of Biochemistry and Molecular Biology, SUNY Health Science Center, Syracuse, NY 13210, USA afknowles@worldnetattnet
TI  - Reconstitution of beef heart mitochondrial F0F1 in reverse phase evaporation vesicles
AB  - Beef heart mitochondrial F0F1 was reconstituted in proteoliposomes by a new procedure. MF0F1 was inserted in preformed reverse phase evaporation vesicles of large diameters prepared from asolectin (MF0F1- REV). Reconstitution was mediated by Triton X-100, which was subsequently removed by treatment with Bio-Beads. Parameters which resulted in optimal reconstitution were described. The MF0F1-REV proteoliposomes catalyzed an exchange between Pi and ATP and were capable of proton pumping. Both reactions were inhibited by oligomycin and uncoupler of oxidative phosphorylation. The range of Pi-ATP exchange activity of the proteoliposomes (70-110 nmol min[-1] mg[-1]) compared favorably with activities obtained in vesicles reconstituted by cholate dialysis or cholate dilution. The most important aspect of this method is that, unlike other reconstitution methods, exogenous F1 and other coupling factors are not required to obtain high Pi-ATP exchange activity by MF0F1-REV. This simple and rapid reconstitution procedure should be useful for future studies dealing with functional analysis of MF0F1
RP  - NOT IN FILE
NT  - UI - 98037474LA - engRN - 0 (Detergents)RN - 0 (Liposomes)RN - 0 (Oligomycins)RN - 0 (Phospholipids)RN - 0 (Proteolipids)RN - 0 (Uncoupling Agents)RN - 0 (proteoliposomes)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM21737/GM/NIGMSDA - 19971210IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:9371423
SO  - Biochim Biophys Acta 1997 Oct 23 ;1329(2):311-320

1444
UI  - 504
AU  - Ko YH
AU  - Bianchet M
AU  - Amzel LM
AU  - Pedersen PL
AD  - Department of Biological Chemistry, The Johns Hopkins University, School of Medicine, Baltimore, Maryland 21205-2185, USA
TI  - Novel insights into the chemical mechanism of ATP synthase. Evidence that in the transition state the gamma-phosphate of ATP is near the conserved alanine within the P-loop of the beta-subunit
AB  - The chemical mechanism by which the F1 moiety of ATP synthase hydrolyzes and synthesizes ATP remains unknown. For this reason, we have carried out studies with orthovanadate (Vi), a phosphate analog which has the potential of "locking" an ATPase, in its transition state by forming a MgADP.Vi complex, and also the potential, in a photochemical reaction resulting in peptide bond cleavage, of identifying an amino acid very near the gamma-phosphate of ATP. Upon incubating purified rat liver F1 with MgADP and Vi for 2 h to promote formation of a MgADP.Vi-F1 complex, the ATPase activity of the enzyme was markedly inhibited in a reversible manner. When the resultant complex was formed in the presence of ultraviolet light inhibition could not be reversed, and SDS-polyacrylamide gel electrophoresis revealed, in addition to the five known subunit bands characteristic of F1 (i.e. alpha, beta, gamma, delta, and epsilon), two new electrophoretic species of 17 and 34 kDa. Western blot and N-terminal sequencing analyses identified both bands as arising from the beta subunit with the site of peptide bond cleavage occurring at alanine 158, a conserved residue within F1-ATPases and the third residue within the nucleotide binding consensus GX4GK(T/S) (P-loop). Quantification of the amount of ADP bound within the MgADP. Vi-F1 complex revealed about 1.0 mol/mol F1, while quantification of the peptide cleavage products revealed that no more than one beta subunit had been cleaved. Consistent with the cleavage reaction involving oxidation of the methyl group of alanine was the finding that [3H] from NaB[3H]4 incorporates into MgADP.Vi-F1 complex following treatment with ultraviolet light. These novel findings provide information about the transition state involved in the hydrolysis of ATP by a single beta subunit within F1- ATPases and implicate alanine 158 as residing very near the gamma- phosphate of ATP during catalysis. When considered with earlier studies on myosin and adenylate kinase, these studies also implicate a special role for the third residue within the GX4GK(T/S) sequence of many other nucleotide-binding proteins
RP  - NOT IN FILE
NT  - UI - 97373590LA - engRN - 0 (Borohydrides)RN - 0 (Phosphoric Acid Esters)RN - 0 (Vanadates)RN - 16940-66-2 (sodium borohydride)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 6898-94-8 (Alanine)RN - 7786-30-3 (Magnesium Chloride)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - CA10951/CA/NCIDA - 19970909IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:9228065
SO  - J Biol Chem 1997 Jul 25 ;272(30):18875-18881

1445
UI  - 815
AU  - Krenn BE
AU  - Strotmann H
AU  - van Walraven HS
AU  - Scholts MJ
AU  - Kraayenhof R
AD  - Institute of Molecular Biological Sciences, BioCentrum Amsterdam, Vrije Universiteit, De Boelelaan 1087, 1081 HV Amsterdam, The Netherlands
TI  - The ATP synthase gamma subunit provides the primary site of activation of the chloroplast enzyme: experiments with a chloroplast-like Synechocystis 6803 mutant
AB  - The activation characteristics of the F1Fo-ATP synthase (where F1 and Fo are the hydrophilic and membrane-bound parts respectively of the enzyme) from Synechocystis 6803 wild-type and a Synechocystis 6803 mutant with a chloroplast-like insertion in the gamma subunit have been studied. Activation of the ATP synthase in wild-type and mutant membrane vesicles was performed by acid-base transition-induced generation of a proton motive force (Delta mu H+). Since the mutant containing the regulatory segment of the chloroplast gamma subunit showed thiol-modulation (typical of the chloroplast enzyme), this segment is indeed involved in the regulation of enzyme activation. It is shown that the ATP synthase from Synechocystis 6803 wild type corresponds functionally to the reduced form of the chloroplast ATP synthase, in view of the low Delta mu H+ required for activation of the enzyme and the high stability of the active state. Both the cyanobacterial wild-type and mutant ATP synthases can be activated by methanol, which apparently does not require the presence of the gamma subunit regulatory segment
RP  - NOT IN FILE
NT  - UI - 97270632LA - engRN - 0 (Bacterial Proteins)RN - 0 (Plant Proteins)RN - 56-65-5 (Adenosine Triphosphate)RN - 56-89-3 (Cystine)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19970627IS - 0264-6021SB - IMCY - ENGLANDJC - 9YO
UR  - PM:9169620
SO  - Biochem J 1997 May 1 ;323 ( Pt 3)():841-845

1446
UI  - 21076
AU  - Krulwich TA
AU  - Ito M
AU  - Gilmour R
AU  - Guffanti AA
AD  - Department of Biochemistry, Mount Sinai School of Medicine of CUNY, New York, NY, USA krulwich@msvaxmssmedu
TI  - Mechanisms of cytoplasmic pH regulation in alkaliphilic strains of Bacillus
AB  - The central challenge for extremely alkaliphilic Bacillus species is the need to establish and sustain a cytoplasmic pH that is over two units lower than the highly alkaline medium. Its centrality is suggested by the strong correlation between the growth rate in the upper range of pH for growth, i.e., at values above pH 10.5, and the cytoplasmic pH. The diminishing growth rate at extremely high pH values correlates better with the rise in cytoplasmic pH than with other energetic parameters. There are also general adaptations of alkaliphiles that are crucial prerequisites for pH homeostasis as well as other cell functions, i.e., the reduced basic amino acid content of proteins or segments thereof that are exposed to the medium, and there are other challenges of alkaliphily that emerge from solution of the cytoplasmic pH problem, i.e., reduction of the chemiosmotic driving force. For cells growing on glucose, strong evidence exists for the importance of acidic cell wall components, teichuronic acid and teichuronopeptides, in alkaliphily. These wall macromolecules may provide a passive barrier to ion flux. For cells growing on fermentable carbon sources, this and other passive mechanisms may have a particularly substantial role, but for cells growing on both fermentable and nonfermentable substrates, an active Na+-dependent cycle is apparently required for alkaliphily and the alkaliphile's remarkable capacity for pH homeostasis. The active cycle involves primary establishment of an electrochemical gradient via proton extrusion, a secondary electrogenic Na+/H+ antiport to achieve net acidification of the cytoplasm relative to the outside pH, and mechanisms for Na+ re-entry. Recent work in several laboratories on the critical antiporters involved in this cycle has begun to clarify the number and characteristics of the porters that support active mechanisms of pH homeostasis
MH  - A
MH  - ACID
MH  - ACTIVE
MH  - Bacillus
MH  - Biochemistry
MH  - Cells
MH  - Cytoplasm
MH  - electrogenic
MH  - function
MH  - Glucose
MH  - Homeostasis
MH  - ion
MH  - mechanism
MH  - MECHANISMS
MH  - pH
MH  - protein
MH  - Proteins
MH  - proton
MH  - regulation
MH  - review
MH  - secondary
RP  - NOT IN FILE
NT  - UI - 98343891LA - engPT - Journal ArticlePT - ReviewPT - Review, TutorialID - GM28454/GM/NIGMSDA - 19980805IS - 1431-0651SB - IMSB - SCY - GERMANY
UR  - PM:9680297
SO  - Extremophiles 1997 Nov ;1(4):163-169

1447
UI  - 985
AU  - Lacapere JJ
AU  - Stokes DL
AU  - Mosser G
AU  - Ranck JL
AU  - Leblanc G
AU  - Rigaud JL
TI  - Two-dimensional crystal formation from solubilized membrane proteins using Bio-Beads to remove detergent
MH  - Bacillus
MH  - Ca(2+)-Transporting ATPase
MH  - Crystallization
MH  - Detergents
MH  - enzymology
MH  - H(+)-Transporting ATP Synthase
MH  - isolation & purification
MH  - Kinetics
MH  - Membrane Proteins
MH  - Methods
MH  - Microscopy,Electron
MH  - Microspheres
MH  - Permeases
MH  - Proteins
MH  - Sarcoplasmic Reticulum
MH  - Support,Non-U.S.Gov't
MH  - ultrastructure
RP  - NOT IN FILE
NT  - Institut Curie, CNRS, Paris, France
SO  - Ann N Y Acad Sci 1997 Nov 3 ;834:9-18.():9-18

1448
UI  - 21239
AU  - Leikin S
AU  - Parsegian VA
AU  - Yang W
AU  - Walrafen GE
AD  - Division of Computer Research and Technology and National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA
TI  - Raman spectral evidence for hydration forces between collagen triple helices
AB  - Hydration forces are thought to result from the energetic cost of water rearrangement near macromolecular surfaces. Raman spectra, collected on the same collagen samples on which these forces were measured, reveal a continuous change in water hydrogen-bonding structure as a function of separation between collagen triple helices. The varying spectral parameters track the force-distance curve. The energetic cost of water "restructuring," estimated from the spectra, is consistent with the measured energy of intermolecular interaction. These correlations support the idea that the change in water structure underlies the exponentially varying forces seen in this system at least over the 13- 18-A range of interaxial separations
MH  - A
MH  - development
MH  - function
MH  - Human
MH  - Hydrogen Bonding
MH  - Macromolecular Systems
MH  - spectra
MH  - structure
MH  - SURFACE
MH  - SYSTEM
MH  - SYSTEMS
MH  - Water
RP  - NOT IN FILE
NT  - UI - 97470963LA - engRN - 0 (Macromolecular Systems)RN - 7732-18-5 (Water)RN - 9007-34-5 (Collagen)PT - Journal ArticleDA - 19971124IS - 0027-8424SB - IMCY - UNITED STATES
UR  - PM:9326606
SO  - Proc Natl Acad Sci U S A 1997 Oct 14 ;94(21):11312-11317

1449
UI  - 417
AU  - Lobau S
AU  - Weber J
AU  - Wilke-Mounts S
AU  - Senior AE
AD  - Department of Biochemistry, University of Rochester Medical Center, Rochester, New York 14642, USA
TI  - F1-ATPase, roles of three catalytic site residues
AB  - Three critical residues, beta-Lys-155, beta-Asp-242, and beta-Glu-181, situated close to the gamma-phosphate of MgATP in F1-ATPase catalytic sites, were investigated. The mutations betaK155Q, betaD242N, and betaE181Q were each combined with the betaY331W mutation; the fluorescence signal of beta-Trp-331 was used to determine MgATP, MgADP, ATP, and ADP binding parameters for the three catalytic sites of the enzyme. The quantitative contribution of side chains to binding energy at all three catalytic sites was calculated. The following conclusions were made. The major functional interaction of beta-Lys-155 is with the gamma-phosphate of MgATP and is of primary importance at site 1 (the site of highest affinity) and site 2. Release of MgATP during oxidative phosphorylation requires conformational re-positioning of this residue. The major functional interaction of beta-Asp-242 is with the magnesium of the magnesium nucleotide at site 1; it has little or no influence at site 2 or 3. In steady-state turnover, the MgATP hydrolysis reaction occurs at site 1. beta-Glu-181 contributes little to nucleotide binding; its major catalytic effect derives apparently from a role in reaction chemistry per se. This work also emphasizes that nucleotide binding cooperativity shown by the three catalytic sites toward MgATP and MgADP is absolutely dependent on the presence of magnesium
RP  - NOT IN FILE
NT  - UI - 97166219LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSDA - 19970402IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:9013618
SO  - J Biol Chem 1997 Feb 7 ;272(6):3648-3656

1450
UI  - 416
AU  - Lobau S
AU  - Weber J
AU  - Senior AE
AD  - Department of Biochemistry and Biophysics, University of Rochester Medical Center, NY 14642, USA
TI  - Nucleotide occupancy of F1-ATPase catalytic sites under crystallization conditions
AB  - Using site-directed tryptophan fluorescence we studied nucleotide occupancy of the catalytic sites of Escherichia coli F1-ATPase, under conditions used previously for crystallization and X-ray structure analysis of the bovine mitochondrial enzyme [Abrahams et al. (1994) Nature 370, 621-628]. We found that only two of the three catalytic sites were filled in the E. coli enzyme under these conditions (250 microM MgAMPPNP plus 5 microM MgADP), consistent with what was reported in the bovine F1 X-ray structure. However, subsequent addition of a physiological concentration of MgATP readily filled the third catalytic site. Therefore the enzyme form seen in the X-ray structure results from the fact that it is obtained under sub-saturating nucleotide conditions. The data show that the X-ray structure is compatible with a catalytic mechanism in which all three F1-ATPase catalytic sites must fill with MgATP to initiate steady-state hydrolysis [e.g. Weber and Senior (1996) Biochim. Biophys. Acta 1275, 101-104]. The data further demonstrate that the site-directed tryptophan fluorescence technique can provide valuable support for F1 crystallography studies
RP  - NOT IN FILE
NT  - UI - 97228568LA - engRN - 0 (Oligodeoxyribonucleotides)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSDA - 19970404IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:9074628
SO  - FEBS Lett 1997 Mar 3 ;404(1):15-18

1451
UI  - 18356
AU  - Maroti P
AU  - Wraight CA
TI  - Kinetics of H+ ion binding by the P+QA-state of bacterial photosynthetic reaction centers: rate limitation within the protein
MH  - A
MH  - ACTIVATION
MH  - BINDING
MH  - Binding Sites
MH  - Biophysics
MH  - Chemistry
MH  - CONSTANT
MH  - DEPENDENCE
MH  - deuterium oxide
MH  - Diffusion
MH  - drug effects
MH  - Electric Conductivity
MH  - electron
MH  - electron transfer
MH  - Electron Transport
MH  - Electron-transfer
MH  - H+
MH  - herbicides,triazine
MH  - Hydrogen-Ion Concentration
MH  - indicator
MH  - ion
MH  - Kinetics
MH  - Light
MH  - metabolism
MH  - Models,Theoretical
MH  - Osmolar Concentration
MH  - Oxidation-Reduction
MH  - pH
MH  - pH-indicator
MH  - pharmacology
MH  - Photosynthetic Reaction Center,Bacterial
MH  - protein
MH  - proton
MH  - Proton transfer
MH  - Proton-transfer
MH  - protonation
MH  - Protons
MH  - quinone
MH  - Quinones
MH  - rate constant
MH  - reaction center
MH  - RESIDUE
MH  - Rhodobacter
MH  - Rhodobacter sphaeroides
MH  - secondary
MH  - sphaeroides
MH  - Support,U.S.Gov't,Non-P.H.S.
MH  - SURFACE
MH  - Temperature
MH  - theory
MH  - Thermodynamics
MH  - TRANSFER
MH  - triazines
MH  - Viscosity
RP  - IN FILE
NT  - The kinetics of flash-induced H+ ion binding by isolated reaction centers (RCs) of Rhodobacter sphaeroides, strain R-26, were measured, using pH indicators and conductimetry, in the presence of terbutryn to block electron transfer between the primary and secondary quinones (QA and QB), and in the absence of exogenous electron donors to the oxidized primary donor, P+, i.e., the P+QA-state. Under these conditions, proton binding by RCs is to the protein rather than to any of the cofactors. After light activation to form P+QA-, the kinetics of proton binding were monoexponential at all pH values studied. At neutral pH, the apparent bimolecular rate constant was close to the diffusional limit for proton transfer in aqueous solution (approximately 10(11) M-1 s-1), but increased significantly in the alkaline pH range (e.g., 2 x 10(13) M-1 s-1 at pH 10). The average slope of the pH dependence was -0.4 instead of -1.0, as might be expected for a H+ diffusion-controlled process. High activation energy (0.54 eV at pH 8.0) and weak viscosity dependence showed that H+ ion uptake by RCs is not limited by diffusion. The salt dependence of the H+ ion binding rate and the pK values of the protonatable amino acid residues of the reaction center implicated surface charge influences, and Gouy-Chapman theory provided a workable description of the ionic effects as arising from modulation of the pH at the surface of the RC. Incubation in D2O caused small increases in the pKs of the protonatable groups and a small, pH (pD)-dependent slowing of the binding rate. The salt, pH, temperature, viscosity, and D2O dependences of the proton uptake by RCs in the P+QA- state were accounted for by three considerations: 1) parallel pathways of H+ delivery to the RC, contributing to the observed (net) H+ disappearance; 2) rate limitation of the protonation of target groups within the protein by conformational dynamics; and 3) electrostatic influences of charged groups in the protein, via the surface pH Center for Biophysics and Computational Biology, University of Illinois, Urbana 81801-3838, USAPMID- 9199801
SO  - Biophys J 1997  ;73(1):367-381

1452
UI  - 714
AU  - Matsui T
AU  - Muneyuki E
AU  - Honda M
AU  - Allison WS
AU  - Dou C
AU  - Yoshida M
AD  - Research Laboratory of Resources Utilization, Tokyo Institute of Technology, 4259 Nagatsuta, Yokohama 226, Japan
TI  - Catalytic activity of the alpha3beta3gamma complex of F1-ATPase without noncatalytic nucleotide binding site
AB  - A mutant alpha3beta3gamma complex of F1-ATPase from thermophilic Bacillus PS3 was generated in which noncatalytic nucleotide binding sites lost their ability to bind nucleotides. It hydrolyzed ATP at an initial rate with cooperative kinetics (Km(1), 4 microM; Km(2), 135 microM) similar to the wild-type complex. However, the initial rate decayed rapidly to an inactivated form. Since the inactivated mutant complex contained 1.5 mol of ADP/mol of complex, this inactivation seemed to be caused by entrapping inhibitory MgADP in a catalytic site. Indeed, the mutant complex was nearly completely inactivated by a 10 min prior incubation with equimolar MgADP. Analysis of the progress of inactivation after initiation of ATP hydrolysis as a function of ATP concentration indicated that the inactivation was optimal at ATP concentrations in the range of Km(1). In the presence of ATP, the wild- type complex dissociated the inhibitory [3H]ADP preloaded onto a catalytic site whereas the mutant complex did not. Lauryl dimethylamineoxide promoted release of preloaded inhibitory [3H]ADP in an ATP-dependent manner and partly restored the activity of the inactivated mutant complex. Addition of ATP promoted single-site hydrolysis of 2',3'-O-(2,4,6-trinitrophenyl)-ATP preloaded at a single catalytic site of the mutant complex. These results indicate that intact noncatalytic sites are essential for continuous catalytic turnover of the F1-ATPase but are not essential for catalytic cooperativity of F1-ATPase observed at ATP concentrations below approximately 300 microM
RP  - NOT IN FILE
NT  - UI - 97236767LA - engRN - 0 (Dimethylamines)RN - 0 (Fluorescent Dyes)RN - 0 (Surface-Active Agents)RN - 1643-20-5 (dodecyldimethylamine oxide)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 61368-63-6 (2',3'-O-(2,4,6-trinitro-cyclohexadienylidine)adenosine 5'- triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19970502IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:9079639
SO  - J Biol Chem 1997 Mar 28 ;272(13):8215-8221

1453
UI  - 36
AU  - Matthey U
AU  - Kaim G
AU  - Dimroth P
AD  - Mikrobiologisches Institut, Eidgenossische Technische Hochschule, Zurich, Switzerland
TI  - Subunit c from the sodium-ion-translocating F1F0-ATPase of Propionigenium modestum--production, purification and properties of the protein in dodecylsulfate solution
AB  - Escherichia coli strain PEF42 produces a sodium-ion-dependent hybrid F1F0-ATPase consisting of the Propionigenium modestum subunits a, b, c and delta, of a hybrid alpha subunit and of the E. coli subunits beta, gamma and epsilon. The gene encoding subunit c of the P. modestum F1F0- ATPase was cloned into the pT7-7 expression vector to yield plasmid pT7c. E. coli PEF42 was transformed with plasmid pT7c together with plasmid pGP1-2, which harbours the gene for the T7 RNA polymerase. The production of the P. modestum subunit c was induced by a temperature shift from 30 degrees C to 42 degrees C for 30 min and led to an increased concentration of this protein in the membrane of the host strain. The c subunit produced in E. coli moved as a monomer in dodecylsulfate electrophoresis. The protein was extracted from the cells with chloroform/methanol, purified and incorporated into sodium dodecylsulfate micelles. Circular dichroism of subunit c in sodium dodecylsulfate showed a temperature-stable spectrum (between 20-60 degrees C) with a high proportion of alpah-helical structure. Upon incubation of subunit c with [14C]dicyclohexylcarbodiimide the protein became labelled in a sodium-ion-dependent manner, similar to the labelling observed if the purified F1F0-ATPase of P. modestum, was treated with the radioactive carbodiimide. The Na+-specific site was therefore retained in the isolated c subunit dissolved in dodecylsulfate
RP  - NOT IN FILE
NT  - UI - 97433263LA - engRN - 0 (Solutions)RN - 151-21-3 (Sodium Dodecyl Sulfate)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 7440-23-5 (Sodium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19971002IS - 0014-2956SB - IMCY - GERMANYJC - EMZ
UR  - PM:9288903
SO  - Eur J Biochem 1997 Aug 1 ;247(3):820-825

1454
UI  - 19856
AU  - Milgrom YM
AU  - Cross RL
AD  - Department of Biochemistry and Molecular Biology State University of New York Health Science Center at Syracuse Syracuse, New York 13210, USA
TI  - Nucleotide-depleted beef heart F1-ATPase exhibits strong positive catalytic cooperativity
AB  - Catalytic cooperativity is a central feature of the binding change mechanism for F0F1-ATP synthases. However, in a recent publication (Reynafarje, B. D., and Pedersen, P. L. (1996) J. Biol. Chem. 271, 32546-32550), Reynafarje and Pedersen claim that cooperative effects are an artifact caused by endogenous nucleotides and that when such nucleotides are removed, the multiple catalytic sites on MF1 behave independently during ATP hydrolysis. In contrast to this conclusion, we show here that when ATP is loaded at a single catalytic site on nucleotide-depleted MF1, the rate of product release is accelerated by up to 5 x 10(4)-fold by the binding of ATP at adjacent catalytic sites. Hence, nucleotide-depleted MF1 is not an exception but does in fact show strong cooperative interactions. In addition, evidence is presented supporting a random order for product release during ATP hydrolysis
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthase
MH  - BINDING
MH  - Biochemistry
MH  - CHANGE MECHANISM
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - H(+)-Transporting ATP Synthase
MH  - Hydrolysis
MH  - mechanism
MH  - Nucleotides
MH  - P
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 98070389LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM 23152/GM/NIGMSDA - 19980122IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:9405423
SO  - J Biol Chem 1997 Dec 19 ;272(51):32211-32214

1455
UI  - 709
AU  - Muneyuki E
AU  - Odaka M
AU  - Yoshida M
AD  - Research Laboratory of Resources Utilization, Tokyo Institute of Technology, Yokohama, Japan
TI  - A single mutation at the catalytic site of TF1-alpha3beta3gamma complex switches the kinetics of ATP hydrolysis from negative to positive cooperativity
AB  - Previously, we reported the substitution of Tyr341 of the F1-ATPase beta subunit from a thermophilic Bacillus strain PS3 with leucine, cysteine, or alanine (M. Odaka et al. J. Biochem., 115 (1994) 789-796). These mutations resulted in a great decrease in the affinity of the isolated beta subunit for ATP-Mg and an increase in the apparent Km of the alpha3beta3gamma complex in ATP hydrolysis when examined above 0.1 mM ATP. Here, we examined the ATPase activity of the mutant complexes in a wide range of ATP concentration and found that the mutants exhibited apparent positive cooperativity in ATP hydrolysis. This is the first clear demonstration that a single mutation in the catalytic sites converts the kinetics from apparent negative cooperativity in the wild-type alpha3beta3gamma complex to apparent positive cooperativity. The conversion of apparent cooperativity could be explained in terms of a simple kinetic scheme based on the binding change model proposed by Boyer
RP  - NOT IN FILE
NT  - UI - 97431611LA - engRN - 52-90-4 (Cysteine)RN - 55520-40-6 (Tyrosine)RN - 56-65-5 (Adenosine Triphosphate)RN - 6898-94-8 (Alanine)RN - 7005-03-0 (Leucine)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19971030IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:9287116
SO  - FEBS Lett 1997 Aug 11 ;413(1):55-59

1456
UI  - 21036
AU  - Murata T
AU  - Takase K
AU  - Yamato I
AU  - Igarashi K
AU  - Kakinuma Y
AD  - Department of Biological Science and Technology, Science University of Tokyo, 2641 Yamazaki, Noda-shi, Chiba 278, Japan
TI  - Purification and reconstitution of Na+-translocating vacuolar ATPase from Enterococcus hirae
AB  - Vacuolar ATPases make up a family of proton pumps distributed widely from bacteria to higher organisms. An unusual member of this family, a sodium-translocating ATPase, has been found in the eubacterium Enterococcus hirae. We report here the purification of enterococcal Na+- ATPase from the plasma membrane of cells, whose ATPase content was highly amplified by expression of the cloned ntp operon that encodes this Na+-ATPase (ntpFIKECGABDHJ). The purified enzyme appears to consist of nine Ntp polypeptides, all the above except for the ntpH and ntpJ gene products. ATPase activity was strictly dependent on the presence of Na+ or Li+ ions and was inhibited by nitrate, N- ethylmaleimide, and the peptide antibiotic destruxin B. When the purified ATPase was reconstituted into liposomes prepared from Enterococcus faecalis phospholipids, ATP-driven Na+ uptake was observed; uptake was blocked by nitrate, destruxin B, and monensin, but it accelerated by carbonyl cyanide m-chlorophenylhydrazone and valinomycin. These data demonstrate that E. hirae Na+-ATPase is an electrogenic sodium pump of the vacuolar type. This is a promising system for research on the fundamental molecular structure and mechanism of vacuolar ATPase
MH  - A
MH  - Adenosinetriphosphatase
MH  - ATPase
MH  - Bacteria
MH  - Cells
MH  - electrogenic
MH  - Enzyme Inhibitors
MH  - Ethylmaleimide
MH  - inhibitor
MH  - ion
MH  - Ions
MH  - liposome
MH  - Liposomes
MH  - mechanism
MH  - membrane
MH  - Molecular Structure
MH  - Peptide Fragments
MH  - Phospholipids
MH  - protein
MH  - Proteins
MH  - Proteolipids
MH  - proteoliposome
MH  - proton
MH  - Proton Pump
MH  - purification
MH  - reconstitution
MH  - Sodium
MH  - structure
MH  - SYSTEM
MH  - Valinomycin
RP  - NOT IN FILE
NT  - UI - 97460069LA - engRN - 0 (Antibodies)RN - 0 (Enzyme Inhibitors)RN - 0 (Liposomes)RN - 0 (Peptide Fragments)RN - 0 (Proteolipids)RN - 0 (Recombinant Proteins)RN - 0 (proteoliposomes)RN - 7439-93-2 (Lithium)RN - 7440-23-5 (Sodium)RN - EC 3.6.1.- (sodium-translocating ATPase)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19971022IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:9312089
SO  - J Biol Chem 1997 Oct 3 ;272(40):24885-24890

1457
UI  - 9889
AU  - Naumann R
AU  - Jonczyk A
AU  - Hampel C
AU  - Ringsdorf H
AU  - Knoll W
AU  - Bunjes N
AU  - Graber P
TI  - Coupling of proton translocation through ATPase incorporated into supported lipid bilayers to an electrochemical process
AB  - H+-ATPase is incorporated into solid-supported lipid bilayers separated from the gold support by a peptide spacer. The translocation of protons across the lipid film to the inner side is coupled to the discharge of protons at the gold surface. The overall process is investigated by square wave voltammetry (SWV) and double potential-pulse chronoamperometry (CA). As a result, the formation of a proton gradient is monitored by SWV whereas currents measured by CA monitor the stationary state when the enzyme activity is directly coupled to the charge transfer at the electrode. These currents markedly depend on the number of ATPases present in the bilayer. (C) 1997 Elsevier Science S.A
MH  - ATPase
MH  - electrochemical coupling
MH  - GOLD
MH  - H+-ATPase
MH  - Hydrolysis
MH  - Lipid Bilayers
MH  - Liposomes
MH  - Membrane Proteins
MH  - Protons
MH  - solid-supported lipid layer
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - JournalArticleELSEVIER SCIENCE SA LAUSANNEMAYXM005LAUSANNENaumann R MERCK KGAA,D-64271 DARMSTADT,GERMANYBIOELECTROCHEM BIOENERGPO BOX 564, 1001 LAUSANNE, SWITZERLAND
AV  - MERCK KGAA,D-64271 DARMSTADT,GERMANY UNIV MAINZ,INST ORGAN CHEM,D-55099 MAINZ,GERMANY MAX PLANCK INST POLYMER RES,D-55021 MAINZ,GERMANY UNIV FREIBURG,INST PHYS CHEM,D-79104 FREIBURG,GERMANY
UR  - ISI:A1997XM00500021
SO  - Bioelectrochemistry and Bioenergetics 1997  ;42(2):241-247

1458
UI  - 716
AU  - Noji H
AU  - Yasuda R
AU  - Yoshida M
AU  - Kinosita K
AD  - Research Laboratory of Resources Utilization, Tokyo Institute of Technology, Yokohama, Japan
TI  - Direct observation of the rotation of F1-ATPase
AB  - Cells employ a variety of linear motors, such as myosin, kinesin and RNA polymerase, which move along and exert force on a filamentous structure. But only one rotary motor has been investigated in detail, the bacterial flagellum (a complex of about 100 protein molecules). We now show that a single molecule of F1-ATPase acts as a rotary motor, the smallest known, by direct observation of its motion. A central rotor of radius approximately 1 nm, formed by its gamma-subunit, turns in a stator barrel of radius approximately 5nm formed by three alpha- and three beta-subunits. F1-ATPase, together with the membrane-embedded proton-conducting unit F0, forms the H+-ATP synthase that reversibly couples transmembrane proton flow to ATP synthesis/hydrolysis in respiring and photosynthetic cells. It has been suggested that the gamma-subunit of F1-ATPase rotates within the alphabeta-hexamer, a conjecture supported by structural, biochemical and spectroscopic studies. We attached a fluorescent actin filament to the gamma-subunit as a marker, which enabled us to observe this motion directly. In the presence of ATP, the filament rotated for more than 100 revolutions in an anticlockwise direction when viewed from the 'membrane' side. The rotary torque produced reached more than 40 pN nm(-1) under high load
RP  - NOT IN FILE
NT  - UI - 97222141LA - engRN - 0 (Actins)RN - 0 (Recombinant Proteins)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19970407IS - 0028-0836SB - IMCY - ENGLANDJC - NSC
UR  - PM:9069291
SO  - Nature 1997 Mar 20 ;386(6622):299-302

1459
UI  - 152
AU  - Ogilvie I
AU  - Aggeler R
AU  - Capaldi RA
AD  - Institute of Molecular Biology, University of Oregon, Eugene, Oregon 97403-1229, USA
TI  - Cross-linking of the delta subunit to one of the three alpha subunits has no effect on functioning, as expected if delta is a part of the stator that links the F1 and F0 parts of the Escherichia coli ATP synthase
AB  - A mutant of the Escherichia coli F1F0-ATPase has been generated (alphaQ2C) in which the glutamine at position 2 of the alpha subunit has been replaced with a cysteine residue. Cu2+ treatment of ECF1 from this mutant cross-linked an alpha subunit to the delta subunit in high yield. Two different sites of disulfide bond formation were involved, i.e. between Cys90 (or the closely spaced Cys47) of alpha with Cys140 of delta, and between Cys2 of alpha and Cys140 of delta. Small amounts of other cross-linked products, including alpha-alpha, delta internal, and alpha-alpha-delta were obtained. In ECF1F0, there was no cross- linking between the intrinsic Cys of alpha and Cys140. Instead, the product generated between Cys2 of alpha and Cys140 of delta was obtained at near 90% yield. Small amounts of alpha-alpha and delta internal were present, and under high Cu2+ concentrations, alpha-alpha- delta was also formed. The ATPase activity of ECF1 and ECF1F0 was not significantly affected by the presence of these cross-links. When Cys140 of delta was first modified with N-ethylmaleimide in ECF1F0, an alpha-delta cross-link was still produced, although in lower yield, between Cys64 of delta and Cys2 of alpha. ATP hydrolysis-linked proton pumping of inner membranes from the mutant alpha2QC was only marginally affected by cross-linking of the alpha to the delta subunit. These results indicate that Cys140 and Cys64 of the delta subunit and Cys2 of the alpha subunit are in close proximity. This places the delta subunit near the top of the alpha-beta hexagon and not in the stalk region. As fixing the delta to the alpha by cross-linking does not greatly impair either the ATPase function of the enzyme, or coupled proton translocation, we argue that the delta subunit forms a portion of the stator linking F1 to F0
RP  - NOT IN FILE
NT  - UI - 97341211LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - 7440-50-8 (Copper)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - HL 24526/HL/NHLBIDA - 19970716IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:9195980
SO  - J Biol Chem 1997 Jun 27 ;272(26):16652-16656

1460
UI  - 713
AU  - Ohta H
AU  - Shirakawa H
AU  - Uchida K
AU  - Yoshida M
AU  - Matuo Y
AU  - Enami I
AD  - Department of Biology, Faculty of Science, Science University of Tokyo, Japan
TI  - Cloning and sequencing of the gene encoding the plasma membrane H(+)- ATPase from an acidophilic red alga, Cyanidium caldarium
AB  - A cDNA containing an open reading frame encoding the putative plasma membrane H(+)-ATPase in an acidophilic red alga, Cyanidium caldarium, was cloned and sequenced by means of PCR and Southern hybridization based on homologous sequences of P-type ATPases found in other organisms. The cloned cDNA is 3300 bp in length, containing a 2865 bp open reading frame encoding a polypeptide of 955 amino acids which has a predicted molecular mass of 105,371. The deduced amino acid sequence was found to be more homologous to those of P-type H(+)-ATPases from higher plants than that from the green alga Dunaliella bioculata
RP  - NOT IN FILE
NT  - UI - 97261359LA - engRN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19970501IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:9107313
SO  - Biochim Biophys Acta 1997 Mar 28 ;1319(1):9-13

1461
UI  - 21245
AU  - Ostermeier C
AU  - Harrenga A
AU  - Ermler U
AU  - Michel H
AD  - Max-Planck-Institut fur Biophysik, Abteilung fur Molekulare Membranbiologie, Heinrich-Hoffmann-Strasse 7, D-60528 Frankfurt, Germany
TI  - Structure at 2.7 A resolution of the Paracoccus denitrificans two- subunit cytochrome c oxidase complexed with an antibody FV fragment
AB  - The aa3 type cytochrome c oxidase consisting of the core subunits I and II only was isolated from the soil bacterium Paracoccus denitrificans and crystallized as complex with a monoclonal antibody Fv fragment. Crystals could be grown in the presence of a number of different nonionic detergents. However, only undecyl-beta-D-maltoside and cyclohexyl-hexyl-beta-D-maltoside yielded well-ordered crystals suitable for high resolution x-ray crystallographic studies. The crystals belong to space group P212121 and diffract x-rays to at least 2.5 A (1 A = 0.1 nm) resolution using synchrotron radiation. The structure was determined to a resolution of 2.7 A using molecular replacement and refined to a crystallographic R-factor of 20.5% (Rfree = 25.9%). The refined model includes subunits I and II and the 2 chains of the Fv fragment, 2 heme A molecules, 3 copper atoms, and 1 Mg/Mn atom, a new metal (Ca) binding site, 52 tentatively identified water molecules, and 9 detergent molecules. Only four of the water molecules are located in the cytoplasmic half of cytochrome c oxidase. Most of them are near the interface of subunits I and II. Several waters form a hydrogen-bonded cluster, including the heme propionates and the Mg/Mn binding site. The Fv fragment binds to the periplasmic polar domain of subunit II and is critically involved in the formation of the crystal lattice. The crystallization procedure is well reproducible and will allow for the analysis of the structures of mechanistically interesting mutant cytochrome c oxidases
MH  - A
MH  - analysis
MH  - Bacteria
MH  - BINDING
MH  - COMPLEX
MH  - Copper
MH  - Crystallization
MH  - cytochrome
MH  - Cytochrome c
MH  - Cytochrome-c Oxidase
MH  - Detergents
MH  - model
MH  - MONOCLONAL-ANTIBODIES
MH  - mutant
MH  - proton
MH  - Protons
MH  - resolution
MH  - Site
MH  - structure
MH  - SUBUNIT
MH  - Water
RP  - NOT IN FILE
NT  - UI - 98021406LA - engRN - 0 (Immunoglobulin Fragments)RN - 0 (Metals)RN - 0 (Protons)RN - EC 1.9.3.1 (Cytochrome-c Oxidase)PT - Journal ArticleDA - 19971110IS - 0027-8424SB - IMCY - UNITED STATES
UR  - PM:9380672
SO  - Proc Natl Acad Sci U S A 1997 Sep 30 ;94(20):10547-10553

1462
UI  - 503
AU  - Pedersen PA
AU  - Rasmussen JH
AU  - Jorgensen PL
AD  - Biomembrane Research Center, August Krogh Institute, University of Copenhagen, Denmark PAPedersen@akikudk
TI  - Increase in affinity for ATP and change in E1-E2 conformational equilibrium after mutations to the phosphorylation site (Asp369) of the alpha subunit of Na,K-ATPase
RP  - NOT IN FILE
NT  - UI - 98068917LA - engRN - 0 (Macromolecular Systems)RN - 0 (Recombinant Proteins)RN - 56-65-5 (Adenosine Triphosphate)RN - 56-84-8 (Aspartic Acid)RN - 630-60-4 (Ouabain)RN - EC 3.6.1.37 (Na(+)-K(+)-Exchanging ATPase)PT - Journal ArticleDA - 19980113IS - 0077-8923SB - IMCY - UNITED STATESJC - 5NM
UR  - PM:9405843
SO  - Ann N Y Acad Sci 1997 Nov 3 ;834():454-456

1463
UI  - 21184
AU  - Radionov AN
AU  - Kaulen AD
AD  - Department of Photobiochemistry, AN Belozersky Institute of Physico- Chemical Biology, Moscow State University, Russia
TI  - Inhibition of the M1-->M2 (M(closed) --> M(open)) transition in the D96N mutant photocycle and its relation to the corresponding transition in wild-type bacteriorhodopsin
AB  - Glutaraldehyde, lutetium ions and glycerol inhibit the blue shift of the difference spectra maximum of the M intermediate in the D96N mutant. The M formed has a spectrum indistinguishable from the M intermediate in wild-type bacteriorhodopsin. It has been concluded that the M(open) form previously described by us is identical to the M2 and Mn intermediates postulated by Zimanyi et al. (Photochem. Photobiol. (1992) 56, 1049-1055) and Sasaki et al. (J. Biol. Chem. (1992) 267, 20782-20786), respectively. It is supposed that its formation is accompanied by the appearance of the cytoplasmic proton half-channel. M(open) in the wild-type protein is present in a very low amount due to the shift of the M(closed) <--> M(open) equilibrium towards the M(closed). The inhibitors used do not prevent the multiphase pattern of the M formation in either mutant or wild-type proteins
MH  - A
MH  - ACID
MH  - Bacteriorhodopsin
MH  - BASE
MH  - inhibitor
MH  - intermediate
MH  - ion
MH  - Ions
MH  - M
MH  - M-intermediate
MH  - mutant
MH  - protein
MH  - Proteins
MH  - proton
MH  - Schiff base
MH  - Schiff Bases
MH  - Schiff-base
MH  - spectra
RP  - NOT IN FILE
NT  - UI - 97345641LA - engRN - 0 (Schiff Bases)RN - 53026-44-1 (Bacteriorhodopsins)RN - 56-84-8 (Aspartic Acid)RN - 7006-34-0 (Asparagine)PT - Journal ArticleDA - 19970714IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:9202133
SO  - FEBS Lett 1997 Jun 9 ;409(2):137-140

1464
UI  - 986
AU  - Rigaud JL
AU  - Mosser G
AU  - Lacapere JJ
AU  - Olofsson A
AU  - Levy D
AU  - Ranck JL
TI  - Bio-Beads: an efficient strategy for two-dimensional crystallization of membrane proteins
AB  - This work establishes the potential of Bio-Beads as a simple alternative to conventional dialysis for removing detergent and for obtaining 2D crystals of integral membrane proteins useful for structure analysis by electron crystallography. Kinetic and equilibrium aspects of removal of different detergents by adsorption onto hydrophobic Bio-Beads SM2 have been systematically investigated and extended to 2D crystallization of different prototypic membrane proteins, including: (a) Ca2+ ATPase from sarcoplasmic reticulum; (b) melibiose permease from Escherichia coli; (c) cytochrome b6f from Chlamydomonas reinhardtii. Different crystals could be produced from all protein preparations, with optical diffraction down to 20-25 A in negative stain
MH  - Animal
MH  - Ca(2+)-Transporting ATPase
MH  - Chemistry
MH  - Chlamydomonas reinhardtii
MH  - Crystallization
MH  - Cytochrome b
MH  - Detergents
MH  - Dialysis
MH  - enzymology
MH  - Escherichia coli
MH  - isolation & purification
MH  - Membrane Proteins
MH  - Microscopy,Electron
MH  - Molecular Structure
MH  - Permeases
MH  - Phospholipids
MH  - Polystyrenes
MH  - Protein Binding
MH  - Proteins
MH  - Sarcoplasmic Reticulum
MH  - Support,Non-U.S.Gov't
MH  - Temperature
MH  - ultrastructure
RP  - NOT IN FILE
NT  - Section de recherche, Institut Curie, UMR-CNRS 168 and LRC-CEA 8, Paris, France rigaud@curiefr
SO  - J Struct Biol 1997 Apr ;118(3):226-235

1465
UI  - 150
AU  - Rodgers AJ
AU  - Wilkens S
AU  - Aggeler R
AU  - Morris MB
AU  - Howitt SM
AU  - Capaldi RA
AD  - Institute of Molecular Biology, University of Oregon, Eugene Oregon 97403-1229, USA
TI  - The subunit delta-subunit b domain of the Escherichia coli F1F0 ATPase. The B subunits interact with F1 as a dimer and through the delta subunit
AB  - The delta and b subunits are both involved in binding the F1 to the F0 part in the Escherichia coli ATP synthase (ECF1F0). The interaction of the purified delta subunit and the isolated hydrophilic domain of the b subunit (bsol) has been studied here. Purified delta binds to bsol weakly in solution, as indicated by NMR studies and protease protection experiments. On F1, i.e. in the presence of ECF1-delta, delta, and bsol interact strongly, and a complex of ECF1.bsol can be isolated by native gel electrophoresis. Both delta subunit and bsol are protected from trypsin cleavage in this complex. In contrast, the delta subunit is rapidly degraded by the protease when bound to ECF1 when bsol is absent. The interaction of bsol with ECF1 involves the C-terminal domain of delta as delta(1-134) cannot replace intact delta in the binding experiments. As purified, bsol is a stable dimer with 80% alpha helix. A monomeric form of bsol can be obtained by introducing the mutation A128D (Howitt, S. M., Rodgers, A. J.,W., Jeffrey, P. D., and Cox, G. B. (1996) J. Biol. Chem. 271, 7038-7042). Monomeric bsol has less alpha helix, i.e. only 58%, is much more sensitive to trypsin cleavage than dimer, and unfolds at much lower temperatures than the dimer in circular dichroism melting studies, indicating a less stable structure. The bsol dimer, but not monomer, binds to delta in ECF1. To examine whether subunit b is a monomor or dimer in intact ECF1F0, CuCl2 was used to induce cross-link formation in the mutants bS60C, bQ104C, bA128C, bG131C, and bS146C. With the exception of bS60C, CuCl2 treatment resulted in formation of b subunit dimers in all mutants. Cross-linking yield was independent of nucleotide conditions and did not affect ATPase activity. These results show the b subunit to be dimeric for a large portion of the C terminus, with residues 124-131 likely forming a pair of parallel alpha helices
RP  - NOT IN FILE
NT  - UI - 98049578LA - engRN - 7440-50-8 (Copper)RN - 7447-39-4 (cupric chloride)RN - EC 3.4.21.5 (Thrombin)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - HL22450/HL/NHLBIDA - 19980108IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:9388256
SO  - J Biol Chem 1997 Dec 5 ;272(49):31058-31064

1466
UI  - 292
AU  - Sabbert D
AU  - Junge W
AD  - Abteilung Biophysik, FB Biologie/Chemie, Universitat Osnabruck, Germany
TI  - Stepped versus continuous rotatory motors at the molecular scale
AB  - Nature invented molecular rotatory devices such as the flagellar motor and ATP synthase. Photoselection techniques have been frequently used to detect the rotational random walk of proteins but only rarely for the rotational drift of subunits in proteins. Pertinent theories predict an oscillatory behavior of the polarization anisotropy, r, for unidirectional rotational drift, as opposed to a monotonic relaxation of r for bidirectional random walk. The underlying assumption of an angular continuum is questionable for intersubunit rotation in proteins. We developed a theory for stepped rotatory devices. It predicts the damped oscillation of r under unidirectional drift. Damping increases with decreasing number of steps. For only three steps a quasi-monotonic relaxation of r is predicted for both random walk and drift. In photoselection experiments with active F-ATPase we observed the relaxation of r when a spectroscopic probe was attached to the central beta-subunit. This behavior is compatible with the expectation for a three-stepped rotatory device
RP  - NOT IN FILE
NT  - UI - 97225949LA - engRN - 0 (Macromolecular Systems)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19970424IS - 0027-8424SB - IMCY - UNITED STATESJC - PV3
UR  - PM:9122191
SO  - Proc Natl Acad Sci U S A 1997 Mar 18 ;94(6):2312-2317

1467
UI  - 291
AU  - Sabbert D
AU  - Engelbrecht S
AU  - Junge W
AD  - Abteilung Biophysik, Fachbereich Biologie/Chemie, Universitat Osnabruck, D-49069 Osnabruck, Germany
TI  - Functional and idling rotatory motion within F1-ATPase
AB  - ATP synthase mediates proton flow through its membrane portion, F0, which drives the synthesis of ATP in its headpiece, F1. The F1-portion contains a hexagonal array of three subunits alpha and three beta encircling a central subunit gamma, that in turn interacts with a smaller epsilon and with F0. Recently we reported that the application of polarized absorption recovery after photobleaching showed the ATP- driven rotation of gamma over at least two, if not three, beta. Here we extend probes of such rotation aided by a new theory for assessing continuous versus stepped, Brownian versus unidirectional molecular motion. The observed relaxation of the absorption anisotropy is fully compatible with a unidirectional and stepping rotation of gamma over three equidistantly spaced angular positions in the hexagon formed by the alternating subunits alpha and beta. The results strongly support a rotational catalysis with equal participation of all three catalytic sites. In addition we report a limited rotation of gamma without added nucleotides, perhaps idling and of Brownian nature, that covers only a narrow angular domain
RP  - NOT IN FILE
NT  - UI - 97272235LA - engRN - 0 (Enzymes, Immobilized)RN - 0 (Molecular Probes)RN - 17372-87-1 (Eosine Yellowish-(YS))RN - 25612-73-1 (Adenylyl Imidodiphosphate)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19970527IS - 0027-8424SB - IMCY - UNITED STATESJC - PV3
UR  - PM:9114001
SO  - Proc Natl Acad Sci U S A 1997 Apr 29 ;94(9):4401-4405

1468
UI  - 289
AU  - Schulenberg B
AU  - Wellmer F
AU  - Lill H
AU  - Junge W
AU  - Engelbrecht S
AD  - Biophysik, Fachbereich Biologie/Chemie, Universitat Osnabruck, Germany
TI  - Cross-linking of chloroplast F0F1-ATPase subunit epsilon to gamma without effect on activity. Epsilon and gamma are parts of the rotor
AB  - Cys residues were directed into positions 17, 28, 41 and 85 of a Cys6-- >Ser mutant of subunit epsilon of spinach chloroplast F0F1 ATP synthase. Wild-type and engineered epsilon were expressed in Escherichia coli, purified in the presence of urea, refolded and reassembled with spinach chloroplast F1 lacking the epsilon subunit [F1(-epsilon)]. Cys-containing epsilon variants were modified with a sulfhydryl-reactive photolabile cross-linker. Photocross-linking of epsilon to F1(-epsilon) yielded the same SDS gel pattern of cross-link products independent of the presence or absence of Mg2+ x ADP, phosphate and Mg2+ x ATP. Epsilon (wild type) [Ser6,Cys28]epsilon and [Ser6,Cys41]epsilon were cross-linked with subunit gamma. With chloroplast F0F1 the same cross-link pattern was obtained, except for one extra cross-link, probably between [Ser6,Cys28]epsilon and F0 subunit III. [Ser6,Cys17]epsilon and [Ser6,Cys85]epsilon did not produce cross-links. Cross-linking of epsilon, [Ser6,Cys28]epsilon, [Ser6,Cys41]epsilon to gamma in soluble chloroplast F1 impaired the ability of epsilon to inhibit Ca2+-ATPase activity. The Mg2+-ATPase activity of soluble F1 (measured in the presence of 30% MeOH) was not affected by cross-linking epsilon with gamma. Functional reconstitution of photophosphorylation in F1-depleted thylakoids was observed with F1 in which gamma was cross-linked to [Ser6,Cys28]epsilon or [Ser6,Cys41]epsilon but not with wild-type epsilon. In view of the intersubunit rotation of gamma relative to (alphabeta)3, which is driven by ATP hydrolysis, gamma and epsilon would seem to act concertedly as parts of the 'rotor' relative to the 'stator' (alphabeta)3
RP  - NOT IN FILE
NT  - UI - 98028394LA - engRN - 0 (Cross-Linking Reagents)RN - 0 (Recombinant Proteins)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19971205IS - 0014-2956SB - IMCY - GERMANYJC - EMZ
UR  - PM:9363764
SO  - Eur J Biochem 1997 Oct 1 ;249(1):134-141

1469
UI  - 814
AU  - Schwarz O
AU  - Schurmann P
AU  - Strotmann H
AD  - Institut fur Biochemie der Pflanzen, Heinrich-Heine-Universitat Dusseldorf, Universitatsstrasse 1, D-40225 Dusseldorf, Federal Republic of Germany
TI  - Kinetics and thioredoxin specificity of thiol modulation of the chloroplast H+-ATPase
AB  - The kinetics of thiol modulation of the chloroplast H+-ATPase (CF0CF1) in membrana were analyzed by employing thioredoxins that were kept reduced by 0.1 mM dithiothreitol. The kinetics of thiol modulation depend on the extent of the proton gradient. The process is an exponential function of the thioredoxin concentration and reaction time and can be described by an irreversible second order reaction. The results indicate that the formation of the complex between thioredoxin and CF0CF1 is slow compared with the subsequent reduction step. Furthermore we have compared the efficiencies of the Escherichia coli thioredoxin Trx and the two chloroplast thioredoxins Tr-m and Tr-f. The second order rate constants are 0.057 (Tr-f), 0.024 (Trx), and 0.010 s- 1 microM-1 (Tr-m) suggesting that Tr-f rather than Tr-m is the physiological reductant for the chloroplast ATPase. The often employed artificial reductant dithiothreitol exhibits a second order rate constant in thiol modulation of 1.02.10(-6) s-1 microM-1
RP  - NOT IN FILE
NT  - UI - 97347496LA - engRN - 0 (Sulfhydryl Compounds)RN - 52500-60-4 (Thioredoxin)RN - 56-40-6 (Glycine)RN - 56-65-5 (Adenosine Triphosphate)RN - 5704-04-1 (tricine)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19970731IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:9202002
SO  - J Biol Chem 1997 Jul 4 ;272(27):16924-16927

1470
UI  - 328
AU  - Shirakihara Y
AU  - Leslie AG
AU  - Abrahams JP
AU  - Walker JE
AU  - Ueda T
AU  - Sekimoto Y
AU  - Kambara M
AU  - Saika K
AU  - Kagawa Y
AU  - Yoshida M
AD  - Department of Physics, Hyogo University of Education, Japan
TI  - The crystal structure of the nucleotide-free alpha 3 beta 3 subcomplex of F1-ATPase from the thermophilic Bacillus PS3 is a symmetric trimer
AB  - BACKGROUND: F1-ATPase, an oligomeric assembly with subunit stoichiometry alpha 3 beta 3 gamma delta epsilon, is the catalytic component of the ATP synthase complex, which plays a central role in energy transduction in bacteria, chloroplasts and mitochondria. The crystal structure of bovine mitochondrial F1-ATPase displays a marked asymmetry in the conformation and nucleotide content of the catalytic beta subunits. The alpha 3 beta 3 subcomplex of F1-ATPase has been assembled from subunits of the moderately thermophilic Bacillus PS3 made in Escherichia coli, and the subcomplex is active but does not show the catalytic cooperativity of intact F1-ATPase. The structure of this subcomplex should provide new information on the conformational variability of F1-ATPase and may provide insights into the unusual catalytic mechanism employed by this enzyme. RESULTS: The crystal structure of the nucleotide-free bacterial alpha 3 beta 3 subcomplex of F1-ATPase, determined at 3.2 A resolution, shows that the oligomer has exact threefold symmetry. The bacterial beta subunits adopt a conformation essentially identical to that of the nucleotide-free beta subunit in mitochondrial F1-ATPase; the alpha subunits have similar conformations in both structures. CONCLUSIONS: The structures of the bacterial F1-ATPase alpha and beta subunits are very similar to their counterparts in the mitochondrial enzyme, suggesting a common catalytic mechanism. The study presented here allows an analysis of the different conformations adopted by the alpha and beta subunits and may ultimately further our understanding of this mechanism
RP  - NOT IN FILE
NT  - UI - 97410391LA - engRN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19970925IS - 0969-2126SB - IMCY - ENGLANDJC - B31
UR  - PM:9261073
SO  - Structure 1997 Jun 15 ;5(6):825-836

1471
UI  - 21274
AU  - Stowell MH
AU  - McPhillips TM
AU  - Rees DC
AU  - Soltis SM
AU  - Abresch E
AU  - Feher G
AD  - Division of Chemistry and Chemical Engineering, 147-75CH, California Institute of Technology, Pasadena, CA 91125, USA
TI  - Light-induced structural changes in photosynthetic reaction center: implications for mechanism of electron-proton transfer
AB  - High resolution x-ray diffraction data from crystals of the Rhodobacter sphaeroides photosynthetic reaction center (RC) have been collected at cryogenic temperature in the dark and under illumination, and the structures were refined at 2.2 and 2.6 angstrom resolution, respectively. In the charge-separated D+
MH  - Chemistry
MH  - mechanism
MH  - proton
MH  - Protons
MH  - reaction center
MH  - resolution
MH  - Rhodobacter
MH  - Rhodobacter sphaeroides
MH  - sphaeroides
MH  - structure
MH  - Temperature
MH  - TRANSFER
MH  - Ubiquinone
MH  - X-Ray Diffraction
RP  - NOT IN FILE
NT  - UI - 97277227LA - engRN - 0 (Photosynthetic Reaction Center, Bacterial)RN - 0 (Protons)RN - 1339-63-5 (Ubiquinone)PT - Journal ArticleID - GM13191/GM/NIGMSID - GM45162/GM/NIGMSDA - 19970519IS - 0036-8075SB - IMCY - UNITED STATES
UR  - PM:9115209
SO  - Science 1997 May 2 ;276(5313):812-816

1472
UI  - 706
AU  - Sumi M
AU  - Yohda M
AU  - Koga Y
AU  - Yoshida M
AD  - Faculty of Pharmaceutical Sciences of Hokkaido University, Japan
TI  - F0F1-ATPase genes from an archaebacterium, Methanosarcina barkeri
AB  - It has been known that an archaebacterium Methanosarcina barkeri strain MS (DSM 800) has a V-type ATPase (Inatomi, K., et al. (1989) J. Biol. Chem. 264, 10954-10959). Here, we report cloning of a cluster of F0F1- ATPase genes from the same organism, the first ever found in archaebacteria. The cluster and encoded subunits exhibit several unusual features such that a gene for delta subunit is lacking, F0-b subunit is unusually large, and gamma subunit is split into two peptide fragments. Attempts to detect F0F1-ATPase proteins and mRNA have been unsuccessful and therefore it is not certain if this gene cluster is really expressed in the cell
RP  - NOT IN FILE
NT  - UI - 98086367LA - engRN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19980126IS - 0006-291XSB - IMCY - UNITED STATESJC - 9Y8
UR  - PM:9425287
SO  - Biochem Biophys Res Commun 1997 Dec 18 ;241(2):427-433

1473
UI  - 711
AU  - Taguchi H
AU  - Amada K
AU  - Murai N
AU  - Yamakoshi M
AU  - Yoshida M
AD  - Research Laboratory of Resources Utilization, R-1, Tokyo Institute of Technology, 4259 Nagatsuta, Yokohama 226, Japan
TI  - ATP-, K+-dependent heptamer exchange reaction produces hybrids between GroEL and chaperonin from Thermus thermophilus
AB  - Chaperonin from Thermus thermophilus (Tcpn6014.Tcpn107) splits at the plane between two Tcpn607 rings into two parts in a solution containing ATP and K+ (Ishii, N., Taguchi, H., Sasabe, H., and Yoshida, M. (1995) FEBS Lett. 362, 121-125). When Escherichia coli GroEL14 was additionally included in the solution described above, hybrid chaperonins GroEL7.Tcpn607 and GroEL7. Tcpn607.Tcpn107 were formed rapidly (<20 s) at 37 degrees C. The hybrid was also formed from Tcpn6014 and GroEL14 but not from a mutant GroEL14 lacking ATPase activity. The hybrid formation was saturated at approximately 300 microM ATP and approximately 300 mM K+. These results imply that GroEL14 also splits and undergoes a heptamer exchange reaction with Thermus chaperonin under nearly physiological conditions. Similar to parent chaperonins, the isolated hybrid chaperonins exhibited ATPase activity that was susceptible to inhibition by Tcpn107 or GroES7 and mediated folding of other proteins. Once formed, the hybrid chaperonins were stable, and the parent chaperonins were not regenerated from the isolated hybrids under the same conditions in which the hybrids had been formed. Only under conditions in which GroEL in the hybrids was selectively destroyed, such as incubation at 70 degrees C, Thermus chaperonin, but not GroEL14, was regenerated from the hybrid. Therefore, the split reaction may not be an obligatory event repeated in each turnover of the chaperonin functional cycles but an event that occurs only when chaperonin is first exposed to ATP/K+
RP  - NOT IN FILE
NT  - UI - 97364740LA - engRN - 0 (Chaperonin 10)RN - 0 (Chaperonin 60)RN - 0 (GroEL Protein)RN - 0 (Recombinant Proteins)RN - 56-65-5 (Adenosine Triphosphate)RN - 7440-09-7 (Potassium)RN - EC 1.1 (Alcohol Oxidoreductases)RN - EC 1.1.1.85 (3-isopropylmalate dehydrogenase)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19970818IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:9218450
SO  - J Biol Chem 1997 Jul 18 ;272(29):18155-18160

1474
UI  - 20843
AU  - Takeyasu K
AU  - Omote H
AU  - Nettikadan S
AU  - Tokumasu F
AU  - Iwamoto-Kihara A
AU  - Futai M
AD  - Department of Natural Environment Science, Faculty of Integrated Human Studies, Kyoto University, Japan takeyasu@gaiahkyoto-uacjp
TI  - Atomic force microscopy of Escherichia coli FoF1-ATPase in reconstituted membranes
MH  - ATPase
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - Human
MH  - Liposomes
MH  - Macromolecular Systems
MH  - membrane
MH  - Membranes
MH  - Microscopy
MH  - SYSTEM
MH  - SYSTEMS
RP  - NOT IN FILE
NT  - UI - 98068875LA - engRN - 0 (Liposomes)RN - 0 (Macromolecular Systems)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19980113IS - 0077-8923SB - IMCY - UNITED STATES
UR  - PM:9405801
SO  - Ann N Y Acad Sci 1997 Nov 3 ;834():149-152

1475
UI  - 107
AU  - Valiyaveetil FI
AU  - Fillingame RH
AD  - Department of Biomolecular Chemistry, University of Wisconsin Medical School, Madison, Wisconsin 53706, USA
TI  - On the role of Arg-210 and Glu-219 of subunit a in proton translocation by the Escherichia coli F0F1-ATP synthase
AB  - A strain of Escherichia coli was constructed which had a complete deletion of the chromosomal uncB gene encoding subunit a of the F0F1- ATP synthase. Gene replacement was facilitated by a selection protocol that utilized the sacB gene of Bacillus subtilis cloned in a kanamycin resistance cartridge (Ried, J. L., and Collmer, A. (1987) Gene (Amst.) 57, 239-246). F0 subunits b and c inserted normally into the membrane in the DeltauncB strain. This observation confirms a previous report (Hermolin, J., and Fillingame, R. H. (1995) J. Biol. Chem. 270, 2815- 2817) that subunit a is not required for the insertion of subunits b and c. The DeltauncB strain has been used to characterize mutations in Arg-210 and Glu-219 of subunit a, residues previously postulated to be essential in proton translocation. The aE219G and aE219K mutants grew on a succinate carbon source via oxidative phosphorylation and membranes from these mutants exhibited ATPase-coupled proton translocation (i.e. ATP driven 9-amino-6-chloromethoxyacridine quenching responses that were 60-80% of wild type membranes). We conclude that the aGlu-219 residue cannot play a critical role in proton translocation. The aR210A mutant did not grow on succinate and membranes exhibited no ATPase-coupled proton translocation. However, on removal of F1 from membrane, the aR210A mutant F0 was active in passive proton translocation, i.e. in dissipating the DeltapH normally established by NADH oxidation with these membrane vesicles. aR210A membranes with F1 bound were also proton permeable. Arg-210 of subunit a may play a critical role in active H+ transport that is coupled to ATP synthesis or hydrolysis, but is not essential for the translocation of protons across the membranes
RP  - NOT IN FILE
NT  - UI - 98070446LA - engRN - 0 (Protons)RN - 56-86-0 (Glutamic Acid)RN - 7004-12-8 (Arginine)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM23105/GM/NIGMSDA - 19980122IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:9405480
SO  - J Biol Chem 1997 Dec 19 ;272(51):32635-32641

1476
UI  - 987
AU  - Villaverde J
AU  - Cladera J
AU  - Padros E
AU  - Rigaud JL
AU  - Dunach M
TI  - Effect of nucleotides on the thermal stability and on the deuteration kinetics of the thermophilic F0F1 ATP synthase
AB  - Differential scanning calorimetry has been used to characterize the influence of specific nucleotide binding on the thermal unfolding of the F0F1-type ATP synthase from the thermophilic Bacillus PS3 (TF0F1). The calorimetric trace shows an irreversible and kinetically controlled endothermic transition for TF0F1 in the absence of nucleotides. The thermal denaturation occurs at a transition temperature (t(m)) of 81.7 degrees C. The remarkable thermostability of this enzyme was decreased upon tight binding of Mg2+ x ATP to noncatalytic sites, whereas binding of Mg2+ x ADP increased the temperature at which thermal denaturation occurred. At high temperatures, an exothermic transition due to aggregation processes was also affected by nucleotide binding. With the aim to correlate these thermal effects with possible structural differences among the various forms of TF0F1, Fourier transform infrared spectroscopy was carried out. Hydrogen/deuterium exchange was clearly affected by specific nucleotide occupancy. As illustrated by the total extent of protons exchanged, our results demonstrate that more peptide groups are exposed to the medium in the presence of Mg2+ x ATP than in the presence of Mg2+ x ADP. Therefore, consistent with microcalorimetric data, binding of Mg2+ x ADP induces conformational changes which shield amide protons to more buried hydrogen-bonded structures, whereas binding of Mg2+ x ATP results in a more open or flexible structure
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - Bacillus
MH  - Calorimetry
MH  - Calorimetry,Differential Scanning
MH  - Chemistry
MH  - Detergents
MH  - Deuterium
MH  - drug effects
MH  - Enzyme Stability
MH  - enzymology
MH  - H(+)-Transporting ATP Synthase
MH  - Hydrogen Bonding
MH  - Kinetics
MH  - Magnesium
MH  - metabolism
MH  - pharmacology
MH  - Protein Conformation
MH  - Protein Denaturation
MH  - Protein Structure,Secondary
MH  - Protons
MH  - Spectroscopy,Fourier Transform Infrared
MH  - Support,Non-U.S.Gov't
MH  - Temperature
MH  - Thermodynamics
RP  - NOT IN FILE
NT  - Departament de Bioquimica i Biologia Molecular, Facultat de Medicina, Universitat Autonoma de Barcelona, Bellaterra, Spain
SO  - Eur J Biochem 1997 Mar 1 ;244(2):441-448

1477
UI  - 153
AU  - Watts SD
AU  - Capaldi RA
AD  - Institute of Molecular Biology, University of Oregon, Eugene, Oregon 97403-1229, USA
TI  - Interactions between the F1 and F0 parts in the Escherichia coli ATP synthase. Associations involving the loop region of C subunits
AB  - The N-ethylmaleimide reactivity of c subunits in Escherichia coli F1F0 ATP synthase (ECF1F0) isolated from five mutants, each with a cysteine at a different position in the polar loop region (positions 39, 40, 42, 43, and 44), has been investigated. The maleimide was found to react with Cys placed at positions 42, 43, and 44 but not at 39 or 40. All copies of the c subunit reacted similarly when the Cys was at position 43 or 44. In contrast, the Cys in the mutant cQ42C reacted as two classes, with 60% reacting relatively rapidly and 40% reacting at a rate 40-fold slower. After removing F1, all copies of the c subunit in this mutant reacted equally fast. Therefore, the slow class in the cQ42C mutant represents c subunits shielded by, and probably involved directly in, the interaction of the F0 with gamma and epsilon subunits of the F1 part. Based on the estimated stoichiometry of c subunits in the ECF1F0 complex, 4 or 5 c subunits are involved in this F1 interaction. N-Ethylmaleimide modification of all of the c subunits reduced ATPase activity by only 30% in ECF1F0 from mutant cQ42C. Modification of the more rapidly reacting class had little effect on ATP hydrolysis-driven proton translocation, and did not alter the DCCD inhibition of ATPase activity. However, as those c subunits involved in the F1 interaction became modified, DCCD inhibition was progressively lost, as was coupling between ATP hydrolysis and proton translocation
RP  - NOT IN FILE
NT  - UI - 97326073LA - engRN - 128-53-0 (Ethylmaleimide)RN - 52-90-4 (Cysteine)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM07759/GM/NIGMSID - HL22450/HL/NHLBIID - HL24526/HL/NHLBIDA - 19970714IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:9182524
SO  - J Biol Chem 1997 Jun 13 ;272(24):15065-15068

1478
UI  - 415
AU  - Weber J
AU  - Senior AE
AD  - Department of Biochemistry, University of Rochester Medical Center, NY 14642, USA
TI  - Catalytic mechanism of F1-ATPase
AB  - The structure of the core catalytic unit of ATP synthase, alpha 3 beta 3 gamma, has been determined by X-ray crystallography, revealing a roughly symmetrical arrangement of alternating alpha and beta subunits around a central cavity in which helical portions of gamma are found. A low-resolution structural model of F0, based on electron spectroscopic imaging, locates subunit a and the two copies of subunit b outside of a subunit c oligomer. The structures of individual subunits epsilon and c (largely) have been solved by NMR spectroscopy, but the oligomeric structure of c is still unknown. The structures of subunits a and delta remain undefined, that of b has not yet been defined but biochemical evidence indicates a credible model. Subunits gamma, epsilon, b, and delta are at the interface between F1 and F0; gamma epsilon complex forms one element of the stalk, interacting with c at the base and alpha and beta at the top. The locations of b and delta are less clear. Elucidation of the structure F0, of the stalk, and of the entire F1F0 remains a challenging goal
RP  - NOT IN FILE
NT  - UI - 97261361LA - engRN - 0 (Bacterial Proteins)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, AcademicID - GM25349/GM/NIGMSDA - 19970501IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:9107315
SO  - Biochim Biophys Acta 1997 Mar 28 ;1319(1):19-58

1479
UI  - 414
AU  - Weber J
AU  - Senior AE
AD  - University of Rochester Medical Center, Department of Biochemistry and Biophysics, NY 14642, USA
TI  - Binding of TNP-ATP and TNP-ADP to the non-catalytic sites of Escherichia coli F1-ATPase
AB  - Using site-directed-tryptophan fluorescence, parameters for equilibrium binding of (Mg)TNP-ATP and (Mg)TNP-ADP to non-catalytic sites of Escherichia coli F1-ATPase were determined. All three non-catalytic sites showed the same affinity for MgTNP-ATP (Kd = 0.2 microM) or MgTNP- ADP (Kd = 6.5 microM) whereas even at concentrations of 100 microM no binding of uncomplexed TNP-ATP or TNP-ADP was observed. The results demonstrate that the three non-catalytic sites bind TNP-nucleotides non- cooperatively, and emphasize the importance of Mg2+ for non-catalytic- site nucleotide binding. Parameters for binding of (Mg)TNP-ADP to the three catalytic sites were also determined, and showed marked cooperativity. This work completes the set of thermodynamic parameters for equilibrium binding of (Mg)TNP-ATP and (Mg)TNP-ADP to all six nucleotide sites of F1, providing essential information to fully exploit the potential of these nucleotide analogs in studies of F1- ATPase
RP  - NOT IN FILE
NT  - UI - 97400220LA - engRN - 0 (Fluorescent Dyes)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 61368-63-6 (2',3'-O-(2,4,6-trinitro-cyclohexadienylidine)adenosine 5'- triphosphate)RN - 7439-95-4 (Magnesium)RN - 84430-17-1 (2',3'-(O-(2,4,6-trinitrocyclohexadienylidine))adenosine 5'- diphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSDA - 19970905IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:9257714
SO  - FEBS Lett 1997 Jul 21 ;412(1):169-172

1480
UI  - 149
AU  - Wilkens S
AU  - Rodgers A
AU  - Ogilvie I
AU  - Capaldi RA
AD  - Institute of Molecular Biology, University of Oregon, Eugene 97403- 1229, USA
TI  - Structure and arrangement of the delta subunit in the E. coli ATP synthase (ECF1F0)
AB  - F1F0 type ATPases are made up of two parts, an F1, which contains three catalytic sites on beta subunits, and an F0 which contains the proton channel. These two domains have been visualized in electron microscopy as linked by a narrow stalk of around 45 A in length. Biochemical studies have provided clear evidence that the gamma and epsilon subunits are components of this stalk. There is an emerging consensus that the gamma and epsilon subunits rotate relative to the alpha 3 beta 3 domain as part of the cooperativity and energy coupling within the complex. Two other subunits are required to link the F1 to F0 in the E. coli enzyme, and these are the delta and b subunits. The structure of a major part of the delta subunit (residues 1-134) has now been obtained by NMR spectroscopy. The main feature is a six alpha-helix bundle, which provides the N-terminal domain of the delta subunit. This domain interacts with the F1 core via the N-terminal part of the alpha subunit. The C-terminal domain of delta is less well defined. This part is required for binding to the F0 part by direct interaction with the b subunits. It is argued that delta and the two copies of the b subunit are components of a second stalk linking the F1 and F0 parts, which acts as a stator to allow the energy-linked rotational movements of delta and epsilon subunits
RP  - NOT IN FILE
NT  - UI - 98129865LA - engRN - 0 (Macromolecular Systems)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - HL24526/HL/NHLBIDA - 19980310IS - 0301-4622SB - IMCY - NETHERLANDSJC - A5T
UR  - PM:9468613
SO  - Biophys Chem 1997 Oct ;68(1-3):95-102

1481
UI  - 71
AU  - Wilkens S
AU  - Dunn SD
AU  - Chandler J
AU  - Dahlquist FW
AU  - Capaldi RA
TI  - Solution structure of the N-terminal domain of the delta subunit of the E. coli ATPsynthase
AB  - NMR studies of the delta subunit of the Escherichia coli F1F0- ATPsynthase reveal that it consists of an N-terminal six alpha-helix bundle and a less well ordered C terminus. Both domains are part of one of two separate connections between F1 and F0
RP  - NOT IN FILE
NT  - UI - 97307253LA - engRN - 0 (Macromolecular Systems)RN - 0 (Peptide Fragments)RN - 0 (Recombinant Proteins)RN - 0 (Solutions)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - LetterDA - 19970616IS - 1072-8368SB - IMCY - UNITED STATESJC - B98
UR  - PM:9164460
SO  - Nat Struct Biol 1997 Mar ;4(3):198-201

1482
UI  - 21262
AU  - Witthuhn VC
AU  - Gao J
AU  - Hong S
AU  - Halls S
AU  - Rott MA
AU  - Wraight CA
AU  - Crofts AR
AU  - Donohue TJ
AD  - Bacteriology Department, University of Wisconsin-Madison 53706, USA
TI  - Reactions of isocytochrome c2 in the photosynthetic electron transfer chain of Rhodobacter sphaeroides
AB  - Rhodobacter sphaeroides strains lacking cytochrome c2 (cyt c2), the normal electron donor to P870+ in light-oxidized reaction center (RC) complexes, are unable to grow photosynthetically. However, spd mutations that suppress the photosynthetic deficiency of cyt c2 mutants elevate levels of the cyt c2 isoform, isocyt c2. We monitored photosynthetic electron transfer in whole cells, in chromatophores, and with purified components to ascertain if and how isocyt c2 reduced light-oxidized RC complexes. These studies revealed that several fundamental aspects of photosynthetic electron transfer were similar in strains that use isocyt c2 and wild-type cells. For example, P870+ reduction accompanied cytochrome c oxidation. In addition, photosynthetic electron transfer was blocked by the well-known cyt bc1 complex inhibitors antimycin and myxothiazol. However, even at the increased isocyt c2 levels present in these strains (approximately 40% that of cyt c2 in wild-type cells), there was little, if any, of the rapid (< 5 microns) electron transfer to P870+ that is characteristic of cytochromes bound to RC complexes at the time of the light flash. Thus, it appears that isocyt c2 function limits the in vivo rate of P870+ reduction. Indeed, at low ionic strength in vitro, the apparent affinity of isocyt c2 for RC complexes (KD approximately 40 microM) is significantly lower than that of cyt c2 (KD approximately 1.0 microM). This reduced affinity does not appear to result from an altered mode of RC binding by isocyt c2 since electrostatic interactions make similar overall contributions to the binding of both cyt c2 and isocyt c2 to this membrane-bound redox partner. Thus, sequence, structural, or local conformational differences between cyt c2 and isocyt c2 significantly alter their apparent affinities for this physiologically relevant redox partner
MH  - BINDING
MH  - Cells
MH  - chromatophore
MH  - chromatophores
MH  - COMPLEX
MH  - cytochrome
MH  - Cytochrome c
MH  - Cytochromes
MH  - electron
MH  - electron transfer
MH  - Electron-transfer
MH  - flash
MH  - function
MH  - In Vitro
MH  - inhibitor
MH  - Light
MH  - mutant
MH  - reaction center
MH  - redox
MH  - Rhodobacter
MH  - Rhodobacter sphaeroides
MH  - sphaeroides
MH  - Time
MH  - TRANSFER
RP  - NOT IN FILE
NT  - UI - 97172922LA - engRN - 0 (Photosynthetic Reaction Center, Bacterial)RN - 0 (iso-2-cytochrome C)RN - 9007-43-6 (Cytochrome c)RN - 9035-43-2 (cytochrome c2)PT - Journal ArticleID - 3-RO1-GM35438/GM/NIGMSID - GM07215/GM/NIGMSDA - 19970313IS - 0006-2960SB - IMCY - UNITED STATES
UR  - PM:9020790
SO  - Biochemistry 1997 Jan 28 ;36(4):903-911

1483
UI  - 708
AU  - Yasuda R
AU  - Noji H
AU  - Kinosita K
AU  - Motojima F
AU  - Yoshida M
AD  - Department of Physics, Faculty of Science and Technology, Keio University, Yokohama, Japan
TI  - Rotation of the gamma subunit in F1-ATPase; evidence that ATP synthase is a rotary motor enzyme
AB  - ATP-dependent, azide-sensitive rotation of the gamma subunit relative to the alpha3beta3 hexagonal ring of ATP synthase was observed with a single molecule imaging system. Thus, ATP synthase is a rotary motor enzyme, the first ever found
RP  - NOT IN FILE
NT  - UI - 97442156LA - engRN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 19971114IS - 0145-479XSB - IMCY - UNITED STATESJC - HIO
UR  - PM:9298705
SO  - J Bioenerg Biomembr 1997 Jun ;29(3):207-209

1484
UI  - 710
AU  - Yoshida M
AU  - Igeta S
AU  - Kawashima R
AU  - Akama Y
AU  - Yoshida K
AD  - Department of Chemistry, St Marianna University School of Medicine, 2- 16-1 Sugao, Miyamae-ku, Kawasaki, 216, Japan
TI  - Changes in adenosine triphosphate (ATP) concentration and its activity in murine tissue after thallium administration
RP  - NOT IN FILE
NT  - UI - 97362175LA - engRN - 0 (Rodenticides)RN - 10031-59-1 (thallium sulfate)RN - 56-65-5 (Adenosine Triphosphate)RN - 7440-28-0 (Thallium)RN - EC 3.6.1.37 (Na(+)-K(+)-Exchanging ATPase)PT - Journal ArticleDA - 19971201IS - 0007-4861SB - IMCY - UNITED STATESJC - BFN
UR  - PM:9211698
SO  - Bull Environ Contam Toxicol 1997 Aug ;59(2):268-273

1485
UI  - 712
AU  - Yoshida M
AU  - Noji H
AU  - Muneyuki E
AD  - Tokyo Institute of Technology, Research Laboratory of Resources Utilization, Yokohama, Japan
TI  - [World smallest motor, ATP synthase]
RP  - NOT IN FILE
NT  - UI - 97359299LA - jpnRN - 0 (Multienzyme Complexes)RN - 0 (Proton Pump)RN - EC 2.7.4 (Phosphotransferases (Phosphate Group Acceptor))RN - EC 2.7.4.- (ATP synthetase complex)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 19971001IS - 0039-9450SB - IMCY - JAPANJC - Q7D
UR  - PM:9216232
SO  - Tanpakushitsu Kakusan Koso 1997 Jul ;42(9):1396-1406

1486
UI  - 19857
AU  - Zhou Y
AU  - Duncan TM
AU  - Cross RL
AD  - Department of Biochemistry and Molecular Biology, State University of New York Health Science Center, 750 East Adams Street, Syracuse, NY 13210, USA
TI  - Subunit rotation in Escherichia coli FoF1-ATP synthase during oxidative phosphorylation
AB  - We report evidence for proton-driven subunit rotation in membrane-bound FoF1-ATP synthase during oxidative phosphorylation. A betaD380C/gammaC87 crosslinked hybrid F1 having epitope-tagged betaD380C subunits (betaflag) exclusively in the two noncrosslinked positions was bound to Fo in F1-depleted membranes. After reduction of the beta-gamma crosslink, a brief exposure to conditions for ATP synthesis followed by reoxidation resulted in a significant amount of betaflag appearing in the beta-gamma crosslinked product. Such a reorientation of gammaC87 relative to the three beta subunits can only occur through subunit rotation. Rotation was inhibited when proton transport through Fo was blocked or when ADP and Pi were omitted. These results establish FoF1 as the second example in nature where proton transport is coupled to subunit rotation
MH  - A
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - ADP
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - BETA-SUBUNIT
MH  - Biochemistry
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1
MH  - H(+)-Transporting ATP Synthase
MH  - membrane
MH  - Membranes
MH  - Oxidative Phosphorylation
MH  - Phosphorylation
MH  - proton
MH  - rotation
MH  - SUBUNIT
MH  - SYNTHASE
MH  - synthesis
MH  - transport
RP  - NOT IN FILE
NT  - UI - 98021412LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM23152/GM/NIGMSDA - 19971110IS - 0027-8424SB - IMCY - UNITED STATESJC - PV3
UR  - PM:9380678
SO  - Proc Natl Acad Sci U S A 1997 Sep 30 ;94(20):10583-10587

1487
UI  - 147
AU  - Aggeler R
AU  - Gruber G
AU  - Capaldi RA
AD  - Institute of Molecular Biology, University of Oregon, Eugene 97403- 1229, USA
TI  - Trapping of conformations of the Escherichia coli F1 ATPase by disulfide bond formation. A state of the enzyme with all three catalytic sites of equal and low affinity for nucleotides
AB  - A mutant of Escherichia coli F1F0-ATPase, alphaS411C/betaY331W/betaE381C/gammaC87S, has been generated. CuCl2 treatment of this mutant led to cross-linking between alpha and beta subunits in yields of up to 90%. This cross-linking across non- catalytic site interfaces inhibited ATP hydrolysis activity. In the absence of cross-linking, MgATP bound in catalytic sites of the mutant with three different affinities of 0.1 microM, 6 microM and 60 microM, respectively, values that are comparable to wild-type. For MgADP, there was one tight site (0.34 microM) and two sites of lower affinity (each 27 microM), again comparable to wild-type enzyme. After cross-linking all three catalytic sites bound MgATP or MgADP with the same relatively low affinity (approximately 60 microM). Thus cross-linking fixed all three catalytic sites in the same conformation. Trypsin cleavage experiments showed that cross-linking fixed the epsilon subunit in the ATP+EDTA conformation
RP  - NOT IN FILE
NT  - UI - 98258921LA - engRN - 0 (Bacterial Proteins)RN - 0 (Disulfides)RN - 0 (Nucleotides)RN - EC 3.4.21.4 (Trypsin)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - HL 24526/HL/NHLBIDA - 19980528IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:9598974
SO  - FEBS Lett 1998 Apr 10 ;426(1):37-40

1488
UI  - 18972
AU  - Avetisyan AV
AU  - Kaulen AD
AU  - Skulachev VP
AU  - Feniouk BA
AD  - Department of Bioenergetics, Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Moscow, 119899, Russia
TI  - Photophosphorylation in alkalophilic halobacterial cells containing halorhodopsin: chloride-ion cycle?
AB  - Light-driven ATP synthesis is found in cells of the alkalophilic bacterium Natronobacterium pharaonis containing halorhodopsin but deficient in H+-pumping bacteriorhodopsin. Photophosphorylation occurs with cyanide-inhibited respiratory chain as well as without cyanide in conditions with low C1- concentration in the incubation medium. Increase in C1- concentration from 0.1 to 2.35 M in the incubation medium leads to inhibition of photophosphorylation. Continuous illumination increases membrane Delta Psi if respiration is inhibited by cyanide. This effect is stimulated by DCCD, an ATPase inhibitor. These data can be explained if one suggests that halorhodopsin pumps C1- into the cells whereas C1- release from the cells through C1--ATP- synthase is coupled with the ATP synthesis (chloride-ion cycle)
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthesis
MH  - ATPase
MH  - Bacteria
MH  - Bacteriorhodopsin
MH  - bioenergetics
MH  - Cells
MH  - membrane
MH  - Photophosphorylation
MH  - PSI
MH  - Respiration
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 98334723LA - engRN - 0 (Chlorides)RN - 0 (halorhodopsin)RN - 53026-44-1 (Bacteriorhodopsin)RN - 56-65-5 (Adenosine Triphosphate)PT - Journal ArticleDA - 19981117IS - 0006-2979SB - IMCY - RUSSIAJC - CSQ
UR  - PM:9668201
SO  - Biochemistry (Mosc ) 1998 Jun ;63(6):625-628

1489
UI  - 705
AU  - Bald D
AU  - Amano T
AU  - Muneyuki E
AU  - Pitard B
AU  - Rigaud JL
AU  - Kruip J
AU  - Hisabori T
AU  - Yoshida M
AU  - Shibata M
AD  - Research Laboratory of Resources Utilization, Tokyo Institute of Technology, 4259 Nagatsuta, Yokohama 226, Japan
TI  - ATP synthesis by F0F1-ATP synthase independent of noncatalytic nucleotide binding sites and insensitive to azide inhibition
AB  - ATP hydrolyzing activity of a mutant alpha3beta3gamma subcomplex of F0F1-ATP synthase (DeltaNC) from the thermophilic Bacillus PS3, which lacked noncatalytic nucleotide binding sites, was inactivated completely soon after starting the reaction (Matsui, T., Muneyuki, E. , Honda, M., Allison, W. S., Dou, C., and Yoshida, M. (1997) J. Biol. Chem. 272, 8215-8221). This inactivation is caused by rapid accumulation of the "MgADP inhibited form" which, in the case of wild- type enzyme, would be relieved by ATP binding to noncatalytic sites. We reconstituted F0F1-ATP synthase into liposomes together with bacteriorhodopsin and measured illumination-driven ATP synthesis. Remarkably, DeltaNC F0F1-ATP synthase catalyzed continuous turnover of ATP synthesis while it could not promote ATP-driven proton translocation. ATP synthesis by DeltaNC F0F1-ATP synthase, as well as wild-type enzyme, proceeded even in the presence of azide, an inhibitor of ATP hydrolysis that stabilizes the MgADP inhibited form. The time course of ATP synthesis by DeltaNC F0F1-ATP synthase was linear, and gradual acceleration to the maximal rate, which was observed for the wild-type enzyme, was not seen. Thus, ATP synthesis can proceed without nucleotide binding to noncatalytic sites even though the rate is sub- maximal. These results indicate that the MgADP inhibited form is not produced in ATP synthesis reaction, and in this regard, ATP synthesis may not be a simple reversal of ATP hydrolysis
RP  - NOT IN FILE
NT  - UI - 98086257LA - engRN - 0 (Azides)RN - 0 (Enzyme Inhibitors)RN - 0 (Protons)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19980209IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:9422743
SO  - J Biol Chem 1998 Jan 9 ;273(2):865-870

1490
UI  - 501
AU  - Bianchet MA
AU  - Hullihen J
AU  - Pedersen PL
AU  - Amzel LM
AD  - Department of Biophysics and Biophysical Chemistry, The Johns Hopkins University School of Medicine, 725 North Wolfe Street, Baltimore, MD 21205-2185, USA mario@nerudamedjhmiedu
TI  - The 2.8-A structure of rat liver F1-ATPase: configuration of a critical intermediate in ATP synthesis/hydrolysis
AB  - During mitochondrial ATP synthesis, F1-ATPase-the portion of the ATP synthase that contains the catalytic and regulatory nucleotide binding sites-undergoes a series of concerted conformational changes that couple proton translocation to the synthesis of the high levels of ATP required for cellular function. In the structure of the rat liver F1- ATPase, determined to 2.8-A resolution in the presence of physiological concentrations of nucleotides, all three beta subunits contain bound nucleotide and adopt similar conformations. This structure provides the missing configuration of F1 necessary to define all intermediates in the reaction pathway. Incorporation of this structure suggests a mechanism of ATP synthesis/hydrolysis in which configurations of the enzyme with three bound nucleotides play an essential role
RP  - NOT IN FILE
NT  - UI - 98409611LA - engRN - 0 (Multienzyme Complexes)RN - 0 (Nucleotides)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 2.7.4 (Phosphotransferases (Phosphate Group Acceptor))RN - EC 2.7.4.- (ATP synthetase complex)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - CA 10951/CA/NCIID - GM 25432/GM/NIGMSDA - 19981026IS - 0027-8424SB - IMCY - UNITED STATESJC - PV3
UR  - PM:9736690
SO  - Proc Natl Acad Sci U S A 1998 Sep 15 ;95(19):11065-11070

1491
UI  - 21285
AU  - Bopp PA
AU  - Kornyshev AA
AU  - Sutmann G
TI  - Frequency and wave-vector dependent dielectric function of water: Collective modes and relaxation spectra
RP  - IN FILE
SO  - J Chem Phys 1998  ;109():1939-1959

1492
UI  - 606
AU  - Borghese R
AU  - Turina P
AU  - Lambertini L
AU  - Melandri BA
AD  - Universita di Bologna, Dipartimento di Biologia, Italy
TI  - The atpIBEXF operon coding for the F0 sector of the ATP synthase from the purple nonsulfur photosynthetic bacterium Rhodobacter capsulatus
AB  - The atpIBEXF operon coding for the F0 sector of the ATP synthase from Rhodobacter capsulatus was cloned and sequenced. The genes for the five subunits were present in the order: atpI (subunit I), atpB (subunit a), atpE (subunit c), atpX (subunit b'), and atpF (subunit b). The transcription initiation site was defined by primer-extension analysis. A duplicated and divergent copy of the b subunit gene (subunit b') was present. This duplication is found only in photosynthetic prokaryotes and in plant chloroplasts. F0 deletion mutants formed tiny colonies during anaerobic growth in the dark but could not sustain continuous growth. Based on the results of the present work, we conclude that a functioning ATP synthase is essential for normal growth under all conditions tested
RP  - NOT IN FILE
NT  - UI - 99035763LA - engRN - 0 (Bacterial Proteins)RN - 0 (DNA, Bacterial)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19981203IS - 0302-8933SB - IMCY - GERMANYJC - 7YN
UR  - PM:9818357
SO  - Arch Microbiol 1998 Oct ;170(5):385-388

1493
UI  - 607
AU  - Borghese R
AU  - Crimi M
AU  - Fava L
AU  - Melandri BA
AD  - Department of Biology, University of Bologna, Italy
TI  - The ATP synthase atpHAGDC (F1) operon from Rhodobacter capsulatus
AB  - The atpHAGDC operon of Rhodobacter capsulatus, containing the five genes coding for the F1 sector of the ATP synthase, has been cloned and sequenced. The promoter region has been defined by primer extension analysis. It was not possible to obtain viable cells carrying atp deletions in the R. capsulatus chromosome, indicating that genes coding for ATP synthase are essential, at least under the growth conditions tested. We were able to circumvent this problem by combining gene transfer agent transduction with conjugation. This method represents an easy way to construct strains carrying mutations in indispensable genes
RP  - NOT IN FILE
NT  - UI - 98101484LA - engRN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19980206IS - 0021-9193SB - IMCY - UNITED STATESJC - HH3
UR  - PM:9440534
SO  - J Bacteriol 1998 Jan ;180(2):416-421

1494
UI  - 9887
AU  - Borsch M
AU  - Turina P
AU  - Eggeling C
AU  - Fries JR
AU  - Seidel CAM
AU  - Labahn A
AU  - Graber P
TI  - Conformational changes of the H+-ATPase from Escherichia coli upon nucleotide binding detected by single molecule fluorescence
AB  - Using a confocal fluorescence microscope with an avalanche photodiode as detector, we studied the fluorescence of the tetramethylrhodamine labeled F-1 part of the H+-ATPase from Escherichia coli, EF1, carrying the gamma T106-C mutation [Aggeler, J.A. and Capaldi, R.A. (1992) J. Biol. Chem. 267, 21355-21359] in aqueous solution upon excitation with a mode- locked argon ion laser at 528 nm. The diffusion of the labeled EF1 through the confocal volume gives rise to photon bursts, which were analyzed with fluorescence correlation spectroscopy, resulting in a diffusion coefficient of 3.3 x 10(-7) cm(2) s(- 1). In the presence of nucleotides the diffusion coefficient increases by about 15%. This effect indicates a change of the shape and/or the volume of the enzyme upon binding of nucleotides, i.e, fluorescence correlation spectroscopy with single EF1 molecules allows the detection of conformational changes. (C) 1998 Federation of European Biochemical Societies
MH  - ADENOSINE-TRIPHOSPHATASE
MH  - conformational change
MH  - CORRELATION SPECTROSCOPY
MH  - Diffusion
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1
MH  - F1-ATPASE
MH  - fluorescence
MH  - GAMMA- SUBUNIT
MH  - H+-ATPase
MH  - nucleotide binding
MH  - Nucleotides
MH  - PS3
MH  - single molecule spectroscopy
RP  - NOT IN FILE
NT  - JournalArticleELSEVIER SCIENCE BVOCT 23133ZPAMSTERDAMBorsch M Univ Freiburg, Inst Phys Chem, Albertstr 23A, D-79104 Freiburg, GermanyFEBS LETTPO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS
AV  - Univ Freiburg, Inst Phys Chem, Albertstr 23A, D-79104 Freiburg, Germany Univ Freiburg, Inst Phys Chem, D-79104 Freiburg, Germany Univ Bologna, Dipartimento Biol, I-40126 Bologna, Italy Max Planck Inst Biophys Chem, D-37077 Gottingen, Germany
UR  - ISI:000076717700019
SO  - Febs Letters 1998  ;437(3):251-254

1495
UI  - 9888
AU  - Bottcher B
AU  - Schwarz L
AU  - Graber P
TI  - Direct indication for the existence of a double stalk in CF0F1
AB  - The H+-ATPase from chloroplasts (CF0F1) was investigated by electron microscopy of negatively stained single molecules followed by image processing. The analysis of about 4700 particles from 72 micrographs gave clear evidence that the membrane-integrated F-0 part is connected by at least two stalks to the F-1 part. One of the two stalks is more prominent and connects a central part of F-1 with a slightly peripheral part of F-0. The other stalk connects a peripheral part of F-1 to a peripheral part of F-1. (C) Academic Press
MH  - analysis
MH  - CF0F1
MH  - chloroplast
MH  - Chloroplasts
MH  - COLI ATP SYNTHASE
MH  - COMPLEX
MH  - CROSS-LINKING
MH  - electron
MH  - electron microscopy
MH  - ELECTRON-MICROSCOPY
MH  - England
MH  - ESCHERICHIA-COLI
MH  - F- ATPASE
MH  - F1
MH  - H+-ATPase
MH  - IMAGE DATA
MH  - image processing
MH  - Microscopy
MH  - rotor-stator-model
MH  - stalk
MH  - SUBUNIT-EPSILON
RP  - NOT IN FILE
NT  - JournalArticleACADEMIC PRESS LTDSEP 4117FXLONDONBottcher B Univ Freiburg, Inst Phys Chem, Albertstr 23A, D-79104 Freiburg, GermanyJ MOL BIOL24-28 OVAL RD, LONDON NW1 7DX, ENGLAND
AV  - Univ Freiburg, Inst Phys Chem, Albertstr 23A, D-79104 Freiburg, Germany Univ Freiburg, Inst Phys Chem, D-79104 Freiburg, Germany
UR  - ISI:000075771200001
SO  - Journal of Molecular Biology 1998  ;281(5):757-762

1496
UI  - 19770
AU  - Boyer PD
AD  - Molecular Biology Institute, University of California at Los Angeles 90077-1570, USA pdboyer@uclaedu
TI  - ATP synthase--past and future
AB  - This paper gives an overview of a lecture scheduled for the opening of the 10th European Bioenergetics Congress. In this lecture I plan to first reflect on the accomplishments of some of the individuals who were involved in research on the ATP synthase during the past 50 years. Then I will give a brief view of the present information about rotational catalysis by the ATP synthase. This will be followed by a discussion of some results from my laboratory that call for additional experimentation. Finally I will direct attention to other questions about the ATP synthase that should be addressed in future studies
MH  - A
MH  - atp
MH  - ATP synthase
MH  - bioenergetics
MH  - Catalysis
MH  - H(+)-Transporting ATP Synthase
MH  - review
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 98358586LA - engRN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 19980824IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:9693715
SO  - Biochim Biophys Acta 1998 Jun 10 ;1365(1-2):3-9

1497
UI  - 19769
AU  - Boyer PD
AD  - Department of Chemistry and Biochemistry, University of California at Los Angeles, 90095-1469, USA
TI  - Energy, life, and ATP
AB  - The mechanism by which ATP is synthesized during oxidative and photophosphorylation has been elucidated by oxygen exchange and other studies: a novel form of catalysis termed rotary catalysis-is involved
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthase
MH  - Biochemistry
MH  - Catalysis
MH  - Chemistry
MH  - H(+)-Transporting ATP Synthase
MH  - mechanism
MH  - Oxygen
MH  - Photophosphorylation
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 99013127LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - 7782-44-7 (Oxygen)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - LecturesDA - 19990114IS - 0144-8463SB - IMCY - UNITED STATESJC - A6D
UR  - PM:9798783
SO  - Biosci Rep 1998 Jun ;18(3):97-117

1498
UI  - 19855
AU  - Bulygin VV
AU  - Duncan TM
AU  - Cross RL
AD  - Department of Biochemistry and Molecular Biology, State University of New York Health Science Center, Syracuse, New York 13210, USA
TI  - Rotation of the epsilon subunit during catalysis by Escherichia coli FOF1-ATP synthase
AB  - We report evidence for catalysis-dependent rotation of the single epsilon subunit relative to the three catalytic beta subunits of functionally coupled, membrane-bound FOF1-ATP synthase. Cysteines substituted at beta380 and epsilon108 allowed rapid formation of a specific beta-epsilon disulfide cross-link upon oxidation. Consistent with a need for epsilon to rotate during catalysis, tethering epsilon to one of the beta subunits resulted in the inhibition of both ATP synthesis and hydrolysis. These activities were fully restored upon reduction of the beta-epsilon cross-link. As a more critical test for rotation, a subunit dissociation/reassociation procedure was used to prepare a beta-epsilon cross-linked hybrid F1 having epitope-tagged betaD380C subunits (betaflag) exclusively in the two noncross-linked positions. This allowed the beta subunit originally aligned with epsilon to form the cross-link to be distinguished from the other two betas. The cross-linked hybrid was reconstituted with FO in F1-depleted membranes. After reduction of the beta-epsilon cross-link and a brief period of catalytic turnover, reoxidation resulted in a significant amount of betaflag in the beta-epsilon cross-linked product. In contrast, exposure to ligands that bind to the catalytic site but do not allow catalysis resulted in the subsequent cross-linking of epsilon to the original untagged beta. Furthermore, catalysis-dependent rotation of epsilon was prevented by prior treatment of membranes with N,N'-dicyclohexylcarbodiimide to block proton translocation through FO. From these results, we conclude that epsilon is part of the rotor that couples proton transport to ATP synthesis
MH  - A
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - BETA-SUBUNIT
MH  - Biochemistry
MH  - Catalysis
MH  - CROSS-LINKING
MH  - EPSILON-SUBUNIT
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1
MH  - H(+)-Transporting ATP Synthase
MH  - Hydrolysis
MH  - Ligands
MH  - Macromolecular Systems
MH  - membrane
MH  - Membranes
MH  - Proteins
MH  - proton
MH  - rotation
MH  - SUBUNIT
MH  - SYNTHASE
MH  - synthesis
MH  - translocation
MH  - transport
RP  - NOT IN FILE
NT  - UI - 99041935LA - engRN - 0 (Cross-Linking Reagents)RN - 0 (Macromolecular Systems)RN - 0 (Recombinant Proteins)RN - 52-90-4 (Cysteine)RN - 69-78-3 (Dithionitrobenzoic Acid)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM 23152/GM/NIGMSDA - 19981223IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:9822640
SO  - J Biol Chem 1998 Nov 27 ;273(48):31765-31769

1499
UI  - 20812
AU  - Caviston TL
AU  - Ketchum CJ
AU  - Sorgen PL
AU  - Nakamoto RK
AU  - Cain BD
AD  - Department of Biochemistry and Molecular Biology, University of Florida, Gainesville 32610, USA
TI  - Identification of an uncoupling mutation affecting the b subunit of F1F0 ATP synthase in Escherichia coli
AB  - A specific b subunit arginine, b(Arg-36) in Escherichia coli, displays evolutionary conservation among bacterial F1F0 ATP synthases. Site- directed mutagenesis was used to generate a collection of mutations affecting b(Arg-36). The phenotype differed depending upon the substitution, and the b(Arg-36-Glu) and b(Arg-36-Ile) substitutions virtually abolished enzyme function. Although the total amounts of F1F0 ATP synthase present in the membranes prepared from mutant strains were reduced, the primary effect of the b(Arg-36) substitutions was on the activities of the intact enzyme complexes. The most interesting result was that the b(Arg-36-Glu) substitution results in the uncoupling of a functional F0 from F1 ATP hydrolysis activity
MH  - A
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Bacterial Proteins
MH  - Biochemistry
MH  - COMPLEX
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - F1
MH  - function
MH  - Hydrolysis
MH  - membrane
MH  - Membranes
MH  - mutagenesis
MH  - mutant
MH  - protein
MH  - Proteins
MH  - Site
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 98312610LA - engRN - 0 (Bacterial Proteins)RN - 0 (uncF protein, Escherichia coli)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleID - GM43495/GM/NIGMSID - GM50957/GM/NIGMSDA - 19980727IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:9650590
SO  - FEBS Lett 1998 Jun 12 ;429(2):201-206

1500
UI  - 28
AU  - Dimroth P
AU  - Kaim G
AU  - Matthey U
AD  - Mikrobiologisches Institut, Eidgenossische Technische Hochschule, Zurich, Switzerland
TI  - The motor of the ATP synthase
AB  - A model is presented in which ion translocation through the F0 part of the ATP synthase drives the rotation of the ring of c subunits (rotor) versus the a subunit (stator). The coupling ion binding sites on the rotor are accessible from the cytoplasm of a bacterial cell except for the c subunit at the interface to the stator. Here, the binding site is accessible from the periplasm through a channel formed by subunit a. In the ATP synthesis mode, a coupling ion is anticipated to pass through the stator channel into the binding site of the adjacent rotor subunit, following the electrical potential. Occupation of this site triggers, probably by electrostatic forces, the rotation of the ring. This makes the binding site accessible to the cytoplasm, where the coupling ion dissociates. Simultaneously, this rotation moves again an empty rotor subunit into the contact site with the stator, where its binding site becomes loaded and rotation continues
RP  - NOT IN FILE
NT  - UI - 98358597LA - engRN - 7440-23-5 (Sodium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 19980824IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:9693726
SO  - Biochim Biophys Acta 1998 Jun 10 ;1365(1-2):87-92

1501
UI  - 19597
AU  - Dioumaev AK
AU  - Richter HT
AU  - Brown LS
AU  - Tanio M
AU  - Tuzi S
AU  - Saito H
AU  - Kimura Y
AU  - Needleman R
AU  - Lanyi JK
TI  - Existence of a proton transfer chain in bacteriorhodopsin: participation of Glu-194 in the release of protons to the extracellular surface.
AB  - Glu-194 near the extracellular surface of bacteriorhodopsin is indispensable for proton release to the medium upon protonation of Asp- 85 during light-driven transport. As for Glu-204, its replacement with glutamine (but not aspartate) abolishes both proton release and the anomalous titration of Asp-85 that originates from coupling between the pKa of this buried aspartate and those of the other acidic groups. Unlike the case of Glu-204, however, replacement of Glu-194 with aspartate raises the pKa for proton release. In Fourier transform infrared spectra of the E194D mutant a prominent positive band is observed at 1720 cm-1. It can be assigned from [4-13C]aspartate and D2O isotope shifts to the C&dbd;O stretch of protonated Asp-194. Its rise correlates with proton transfer from the retinal Schiff base to Asp-85. Its decay coincides with the appearance of a proton at the surface, detected under similar conditions with fluorescein covalently bound to Lys-129 and with pyranine. Its amplitude decreases with increasing pH, with a pKa of about 9. We show that this pKa is likely to be that of the internal proton donor to Asp-194, the Glu-204 site, before photoexcitation, while 13C NMR titration indicates that Asp-194 has an initial pKa of about 3. We propose that there is a chain of interacting residues between the retinal Schiff base and the extracellular surface. After photoisomerization of the retinal the pKa's change so as to allow (i) Asp-85 to become protonated by the Schiff base, (ii) the Glu-204 site to transfer its proton to Asp-194 in E194D, and therefore to Glu- 194 in the wild type, and (iii) residue 194 to release the proton to the medium.
MH  - A
MH  - Asp-85
MH  - Bacteriorhodopsin
MH  - BASE
MH  - Binding Sites
MH  - Biophysics
MH  - Cell Membrane
MH  - Chemistry
MH  - fluorescein
MH  - FOURIER TRANSFORM
MH  - genetics
MH  - glutamic acid
MH  - Glutamine
MH  - Halobacterium salinarium
MH  - Hydrogen-Ion Concentration
MH  - isotope
MH  - Kinetics
MH  - metabolism
MH  - Models,Molecular
MH  - mutant
MH  - NMR
MH  - pH
MH  - photochemistry
MH  - photoisomerization
MH  - physiology
MH  - point mutation
MH  - Protein Conformation
MH  - proton
MH  - proton release
MH  - Proton transfer
MH  - Proton-transfer
MH  - protonation
MH  - Protons
MH  - RESIDUE
MH  - retinal
MH  - Retinaldehyde
MH  - Schiff base
MH  - Schiff Bases
MH  - Schiff-base
MH  - Site
MH  - spectra
MH  - Spectroscopy,Fourier Transform Infrared
MH  - Support,U.S.Gov't,Non-P.H.S.
MH  - Support,U.S.Gov't,P.H.S.
MH  - SURFACE
MH  - TRANSFER
MH  - transport
RP  - IN FILE
NT  - Department of Physiology & Biophysics, University of California, Irvine, California 92697-4056, USAPMID- 0009485398
SO  - Biochemistry 1998 Feb 24 ;37(8):2496-2506

1502
UI  - 19908
AU  - Elston T
AU  - Wang H
AU  - Oster G
AD  - Department of Molecular and Cellular Biology, University of California, Berkeley 94720-3112, USA
TI  - Energy transduction in ATP synthase
AB  - Mitochondria, bacteria and chloroplasts use the free energy stored in transmembrane ion gradients to manufacture ATP by the action of ATP synthase. This enzyme consists of two principal domains. The asymmetric membrane-spanning F0 portion contains the proton channel, and the soluble F1 portion contains three catalytic sites which cooperate in the synthetic reactions. The flow of protons through F0 is thought to generate a torque which is transmitted to F1 by an asymmetric shaft, the coiled-coil gamma-subunit. This acts as a rotating 'cam' within F1, sequentially releasing ATPs from the three active sites. The free- energy difference across the inner membrane of mitochondria and bacteria is sufficient to produce three ATPs per twelve protons passing through the motor. It has been suggested that this proton motive force biases the rotor's diffusion so that F0 constitutes a rotary motor turning the gamma shaft. Here we show that biased diffusion, augmented by electrostatic forces, does indeed generate sufficient torque to account for ATP production. Moreover, the motor's reversibility- supplying torque from ATP hydrolysis in F1 converts the motor into an efficient proton pump-can also be explained by our model
MH  - A
MH  - ACTIVE
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP production
MH  - ATP synthase
MH  - Bacteria
MH  - chloroplast
MH  - Chloroplasts
MH  - Diffusion
MH  - England
MH  - F0
MH  - F1
MH  - H(+)-Transporting ATP Synthase
MH  - Hydrolysis
MH  - ion
MH  - membrane
MH  - Mitochondria
MH  - model
MH  - proton
MH  - Protons
MH  - Site
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 98121120LA - engRN - 0 (Protons)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19980217IS - 0028-0836SB - IMCY - ENGLANDJC - NSC
UR  - PM:9461222
SO  - Nature 1998 Jan 29 ;391(6666):510-513

1503
UI  - 963
AU  - Embley TM
AU  - Martin W
TI  - A hydrogen-producing mitochondrion
MH  - Adenosine Triphosphate
MH  - Anaerobiosis
MH  - Animal
MH  - biosynthesis
MH  - Ciliophora
MH  - Cockroaches
MH  - DNA,Mitochondrial
MH  - DNA,Protozoan
MH  - Energy Metabolism
MH  - Evolution
MH  - genetics
MH  - Hydrogen
MH  - metabolism
MH  - Mitochondria
MH  - Organelles
MH  - parasitology
MH  - ultrastructure
RP  - NOT IN FILE
SO  - Nature 1998 Dec 10 ;396(6711):517-519

1504
UI  - 19758
AU  - Feniouk BA
AU  - Junge W
AU  - Mulkidjanian AY
TI  - Tracking of proton flow across the active ATP-synthase of Rhodobacter capsulatus in response to a series of light flashes.
MH  - A
MH  - ACTIVE
MH  - ATP synthase
MH  - capsulatus
MH  - electrogenic
MH  - ion
MH  - Light
MH  - proton
MH  - Rhodobacter
MH  - rhodobacter capsulatus
MH  - RHODOBACTER-CAPSULATUS
RP  - NOT IN FILE
SO  - EBEC Short Reports 1998  ;10():112-112

1505
UI  - 19757
AU  - Feniouk BA
AU  - Junge W
AU  - Mulkidjanian AY
TI  - ATP-synthase of Rhodobacter capsulatus: Proton flow and ATP synthesis in response to light flashes.
MH  - A
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - capsulatus
MH  - electrogenic
MH  - ion
MH  - Light
MH  - proton
MH  - Rhodobacter
MH  - rhodobacter capsulatus
MH  - RHODOBACTER-CAPSULATUS
MH  - synthesis
RP  - NOT IN FILE
SO  - Photosynthesis: Mechanisms and Effects ( G Garab, editor) 1998  ;():1679-1682

1506
UI  - 15
AU  - Fillingame RH
AU  - Girvin ME
AU  - Jiang W
AU  - Valiyaveetil F
AU  - Hermolin J
AD  - Department of Biomolecular Chemistry, University of Wisconsin Medical School, Madison 53705, USA
TI  - Subunit interactions coupling H+ transport and ATP synthesis in F1F0 ATP synthase
RP  - NOT IN FILE
NT  - UI - 99005986LA - engRN - 0 (Isoenzymes)RN - 0 (Protons)RN - 1333-74-0 (Hydrogen)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialID - GM23105/GM/NIGMSDA - 19990112IS - 0302-2994SB - IMCY - ENGLANDJC - 1UF
UR  - PM:9789557
SO  - Acta Physiol Scand Suppl 1998 Aug ;643():163-168

1507
UI  - 104
AU  - Fillingame RH
AU  - Jones PC
AU  - Jiang W
AU  - Valiyaveetil FI
AU  - Dmitriev OY
AD  - Department of Biomolecular Chemistry, University of Wisconsin Medical School, Madison 53705, USA
TI  - Subunit organization and structure in the F0 sector of Escherichia coli F1F0 ATP synthase
AB  - In this review, we summarize recent work from our laboratory which establishes the topology and nearest neighbor organization of subunits in the F0 sector of the H+ transporting ATP synthase of Escherichia coli. The E. coli F0 sector is composed of three subunits in an a1b2c12 stoichiometric ratio. Crosslinking experiments with genetically introduced Cys establish a ring-like organization of the 12 c subunits with subunits a and b lying to the outside of the ring. The results are interpreted using an atomic resolution structural model of monomeric subunit c in a chloroform-methanol-water (4:4:1, v/v/v) solution, derived by heteronuclear NMR (M.E. Girvin, F. Abildgaard, V. Rastogi, J. Markley, R.H. Fillingame, in press). The crosslinking results validate many predictions of the structural model and confirm a front- to-back-type packing of two subunit c into a functional dimer, as was first predicted from genetic studies. Aspartyl-61, the proton translocating residue, lies at the center of the four transmembrane helices of the functional dimer, rather than at the periphery of the subunit c ring. Subunit a is shown to fold with five transmembrane helices, and a functionally important interaction of transmembrane helix-4 with transmembrane helix-2 of subunit c is established. The single transmembrane helices of the two subunit b dimerize in the membrane. The structure of the transmembrane segment of subunit b is predicted from the NMR structure of the monomeric peptide
RP  - NOT IN FILE
NT  - UI - 98358603LA - engRN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialID - GM-23105/GM/NIGMSDA - 19980824IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:9693732
SO  - Biochim Biophys Acta 1998 Jun 10 ;1365(1-2):135-142

1508
UI  - 146
AU  - Garcia JJ
AU  - Capaldi RA
AD  - Institute of Molecular Biology, University of Oregon, Eugene, Oregon 97403-1229, USA
TI  - Unisite catalysis without rotation of the gamma-epsilon domain in Escherichia coli F1-ATPase
AB  - Unisite [gamma-32P]ATP hydrolysis was studied in ECF1 from the mutant betaE381C after generating a single disulfide bond between beta and gamma subunits to prevent the rotation of the gamma/epsilon domain. The single beta-gamma cross-link was obtained by removal of the delta subunit from F1 and then treating with CuCl2 as described previously (Aggeler, R., Haughton, M. A., and Capaldi, R. A. (1996) J. Biol. Chem. 270, 9185-9191). The mutant enzyme, betaE381C, had an increased overall rate of unisite hydrolysis of [gamma-32P]ATP compared with the wild type ECF1 due to increases in the rate of ATP binding (k+1), Pi release (k+3), and ADP release (k+4). Release of bound substrate ([gamma- 32P]ATP) was also increased in the betaE381C mutant. Cross-linking between Cys-381 and the intrinsic Cys-87 of gamma caused a further increase in the rate of unisite catalysis, mainly by additional effects on nucleotide binding in the high affinity catalytic site (k+1 and k+4). In delta-subunit-free ECF1 from wild type or betaE381C F1, addition of an excess of ATP accelerated unisite catalysis. After cross- linking, unisite catalysis of betaE381C was not enhanced by the cold chase. The covalent linkage of gamma to beta increased the rate of unisite catalysis to that obtained by cold chase of ATP of the noncross- linked enzyme. It is concluded that the conversion of Glu-381 of beta to Cys induces an activated conformation of the high affinity catalytic site with low affinity for substrate and products. This state is stabilized by cross-linking the Cys at beta381 to Cys-87 of gamma. We infer from the data that rotation of the gamma/epsilon rotor in ECF1 is not linked to unisite hydrolysis of ATP at the high affinity catalytic site but to ATP binding to a second or third catalytic site on the enzyme
RP  - NOT IN FILE
NT  - UI - 98298094LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - 7440-50-8 (Copper)RN - 7758-89-6 (cuprous chloride)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - HL24526/HL/NHLBIDA - 19980803IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:9632641
SO  - J Biol Chem 1998 Jun 26 ;273(26):15940-15945

1509
UI  - 105
AU  - Girvin ME
AU  - Rastogi VK
AU  - Abildgaard F
AU  - Markley JL
AU  - Fillingame RH
AD  - Department of Biomolecular Chemistry, University of Wisconsin Medical School, Madison 53706, USA girvin@aecomyuedu
TI  - Solution structure of the transmembrane H+-transporting subunit c of the F1F0 ATP synthase
AB  - Subunit c is the H+-translocating component of the F1F0 ATP synthase complex. H+ transport is coupled to conformational changes that ultimately lead to ATP synthesis by the enzyme. The properties of the monomeric subunit in a single-phase solution of chloroform-methanol- water (4:4:1) have been shown to mimic those of the protein in the native complex. Triple resonance NMR experiments were used to determine the complete structure of monomeric subunit c in this solvent mixture. The structure of the protein was defined by >2000 interproton distances, 64 (3)JN alpha, and 43 hydrogen-bonding NMR-derived restraints. The root mean squared deviation for the backbone atoms of the two transmembrane helices was 0.63 A. The protein folds as a hairpin of two antiparallel helical segments, connected by a short structured loop. The conserved Arg41-Gln42-Pro43 form the top of this loop. The essential H+-transporting Asp61 residue is located at a slight break in the middle of the C-terminal helix, just prior to Pro64. The C-terminal helix changes direction by 30 +/- 5 degrees at the conserved Pro64. In its protonated form, the Asp61 lies in a cavity created by the absence of side chains at Gly23 and Gly27 in the N- terminal helix. The shape and charge distribution of the molecular surface of the monomeric protein suggest a packing arrangement for the oligomeric protein in the F0 complex, with the front face of one monomer packing favorably against the back face of a second monomer. The packing suggests that the proton (cation) binding site lies between packed pairs of adjacent subunit c
RP  - NOT IN FILE
NT  - UI - 98301362LA - engRN - 0 (Carbon Isotopes)RN - 0 (Nitrogen Isotopes)RN - 0 (Solutions)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM23105/GM/NIGMSID - GM55371/GM/NIGMSID - R02301/PHSID - etcDA - 19980720IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:9636021
SO  - Biochemistry 1998 Jun 23 ;37(25):8817-8824

1510
UI  - 500
AU  - Golden TR
AU  - Pedersen PL
AD  - Department of Biological Chemistry, Johns Hopkins University, School of Medicine, Baltimore, Maryland 21205-2185, USA
TI  - The oligomycin sensitivity conferring protein of rat liver mitochondrial ATP synthase: arginine 94 is important for the binding of OSCP to F1
AB  - The oligomycin sensitivity conferring protein (OSCP) is an essential subunit of the mitochondrial ATP synthase (F0F1) long regarded as being directly involved in the energetic coupling of proton transport to ATP synthesis. To gain insight into the function of OSCP, mutations were made in a highly conserved central region of the subunit, and the recombinant proteins were studied using several biochemical assays. Rat liver OSCP was expressed to high levels in Escherichia coli, solubilized from inclusion bodies, renatured, and purified to homogeneity. The recombinant protein was able to reconstitute oligomycin-sensitive ATPase activity to inner membrane vesicles depleted of F1 and OSCP, and bound to F1 with a stoichiometry of 1:1. A novel fluorescence anisotropy assay was developed to study the affinity of binding of F1 to OSCP, providing a Kd value of 51 +/- 11 nM. Two highly conserved, charged residues (E91 and R94) which lie within the central region of OSCP were mutated, and the recombinant proteins (E91Q, R94Q, and R94A) were purified to homogeneity and judged by CD spectroscopy to have structures similar to that of the wild-type protein. Both R94 mutants demonstrated little or no binding to F1, while the E91Q bound in a manner identical to that of wild-type OSCP. Significantly, all three mutant proteins were able to reconstitute F1 with membranes and to confer oligomycin sensitivity to the same extent as wild-type OSCP. These results demonstrate that a single tight binding site exists on isolated rat liver F1 for OSCP, and implicate arginine 94 as playing a critical role in this site. In addition, these results indicate that this tight binding site is not required for conferral of oligomycin sensitivity to the reconstituted F0F1 complex
RP  - NOT IN FILE
NT  - UI - 98426086LA - engRN - 0 (Membrane Proteins)RN - 0 (Oligomycins)RN - 0 (Recombinant Proteins)RN - 0 (oligomycin sensitivity-conferring protein)RN - 7004-12-8 (Arginine)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - CA 10957/CA/NCIDA - 19981028IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:9753477
SO  - Biochemistry 1998 Sep 29 ;37(39):13871-13881

1511
UI  - 18971
AU  - Gopta OA
AU  - Feniouk BA
AU  - Junge W
AU  - Mulkidjanian AY
AD  - AN Belozersky Institute of Physico-Chemical Biology, Moscow University, Russia
TI  - The cytochrome bc1 complex of Rhodobacter capsulatus: ubiquinol oxidation in a dimeric Q-cycle?
AB  - We studied the cytochrome bc1 complex (hereafter bc) by flash excitation of Rhodobacter capsulatis chromatophores. The reduction of the high-potential heme b(h), of cytochrome b (at 561 nm) and of cytochromes c (at 552 nm) and the electrochromic absorption transients (at 524 nm) were monitored after the first and second flashes of light, respectively. We kept the ubiquinone pool oxidized in the dark and concerned for the ubiquinol formation in the photosynthetic reaction center only after the second flash. Surprisingly, the first flash caused the oxidation of about one ubiquinol per bc dimer. Based on these and other data we propose a dimeric Q-cycle where the energetically unfavorable oxidation of the first ubiquinol molecule by one of the bc monomers is driven by the energetically favorable oxidation of the second ubiquinol by the other bc monomer resulting in a pairwise oxidation of ubiquinol molecules by the dimeric bc in the dark. The residual unpaired ubiquinol supposedly remains on the enzyme and is then oxidized after the first flash
MH  - A
MH  - absorption
MH  - capsulatus
MH  - COMPLEX
MH  - cytochrome
MH  - Cytochrome b
MH  - Cytochromes
MH  - Light
MH  - Rhodobacter
MH  - RHODOBACTER-CAPSULATUS
MH  - Ubiquinol-Cytochrome-c Reductase
MH  - Ubiquinone
RP  - NOT IN FILE
NT  - UI - 98372653LA - engRN - 1339-63-5 (Ubiquinone)RN - 56275-39-9 (ubiquinol)RN - EC 1.10.2.2 (Ubiquinol-Cytochrome-c Reductase)PT - Journal ArticleDA - 19980908IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:9708922
SO  - FEBS Lett 1998 Jul 17 ;431(2):291-296

1512
UI  - 9947
AU  - Groth G
AU  - Tilg Y
AU  - Schirwitz K
AD  - Biochemie der Pflanzen, Heinrich-Heine Universitat Dussseldorf, Universitatsstr1, Dusseldorf, 40225, Germany georggroth@uni- duesseldorfde
TI  - Molecular architecture of the c-subunit oligomer in the membrane domain of F-ATPases probed by tryptophan substitution mutagenesis
AB  - Subunit c of the proton-transporting ATP synthase of Escherichia coli forms an oligomeric complex in the membrane domain that functions in transmembrane proton conduction. In order to gain some insight into the architecture of this oligomeric complex, the transmembrane region in the C-terminal membrane-spanning segment was analysed by a site- directed mutagenesis approach. Tryptophan substitution of consecutive residues in positions 61 to 72 of subunit c was used to identify residues oriented towards a helix-helix surface or an accessible phase in the oligomeric complex. Mutants were analysed in functional assays of ATP hydrolysis, ATP synthesis and ATP-dependent proton transport. Function was disrupted according to a pattern that identified inter- and intramolecular contacts in the c-subunit oligomer. Screening experiments on minimal medium support the helix-helix contacts found in the functional assays. The results add strong constraints to the potential orientation of the monomers in the oligomeric complex and are discussed against the background of different structural models that have been proposed for the c-subunit oligomer
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - COMPLEX
MH  - England
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F-ATPASE
MH  - H(+)-Transporting ATP Synthase
MH  - Hydrolysis
MH  - model
MH  - proton
MH  - Protons
MH  - SUBUNIT
MH  - SYNTHASE
MH  - transport
RP  - NOT IN FILE
NT  - UI - 98346992LA - engRN - 0 (Culture Media)RN - 0 (DNA, Bacterial)RN - 0 (Protons)RN - 56-65-5 (Adenosine Triphosphate)RN - 73-22-3 (Tryptophan)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19980820IS - 0022-2836SB - IMCY - ENGLANDJC - J6V
UR  - PM:9680474
SO  - J Mol Biol 1998 Aug 7 ;281(1):49-59

1513
UI  - 287
AU  - Hasler K
AU  - Engelbrecht S
AU  - Junge W
AD  - Dept Biology/Chemistry, Universitat Osnabruck, Germany
TI  - Three-stepped rotation of subunits gamma and epsilon in single molecules of F-ATPase as revealed by polarized, confocal fluorometry
AB  - The proton translocating ATP synthase is conceived as a rotatory molecular engine. ATP hydrolysis by its headpiece, CF1, drives the rotation of subunit gamma relative to the hexagonally arranged large subunits, (alphabeta)3. We investigated transition states of the rotatory drive by polarized confocal fluorometry (POCOF) as applied to single molecules of engineered, immobilized and load-free spinach-CF1. We found that the hydrolysis of ATP caused the stepped and sequential progression of subunit gamma through three discrete angular positions, with the transition states of gamma being too shortlived for detection. We also observed the stepped motion of epsilon, whereas delta was immobile as (alphabeta)3
RP  - NOT IN FILE
NT  - UI - 98260882LA - engRN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19980610IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:9600255
SO  - FEBS Lett 1998 Apr 24 ;426(3):301-304

1514
UI  - 811
AU  - Hisabori T
AU  - Motohashi K
AU  - Kroth P
AU  - Strotmann H
AU  - Amano T
AD  - Research Laboratory of Resources Utilization, Tokyo Institute of Technology, Nagatsuta 4259, Midori-ku, Yokohama 226-8503, Japan thisabor@restitechacjp
TI  - The formation or the reduction of a disulfide bridge on the gamma subunit of chloroplast ATP synthase affects the inhibitory effect of the epsilon subunit
AB  - We have studied the change of the catalytic activity of chimeric complexes that were formed by chloroplast coupling factor 1 (CF1) - gamma, alpha and beta subunits of thermophilic bacterial F1 after formation or reduction of the disulfide bridge of different gamma subunits modified by oligonucleotide-directed mutagenesis techniques. For this purpose, three mutant gamma subunits were produced: gamma Delta194-230, here 37 amino acids from Pro-194 to Ile-230 are deleted, gammaC199A, Cys-199 is changed to Ala, and gamma Delta200-204, amino acids from Asp-200 to Lys-204 are deleted. All of the chimeric subunit complexes produced from each of these mutant CF1-gamma subunits and alpha and beta subunits from thermophilic bacterial F1 lost the sensitivity against thiol reagents when compared with the complex containing wild-type CF1-gamma. The pH optimum (pH 8.5-9.0) and the concentration of methanol to stimulate ATPase activities were not affected by these mutations. These indicate that the introduction of the mutations did not change the main features of ATPase activity of the chimeric complex. However, the interaction between gamma subunit and epsilon subunit was strongly influenced by the type of gamma subunit itself. Although the ATPase activity of the chimeric complex that contained gamma Delta200-204 or gammaC199A was inhibited by the addition of recombinant epsilon subunit from CF1 similarly to complexes containing the reduced wild-type gamma subunit, the recombinant epsilon subunit did not inhibit the ATPase of the complex, which contained the oxidized form of gamma subunit. Therefore the affinity of the epsilon subunit to the gamma subunit may be dependent on the state of the gamma subunit or the epsilon subunit may bind to the oxidized form of gamma subunit in a mode that does not inhibit the activity. The ATPase activity of the complex that contains gamma Delta194-230 was not efficiently inhibited by epsilon subunit. These results show that the formation or reduction of the disulfide bond on the gamma subunit may induce a conformational change in the region that directly affects the interaction of this subunit with the adjacent epsilon subunit
RP  - NOT IN FILE
NT  - UI - 98298088LA - engRN - 0 (Chimeric Proteins)RN - 0 (Disulfides)RN - 67-56-1 (Methanol)RN - EC 3.6.1.- (Ca(2+) Mg(2+)-ATPase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19980803IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:9632635
SO  - J Biol Chem 1998 Jun 26 ;273(26):15901-15905

1515
UI  - 812
AU  - Hu D
AU  - Strotmann H
AU  - Shavit N
AU  - Leu S
AD  - The Doris and Bertie Black Center for Bioenergetics in Life Sciences, Ben Gurion University of the Negev, Beer Sheva, Israel
TI  - The C. reinhardtii CF1 with the mutation betaT168S has high ATPase activity
AB  - We have generated the mutation T168S in the beta subunit of the chloroplast ATP synthase complex of Chlamydomonas reinhardtii by site directed mutagenesis and chloroplast transformation. CF1 and the alpha3beta3gamma complex of this mutant strain were isolated and their enzymatic activities were characterized and compared to those of the corresponding wild type complexes. Without activation the mutant CF1 exhibits MgATPase activity with at least 10 times higher rates than the wild type enzyme. The MgATPase activity could be stimulated to some extent by methanol, but less by ethanol and octylglucoside. The alpha3beta3gamma complex had an even higher MgATPase activity, which was only slightly enhanced by ethanol or methanol. The ATPase activities of the mutant complexes, like those of the wild type complexes, displayed a sharp concentration optimum for Mg2+. Free ADP inhibited neither the mutant nor the wild type ATPase significantly. Azide, which strongly inhibited the ATPase activity of the wild type enzyme, inhibited the mutant enzyme only at an about 30 times higher concentration suggesting that the mutation T168S prevents trapping of a tightly bound MgADP by a catalytic site that regulates chloroplast ATPase activity. The mutant cells grew photoautotrophically at a growth rate of about 50%. Similar to the wild type the cells survived on minimal medium in the dark. Under heterotrophic conditions with acetate as energy and carbon source the mutant cells grew much faster than the wild type cells, but the chlorophyll content per cell decreased dramatically
RP  - NOT IN FILE
NT  - UI - 98122673LA - engRN - 0 (Recombinant Proteins)RN - 1406-65-1 (Chlorophyll)RN - 64-17-5 (Ethanol)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)RN - EC 3.6.1.38 (Ca(2+)-Transporting ATPase)PT - Journal ArticleDA - 19980224IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:9462841
SO  - FEBS Lett 1998 Jan 2 ;421(1):65-68

1516
UI  - 21243
AU  - Iwata S
AU  - Lee JW
AU  - Okada K
AU  - Lee JK
AU  - Iwata M
AU  - Rasmussen B
AU  - Link TA
AU  - Ramaswamy S
AU  - Jap BK
AD  - Life Sciences Division, Lawrence Berkeley National Laboratory, University of California, Berkeley, CA 94720, USA iwata@xraybmcuuse
TI  - Complete structure of the 11-subunit bovine mitochondrial cytochrome bc1 complex
AB  - Mitochondrial cytochrome bc1 complex performs two functions: It is a respiratory multienzyme complex and it recognizes a mitochondrial targeting presequence. Refined crystal structures of the 11-subunit bc1 complex from bovine heart reveal full views of this bifunctional enzyme. The "Rieske" iron-sulfur protein subunit shows significant conformational changes in different crystal forms, suggesting a new electron transport mechanism of the enzyme. The mitochondrial targeting presequence of the "Rieske" protein (subunit 9) is lodged between the two "core" subunits at the matrix side of the complex. These "core" subunits are related to the matrix processing peptidase, and the structure unveils how mitochondrial targeting presequences are recognized
MH  - A
MH  - COMPLEX
MH  - conformational change
MH  - conformational changes
MH  - cytochrome
MH  - Cytochrome b
MH  - Cytochrome c1
MH  - electron
MH  - Electron Transport
MH  - Enzyme Inhibitors
MH  - function
MH  - inhibitor
MH  - mechanism
MH  - Multienzyme Complexes
MH  - protein
MH  - Proteins
MH  - structure
MH  - SUBUNIT
MH  - Thiazoles
MH  - transport
MH  - Ubiquinol-Cytochrome-c Reductase
RP  - NOT IN FILE
NT  - UI - 98316377LA - engRN - 0 (Enzyme Inhibitors)RN - 0 (Hydroquinones)RN - 0 (Iron-Sulfur Proteins)RN - 0 (Rieske iron-sulfur protein)RN - 0 (Thiazoles)RN - 123-31-9 (hydroquinone)RN - 76706-55-3 (myxothiazol)RN - 9035-37-4 (Cytochrome b)RN - 9035-42-1 (Cytochrome c1)RN - EC 1.10.2.2 (Ubiquinol-Cytochrome-c Reductase)PT - Journal ArticleDA - 19980723IS - 0036-8075SB - IMCY - UNITED STATES
UR  - PM:9651245
SO  - Science 1998 Jul 3 ;281(5373):64-71

1517
UI  - 20936
AU  - Jager H
AU  - Birkenhager R
AU  - Stalz WD
AU  - Altendorf K
AU  - Deckers-Hebestreit G
AD  - Universitat Osnabruck, Fachbereich Biologie/Chemie, Abteilung Mikrobiologie, Germany
TI  - Topology of subunit a of the Escherichia coli ATP synthase
AB  - The antigenic determinants of mAbs against subunit a of the Escherichia coli ATP synthase were mapped by ELISA using overlapping synthetic decapeptides. For two of the mAbs the epitopes are E4NMTPQD10 (GDH 14- 5C6) and V29DPQ32 (GDH 8-8B3). Binding of these mAbs to membrane vesicles of different orientation revealed that both epitopes are accessible in vesicles with inside-out orientation. These results demonstrate that at least the N-terminal amino acids 1-32 of subunit a are located at the cytoplasmic side of the membrane. A further determination of the topology of subunit a was performed by inserting the reporter epitope DYKDDDDK (FLAG epitope) at different positions of the polypeptide chain. 10 of 13 insertions led to a functional F0F1 ATP synthase and allowed specific detection of the modified subunit a by immunoblotting using an mAb against the FLAG epitope. In addition, polyclonal anti-FLAG IgG was applied for the recognition of the mutant FLAG epitope DYKDDVDK. Cells carrying this mutant FLAG epitope at the C terminus of subunit a were able to grow on succinate as sole carbon and energy source, revealing a functional ATP synthase, in contrast to those carrying the original FLAG epitope at the same position. Binding studies with membrane vesicles of different orientation and anti-FLAG Ig demonstrated that both termini of the protein are located at the cytoplasmic side of the membrane, indicating that an even number of membrane-spanning segments is present in subunit a. In addition, insertion of two FLAG epitopes in tandem after K66, or one epitope after H95, and Q181 revealed that the polypeptide regions including these residues are accessible from the cytoplasmic surface of the membrane. These results support the view that the polypeptide chain of subunit a traverses the membrane six times
MH  - A
MH  - ACID
MH  - Amino Acids
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BINDING
MH  - Cells
MH  - COLI ATP SYNTHASE
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0F1
MH  - Histidine
MH  - membrane
MH  - membrane vesicles
MH  - mutant
MH  - protein
MH  - Proteins
MH  - RESIDUE
MH  - SUBUNIT
MH  - succinate
MH  - SURFACE
MH  - SYNTHASE
MH  - Time
RP  - NOT IN FILE
NT  - UI - 98151240LA - engRN - 0 (Antibodies, Monoclonal)RN - 0 (Epitopes)RN - 0 (Recombinant Proteins)RN - 71-00-1 (Histidine)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19980323IS - 0014-2956SB - IMCY - GERMANY
UR  - PM:9492276
SO  - Eur J Biochem 1998 Jan 15 ;251(1-2):122-132

1518
UI  - 106
AU  - Jiang W
AU  - Fillingame RH
AD  - Department of Biomolecular Chemistry, University of Wisconsin Medical School, Madison, WI 53706, USA
TI  - Interacting helical faces of subunits a and c in the F1Fo ATP synthase of Escherichia coli defined by disulfide cross-linking
AB  - Subunits a and c of Fo are thought to cooperatively catalyze proton translocation during ATP synthesis by the Escherichia coli F1Fo ATP synthase. Optimizing mutations in subunit a at residues A217, I221, and L224 improves the partial function of the cA24D/cD61G double mutant and, on this basis, these three residues were proposed to lie on one face of a transmembrane helix of subunit a, which then interacted with the transmembrane helix of subunit c anchoring the essential aspartyl group. To test this model, in the present work Cys residues were introduced into the second transmembrane helix of subunit c and the predicted fourth transmembrane helix of subunit a. After treating the membrane vesicles of these mutants with Cu(1, 10-phenanthroline)2SO4 at 0 degrees, 10 degrees, or 20 degreesC, strong a-c dimer formation was observed at all three temperatures in membranes of 7 of the 65 double mutants constructed, i.e., in the aS207C/cI55C, aN214C/cA62C, aN214C/cM65C, aI221C/cG69C, aI223C/cL72C, aL224C/cY73C, and aI225C/cY73C double mutant proteins. The pattern of cross-linking aligns the helices in a parallel fashion over a span of 19 residues with the aN214C residue lying close to the cA62C and cM65C residues in the middle of the membrane. Lesser a-c dimer formation was observed in nine other double mutants after treatment at 20 degreesC in a pattern generally supporting that indicated by the seven landmark residues cited above. Cross-link formation was not observed between helix-1 of subunit c and helix-4 of subunit a in 19 additional combinations of doubly Cys-substituted proteins. These results provide direct chemical evidence that helix-2 of subunit c and helix-4 of subunit a pack close enough to each other in the membrane to interact during function. The proximity of helices supports the possibility of an interaction between Arg210 in helix-4 of subunit a and Asp61 in helix-2 of subunit c during proton translocation, as has been suggested previously
RP  - NOT IN FILE
NT  - UI - 98283978LA - engRN - 0 (Cross-Linking Reagents)RN - 0 (Sulfides)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM23105/GM/NIGMSDA - 19980709IS - 0027-8424SB - IMCY - UNITED STATESJC - PV3
UR  - PM:9618459
SO  - Proc Natl Acad Sci U S A 1998 Jun 9 ;95(12):6607-6612

1519
UI  - 14
AU  - Jones PC
AU  - Fillingame RH
AD  - Department of Biomolecular Chemistry, University of Wisconsin Medical School, Madison, Wisconsin 53706, USA
TI  - Genetic fusions of subunit c in the F0 sector of H+-transporting ATP synthase. Functional dimers and trimers and determination of stoichiometry by cross-linking analysis
AB  - The multicopy c subunit of the H+-transporting ATP synthase of Escherichia coli folds through the transmembrane F0 sector as a hairpin of two hydrophobic alpha-helices with the proton-translocating aspartyl- 61 side chain centered in the second transmembrane helix. The number of subunits c in the F0 complex, which is thought to determine the H+- pumping/ATP stoichiometry, was previously not determined with exactness but thought to range from 9-12. The studies described here indicate that the exact number is 12. Based upon the precedent of the subunit c in vacuolar-type ATPases, which are composed of four transmembrane helices and seem to have evolved by gene duplication of an F0-type progenitor gene, we constructed genetically fused dimers and trimers of E. coli subunit c. Both the dimeric and trimeric forms proved to be functional. These results indicate that the total number of subunit c in F0 should be a multiple of 2 and 3. Based upon a previous study in which the oligomeric organization of c subunits in F0 was determined by cross-linking of Cys-substituted subunits (Jones, P. C. , Jiang, W., and Fillingame, R. H. (1998) J. Biol. Chem. 273, 17178-17185), we introduced Cys into the first and last transmembrane helices of subunit c monomers, dimers, and trimers and attempted to generate cross-linked products by oxidation with Cu(II)-(1,10-phenanthroline)2. Double Cys substitutions at two sets of positions gave rise to extensive cross- linked multimers. Multimers of the monomer that extended up to the position of c12 were correlated and calibrated with distinct cross- linked species of the appropriate doubly Cys-substituted dimers (i.e. c2, c4, . c12) and doubly Cys-substituted trimers (i.e. c3, c6, c9, c12). The results show that there are 12 copies of subunit c per F0 in E. coli, the exact number having both mechanistic and structural significance
RP  - NOT IN FILE
NT  - UI - 99009083LA - engRN - 0 (DNA Primers)RN - 0 (Recombinant Proteins)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM23105/GM/NIGMSDA - 19981210IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:9792682
SO  - J Biol Chem 1998 Nov 6 ;273(45):29701-29705

1520
UI  - 17
AU  - Jones PC
AU  - Jiang W
AU  - Fillingame RH
AD  - Department of Biomolecular Chemistry, University of Wisconsin Medical School, Madison, Wisconsin 53706, USA
TI  - Arrangement of the multicopy H+-translocating subunit c in the membrane sector of the Escherichia coli F1F0 ATP synthase
AB  - The multicopy subunit c of the H+-transporting F1F0 ATP synthase of Escherichia coli is thought to fold across the membrane as a hairpin of two hydrophobic alpha-helices. The conserved Asp61, centered in the second transmembrane helix, is essential for H+ transport. In this study, we have made sequential Cys substitutions across both transmembrane helices and used disulfide cross-link formation to determine the oligomeric arrangement of the c subunits. Cross-link formation between single Cys substitutions in helix 1 provided initial limitations on how the subunits could be arranged. Double Cys substitutions at positions 14/16, 16/18, and 21/23 in helix 1 and 70/72 in helix 2 led to the formation of cross-linked multimers upon oxidation. Double Cys substitutions in helix 1 and helix 2, at residues 14/72, 21/65, and 20/66, respectively, also formed cross-linked multimers. These results indicate that at least 10 and probably 12 subunits c interact in a front-to-back fashion to form a ring-like arrangement in F0. Helix 1 packs at the interior and helix 2 at the periphery of the ring. The model indicates that the Asp61 carboxylate is centered between the helical faces of adjacent subunit c at the center of a four-helix bundle
RP  - NOT IN FILE
NT  - UI - 98307962LA - engRN - 0 (Disulfides)RN - 0 (Oligonucleotides, Antisense)RN - 52-90-4 (Cysteine)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM23105/GM/NIGMSDA - 19980806IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:9642286
SO  - J Biol Chem 1998 Jul 3 ;273(27):17178-17185

1521
UI  - 25
AU  - Kaim G
AU  - Dimroth P
AD  - Mikrobiologisches Institut, Eidgenossische Technische Hochschule, ETH- Zentrum, Schmelzbergstrasse 7, CH-8092 Zurich, Switzerland
TI  - Voltage-generated torque drives the motor of the ATP synthase
AB  - The mechanism by which ion-flux through the membrane-bound motor module (F0) induces rotational torque, driving the rotation of the gamma subunit, was probed with a Na+-translocating hybrid ATP synthase. The ATP-dependent occlusion of 1 (22)Na+ per ATP synthase persisted after modification of the c subunit ring with dicyclohexylcarbodiimide (DCCD), when 22Na+ was added first and ATP second, but not if the order of addition was reversed. These results support the model of ATP-driven rotation of the c subunit oligomer (rotor) versus subunit a (stator) that stops when either a 22Na+-loaded or a DCCD-modified rotor subunit reaches the Na+-impermeable stator. The ATP synthase with a Na+- permeable stator catalyzed 22Na+out/Na+in-exchange after reconstitution into proteoliposomes, which was not significantly affected by DCCD modification of the c subunit oligomer, but was abolished by the additional presence of ATP or by a membrane potential (DeltaPsi) of 90 mV. We propose that in the idling mode of the motor, Na+ ions are shuttled across the membrane by limited back and forth movements of the rotor against the stator. This motional flexibility is arrested if either ATP or DeltaPsi induces the switch from idling into a directed rotation. The Propionigenium modestum ATP synthase catalyzed ATP formation with DeltaPsi of 60-125 mV but not with DeltapNa+ of 195 mV. These results demonstrate that electric forces are essential for ATP synthesis and lead to a new concept of rotary-torque generation in the ATP synthase motor
RP  - NOT IN FILE
NT  - UI - 98447591LA - engRN - 0 (Chimeric Proteins)RN - 0 (Liposomes)RN - 0 (Sodium Radioisotopes)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - 7440-23-5 (Sodium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)RN - EC 3.6.1.37 (Na(+)-K(+)-Exchanging ATPase)PT - Journal ArticleDA - 19981210IS - 0261-4189SB - IMCY - ENGLANDJC - EMB
UR  - PM:9774333
SO  - EMBO J 1998 Oct 15 ;17(20):5887-5895

1522
UI  - 32
AU  - Kaim G
AU  - Dimroth P
AD  - Mikrobiologisches Institut, Eidgenossische Technische Hochschule, ETH- Zentrum, Schmelzbergstrasse 7, CH-8092 Zurich, Switzerland
TI  - A triple mutation in the a subunit of the Escherichia coli/Propionigenium modestum F1Fo ATPase hybrid causes a switch from Na+ stimulation to Na+ inhibition
AB  - Previously we have shown that the Na+-translocating Escherichia coli (F1-delta)/Propionigenium modestum (Fo+delta) hybrid ATPase acquires a Na+-independent phenotype by the c subunit double mutation F84L, L87V that is reflected by Na+-independent growth of the mutant strain MPC8487 on succinate [Kaim, G., and Dimroth, P. (1995) J. Mol. Biol. 253, 726-738]. Here we describe a new class of mutants that were obtained by random mutagenesis and screening for Na+-independent growth on succinate. All six mutants of the new class contained four mutations in the a subunit (S89P, K220R, V264E, I278N). Results from site- specific mutagenesis revealed that the substitutions K220R, V264E, and I278N were sufficient to create the new phenotype. The resulting E. coli mutant strain MPA762 could only grow in the absence but not in the presence of Na+ ions on succinate minimal medium. This effect of Na+ ions on growth correlated with a Na+-specific inhibition of the mutant ATPase. The Ki for NaCl was 1. 5 mM at pH 6.5, similar to the Km for NaCl in activating the parent hybrid ATPase at this pH. On the other hand, activation by Li+ ions was retained in the new mutant ATPase. In the absence of Na+ or Li+, the mutant enzyme had the same pH optimum at pH 6.5 and twice the specific activity as the parent hybrid ATPase. In accordance with the kinetic data, the reconstituted mutant ATPase catalyzed H+ or Li+ transport but no Na+ transport. These results show for the first time that the coupling ion selectivity of F1Fo ATPases is determined by structural elements not only of the c subunit but also of the a subunit
RP  - NOT IN FILE
NT  - UI - 98190038LA - engRN - 0 (Bacterial Proteins)RN - 0 (Culture Media)RN - 0 (Plasmids)RN - 0 (Recombinant Proteins)RN - 110-15-6 (Succinic Acid)RN - 7439-93-2 (Lithium)RN - 7440-23-5 (Sodium)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19980423IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:9521783
SO  - Biochemistry 1998 Mar 31 ;37(13):4626-4634

1523
UI  - 26
AU  - Kaim G
AU  - Dimroth P
AD  - Mikrobiologisches Institut, Eidgenossische Technische Hochschule, ETH- Zentrum, Zurich, Switzerland
TI  - ATP synthesis by the F1Fo ATP synthase of Escherichia coli is obligatorily dependent on the electric potential
AB  - The H+-translocating F1Fo ATP synthase of Escherichia coli was purified and reconstituted into proteoliposomes. This system catalyzed ATP synthesis when energized by an acid/base transition (pHin = 5.0; pHout = 8.3) with succinate, malonate or maleinate but not with MES as the acidic buffer. Under these experimental conditions an electric potential of 125-130 mV is generated by the diffusion of succinate, probably the monoanionic species, whereas with MES buffer the measured potential was at background level (approximately 5 mV). ATP was also synthesized at pH 7.2 in the absence of a delta pH by applying a K+/valinomycin diffusion potential. The rate of ATP synthesis increased with the potential in an exponential manner with an inflection point at about 70 mV. We conclude from these results that delta pH and delta psi are kinetically unequivalent driving forces for ATP synthesis by the E. coli ATP synthase and that delta psi is a mandatory force for this synthesis. The significance of these findings for the mechanism of ATP synthesis in general is discussed
RP  - NOT IN FILE
NT  - UI - 98408831LA - engRN - 0 (Bacterial Proteins)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19980929IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:9738451
SO  - FEBS Lett 1998 Aug 28 ;434(1-2):57-60

1524
UI  - 33
AU  - Kaim G
AU  - Matthey U
AU  - Dimroth P
AD  - Mikrobiologisches Institut, Eidgenossische Technische Hochschule, ETH- Zentrum, Schmelzbergstr 7, CH-8092 Zurich, Switzerland
TI  - Mode of interaction of the single a subunit with the multimeric c subunits during the translocation of the coupling ions by F1F0 ATPases
AB  - We have recently isolated a mutant (aK220R, aV264E, aI278N) of the Na+- translocating Escherichia coli/Propionigenium modestum ATPase hybrid with a Na+-inhibited growth phenotype on succinate. ATP hydrolysis by the reconstituted mutant ATPase was inhibited by external (N side) NaCl but not by internal (P side) NaCl. In contrast, LiCl activated the ATPase from the N side and inhibited it from the P side. A similar pattern of activation and inhibition was observed with NaCl and the ATPase from the parent strain PEF42. We conclude from these results that the binding sites for the coupling ions on the c subunits are freely accessible from the N side. Upon occupation of these sites, the ATPase becomes more active, provided that the ions can be further translocated to the P side through a channel of the a subunit. If by mutation of the a subunit this channel becomes impermeable for Na+, N side Na+ ions specifically inhibit the ATPase activity. These conclusions were corroborated by the observation that proton transport into proteoliposomes containing the mutant ATPase was abolished by N side but not by P side Na+ ions. In contrast, LiCl affected proton translocation from either side, similar to the sidedness effect of Na+ ions on H+ transport by the parent hybrid ATPase. If the ATPase carrying the mutated a subunit was incubated with 22NaCl and ATP, 1 mol 22Na+/mol enzyme was occluded. With the parent hybrid ATPase, 22Na+ occlusion was not observed. The occluded 22Na+ could be removed from its tight binding site by 20 mM LiCl, while incubation with 20 mM NaCl was without effect. Li+ but not Na+ is therefore apparently able to pass through the mutated a subunit and make the entrapped Na+ ions accessible again to the aqueous environment. These results suggest an ion translocation mechanism through F0 that in the ATP hydrolysis mode involves binding of the coupling ions from the cytoplasm to the multiple c subunits, ATP-driven rotation to bring a Na+, Li+, or H+- loaded c subunit into a contact site with the a subunit and release of the coupling ions through the a subunit channel to the periplasmic surface of the membrane
RP  - NOT IN FILE
NT  - UI - 98119812LA - engRN - 0 (Proteolipids)RN - 0 (Recombinant Proteins)RN - 0 (Sodium Radioisotopes)RN - 0 (proteoliposomes)RN - 7439-93-2 (Lithium)RN - 7440-23-5 (Sodium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19980306IS - 0261-4189SB - IMCY - ENGLANDJC - EMB
UR  - PM:9450994
SO  - EMBO J 1998 Feb 2 ;17(3):688-695

1525
UI  - 21035
AU  - Kakinuma Y
AD  - Faculty of Pharmaceutical Sciences, Chiba University, 1-33 Yayoi-cho, Inage-ku, Chiba 263-8522, Japan yoshimi@athenaeumpchiba-uacjp
TI  - Inorganic cation transport and energy transduction in Enterococcus hirae and other streptococci
AB  - Energy metabolism by bacteria is well understood from the chemiosmotic viewpoint. We know that bacteria extrude protons across the plasma membrane, establishing an electrochemical potential that provides the driving force for various kinds of physiological work. Among these are the uptake of sugars, amino acids, and other nutrients with the aid of secondary porters and the regulation of the cytoplasmic pH and of the cytoplasmic concentration of potassium and other ions. Bacteria live in diverse habitats and are often exposed to severe conditions. In some circumstances, a proton circulation cannot satisfy their requirements and must be supplemented with a complement of primary transport systems. This review is concerned with cation transport in the fermentative streptococci, particularly Enterococcus hirae. Streptococci lack respiratory chains, relying on glycolysis or arginine fermentation for the production of ATP. One of the major findings with E. hirae and other streptococci is that ATP plays a much more important role in transmembrane transport than it does in nonfermentative organisms, probably due to the inability of this organism to generate a large proton potential. The movements of cations in streptococci illustrate the interplay between a variety of primary and secondary modes of transport
MH  - A
MH  - ACID
MH  - Amino Acids
MH  - atp
MH  - Bacteria
MH  - Cations
MH  - Energy Metabolism
MH  - Fermentation
MH  - Glycolysis
MH  - ion
MH  - Ions
MH  - membrane
MH  - metabolism
MH  - Movement
MH  - pH
MH  - Potassium
MH  - proton
MH  - Protons
MH  - regulation
MH  - review
MH  - secondary
MH  - SYSTEM
MH  - SYSTEMS
MH  - transport
RP  - NOT IN FILE
NT  - UI - 99058129LA - engRN - 0 (Cations)PT - Journal ArticlePT - ReviewPT - Review, AcademicDA - 19990204IS - 1092-2172SB - IMCY - UNITED STATES
UR  - PM:9841664
SO  - Microbiol Mol Biol Rev 1998 Dec ;62(4):1021-1045

1526
UI  - 21181
AU  - Kalaidzidis IV
AU  - Belevich IN
AU  - Kaulen AD
AD  - Department of Photobiochemistry, AN Belozersky Institute of Physico- Chemical Biology, Moscow State University, Russia
TI  - Photovoltage evidence that Glu-204 is the intermediate proton donor rather than the terminal proton release group in bacteriorhodopsin
AB  - Electrogenic events in the E204Q bacteriorhodopsin mutant have been studied. A two-fold decrease in the magnitude of microsecond photovoltage generation coupled to M intermediate formation in the E204Q mutant is shown. This means that deprotonation of E204 is an electrogenic process and its electrogenicity is comparable to that of the proton transfer from the Schiff base to D85. pH dependence of the electrogenicity of M intermediate formation in the wild-type bacteriorhodopsin reveals only one component corresponding to the protonation of D85 in the bacteriorhodopsin ground state and transition of the purple neutral form into the blue acid form. Thus, the pK of E204 in the M state is close to the pK of D85 in the bacteriorhodopsin ground state (< 3) and far below the pK of the terminal proton release group (approximately 6). It is concluded that E204 functions as the intermediate proton donor rather than the terminal proton release group in the bacteriorhodopsin proton pump
MH  - A
MH  - ACID
MH  - Bacteriorhodopsin
MH  - BASE
MH  - DEPENDENCE
MH  - electrogenic
MH  - function
MH  - intermediate
MH  - M
MH  - M-intermediate
MH  - microsecond
MH  - mutant
MH  - pH
MH  - proton
MH  - Proton Pump
MH  - proton release
MH  - Proton transfer
MH  - Proton-transfer
MH  - protonation
MH  - Protons
MH  - Schiff base
MH  - Schiff-base
MH  - TRANSFER
RP  - NOT IN FILE
NT  - UI - 98408857LA - engRN - 0 (Protons)RN - 53026-44-1 (Bacteriorhodopsins)PT - Journal ArticleDA - 19980929IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:9738477
SO  - FEBS Lett 1998 Aug 28 ;434(1-2):197-200

1527
UI  - 21182
AU  - Kalaidzidis IV
AU  - Kalaidzidis YL
AU  - Kaulen AD
AD  - Department of Photobiochemistry, AA Belozersky Institute of Physico- Chemical Biology, Moscow State University, Russia
TI  - Flash-induced voltage changes in halorhodopsin from Natronobacterium pharaonis
AB  - The flash-induced voltage response of halorhodopsin at high NaCl concentration comprises two main kinetic components. The first component with tau approximately 1 micros does not exceed 4% of the overall response amplitude and is probably associated with the formation of the L (hR520) intermediate. The second main component with tau approximately 1-2.5 ms which is independent of Cl- concentration can be ascribed to the transmembrane Cl- translocation during the L intermediate decay. The photoelectric response in the absence of Cl- has the opposite polarity and does not exceed 6% of the overall response amplitude at high NaCl concentration. A pH decrease results in substitution of the Cl(-)-dependent components by the photoresponse which is similar to that in the absence of Cl-. Thus, the difference between photoresponses of chloride-binding and chloride-free halorhodopsin forms resembles that of bacteriorhodopsin purple neutral and blue acid forms, respectively. The photovoltage data obtained can hardly be explained within the framework of the photocycle scheme suggested by Varo et al. [Biochemistry 34 (1995), 14490-14499]. We suppose that the O-type intermediate belongs to some form of halorhodopsin incapable of Cl- transport
MH  - A
MH  - ACID
MH  - Bacterial Proteins
MH  - Bacteriorhodopsin
MH  - intermediate
MH  - pH
MH  - protein
MH  - Proteins
MH  - Sodium
MH  - translocation
MH  - transport
MH  - voltage
RP  - NOT IN FILE
NT  - UI - 98273656LA - engRN - 0 (Bacterial Proteins)RN - 0 (Chlorides)RN - 0 (Halorhodopsins)RN - 53026-44-1 (Bacteriorhodopsins)RN - 7647-14-5 (Sodium Chloride)PT - Journal ArticleDA - 19980618IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:9613600
SO  - FEBS Lett 1998 May 1 ;427(1):59-63

1528
UI  - 702
AU  - Kato-Yamada Y
AU  - Noji H
AU  - Yasuda R
AU  - Kinosita K
AU  - Yoshida M
AD  - Research Laboratory of Resources Utilization, Tokyo Institute of Technology, 4259 Nagatsuta, Yokohama, 226-8503, Japan
TI  - Direct observation of the rotation of epsilon subunit in F1-ATPase
AB  - Rotation of the epsilon subunit in F1-ATPase from thermophilic Bacillus strain PS3 (TF1) was observed under a fluorescence microscope by the method used for observation of the gamma subunit rotation (Noji, H., Yasuda, R., Yoshida, M., and Kinosita, K., Jr. (1997) Nature 386, 299- 302). The alpha3 beta3 gamma epsilon complex of TF1 was fixed to a solid surface, and fluorescently labeled actin filament was attached to the epsilon subunit through biotin-streptavidin. In the presence of ATP, the filament attached to epsilon subunit rotated in a unidirection. The direction of the rotation was the same as that observed for the gamma subunit. The rotational velocity was slightly slower than the filament attached to the gamma subunit, probably due to the experimental setup used. Thus, as suggested from biochemical studies (Aggeler, R., Ogilvie, I. , and Capaldi, R. A. (1997) J. Biol. Chem. 272, 19621-19624), the epsilon subunit rotates with the gamma subunit in F1-ATPase during catalysis
RP  - NOT IN FILE
NT  - UI - 98344020LA - engRN - 0 (Actins)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19980910IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:9677353
SO  - J Biol Chem 1998 Jul 31 ;273(31):19375-19377

1529
UI  - 20923
AU  - Ketchum CJ
AU  - al Shawi MK
AU  - Nakamoto RK
AD  - Department of Molecular Physiology and Biological Physics, University of Virginia, PO Box 10011, Charlottesville, VA 22906-0011, USA
TI  - Intergenic suppression of the gammaM23K uncoupling mutation in F0F1 ATP synthase by betaGlu-381 substitutions: the role of the beta380DELSEED386 segment in energy coupling
AB  - We previously demonstrated that the Escherichia coli F0F1-ATP synthase mutation, gammaM23K, caused increased energy of interaction between gamma- and beta-subunits which was correlated to inefficient coupling between catalysis and transport [Al-Shawi, Ketchum and Nakamoto (1997) J. Biol. Chem. 272, 2300-2306]. Based on these results and the X-ray crystallographic structure of bovine F1-ATPase [Abrahams, Leslie, Lutter and Walker (1994) Nature (London) 370, 621-628] gammaM23K is believed to form an ionized hydrogen bond with betaGlu-381 in the conserved beta380DELSEED386 segment. In this report, we further test the role of gamma-beta-subunit interactions by introducing a series of substitutions for betaGlu-381 and gammaArg-242, the residue which forms a hydrogen bond with betaGlu-381 in the wild-type enzyme. betaE381A, D, and Q were able to restore efficient coupling when co-expressed with gammaM23K. All three mutations reversed the increased transition state thermodynamic parameters for steady state ATP hydrolysis caused by gammaM23K. betaE381K by itself caused inefficient coupling, but opposite from the effect of gammaM23K, the transition state thermodynamic parameters were lower than wild-type. These results suggest that the betaE381K mutation perturbs the gamma-beta-subunit interaction and the local conformation of the beta380DELSEED386 segment in a specific way that disrupts the communication of coupling information between transport and catalysis. betaE381A, L, K, and R, and gammaR242L and E mutations perturbed enzyme assembly and stability to varying degrees. These results provide functional evidence that the beta380DELSEED386 segment and its interactions with the gamma-subunit are involved in the mechanism of coupling
MH  - A
MH  - ACID
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BETA-SUBUNIT
MH  - Catalysis
MH  - conformation
MH  - coupling
MH  - England
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - glutamic acid
MH  - Hydrogen
MH  - Hydrolysis
MH  - mechanism
MH  - physiology
MH  - RESIDUE
MH  - SYNTHASE
MH  - transport
RP  - NOT IN FILE
NT  - UI - 98149741LA - engRN - 56-86-0 (Glutamic Acid)RN - 74-79-3 (Arginine)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleID - GM50957/GM/NIGMSID - GM52502/GM/NIGMSID - HL07284/HL/NHLBIDA - 19980416IS - 0264-6021SB - IMCY - ENGLAND
UR  - PM:9480879
SO  - Biochem J 1998 Mar 1 ;330 ( Pt 2)():707-712

1530
UI  - 21180
AU  - Khitrina LV
AU  - Eremin SV
AU  - Khodonov AA
AU  - Shvets VI
AU  - Kaulen AD
AD  - Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Russia khitr@phtbiogenebeemsusu
TI  - 13-Demethylbacteriorhodopsin: formation and some properties
AB  - Incorporation of 9-cis-13-demethylretinal into bacterioopsin was shown to form the covalent purple complex. This result was predicted by our hypothesis about the structure of the BR chromophore cavity (Mol. Biologiya 29:1398-1407 (1995) (in Russian)). It supports the hypothesis and eliminates the main objection known from the literature
MH  - Bacteriorhodopsin
MH  - COMPLEX
MH  - Retinaldehyde
MH  - structure
RP  - NOT IN FILE
NT  - UI - 99046705LA - engRN - 0 (13-demethylbacteriorhodopsin)RN - 0 (9,13-demethylretinal)RN - 116-31-4 (Retinaldehyde)RN - 1405-97-6 (Gramicidin)RN - 53026-44-1 (Bacteriorhodopsins)PT - Journal ArticleDA - 19990323IS - 1023-6597SB - IMCY - SWITZERLAND
UR  - PM:9829265
SO  - Membr Cell Biol 1998  ;12(1):121-123

1531
UI  - 704
AU  - Kinosita K
AU  - Yasuda R
AU  - Noji H
AU  - Ishiwata S
AU  - Yoshida M
AD  - Department of Physics, Faculty of Science and Technology, Keio University, Yokohama, Japan
TI  - F1-ATPase: a rotary motor made of a single molecule
RP  - NOT IN FILE
NT  - UI - 98206464LA - engRN - 0 (Macromolecular Systems)RN - 0 (Myosin)RN - EC 3.6.1.- (Kinesin)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 19980501IS - 0092-8674SB - IMCY - UNITED STATESJC - CQ4
UR  - PM:9546388
SO  - Cell 1998 Apr 3 ;93(1):21-24

1532
UI  - 21144
AU  - Krasinskaya IP
AU  - Lapin MV
AU  - Yaguzhinsky LS
AD  - AN Belozersky Institute of Physico-Chemical Biology, Moscow State University, Russia
TI  - Detection of the local H+ gradients on the internal mitochondrial membrane
AB  - Respiration-dependent responses of a pH probe (fluorescein isothiocyanate, FITC), covalently bound to the membrane proteins of mitochondria and submitochondrial particles (SMP) have been studied. A spectral shift indicating FITC deprotonation was observed when respiration was activated in coupled mitochondria. Such a response was increased by valinomycin and reduced by uncoupler. Some FITC deprotonation was detected in the presence of excess of an uncoupler, but the response was smaller and insensitive to valinomycin. FITC deprotonation was also observed in submitochondrial particles after succinate addition. In this case it was not affected by uncoupler. Increase in the buffer concentration was found to (i) decrease the FITC response and (ii) increase the rate of uncoupled respiration in both mitochondria and submitochondrial particles. The results are consistent with the assumption that respiration initiates appearance of local H+ activity gradients on the inner side of the internal mitochondrial membrane during the steady-state H+ pumping. We suggest that the formation of this gradient is due to kinetic barrier to proton transfer from the bulk phase to the respiratory proton pump vicinity
MH  - A
MH  - ACID
MH  - buffer
MH  - Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone
MH  - fluorescein
MH  - H+
MH  - membrane
MH  - Membrane Proteins
MH  - Mitochondria
MH  - pH
MH  - Potassium
MH  - protein
MH  - Proteins
MH  - proton
MH  - Proton Pump
MH  - Proton transfer
MH  - Proton-transfer
MH  - Protons
MH  - Respiration
MH  - Submitochondrial Particles
MH  - succinate
MH  - Succinic Acid
MH  - TRANSFER
MH  - Uncoupling Agents
MH  - Valinomycin
RP  - NOT IN FILE
NT  - UI - 99077275LA - engRN - 0 (Membrane Proteins)RN - 0 (Proton Pumps)RN - 0 (Protons)RN - 0 (Uncoupling Agents)RN - 110-15-6 (Succinic Acid)RN - 2001-95-8 (Valinomycin)RN - 3326-32-7 (Fluorescein-5-isothiocyanate)RN - 370-86-5 (Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone)RN - 67-64-1 (Acetone)RN - 7440-09-7 (Potassium)PT - Journal ArticleDA - 19990111IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:9862459
SO  - FEBS Lett 1998 Nov 27 ;440(1-2):223-225

1533
UI  - 21074
AU  - Krulwich TA
AU  - Ito M
AU  - Gilmour R
AU  - Hicks DB
AU  - Guffanti AA
AD  - Department of Biochemistry, Mount Sinai School of Medicine of CUNY, New York, USA
TI  - Energetics of alkaliphilic Bacillus species: physiology and molecules
AB  - The challenge of maintaining a cytoplasmic pH that is much lower than the external pH is central to the adaptation of extremely alkaliphilic Bacillus species to growth at pH values above 10. The success with which this challenge is met may set the upper limit of pH for growth in these bacteria, all of which also exhibit a low content of basic amino acids in proteins or protein segments that are exposed to the outside bulk phase liquid. The requirement for an active Na(+)-dependent cycle and possible roles of acidic cell wall components in alkaliphile pH homeostasis are reviewed. The gene loci that encode Na+/H+ antiporters that function in the active cycle are described and compared with the less Na(+)-specific homologues thus far found in non-alkaliphilic Gram- positive prokaryotes. Alkaliphilic Bacillus species carry out oxidative phosphorylation using an exclusively H(+)-coupled ATPase (synthase). Nonetheless, ATP synthesis is more rapid and reaches a higher phosphorylation potential at highly alkaline pH than at near-neutral pH even though the bulk electrochemical proton gradient across the coupling membrane is lower at highly alkaline pH. It is possible that some of the protons extruded by the respiratory chain are conveyed to the ATP synthase without first equilibrating with the external bulk phase. Mechanisms that might apply to oxidative phosphorylation in this type of extensively studied alkaliphile are reviewed, and note is made of the possibility of different kinds of solutions to the problem that may be found in new alkaliphilic bacteria that are yet to be isolated or characterized
MH  - A
MH  - ACID
MH  - ACTIVE
MH  - Amino Acids
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - ATPase
MH  - Bacillus
MH  - Bacteria
MH  - Biochemistry
MH  - coupling
MH  - England
MH  - function
MH  - Homeostasis
MH  - LIQUID
MH  - mechanism
MH  - MECHANISMS
MH  - membrane
MH  - Oxidative Phosphorylation
MH  - pH
MH  - Phosphorylation
MH  - physiology
MH  - protein
MH  - Proteins
MH  - proton
MH  - Proton-Translocating ATPases
MH  - Protons
MH  - review
MH  - Solutions
MH  - SYNTHASE
MH  - synthesis
RP  - NOT IN FILE
NT  - UI - 99106657LA - engRN - 0 (Sodium-Hydrogen Antiporter)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticlePT - ReviewPT - Review, AcademicID - GM28454/GM/NIGMSDA - 19990202IS - 0065-2911SB - IMCY - ENGLAND
UR  - PM:9889983
SO  - Adv Microb Physiol 1998  ;40():401-438

1534
UI  - 21075
AU  - Krulwich TA
AU  - Ito M
AU  - Hicks DB
AU  - Gilmour R
AU  - Guffanti AA
AD  - Department of Biochemistry, Mount Sinai School of Medicine, New York, NY 10029, USA krulwich@msvaxmssmedu
TI  - pH homeostasis and ATP synthesis: studies of two processes that necessitate inward proton translocation in extremely alkaliphilic Bacillus species
AB  - Alkaliphilic Bacillus species that are isolated from nonmarine, moderate salt, and moderate temperature environments offer the opportunity to explore strategies that have developed for solving the energetic challenges of aerobic growth at pH values between 10 and 11. Such bacteria share many structural, metabolic, genomic, and regulatory features with nonextremophilic species such as Bacillus subtilis. Comparative studies can therefore illuminate the specific features of gene organization and special features of gene products that are homologs of those found in non-extremophiles, and potentially identify novel gene products of importance in alkaliphily. We have focused our studies on the facultative alkaliphile Bacillus firmus OF4, which is routinely grown on malate-containing medium at either pH 7.5 or 10.5. Current work is directed toward clarification of the characteristics and energetics of membrane-associated proteins that must catalyze inward proton movements. One group of such proteins are the Na+/H+ antiporters that enable cells to adapt to a sudden upward shift in pH and to maintain a cytoplasmic pH that is 2-2.3 units below the external pH in the most alkaline range of pH for growth. Another is the proton- translocating ATP synthase that catalyzes robust production of ATP under conditions in which the external proton concentration and the bulk chemiosmotic driving force are low. Three gene loci that are candidates for Na+/H+ antiporter encoding genes with roles in Na(+)- dependent pH homeostasis have been identified. All of them have homologs in B. subtilis, in which pH homeostasis can be carried out with either K+ or Na+. The physiological importance of one of the B. firmus OF4 loci, nhaC, has been studied by targeted gene disruption, and the same approach is being extended to the others. The atp genes that encode the alkaliphile's F1F0-ATP synthase are found to have interesting motifs in areas of putative importance for proton translocation. As an initial step in studies that will probe the importance and possible roles of these motifs, the entire atp operon from B. firmus OF4 has been cloned and functionally expressed in an Escherichia coli mutant that has a full deletion of its atp genes. The transformant does not exhibit growth on succinate, but shows reproducible, modest increases in the aerobic growth yields on glucose as well as membrane ATPase activity that exhibits characteristics of the alkaliphile enzyme
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - ATPase
MH  - Bacillus
MH  - Bacteria
MH  - Biochemistry
MH  - Cells
MH  - Comparative Study
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1F0-ATP SYNTHASE
MH  - Glucose
MH  - Homeostasis
MH  - membrane
MH  - Movement
MH  - mutant
MH  - pH
MH  - protein
MH  - Proteins
MH  - proton
MH  - Proton-Translocating ATPases
MH  - Protons
MH  - review
MH  - succinate
MH  - SYNTHASE
MH  - synthesis
MH  - Temperature
MH  - translocation
RP  - NOT IN FILE
NT  - UI - 98456497LA - engRN - 0 (Protons)RN - 0 (Sodium-Hydrogen Antiporter)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticlePT - ReviewPT - Review, TutorialID - GM28454/GM/NIGMSDA - 19981230IS - 1431-0651SB - IMSB - SCY - GERMANY
UR  - PM:9783168
SO  - Extremophiles 1998 Aug ;2(3):217-222

1535
UI  - 982
AU  - Lacapere JJ
AU  - Stokes DL
AU  - Olofsson A
AU  - Rigaud JL
TI  - Two-dimensional crystallization of Ca-ATPase by detergent removal
AB  - By using Bio-Beads as a detergent-removing agent, it has been possible to produce detergent-depleted two-dimensional crystals of purified Ca-ATPase. The crystallinity and morphology of these different crystals were analyzed by electron microscopy under different experimental conditions. A lipid-to-protein ratio below 0.4 w/w was required for crystal formation. The rate of detergent removal critically affected crystal morphology, and large multilamellar crystalline sheets or wide unilamellar tubes were generated upon slow or fast detergent removal, respectively. Electron crystallographic analysis indicated unit cell parameters of a = 159 A, b = 54 A, and gamma = 90 degrees for both types of crystals, and projection maps at 15-A resolution were consistent with Ca-ATPase molecules alternately facing the two sides of the membrane. Crystal formation was also affected by the protein conformation. Indeed, tubular and multilamellar crystals both required the presence of Ca2+; the presence of ADP gave rise to another type of packing within the unit cell (a = 86 A, b = 77 A, and gamma = 90 degrees), while maintaining a bipolar orientation of the molecules within the bilayer. All of the results are discussed in terms of nucleation and crystal growth, and a model of crystallogenesis is proposed that may be generally true for asymmetrical proteins with a large hydrophilic cytoplasmic domain
MH  - Animal
MH  - Biophysics
MH  - Ca(2+)-Transporting ATPase
MH  - Chemistry
MH  - Crystallization
MH  - Detergents
MH  - enzymology
MH  - isolation & purification
MH  - Lipids
MH  - Micelles
MH  - Microscopy
MH  - Microscopy,Electron
MH  - Polystyrenes
MH  - Protein Conformation
MH  - Proteins
MH  - Sarcoplasmic Reticulum
MH  - Support,Non-U.S.Gov't
MH  - Support,U.S.Gov't,P.H.S.
MH  - ultrastructure
RP  - NOT IN FILE
NT  - Institut Curie, Section de Recherche, UMR-CNRS 168, Paris, France lacapere@curiefr
SO  - Biophys J 1998 Sep ;75(3):1319-1329

1536
UI  - 19901
AU  - Leite VBP
AU  - Cavalli A
AU  - Oliveira ON
TI  - Hydrogen-bond control of structure and conductivity of Langmur films
RP  - IN FILE
SO  - Physical Review e 1998  ;57():6835-6839

1537
UI  - 412
AU  - Lobau S
AU  - Weber J
AU  - Senior AE
AD  - Department of Biochemistry and Biophysics, University of Rochester Medical Center, New York 14642, USA
TI  - Catalytic site nucleotide binding and hydrolysis in F1F0-ATP synthase
AB  - F1F0-ATP synthase was purified from Escherichia coli beta Y331W mutant. The beta-Trp-331 provided a specific fluorescent probe of catalytic site nucleotide binding. Physiological (mM) concentration of substrate MgATP filled all three catalytic sites. With MgATP or MgADP the catalytic sites showed marked binding cooperativity and asymmetry, which was dependent on Mg2+. Nucleotide binding was fast, with kon = approximately 6 x 10(5) M-1 s-1. Pi at physiological concentration (5 mM) did not bind to catalytic sites. Measurement of MgATP hydrolysis and binding under identical conditions as a function of MgATP concentration revealed that Vmax was achieved only when all three catalytic sites were filled in every enzyme molecule. The enzyme species with two catalytic sites occupied and one site empty displayed low, nonphysiological catalytic rate. This is the first characterization of nucleotide binding parameters in F1F0. The fact that the behavior of purified F1F0 was similar in most respects to that of isolated F1 demonstrated that the presence of the additional F0 subunits a, b, and c, and also fixed stoichiometric amounts of epsilon and delta, does not affect catalytic site properties. The results impact on possible catalytic mechanisms, namely, they emphasize that Pi cannot simply bind spontaneously, that an enzyme species with all three sites occupied is the only catalytically competent species, and that release of product and binding of substrate cannot be simultaneous, rather the former must precede the latter
RP  - NOT IN FILE
NT  - UI - 98367527LA - engRN - 0 (Proteolipids)RN - 0 (Proton Pump)RN - 0 (proteoliposomes)RN - 55520-40-6 (Tyrosine)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 73-22-3 (Tryptophan)RN - 7439-95-4 (Magnesium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSDA - 19980821IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:9692975
SO  - Biochemistry 1998 Jul 28 ;37(30):10846-10853

1538
UI  - 19771
AU  - Milgrom YM
AU  - Murataliev MB
AU  - Boyer PD
AD  - Department of Biochemistry and Molecular Biology, State University of New York Health Science Center at Syracuse, 750 E Adams St, Syracuse, NY 13210, USA
TI  - Bi-site activation occurs with the native and nucleotide-depleted mitochondrial F1-ATPase
AB  - Experiments are reported on the uni-site catalysis and the transition from uni-site to multi-site catalysis with bovine heart mitochondrial F1-ATPase. The very slow uni-site ATP hydrolysis is shown to occur without tightly bound nucleotides present and with or without Pi in the buffer. Measurements of the transition to higher rates and the amount of bound ATP committed to hydrolysis as the ATP concentration is increased at different fixed enzyme concentrations give evidence that the filling of a second site can initiate near maximal turnover rates. They provide rate constant information, and show that an apparent Km for a second site of about 2 microM and Vmax of 10 s-1, as suggested by others, is not operative. Careful initial velocity measurements also eliminate other suggested Km values and are consistent with bi-site activation to near maximal hydrolysis rates, with a Km of about 130 microM and Vmax of about 700 s-1. However, the results do not eliminate the possibility of additional 'hidden' Km values with similar Vmax:Km ratios. Recent data on competition between TNP-ATP and ATP revealed a third catalytic site for ATP in the millimolar concentration range. This result, and those reported in the present paper, allow the conclusion that the mitochondrial F1-ATPase can attain near maximal activity in bi-site catalysis. Our data also add to the evidence that a recent claim, that the mitochondrial F1-ATPase does not show catalytic site cooperativity, is invalid
MH  - A
MH  - ACTIVATION
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthase
MH  - Biochemistry
MH  - BOUND NUCLEOTIDES
MH  - Catalysis
MH  - England
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - H(+)-Transporting ATP Synthase
MH  - Hydrolysis
MH  - MITOCHONDRIAL F1-ATPASE
MH  - Nucleotides
MH  - Phosphates
MH  - SYNTHASE
MH  - uni-site catalysis
MH  - UNISITE CATALYSIS
RP  - NOT IN FILE
NT  - UI - 98149789LA - engRN - 0 (Phosphates)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM 23152/GM/NIGMSDA - 19980416IS - 0264-6021SB - IMCY - ENGLANDJC - 9YO
UR  - PM:9480927
SO  - Biochem J 1998 Mar 1 ;330 ( Pt 2)():1037-1043

1539
UI  - 30
AU  - Neumann S
AU  - Matthey U
AU  - Kaim G
AU  - Dimroth P
AD  - Mikrobiologisches Institut, Eidgenossische Technische Hochschule Zurich, Switzerland
TI  - Purification and properties of the F1F0 ATPase of Ilyobacter tartaricus, a sodium ion pump
AB  - The ATPase of Ilyobacter tartaricus was solubilized from the bacterial membranes and purified. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the purified enzyme revealed the usual subunit pattern of a bacterial F1F0 ATPase. The polypeptides with apparent molecular masses of 56, 52, 35, 16.5, and 6.5 kDa were identified as the alpha, beta, gamma, epsilon, and c subunits, respectively, by N- terminal protein sequencing and comparison with the sequences of the corresponding subunits from the Na(+)-translocating ATPase of Propionigenium modestum. Two overlapping sequences were obtained for the polypeptides moving with an apparent molecular mass of 22 kDa (tentatively assigned as b and delta subunits). No sequence could be determined for the putative a subunit (apparent molecular mass, 25 kDa). The c subunits formed a strong aggregate with the apparent molecular mass of 50 kDa which required treatment with trichloroacetic acid for dissociation. The ATPase was inhibited by dicyclohexyl carbodiimide, and Na+ ions protected the enzyme from this inhibition. The ATPase was specifically activated by Na+ or Li+ ions, markedly at high pH. After reconstitution into proteoliposomes, the enzyme catalyzed the ATP-dependent transport of Na+, Li+, or Hi+. Proton transport was specifically inhibited by Na+ or Li+ ions, indicating a competition between these alkali ions and protons for binding and translocation across the membrane. These experiments characterize the I. tartaricus ATPase as a new member of the family of FS-ATPases, which use Na+ as the physiological coupling ion for ATP synthesis
RP  - NOT IN FILE
NT  - UI - 98317267LA - engRN - 0 (Macromolecular Systems)RN - 0 (Peptide Fragments)RN - 7439-93-2 (Lithium)RN - 7440-23-5 (Sodium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)RN - EC 3.6.1.37 (Na(+)-K(+)-Exchanging ATPase)PT - Journal ArticleDA - 19980727IS - 0021-9193SB - IMCY - UNITED STATESJC - HH3
UR  - PM:9642181
SO  - J Bacteriol 1998 Jul ;180(13):3312-3316

1540
UI  - 143
AU  - Ogilvie I
AU  - Wilkens S
AU  - Rodgers AJ
AU  - Aggeler R
AU  - Capaldi RA
AD  - Institute of Molecular Biology, University of Oregon, Eugene 97403- 1229, USA
TI  - The second stalk: the delta-b subunit connection in ECF1F0
AB  - The ATP synthase F1F0 is the smallest molecular motor yet studied. ATP hydrolysis drives the rotary motion of the primary stalk subunits gamma and epsilon relative to the alpha 3 beta 3 part of F1. Evidence is reviewed to show that the delta and b subunits provide a second stalk that can act as a stator to facilitate these rotational movements
RP  - NOT IN FILE
NT  - UI - 99005987LA - engRN - 0 (Isoenzymes)RN - 0 (Molecular Motors)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 19990112IS - 0302-2994SB - IMCY - ENGLANDJC - 1UF
UR  - PM:9789558
SO  - Acta Physiol Scand Suppl 1998 Aug ;643():169-175

1541
UI  - 20842
AU  - Omote H
AU  - Tainaka K
AU  - Fujie K
AU  - Iwamoto-Kihara A
AU  - Wada Y
AU  - Futai M
AD  - Institute of Scientific and Industrial Research, Osaka University, Osaka, Ibaraki, 567-0047, Japan
TI  - Stability of the Escherichia coli ATP synthase F0F1 complex is dependent on interactions between gamma Gln-269 and the beta subunit loop beta Asp-301-beta Asp-305
AB  - The role of the conserved sequence motif 301DDLTDP306 in the F0F1 ATP synthase beta subunit was assessed by mutagenic analysis in the Escherichia coli enzyme. Mutations gave variable effects on F1 sector activity, stability, and membrane binding to the F0 sector. Upon solubilization, F1 sectors of the betaD302E and betaD305E mutants (betaAsp-302 and betaAsp-305 replaced by glutamate) dissociated into subunits, while mutants with other beta305 substitutions failed to assemble. Membrane ATPase activities of beta301 and 302 mutants were 20- 70% of wild type. Replacements of the gamma subunit Gln-269 had similar effects. The membrane ATPase activities of the gammaQ269E or gammaQ269D mutants were significantly lower and their F1 sectors dissociated into subunits upon solubilization. These results suggest that the beta301- 305 loop and the gamma subunit region around Gln-269 form a key region for the assembly of alpha3 beta3 gamma complex. These results are consistent with the X-ray crystallographic structure of bovine F1 (J. P. Abrahams, A. G. W. Leslie, R. Lutter, and J. E. Walker (1994) Nature 370, 621-628) where the beta301DDLTD305 loop directly interacts with gammaGln-269
MH  - A
MH  - ACID
MH  - analysis
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BETA-SUBUNIT
MH  - BINDING
MH  - COLI ATP SYNTHASE
MH  - COMPLEX
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - F0F1
MH  - F1
MH  - Glutamine
MH  - membrane
MH  - mutant
MH  - P
MH  - Peptide Fragments
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 99003474LA - engRN - 0 (Peptide Fragments)RN - 56-84-8 (Aspartic Acid)RN - 56-85-9 (Glutamine)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19981113IS - 0003-9861SB - IMCY - UNITED STATES
UR  - PM:9784240
SO  - Arch Biochem Biophys 1998 Oct 15 ;358(2):277-282

1542
UI  - 20841
AU  - Omote H
AU  - Futai M
AD  - Division of Biological Sciences, Osaka University, Ibaraki, Japan
TI  - Mutational analysis of F1F0 ATPase: catalysis and energy coupling
AB  - Escherichia coli ATP synthase has eight subunits and functions through transmission of conformational changes between subunits. Extensive mutational analyses identified essential residues for catalysis and conformation transmission. Pseudorevertant studies revealed that beta/alpha and beta/gamma subunits interactions are important for the energy coupling between catalysis and H+ translocation. In this article, we discuss mechanism of catalysis and energy coupling based on our recent mutation studies
MH  - analysis
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Catalysis
MH  - COLI ATP SYNTHASE
MH  - conformation
MH  - conformational change
MH  - coupling
MH  - England
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - function
MH  - H+
MH  - mechanism
MH  - protein
MH  - RESIDUE
MH  - review
MH  - SUBUNIT
MH  - SYNTHASE
MH  - translocation
RP  - NOT IN FILE
NT  - UI - 99005988LA - engRN - EC 3.6.1.- (ATP17 protein, S cerevisiae)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 19990112IS - 0302-2994SB - IMCY - ENGLAND
UR  - PM:9789559
SO  - Acta Physiol Scand Suppl 1998 Aug ;643():177-183

1543
UI  - 326
AU  - Orriss GL
AU  - Leslie AG
AU  - Braig K
AU  - Walker JE
AD  - Medical Research Council, Laboratory of Molecular Biology, Cambridge, UK
TI  - Bovine F1-ATPase covalently inhibited with 4-chloro-7- nitrobenzofurazan: the structure provides further support for a rotary catalytic mechanism
AB  - BACKGROUND: F1-ATPase is the globular domain of F1F0-ATP synthase that catalyses the hydrolysis of ATP to ADP and phosphate. The crystal structure of bovine F1-ATPase has been determined previously to 2.8 A resolution. The enzyme comprises five different subunits in the stoichiometry alpha 3 beta 3 gamma delta epsilon; the three catalytic beta subunits alternate with the three alpha subunits around the centrally located single gamma subunit. To understand more about the catalytic mechanisms, F1-ATPase was inhibited by reaction with 4-chloro- 7-nitrobenzofurazan (NBD-Cl) and the structure of the inhibited complex (F1-NBD) determined by X-ray crystallography. RESULTS: In the structure the three beta subunits adopt a different conformation with different nucleotide occupancy. NBD-Cl reacts with the phenolic oxygen of Tyr311 of the beta E subunit, which contains no bound nucleotide. The two other catalytic subunits beta TP and beta DP contain bound adenylyl- imidodiphosphate (AMP-PNP) and ADP, respectively. The binding site of the NBD moiety does not overlap with the regions of beta E that form the nucleotide-binding pocket in subunits beta TP and beta DP nor does it occlude the nucleotide-binding site. Catalysis appears to be inhibited because neither beta TP nor beta DP can accommodate a Tyr311 residue bearing an NBD group. CONCLUSIONS: The results presented here are consistent with a rotary catalytic mechanism of ATP synthesis and hydrolysis, which requires the sequential and concerted participation of all three catalytic sites. NBD-Cl inhibits the enzyme by preventing the modified subunit from adopting a conformation that is essential for catalysis to proceed
RP  - NOT IN FILE
NT  - UI - 98362587LA - engRN - 0 (Enzyme Inhibitors)RN - 10199-89-0 (4-Chloro-7-nitrobenzofurazan)RN - 55520-40-6 (Tyrosine)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19981022IS - 0969-2126SB - IMCY - ENGLANDJC - B31
UR  - PM:9687365
SO  - Structure 1998 Jul 15 ;6(7):831-837

1544
UI  - 502
AU  - Pan W
AU  - Ko YH
AU  - Pedersen PL
AD  - Department of Biological Chemistry, Johns Hopkins University, School of Medicine, Baltimore 21205-2185, USA
TI  - Delta subunit of rat liver mitochondrial ATP synthase: molecular description and novel insights into the nature of its association with the F1-moiety
AB  - The F1 moiety of ATP synthase complexes consists of five subunit types in the stoichiometric ratio alpha 3, beta 3, gamma, delta epsilon. Of these, the delta subunit has received very little attention in the study of F1 preparations from eukaryotic cells. Although recently shown to associate tightly with the beta subunit [Pedersen, P. L., Hullihen, J., Bianchet, M., Amzel, L. M., and Lebowitz, M. S. (1995) J. Biol. Chem. 270, 1775-1784], the delta subunit is not resolved in the X-ray structure of either the rat liver or bovine heart enzyme. For these reasons, the novel studies reported here were designed both to provide a molecular description of the rat liver delta subunit and to gain insight into the nature of its interaction with F1. The rat liver delta subunit was cloned from a lambda gt11 library, sequenced, overexpressed in Escherichia coli (E. coli) in fusion with the maltose binding protein, and, after cleavage of the latter protein, purified to homogeneity. The purified delta subunit (MW = 14.7 kDa) was shown by circular dichroism spectroscopy to be highly structured and to exhibit about 25% sequence identity to the chloroplast and E. coli epsilon subunits, frequently regarded as homologues of higher eukaryotic delta subunits. Significantly, and in contrast to the chloroplast and E. coli epsilon subunits, which are readily removed from their parent F1 moieties after treatment respectively with ethanol and lauryldimethylamine oxide, the rat liver delta subunit remained tightly bound to F1 under these relatively mild conditions. For the above reasons, four types of experiments were carried out on rat liver F1 in order to (1) determine the accessibility of the delta subunit to both specific antibodies and to proteases, (2) establish the effect of nucleotides on this subunit's accessibility, (3) identify in cross- linking studies with disuccinimidyl glutarate this subunit's most reactive neighbor, and (4) determine whether this subunit can be dissociated from F1 by using ionic detergents while leaving the remaining complex intact. The data derived from this detailed set of studies (a) supports the view that the rat liver F1-delta subunit is in very close proximity to the gamma subunit near the bottom of the F1 molecule but does not penetrate deeply into the central core, (b) shows that within F1 the delta subunit's N-terminus is exposed while its C- terminus is masked, (c) indicates that access to the delta subunit is shielded in part by the alpha, beta, and gamma subunits and changes during the catalytic cycle of F1, and (d) implicates the delta subunit as important for the structural stability of the F1 unit. These novel findings on a higher eukaryotic F1-delta subunit are discussed in relationship to earlier studies on the related epsilon subunits from both chloroplasts and E. coli
RP  - NOT IN FILE
NT  - UI - 98244872LA - engRN - 0 (Antibodies)RN - 0 (Cross-Linking Reagents)RN - 0 (Dimethylamines)RN - 0 (Macromolecular Systems)RN - 0 (Succinimides)RN - 0 (Surface-Active Agents)RN - 0 (disuccinimidyl glutarate)RN - 1643-20-5 (dodecyldimethylamine oxide)RN - 64-17-5 (Ethanol)RN - EC 3.4.- (Carboxypeptidases)RN - EC 3.4.16.5 (carboxypeptidase C)RN - EC 3.4.21.4 (Trypsin)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - CA 10951/CA/NCIDA - 19980604IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:9578578
SO  - Biochemistry 1998 May 12 ;37(19):6911-6923

1545
UI  - 605
AU  - Papageorgiou S
AU  - Melandri AB
AU  - Solaini G
AD  - Scuola Superiore di Studi Universitari e di Perfezionamento S Anna, Pisa, Italy
TI  - Relevance of divalent cations to ATP-driven proton pumping in beef heart mitochondrial F0F1-ATPase
AB  - The ATP hydrolysis rate and the ATP hydrolysis-linked proton translocation by the F0F1-ATPase of beef heart submitochondrial particles were examined in the presence of several divalent metal cations. All Me-ATP complexes tested sustained ATP hydrolysis, although to a different extent. However, only Mg- and Mn-ATP-dependent hydrolysis could sustain a high level of proton pumping activity, as determined by acridine fluorescence quenching. Moreover, the Km of the Me-ATP hydrolysis-induced proton pumping activity was very similar to the Km value of Me-ATP hydrolysis. Both oligomycin and DCCD caused the full recovery of the fluorescence, providing clear evidence for the association of Mg-ATP hydrolysis with proton translocation through the F0F1-ATPase complex. In contrast, with other Me-ATP complexes, including Ca-ATP as substrate, the proton pumping activity was undetectable, implicating an uncoupling nature for these substrates. Attempts to demonstrate the involvement of the epsilon subunit of the enzyme in the coupling mechanism failed, suggesting that the participation of at least the N-terminal segment of the subunit in the coupling mechanism of the mitochondrial enzyme is unlikely
RP  - NOT IN FILE
NT  - UI - 99221314LA - engRN - 0 (Cations, Divalent)RN - 0 (Proton Pump)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19990617IS - 0145-479XSB - IMCY - UNITED STATESJC - HIO
UR  - PM:10206473
SO  - J Bioenerg Biomembr 1998 Dec ;30(6):533-541

1546
UI  - 142
AU  - Rodgers AJ
AU  - Capaldi RA
AD  - Institute of Molecular Biology, University of Oregon, Eugene, Oregon 97403-1229, USA
TI  - The second stalk composed of the b- and delta-subunits connects F0 to F1 via an alpha-subunit in the Escherichia coli ATP synthase
AB  - The b- and delta-subunits of the Escherichia coli ATP synthase are critical for binding ECF1 to the F0 part, and appear to constitute the stator necessary for holding the alpha3beta3 hexamer as the c-epsilon- gamma domain rotates during catalysis. Previous studies have determined that the b-subunits are dimeric for a large part of their length, and interact with the F1 part through the delta-subunit (Rodgers, A. J. W., Wilkens, S., Aggeler, R., Morris, M. B., Howitt, S. M., and Capaldi, R. A. (1997) J. Biol. Chem. 272, 31058-31064). To further study b-subunit interactions, three mutants were constructed in which Ser-84, Ala-144, and Leu-156, respectively, were replaced by Cys. Treatment of purified ECF1F0 from all three mutants with CuCl2 induced disulfide formation resulting in b-subunit dimer cross-link products. In addition, the mutant bL156C formed a cross-link from a b-subunit to an alpha-subunit via alphaCys90. Neither b-b nor b-alpha cross-linking had significant effect on ATPase activities in any of the mutants. Proton pumping activities were measured in inner membranes from the three mutants. Dimerization of the b-subunit did not effect proton pumping in mutants bS84C or bA144C. In the mutant bL156C, CuCl2 treatment reduced proton pumping markedly, probably because of uncoupling caused by the b-alpha cross-link formation. The results show that the alpha-subunit forms part of the binding site on ECF1 for the b2delta domain and that the b- subunit extends all the way from the membrane to the top of the F1 structure. Some conformational flexibility in the connection between the second stalk and F1 appears to be required for coupled catalysis
RP  - NOT IN FILE
NT  - UI - 99009044LA - engRN - 0 (DNA Primers)RN - 7440-50-8 (Copper)RN - 7758-89-6 (cuprous chloride)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - HL58671/HL/NHLBIDA - 19981210IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:9792643
SO  - J Biol Chem 1998 Nov 6 ;273(45):29406-29410

1547
UI  - 21156
AU  - Schemidt RA
AU  - Qu J
AU  - Williams JR
AU  - Brusilow WS
AD  - Department of Biochemistry and Molecular Biology, Wayne State University School of Medicine, Detroit, Michigan 48201, USA
TI  - Effects of carbon source on expression of F0 genes and on the stoichiometry of the c subunit in the F1F0 ATPase of Escherichia coli
AB  - Expression of the genes for the membrane-bound F0 sector of the Escherichia coli F1F0 proton-translocating ATPase can respond to changes in metabolic conditions, and these changes are reflected in alterations in the subunit stoichiometry of the oligomeric F0 proton channel. Transcriptional and translational lacZ fusions to the promoter and to two F0 genes show that, during growth on the nonfermentable carbon source succinate, transcription of the operon and translation of uncB, encoding the a subunit of F0, are higher than during growth on glucose. In contrast, translation of the uncE gene, encoding the c subunit of F0, is higher during growth on glucose than during growth on succinate. Translation rates of both uncB and uncE change as culture density increases, but transcription rates do not. Quantitation of the c stoichiometry shows that more c subunits are assembled into the F1F0 ATPase in cells grown on glucose than in cells grown on succinate. E. coli therefore appears to have a mechanism for regulating the composition and, presumably, the function of the ATPase in response to metabolic circumstances
MH  - A
MH  - ACID
MH  - ATPase
MH  - Biochemistry
MH  - Cells
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - function
MH  - Glucose
MH  - mechanism
MH  - proton
MH  - Proton-Translocating ATPases
MH  - SUBUNIT
MH  - succinate
MH  - Succinic Acid
RP  - NOT IN FILE
NT  - UI - 98292755LA - engRN - 110-15-6 (Succinic Acid)RN - 50-99-7 (Glucose)RN - 7440-44-0 (Carbon)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19980706IS - 0021-9193SB - IMCY - UNITED STATES
UR  - PM:9620972
SO  - J Bacteriol 1998 Jun ;180(12):3205-3208

1548
UI  - 21347
AU  - Shi J
AU  - Wei JM
AU  - Shen YK
TI  - Site-directed Mutagenesis of epsilon Subunit of ATP Synthase of Maize Chloroplast
AB  - The effects of some residues in maize chloroplast epsilon subunit on its activity have been studied by site-directed mutagenesis. After replacing Thr-42 of epsilon subunit with Cys, Arg, Ile or Pro, and forming the mutant epsilonT42C, mutant epsilonT42R, mutant epsilonT42I and mutant epsilonT42P respectively, it was found that the mutant epsilonT42P protein could no longer be expressed, but expression of other epsilon subunit mutants was similar to that of wild type. Comparing the inhibitory potency of different mutants of epsilon subunit with that of the wild type, it was found that the inhibitory effects of epsilon subunit mutants epsilonT42C and epsilonT42R on ATPase activity were slightly higher than that of wild type, but the epsilonT42I protein strongly inhibited the Ca(2+)-ATPase activity
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - chloroplast
MH  - EPSILON-SUBUNIT
MH  - mutagenesis
MH  - mutant
MH  - physiology
MH  - plant
MH  - protein
MH  - RESIDUE
MH  - site-directed
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - Shanghai Institute of Plant Physiology, the Chinese Academy of Sciences, Shanghai 200032, China wjm@irissippaccn
SO  - Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai ) 1998  ;30(5):483-487

1549
UI  - 20813
AU  - Sorgen PL
AU  - Bubb MR
AU  - McCormick KA
AU  - Edison AS
AU  - Cain BD
AD  - Department of Biochemistry and Molecular Biology, University of Florida, Gainesville 32610, USA
TI  - Formation of the b subunit dimer is necessary for interaction with F1- ATPase
AB  - In earlier work, we [McCormick, K. A., et al. (1993) J. Biol. Chem. 268, 24683-24691] observed that mutations at Ala-79 of the b subunit affect assembly of F1F0 ATP synthase. Polypeptides modeled on the soluble portion of the b subunit (bsol) with substitutions at the position corresponding to Ala-79 have been used to investigate secondary structure and dimerization of the b subunit. Circular dichroism spectra and chymotrypsin digestion experiments suggested that the recombinant polypeptides with Ala-79 substitutions assumed conformations similar to the bsol polypeptide. However, cross-linking studies of the Ala-79 substitution bsol polypeptides revealed defects in dimerization. The efficiency of dimer formation appeared to be related to the capacity of the altered bsol polypeptides for competing with F1-ATPase for binding to F1-depleted membrane vesicles. Ala-79 substitution polypeptides displaying limited dimerization, such as bsol Ala-79-->Leu, were shown to elute with F1-ATPase during size exclusion chromatography, suggesting a specific interaction. Sedimentation equilibrium studies indicated that 8% of the bsol Ala-79-->Leu polypeptide was in the form of a 30.6 kDa dimer and 92% a 15.3 kDa monomer. When the dimer concentration of bsol Ala-79-->Leu was normalized to the concentration of bsol, both had virtually identical capacities for competing with F1-depleted membrane vesicles for binding F1-ATPase. The result indicated that the amount of dimer formed is directly proportional to its ability to bind F1-ATPase. This suggests that formation of the b subunit dimer may be a necessary step preceding F1-ATPase binding in the assembly of the enzyme complex
MH  - A
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BINDING
MH  - Biochemistry
MH  - COMPLEX
MH  - conformation
MH  - CROSS-LINKING
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - membrane
MH  - membrane vesicles
MH  - protein
MH  - Proteins
MH  - secondary
MH  - spectra
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 98118179LA - engRN - 0 (Peptides)RN - 0 (Recombinant Proteins)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleID - GM43495/GM/NIGMSDA - 19980219IS - 0006-2960SB - IMCY - UNITED STATES
UR  - PM:9454582
SO  - Biochemistry 1998 Jan 20 ;37(3):923-932

1550
UI  - 20811
AU  - Sorgen PL
AU  - Caviston TL
AU  - Perry RC
AU  - Cain BD
AD  - Department of Biochemistry and Molecular Biology, University of Florida, Gainesville, Florida 32610, USA
TI  - Deletions in the second stalk of F1F0-ATP synthase in Escherichia coli
AB  - In Escherichia coli F1F0-ATP synthase, the two b subunits form the second stalk spanning the distance between the membrane F0 sector and the bulk of F1. Current models predict that the stator should be relatively rigid and engaged in contact with F1 at fixed points. To test this hypothesis, we constructed a series of deletion mutations in the uncF(b) gene to remove segments from the middle of the second stalk of the subunit. Mutants with deletions of 7 amino acids were essentially normal, and those with deletions of up to 11 amino acids retained considerable activity. Membranes prepared from these strains had readily detectable levels of F1-ATPase activity and proton pumping activity. Removal of 12 or more amino acids resulted in loss of oxidative phosphorylation. Levels of membrane-associated F1-ATPase dropped precipitously for the longer deletions, and immunoblot analysis indicated that reductions in activity correlated with reduced levels of b subunit in the membranes. Assuming the likely alpha-helical conformation for this area of the b subunit, the 11-amino acid deletion would result in shortening the subunit by approximately 16 A. Since these deletions did not prevent the b subunit from participating in productive interactions with F1, we suggest that the b subunit is not a rigid rodlike structure, but has an inherent flexibility compatible with a dynamic role in coupling
MH  - A
MH  - ACID
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - Amino Acids
MH  - analysis
MH  - ATPase
MH  - Bacterial Proteins
MH  - Biochemistry
MH  - conformation
MH  - coupling
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - F1F0-ATP SYNTHASE
MH  - membrane
MH  - Membranes
MH  - model
MH  - mutant
MH  - Oxidative Phosphorylation
MH  - Phosphorylation
MH  - protein
MH  - Proteins
MH  - proton
MH  - Proton Pump
MH  - stalk
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 98447629LA - engRN - 0 (Bacterial Proteins)RN - 0 (Proton Pumps)RN - 0 (uncF protein, Escherichia coli)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleID - GM43495/GM/NIGMSDA - 19981112IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:9774398
SO  - J Biol Chem 1998 Oct 23 ;273(43):27873-27878

1551
UI  - 983
AU  - Steinberg-Yfrach G
AU  - Rigaud JL
AU  - Durantini EN
AU  - Moore AL
AU  - Gust D
AU  - Moore TA
TI  - Light-driven production of ATP catalysed by F0F1-ATP synthase in an artificial photosynthetic membrane
AB  - Energy-transducing membranes of living organisms couple spontaneous to non-spontaneous processes through the intermediacy of protonmotive force (p.m.f.)--an imbalance in electrochemical potential of protons across the membrane. In most organisms, p.m.f. is generated by redox reactions that are either photochemically driven, such as those in photosynthetic reaction centres, or intrinsically spontaneous, such as those of oxidative phosphorylation in mitochondria. Transmembrane proteins (such as the cytochromes and complexes I, III and IV in the electron-transport chain in the inner mitochondrial membrane) couple the redox reactions to proton translocation, thereby conserving a fraction of the redox chemical potential as p.m.f. Many transducer proteins couple p.m.f. to the performance of biochemical work, such as biochemical synthesis and mechanical and transport processes. Recently, an artificial photosynthetic membrane was reported in which a photocyclic process was used to transport protons across a liposomal membrane, resulting in acidification of the liposome's internal volume. If significant p.m.f. is generated in this system, then incorporating an appropriate transducer into the liposomal bilayer should make it possible to drive a non-spontaneous chemical process. Here we report the incorporation of F0F1-ATP synthase into liposomes containing the components of the proton-pumping photocycle. Irradiation of this artificial membrane with visible light results in the uncoupler- and inhibitor-sensitive synthesis of adenosine triphosphate (ATP) against an ATP chemical potential of approximately 12 kcal mol(-1), with a quantum yield of more than 7%. This system mimics the process by which photosynthetic bacteria convert light energy into ATP chemical potential
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - Bacteria
MH  - Biochemistry
MH  - Catalysis
MH  - Chemistry
MH  - Electron Transport
MH  - H(+)-Transporting ATP Synthase
MH  - Light
MH  - Liposomes
MH  - Membranes
MH  - metabolism
MH  - Mitochondria
MH  - Oxidative Phosphorylation
MH  - Phosphorylation
MH  - Photosynthesis
MH  - Proteins
MH  - Proton Pump
MH  - Protons
MH  - radiation effects
MH  - Spinach
MH  - Support,U.S.Gov't,Non-P.H.S.
RP  - NOT IN FILE
NT  - Department of Chemistry and Biochemistry, Arizona State University, Tempe 85287-1604, USA
SO  - Nature 1998 Apr 2 ;392(6675):479-482

1552
UI  - 20935
AU  - Svergun DI
AU  - Aldag I
AU  - Sieck T
AU  - Altendorf K
AU  - Koch MH
AU  - Kane DJ
AU  - Kozin MB
AU  - Gruber G
AD  - European Molecular Biology Laboratory, Hamburg Outstation, D-22603 Hamburg, Germany
TI  - A model of the quaternary structure of the Escherichia coli F1 ATPase from X-ray solution scattering and evidence for structural changes in the delta subunit during ATP hydrolysis
AB  - The shape and subunit arrangement of the Escherichia coli F1 ATPase (ECF1 ATPase) was investigated by synchrotron radiation x-ray solution scattering. The radius of gyration and the maximum dimension of the enzyme complex are 4.61 +/- 0.03 nm and 15.5 +/- 0.05 nm, respectively. The shape of the complex was determined ab initio from the scattering data at a resolution of 3 nm, which allowed unequivocal identification of the volume occupied by the alpha3beta3 subassembly and further positioning of the atomic models of the smaller subunits. The delta subunit was positioned near the bottom of the alpha3beta3 hexamer in a location consistent with a beta-delta disulfide formation in the mutant ECF1 ATPase, betaY331W:betaY381C:epsilonS108C, when MgADP is bound to the enzyme. The position and orientation of the epsilon subunit were found by interactively fitting the solution scattering data to maintain connection of the two-helix hairpin with the alpha3beta3 complex and binding of the beta-sandwich domain to the gamma subunit. Nucleotide- dependent changes of the delta subunit were investigated by stopped- flow fluorescence technique at 12 degrees C using N-[4-[7- (dimethylamino)-4-methyl]coumarin-3-yl]maleimide (CM) as a label. Fluorescence quenching monitored after addition of MgATP was rapid [k = 6.6 s-1] and then remained constant. Binding of MgADP and the noncleavable nucleotide analog AMP . PNP caused an initial fluorescent quenching followed by a slower decay back to the original level. This suggests that the delta subunit undergoes conformational changes and/or rearrangements in the ECF1 ATPase during ATP hydrolysis
MH  - A
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATPase
MH  - BINDING
MH  - COLI F1 ATPASE
MH  - COMPLEX
MH  - conformational change
MH  - CONSTANT
MH  - Copper
MH  - CROSS-LINKING
MH  - DELTA-SUBUNIT
MH  - DYE
MH  - dyes
MH  - EPSILON-SUBUNIT
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - fluorescence
MH  - Fluorescent Dyes
MH  - Hydrolysis
MH  - model
MH  - mutant
MH  - resolution
MH  - SUBUNIT
RP  - NOT IN FILE
NT  - UI - 99007340LA - engRN - 0 (Coumarins)RN - 0 (Cross-Linking Reagents)RN - 0 (Fluorescent Dyes)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 7440-50-8 (Copper)RN - 7447-39-4 (cupric chloride)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19981204IS - 0006-3495SB - IMCY - UNITED STATES
UR  - PM:9788916
SO  - Biophys J 1998 Nov ;75(5):2212-2219

1553
UI  - 20934
AU  - Svergun DI
AU  - Konrad S
AU  - Huss M
AU  - Koch MH
AU  - Wieczorek H
AU  - Altendorf K
AU  - Volkov VV
AU  - Gruber G
AD  - European Molecular Biology Laboratory, Hamburg Outstation, Germany
TI  - Quaternary structure of V1 and F1 ATPase: significance of structural homologies and diversities
AB  - The V1 ATPase from the tobacco hornworm Manduca sexta and the Escherichia coli F1 ATPase were characterized by small-angle X-ray scattering (SAXS). The radii of gyration (Rg) of the complexes were 6.2 +/- 0.1 and 4.7 +/- 0.02 nm, respectively. The shape of the M. sexta V1 ATPase was determined ab initio from the scattering data showing six masses, presumed to be the A and B subunits, arranged in an alternating manner about a 3-fold axis. A seventh mass with a length of about 11.0 nm extends perpendicularly to the center of the hexameric unit. This central mass is presumed to be the stalk that connects V1 with the membrane domain (V(O)) in the intact V1V(O)-ATPase. In comparison, the shape of the F1 ATPase from E. coli possesses a quasi-3-fold symmetry over the major part of the enzyme. The overall asymmetry of the structure is given by a stem, assumed to include the central stalk subunits. The features of the V1 and F1 ATPase reveal structural homologies and diversities of the key components of the complexes
MH  - A
MH  - ATPase
MH  - COLI F1 ATPASE
MH  - COMPLEX
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - H+-ATPase
MH  - M
MH  - membrane
MH  - Solutions
MH  - stalk
MH  - SUBUNIT
RP  - NOT IN FILE
NT  - UI - 99119193LA - engRN - 0 (Solutions)RN - EC 3.6.1.- (vacuolar H+-ATPase)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleID - A1 22444/PHSDA - 19990211IS - 0006-2960SB - IMCY - UNITED STATES
UR  - PM:9922131
SO  - Biochemistry 1998 Dec 22 ;37(51):17659-17663

1554
UI  - 21028
AU  - Syroeshkin AV
AU  - Bakeeva LE
AU  - Cherepanov DA
AD  - Institute of Immunology, Moscow, Russia
TI  - Contraction transitions of F1-F0 ATPase during catalytic turnover
AB  - Strong acoustic pressure was applied to submitochondrial particles (SMP) from bovine heart in order to drive ATP synthesis by F1-F0 complex for the account of sound waves. We observed a net ATP production at two narrow frequency ranges, about 170 Hz and about 340 Hz, that corresponds to the resonance oscillations of experimental cuvette when the acoustic pressure had a magnitude of 100 kPa. The results can be explained quantitatively by contractive conformational changes of F1-F0 complex during catalytic turnover. Negative staining electron microscopy of SMP preparations was used to visualize the ADP(Mg2+)-induced conformational changes of F1-F0 complex. In the particles with high ATPase activity in the presence of phosphate the factors F1 and F0 formed a congregated domain plunged into the membrane without any observable stalk in between. The presence of ADP(Mg2+) caused a structural rearrangement of F1-F0 to the essentially different conformation: the domains F1 and F0 were dislodged distinctly from each other and connected by a long thin stalk. The latter conformation resembled well the usual bipartite profile of ATPase. The data indicate that besides rotation, the catalytic turnover of ATP synthase is also accompanied by stretch transitions of F1-F0 complex
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP production
MH  - ATP synthase
MH  - ATP synthesis
MH  - ATPase
MH  - COMPLEX
MH  - conformation
MH  - conformational change
MH  - conformational changes
MH  - electron
MH  - electron microscopy
MH  - ELECTRON-MICROSCOPY
MH  - F0
MH  - F1
MH  - membrane
MH  - Microscopy
MH  - rotation
MH  - stalk
MH  - Submitochondrial Particles
MH  - SYNTHASE
MH  - synthesis
RP  - NOT IN FILE
NT  - UI - 99057630LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19990120IS - 0006-3002SB - IMCY - NETHERLANDS
UR  - PM:9838045
SO  - Biochim Biophys Acta 1998 Dec 1 ;1409(2):59-71

1555
UI  - 19
AU  - Valiyaveetil FI
AU  - Fillingame RH
AD  - Department of Biomolecular Chemistry, University of Wisconsin Medical School, Madison, Wisconsin 53706, USA
TI  - Transmembrane topography of subunit a in the Escherichia coli F1F0 ATP synthase
AB  - Subunit a is the least understood of the three subunits that compose the F0 sector in the Escherichia coli F0F1 ATP synthase. In this study, we have substituted Cys into predicted extramembranous loops of the protein and used chemical modification to obtain topographical information on the folding of subunit a. The extent of labeling of the substituted Cys residues by fluorescein-5'-maleimide was determined. The localization of reactive Cys residues was inferred from differences in the extent of labeling in inside out and right side out membrane vesicles. The NH2-terminal segment of subunit a was localized to the outside (periplasmic) surface and the COOH terminus to the cytoplasmic surface by these procedures. Loop residues in two periplasmic extramembranous loops and in two cytoplasmic extramembranous loops were also localized. The localization of two cytoplasmic Cys residues was confirmed by using 4-acetamido-4'-maleimidylstilbene-2,2'-disulfonic acid to block fluorescein-5'-maleimide labeling. From the localization of the Cys residues, a model for the topography is proposed that consists of five transmembrane segments with the NH2 terminus periplasmic and the COOH terminus cytoplasmic. The positions of second site suppressors, including several isolated here to the nonfunctional E219C and H245C substitutions, provide support for the topographical model proposed
RP  - NOT IN FILE
NT  - UI - 98298136LA - engRN - 0 (Sulfhydryl Reagents)RN - 52-90-4 (Cysteine)RN - 56-45-1 (Serine)RN - 56-65-5 (Adenosine Triphosphate)RN - 56-85-9 (Glutamine)RN - 7006-35-1 (Histidine)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19980803IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:9632683
SO  - J Biol Chem 1998 Jun 26 ;273(26):16241-16247

1556
UI  - 19907
AU  - Wang H
AU  - Oster G
AD  - Department of Molecular and Cellular Biology, College of Natural Resources, University of California, Berkeley 94720-3112, USA
TI  - Energy transduction in the F1 motor of ATP synthase
AB  - ATP synthase is the universal enzyme that manufactures ATP from ADP and phosphate by using the energy derived from a transmembrane protonmotive gradient. It can also reverse itself and hydrolyse ATP to pump protons against an electrochemical gradient. ATP synthase carries out both its synthetic and hydrolytic cycles by a rotary mechanism. This has been confirmed in the direction of hydrolysis after isolation of the soluble F1 portion of the protein and visualization of the actual rotation of the central 'shaft' of the enzyme with respect to the rest of the molecule, making ATP synthase the world's smallest rotary engine. Here we present a model for this engine that accounts for its mechanochemical behaviour in both the hydrolysing and synthesizing directions. We conclude that the F1 motor achieves its high mechanical torque and almost 100% efficiency because it converts the free energy of ATP binding into elastic strain, which is then released by a coordinated kinetic and tightly coupled conformational mechanism to create a rotary torque
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - ADP
MH  - atp
MH  - ATP synthase
MH  - BINDING
MH  - England
MH  - F1
MH  - H(+)-Transporting ATP Synthase
MH  - Hydrolysis
MH  - mechanism
MH  - model
MH  - protein
MH  - proton
MH  - Protons
MH  - rotation
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 99049782LA - engRN - 0 (Molecular Motors)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19981208IS - 0028-0836SB - IMCY - ENGLANDJC - NSC
UR  - PM:9834036
SO  - Nature 1998 Nov 19 ;396(6708):279-282

1557
UI  - 411
AU  - Weber J
AU  - Wilke-Mounts S
AU  - Hammond ST
AU  - Senior AE
AD  - Department of Biochemistry and Biophysics, University of Rochester Medical Center, New York 14642, USA
TI  - Tryptophan substitutions surrounding the nucleotide in catalytic sites of F1-ATPase
AB  - Novel tryptophan substitutions, surrounding the nucleotide bound in catalytic sites, were introduced into Escherichia coli F1-ATPase. The mutant enzymes were purified and studied by fluorescence spectroscopy. One cluster of Trp substitutions, consisting of beta-Trp-404, beta-Trp- 410, beta-Asp-158 (lining the adenine-binding pocket), and beta-Trp-153 (close to the alpha/beta-phosphates), showed the same fluorescence responses to MgADP, MgAMPPNP, and MgATP and the same nucleotide binding pattern with MgADP and MgAMPPNP, with one site of higher and two sites of lower affinity. Therefore, in absence of catalytic turnover (and of gamma-subunit rotation), sites 2 and 3 appeared similar in affinity, and the region of the catalytic site sensed by these Trp substitutions did not change conformation with different nucleotides. In contrast, alpha-Trp-291 and beta-Trp-297, both close to the gamma-phosphate, showed very different fluorescence responses to MgADP versus MgAMPPNP, and in these cases the response was due exclusively or predominantly to nucleotide binding at the first, high-affinity catalytic site, thus allowing specific detection of this site. Titration with MgATP showed that the high-affinity site was present under conditions of steady- state, Vmax MgATP hydrolysis
RP  - NOT IN FILE
NT  - UI - 98393455LA - engRN - 0 (Adenine Nucleotides)RN - 0 (Ligands)RN - 25612-73-1 (Adenylyl Imidodiphosphate)RN - 73-22-3 (Tryptophan)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSDA - 19980924IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:9724515
SO  - Biochemistry 1998 Sep 1 ;37(35):12042-12050

1558
UI  - 413
AU  - Weber J
AU  - Hammond ST
AU  - Wilke-Mounts S
AU  - Senior AE
AD  - Department of Biochemistry and Biophysics, University of Rochester Medical Center, Box 712, Rochester, New York 14642, USA
TI  - Mg2+ coordination in catalytic sites of F1-ATPase
AB  - Coordination of the Mg2+ ion in Mg-nucleotide substrates by amino acid residue side chains in the catalytic site of Escherichia coli F1-ATPase was investigated. From the X-ray structure of the mitochondrial enzyme [Abrahams, J. P., Leslie, A. G. W., Lutter, R., and Walker, J. E. (1994) Nature 370, 621-628], it may be inferred that the hydroxyl of betaThr-156 is a direct ligand of Mg2+, whereas the carboxyls of betaGlu-181, betaGlu-185, and betaAsp-242 might contribute via intervening water molecules. Elimination of each respective functional group by site-directed mutagenesis, followed by determination of Mg- nucleotide and uncomplexed nucleotide binding affinities using a tryptophan probe, showed that betaThr-156, betaGlu-185, and betaAsp-242 are all involved in Mg2+ coordination, whereas betaGlu-181 is not. A derived structural model for the octahedral coordination around the Mg2+ ion is presented. The results indicate that the ADP-containing site in the X-ray structure is the catalytic site of highest affinity. Correct Mg2+ coordination is required for catalytic activity at physiological rates. Elimination of any one of the Mg2+-coordinating residues led to complete loss of Mg2+-dependent nucleotide binding cooperativity of the catalytic sites
RP  - NOT IN FILE
NT  - UI - 98087534LA - engRN - 0 (Adenine Nucleotides)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 7439-95-4 (Magnesium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSDA - 19980213IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:9425083
SO  - Biochemistry 1998 Jan 13 ;37(2):608-614

1559
UI  - 19209
AU  - Weber J
AU  - Senior AE
TI  - Effects of the inhibitors azide, dicyclohexylcarbodiimide, and aurovertin on nucleotide binding to the three F1-ATPase catalytic sites measured using specific tryptophan probes.
AB  - Equilibrium nucleotide binding to the three catalytic sites of Escherichia coli F1-ATPase was measured in the presence of the inhibitors azide, dicyclohexylcarbodiimide, and aurovertin to elucidate mechanisms of inhibition. Fluorescence signals of beta-Trp-331 and beta- Trp-148 substituted in catalytic sites were used to determine nucleotide binding parameters. Azide brought about small decreases in Kd(MgATP) and Kd(MgADP). Notably, under MgATP hydrolysis conditions, it caused all enzyme molecules to assume a state with three catalytic site- bound MgATP and zero bound MgADP. These results rule out the idea that azide inhibits by "trapping" MgADP. Rather, azide blocks the step at which signal transmission between catalytic sites promotes multisite hydrolysis. Aurovertin bound with stoichiometry of 1.8 (mol/mol of F1) and allowed significant residual turnover. Cycling of the aurovertin- free beta-subunit catalytic site through three normal conformations was indicated by MgATP binding data. Aurovertin did not change the normal ratio of 1 bound MgATP/2 bound MgADP in catalytic sites. The results indicate that it acts to slow the switch of catalytic site affinities ("binding change step") subsequent to MgATP hydrolysis. Dicyclohexylcarbodiimide shifted the ratio of catalytic site-bound MgATP/MgADP from 1:2 to 1.6:1.4, without affecting Kd(MgATP) values. Like azide, it also appears to affect activity at the step after MgATP binding, in which signal transmission between catalytic sites promotes MgATP hydrolysis.
MH  - A
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - affinity
MH  - antagonists & inhibitors
MH  - aurovertin
MH  - Aurovertins
MH  - azide
MH  - BETA
MH  - BETA-SUBUNIT
MH  - BINDING
MH  - Biochemistry
MH  - Biophysics
MH  - catalytic
MH  - catalytic domain
MH  - Chemistry
MH  - COLI F1 ATPASE
MH  - conformation
MH  - data
MH  - Dicyclohexylcarbodiimide
MH  - enzyme
MH  - Enzyme Inhibitors
MH  - equilibrium
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - fluorescence
MH  - H(+)-Transporting ATP Synthase
MH  - Hydrolysis
MH  - inhibitor
MH  - inhibitors
MH  - mechanism
MH  - MECHANISMS
MH  - metabolism
MH  - molecular probes
MH  - nucleotide
MH  - nucleotide binding
MH  - pharmacology
MH  - Protein Binding
MH  - Site
MH  - sodium azide
MH  - Spectrometry,Fluorescence
MH  - stoichiometry
MH  - Support,U.S.Gov't,P.H.S.
MH  - switch
MH  - tryptophan
MH  - turnover
MH  - universities
RP  - NOT IN FILE
NT  - Department of Biochemistry and Biophysics, University of Rochester Medical Center, Rochester, New York 14642, USAPMID- 0009837890
SO  - J Biol Chem 1998 Dec 11 ;273(50):33210-33215

1560
UI  - 410
AU  - Weber J
AU  - Senior AE
AD  - Department of Biochemistry and Biophysics, University of Rochester Medical Center, Rochester, New York 14642, USA
TI  - Effects of the inhibitors azide, dicyclohexylcarbodiimide, and aurovertin on nucleotide binding to the three F1-ATPase catalytic sites measured using specific tryptophan probes
AB  - Equilibrium nucleotide binding to the three catalytic sites of Escherichia coli F1-ATPase was measured in the presence of the inhibitors azide, dicyclohexylcarbodiimide, and aurovertin to elucidate mechanisms of inhibition. Fluorescence signals of beta-Trp-331 and beta- Trp-148 substituted in catalytic sites were used to determine nucleotide binding parameters. Azide brought about small decreases in Kd(MgATP) and Kd(MgADP). Notably, under MgATP hydrolysis conditions, it caused all enzyme molecules to assume a state with three catalytic site- bound MgATP and zero bound MgADP. These results rule out the idea that azide inhibits by "trapping" MgADP. Rather, azide blocks the step at which signal transmission between catalytic sites promotes multisite hydrolysis. Aurovertin bound with stoichiometry of 1.8 (mol/mol of F1) and allowed significant residual turnover. Cycling of the aurovertin- free beta-subunit catalytic site through three normal conformations was indicated by MgATP binding data. Aurovertin did not change the normal ratio of 1 bound MgATP/2 bound MgADP in catalytic sites. The results indicate that it acts to slow the switch of catalytic site affinities ("binding change step") subsequent to MgATP hydrolysis. Dicyclohexylcarbodiimide shifted the ratio of catalytic site-bound MgATP/MgADP from 1:2 to 1.6:1.4, without affecting Kd(MgATP) values. Like azide, it also appears to affect activity at the step after MgATP binding, in which signal transmission between catalytic sites promotes MgATP hydrolysis
RP  - NOT IN FILE
NT  - UI - 99057874LA - engRN - 0 (Aurovertins)RN - 0 (Enzyme Inhibitors)RN - 0 (Molecular Probes)RN - 26628-22-8 (Sodium Azide)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 73-22-3 (Tryptophan)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSDA - 19990114IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:9837890
SO  - J Biol Chem 1998 Dec 11 ;273(50):33210-33215

1561
UI  - 148
AU  - Wilkens S
AU  - Capaldi RA
TI  - ATP synthase's second stalk comes into focus
RP  - NOT IN FILE
NT  - UI - 98250570LA - engRN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - LetterDA - 19980527IS - 0028-0836SB - IMCY - ENGLANDJC - NSC
UR  - PM:9590688
SO  - Nature 1998 May 7 ;393(6680):29

1562
UI  - 145
AU  - Wilkens S
AU  - Capaldi RA
AD  - Institute of Molecular Biology, University of Oregon, Eugene 97403- 1229, USA
TI  - Electron microscopic evidence of two stalks linking the F1 and F0 parts of the Escherichia coli ATP synthase
AB  - The structure of monodisperse ATP synthase from Escherichia coli (ECF1F0) has been examined by electron microscopy after negative staining of specimens. The F1 part is seen to be connected by two stalks. One is more centrally located and includes the gamma and epsilon subunits. The second stalk, observed here in ECF1F0, is arranged peripherally. It probably contains the delta and b subunits which, in addition to gamma and epsilon, are required for binding of the F1 and F0 parts of the complex. Other novel features of the F1F0 complex can be discerned. There is a cap at the top of the F1 part at which the second stalk may bind. This likely includes N-terminal stretches of the three copies of the alpha subunit and a part of the delta subunit. The F0 part is clearly asymmetric. The presence of two stalks in the complex has important functional implications. There is good evidence that the more central stalk of gamma and epsilon subunits is a mobile domain that rotates to link the three catalytic sites on beta subunits in turn, with the proton channel of the F0 part. The second stalk of delta and b subunits is then the stator which makes this rotation possible
RP  - NOT IN FILE
NT  - UI - 98358598LA - engRN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialID - HL58671/HL/NHLBIDA - 19980824IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:9693727
SO  - Biochim Biophys Acta 1998 Jun 10 ;1365(1-2):93-97

1563
UI  - 144
AU  - Wilkens S
AU  - Capaldi RA
AD  - University of Oregon, Institute of Molecular Biology, Eugene, Oregon 97403, USA
TI  - Solution structure of the epsilon subunit of the F1-ATPase from Escherichia coli and interactions of this subunit with beta subunits in the complex
AB  - The solution structure of the epsilon subunit of the Escherichia coli F1-ATPase has been determined by NMR spectroscopy. This subunit has a two-domain structure with an N-terminal 10-stranded beta sandwich and a C-terminal antiparallel two alpha-helix hairpin, as described previously (Wilkens, S., Dahlquist, F. W., McIntosh, L. P., Donaldson, L. W., and Capaldi, R. A. (1995) Nat. Struct. Biol. 2, 961-967). New data show that the two domains interact in solution in an interface formed by beta strand 7 and the very C-terminal alpha-helix. This interface involves only hydrophobic interactions. The dynamics of the epsilon subunit in solution were examined. The two domains are relatively tightly associated with little or no flexibility relative to one another. The epsilon subunit can exist in two states in the ECF1F0 complex depending on whether ATP or ADP occupies catalytic sites. Proteolysis of the epsilon subunit in solution and when bound to the core F1 complex indicates that the conformation of the polypeptide in solution closely resembles the conformation of epsilon when bound to the F1 in the ADP state. Chemical and photo-cross-linking show that the epsilon subunit spans and interacts with two beta subunits in the ADP state. These interactions are disrupted on binding of ATP + Mg2+, as is the interaction between the N- and C-terminal domains of the epsilon subunit
RP  - NOT IN FILE
NT  - UI - 98434577LA - engRN - 0 (Solutions)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - 58671/PHSDA - 19981102IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:9756905
SO  - J Biol Chem 1998 Oct 9 ;273(41):26645-26651

1564
UI  - 703
AU  - Yasuda R
AU  - Noji H
AU  - Kinosita K
AU  - Yoshida M
AD  - Department of Physics, Faculty of Science and Technology, Keio University, Yokohama, Japan
TI  - F1-ATPase is a highly efficient molecular motor that rotates with discrete 120 degree steps
AB  - A single molecule of F1-ATPase, a portion of ATP synthase, is by itself a rotary motor in which a central gamma subunit rotates against a surrounding cylinder made of alpha3beta3 subunits. Driven by three catalytic betas, each fueled with ATP, gamma makes discrete 120 degree steps, occasionally stepping backward. The work done in each step is constant over a broad range of imposed load and is close to the free energy of hydrolysis of one ATP molecule
RP  - NOT IN FILE
NT  - UI - 98319227LA - engRN - 0 (Actins)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19980727IS - 0092-8674SB - IMCY - UNITED STATESJC - CQ4
UR  - PM:9657145
SO  - Cell 1998 Jun 26 ;93(7):1117-1124

1565
UI  - 701
AU  - Yokoyama K
AU  - Muneyuki E
AU  - Amano T
AU  - Mizutani S
AU  - Yoshida M
AU  - Ishida M
AU  - Ohkuma S
AD  - Department of Biochemistry, Faculty of Pharmaceutical Science, Kanazawa University, Takara-machi 13-1, Kanazawa, Ishikawa 920, Japan yokoken@kenrokukanazawa-uacjp
TI  - V-ATPase of Thermus thermophilus is inactivated during ATP hydrolysis but can synthesize ATP
AB  - The ATP hydrolysis of the V1-ATPase of Thermus thermophilus have been investigated with an ATP-regenerating system at 25 degreesC. The ratio of ATPase activity to ATP concentration ranged from 40 to 4000 microM; from this, an apparent Km of 240 +/- 24 microM and a Vmax of 5.2 +/- 0.5 units/mg were deduced. An apparent negative cooperativity, which is frequently observed in case of F1-ATPases, was not observed for the V1- ATPase. Interestingly, the rate of hydrolysis decayed rapidly during ATP hydrolysis, and the ATP hydrolysis finally stopped. Furthermore, the inactivation of the V1-ATPase was attained by a prior incubation with ADP-Mg. The inactivated V1-ATPase contained 1.5 mol of ADP/mol of enzyme. Difference absorption spectra generated from addition of ATP-Mg to the isolated subunits revealed that the A subunit can bind ATP-Mg, whereas the B subunit cannot. The inability to bind ATP-Mg is consistent with the absence of Walker motifs in the B subunit. These results indicate that the inactivation of the V1-ATPase during ATP hydrolysis is caused by entrapping inhibitory ADP-Mg in a catalytic site. Light-driven ATP synthesis by bacteriorhodopsin-VoV1-ATPase proteoliposomes was observed, and the rate of ATP synthesis was approximately constant. ATP synthesis occurred in the presence of an ADP-Mg of which concentration was high enough to induce complete inactivation of ATP hydrolysis of VoV1-ATPase. This result indicates that the ADP-Mg-inhibited form is not produced in ATP synthesis reaction
RP  - NOT IN FILE
NT  - UI - 98352093LA - engRN - 0 (Bacterial Proteins)RN - 0 (Enzyme Inhibitors)RN - 0 (Liposomes)RN - 0 (Recombinant Proteins)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.1.- (V-type ATPase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19980910IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:9685406
SO  - J Biol Chem 1998 Aug 7 ;273(32):20504-20510

1566
UI  - 21071
AU  - Zhang Z
AU  - Huang L
AU  - Shulmeister VM
AU  - Chi YI
AU  - Kim KK
AU  - Hung LW
AU  - Crofts AR
AU  - Berry EA
AU  - Kim SH
AD  - E O Lawrence Berkeley National Laboratory, University of California, 94720, USA
TI  - Electron transfer by domain movement in cytochrome bc1
AB  - The cytochrome bc1 is one of the three major respiratory enzyme complexes residing in the inner mitochondrial membrane. Cytochrome bc1 transfers electrons from ubiquinol to cytochrome c and uses the energy thus released to form an electrochemical gradient across the inner membrane. Our X-ray crystal structures of the complex from chicken, cow and rabbit in both the presence and absence of inhibitors of quinone oxidation, reveal two different locations for the extrinsic domain of one component of the enzyme, an iron-sulphur protein. One location is close enough to the supposed quinol oxidation site to allow reduction of the Fe-S protein by ubiquinol. The other site is close enough to cytochrome c1 to allow oxidation of the Fe-S protein by the cytochrome. As neither location will allow both reactions to proceed at a suitable rate, the reaction mechanism must involve movement of the extrinsic domain of the Fe-S component in order to shuttle electrons from ubiquinol to cytochrome c1. Such a mechanism has not previously been observed in redox protein complexes
MH  - A
MH  - Antimycin A
MH  - COMPLEX
MH  - cytochrome
MH  - Cytochrome c
MH  - Cytochrome c1
MH  - electron
MH  - electron transfer
MH  - Electron-transfer
MH  - Electrons
MH  - England
MH  - inhibitor
MH  - mechanism
MH  - membrane
MH  - Movement
MH  - protein
MH  - Proteins
MH  - quinone
MH  - redox
MH  - Site
MH  - structure
MH  - Thiazoles
MH  - TRANSFER
MH  - Ubiquinol-Cytochrome-c Reductase
RP  - NOT IN FILE
NT  - UI - 98224700LA - engRN - 0 (Iron-Sulfur Proteins)RN - 0 (Polyenes)RN - 0 (Thiazoles)RN - 11118-72-2 (antimycin)RN - 642-15-9 (Antimycin A)RN - 76706-55-3 (myxothiazol)RN - 9007-43-6 (Cytochrome c)RN - 91682-96-1 (stigmatellin)RN - EC 1.10.2.2 (Ubiquinol-Cytochrome-c Reductase)PT - Journal ArticleDA - 19980514IS - 0028-0836SB - IMCY - ENGLAND
UR  - PM:9565029
SO  - Nature 1998 Apr 16 ;392(6677):677-684

1567
UI  - 697
AU  - Amano T
AU  - Matsui T
AU  - Muneyuki E
AU  - Noji H
AU  - Hara K
AU  - Yoshida M
AU  - Hisabori T
AD  - Research Laboratory of Resources Utilization, Tokyo Institute of Technology, Nagatsuta 4259, Midori-ku, Yokohama 226-8503, Japan
TI  - alpha3beta3gamma complex of F1-ATPase from thermophilic Bacillus PS3 can maintain steady-state ATP hydrolysis activity depending on the number of non-catalytic sites
AB  - Homogeneous preparations of alpha(3)beta(3)gamma complexes with one, two or three non-competent non-catalytic site(s) were performed as described [Amano, Hisabori, Muneyuki, and Yoshida (1996) J. Biol. Chem. 271, 18128-18133] and their properties were compared with those of the wild-type complex. The ATPase activity of the complex with three non- competent non-catalytic sites decayed rapidly to an inactivated state, as reported previously [Matsui, Muneyuki, Honda, Allison, Dou, and Yoshida (1997) J. Biol. Chem. 272, 8215-8221]. In contrast, the complex with one or two non-competent non-catalytic sites displayed a substantial steady-state phase activity depending on the number of non- competent non-catalytic sites in the complex. This result indicates that one competent non-catalytic site can maintain the continuous catalytic turnover of the enzyme and can potentially relieve all three catalytic sites from inhibition by MgADP(-). Furthermore, the results suggest that the interaction between three non-catalytic sites might not be as strong as that between catalytic sites, which are all strictly required for a continuous catalytic turnover
RP  - NOT IN FILE
NT  - UI - 99425078LA - engRN - 0 (DNA Primers)RN - 0 (Dimethylamines)RN - 0 (Enzyme Activators)RN - 1643-20-5 (dodecyldimethylamine oxide)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19991207IS - 0264-6021SB - IMCY - ENGLANDJC - 9YO
UR  - PM:10493921
SO  - Biochem J 1999 Oct 1 ;343 Pt 1():135-138

1568
UI  - 698
AU  - Bald D
AU  - Muneyuki E
AU  - Amano T
AU  - Kruip J
AU  - Hisabori T
AU  - Yoshida M
AD  - Tokyo Institute of Technology, Research Laboratory for Resources Utilization, Midori-ku, Yokohama, Japan
TI  - The noncatalytic site-deficient alpha3beta3gamma subcomplex and FoF1- ATP synthase can continuously catalyse ATP hydrolysis when Pi is present
AB  - We investigated ATP hydrolysis by a mutant (DeltaNC) alpha3beta3gamma subcomplex of F0F1-ATP synthase from the thermophilic Bacillus PS3 that is defective in the noncatalytic nucleotide binding sites. This mutant subcomplex was activated by inorganic phosphate ions (Pi) and did not show continuous ATP hydrolysis activity in the absence of Pi. Pi also activated the wild-type alpha3beta3gamma subcomplex in a similar manner. Sulphate activated wild-type alpha3beta3gamma but not DeltaNC alpha3beta3gamma, indicating that Pi activation did not involve noncatalytic sites but that sulphate activation did. Pi also activated ATP hydrolysis and coupled proton translocation by the wild-type and DeltaNC F0F1-ATP synthases reconstituted into vesicle membranes
RP  - NOT IN FILE
NT  - UI - 99269031LA - engRN - 0 (Phosphates)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19990624IS - 0014-2956SB - IMCY - GERMANYJC - EMZ
UR  - PM:10336643
SO  - Eur J Biochem 1999 Jun ;262(2):563-568

1569
UI  - 799
AU  - Berry S
AU  - Rumberg B
AD  - Lehrstuhl Biochemie der Pflanzen, Ruhr-Universitat Bochum, D-44780, Bochum, Germany
TI  - Proton to electron stoichiometry in electron transport of spinach thylakoids
AB  - According to the concept of the Q-cycle, the H+/e- ratio of the electron transport chain of thylakoids can be raised from 2 to 3 by means of the rereduction of plastoquinone across the cytochrome b6f complex. In order to investigate the H+/e- ratio we compared stationary rates of electron transport and proton translocation in spinach thylakoids both in the presence of the artificial electron acceptor ferricyanide and in the presence of the natural acceptor system ferredoxin+NADP. The results may be summarised as follows: (1) a variability of the H+/e- ratio occurs with either acceptor. H+/e- ratios of 3 (or even higher in the case of the natural acceptor system, see below) are decreased towards 2 if strong light intensity and low membrane permeability are employed. Mechanistically this could be explained by proton channels connecting the plastoquinol binding site alternatively to the lumenal or stromal side of the cytochrome b6f complex, giving rise to a proton slip reaction at high transmembrane DeltapH. In this slip reaction protons are deposited on the stromal instead of the lumenal side. In addition to the pH effect there seems to be a contribution of the redox state of the plastoquinone pool to the control of proton translocation; switching over to stromal proton deposition is favoured when the reduced state of plastoquinone becomes dominant. (2) In the presence of NADP a competition of both NADP and oxygen for the electrons supplied by photosystem I takes place, inducing a general increase of the H+/e- ratios above the values obtained with ferricyanide. The implications with respect to the adjustment of a proper ATP/NADPH ratio for CO2 reduction are discussed
RP  - NOT IN FILE
NT  - UI - 0LA - EngPT - JOURNAL ARTICLEDA - 19990401IS - 0006-3002SB - IMJC - A0W
UR  - PM:0010082791
SO  - Biochim Biophys Acta 1999 Mar 9 ;1410(3):248-261

1570
UI  - 20933
AU  - Birkenhager R
AU  - Greie JC
AU  - Altendorf K
AU  - Deckers-Hebestreit G
AD  - Universitat Osnabruck, Fachbereich Biologie/Chemie, Abteilung Mikrobiologie, Osnabruck, Germany
TI  - F0 complex of the Escherichia coli ATP synthase. Not all monomers of the subunit c oligomer are involved in F1 interaction
AB  - The antigenic determinants of mAbs against subunit c of the Escherichia coli ATP synthase were mapped by ELISA using overlapping synthetic heptapeptides. All epitopes recognized are located in the hydrophilic loop region and are as follows: 31-LGGKFLE-37, 35-FLEGAAR-41, 36-LEGAAR- 41 and 36-LEGAARQ-42. Binding studies with membrane vesicles of different orientation revealed that all mAbs bind to everted membrane vesicles independent of the presence or absence of the F1 part. Although the hydrophilic region of subunit c and particularly the highly conserved residues A40, R41, Q42 and P43 are known to interact with subunits gamma and epsilon of the F1 part, the mAb molecules have no effect on the function of F0. Furthermore, it could be demonstrated that the F1 part and the mAb molecule(s) are bound simultaneously to the F0 complex suggesting that not all c subunits are involved in F1 interaction. From the results obtained, it can be concluded that this interaction is fixed, which means that subunits gamma and epsilon do not switch between the c subunits during catalysis and furthermore, a complete rotation of the subunit c oligomer modified with mAb(s) along the stator of the F1F0 complex, proposed to be composed of at least subunits b and delta, seems to be unlikely
MH  - A
MH  - acceptor
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BINDING
MH  - Catalysis
MH  - COLI ATP SYNTHASE
MH  - COMPLEX
MH  - DYE
MH  - dyes
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - F1
MH  - Fluorescent Dyes
MH  - function
MH  - membrane
MH  - membrane vesicles
MH  - Multienzyme Complexes
MH  - Peptide Fragments
MH  - Phosphotransferases
MH  - RESIDUE
MH  - rotation
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 99421642LA - engRN - 0 (Aminoacridines)RN - 0 (Antibodies, Monoclonal)RN - 0 (Fluorescent Dyes)RN - 0 (Multienzyme Complexes)RN - 0 (Peptide Fragments)RN - 3548-09-2 (9-amino-6-chloro-2-methoxyacridine)RN - EC 2.7.4 (Phosphotransferases (Phosphate Group Acceptor))RN - EC 2.7.4.- (ATP Synthetase Complexes)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19991014IS - 0014-2956SB - IMCY - GERMANY
UR  - PM:10491083
SO  - Eur J Biochem 1999 Sep ;264(2):385-396

1571
UI  - 19768
AU  - Boyer PD
TI  - What makes ATP synthase spin?
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthase
MH  - England
MH  - H(+)-Transporting ATP Synthase
MH  - proton
MH  - Protons
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 20046436LA - engRN - 0 (Protons)RN - 56-65-5 (Adenosine Triphosphate)RN - 56-84-8 (Aspartic Acid)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - CommentPT - NewsDA - 19991210IS - 0028-0836SB - IMCY - ENGLANDJC - NSC
UR  - PM:10580491
SO  - Nature 1999 Nov 18 ;402(6759):247, 249

1572
UI  - 283
AU  - Cherepanov DA
AU  - Mulkidjanian AY
AU  - Junge W
AD  - Division of Biophysics, Faculty of Biology/Chemistry, University of Osnabruck, Germany
TI  - Transient accumulation of elastic energy in proton translocating ATP synthase
AB  - ATP synthase is conceived as a rotatory engine with two reversible drives, the proton-transporting membrane portion, F0, and the catalytic peripheral portion, F1. They are mounted on a central shaft (subunit gamma) and held together by an eccentric bearing. It is established that the hydrolysis of three molecules of ATP in F1 drives the shaft over a full circle in three steps of 120 degrees each. Proton flow through F0 probably generates a 12-stepped rotation of the shaft so that four proton-translocating steps of 30 degrees each drive the synthesis of one molecule of ATP. We addressed the elasticity of the transmission between F0 and F1 in a model where the four smaller steps in F0 load a torsional spring which is only released under liberation of ATP from F1. The kinetic model of an elastic ATP synthase described a wealth of published data on the synthesis/hydrolysis of ATP by F0F1 and on proton conduction by F0 as function of the pH and the protonmotive force. The pK values of the proton-carrying group interacting with the acidic and basic sides of the membrane were estimated as 5.3-6.4 and 8.0-8.3, respectively
RP  - NOT IN FILE
NT  - UI - 99239922LA - engRN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19990601IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:10225416
SO  - FEBS Lett 1999 Apr 16 ;449(1):1-6

1573
UI  - 21298
AU  - Cheung MS
AU  - Daizadeh I
AU  - Stuchebrukhov AA
AU  - Heelis PF
AD  - Department of Chemistry, University of California, Davis, California 95616, USA
TI  - Pathways of electron transfer in Escherichia coli DNA photolyase: Trp306 to FADH
AB  - We describe the results of a series of theoretical calculations of electron transfer pathways between Trp306 and *FADH. in the Escherichia coli DNA photolyase molecule, using the method of interatomic tunneling currents. It is found that there are two conformationally orthogonal tryptophans, Trp359 and Trp382, between donor and acceptor that play a crucial role in the pathways of the electron transfer process. The pathways depend vitally on the aromaticity of tryptophans and the flavin molecule. The results of this calculation suggest that the major pathway of the electron transfer is due to a set of overlapping orthogonal pi-rings, which starts from the donor Trp306, runs through Trp359 and Trp382, and finally reaches the flavin group of the acceptor complex, FADH
MH  - A
MH  - acceptor
MH  - Chemistry
MH  - COMPLEX
MH  - electron
MH  - electron transfer
MH  - Electron-transfer
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - method
MH  - TRANSFER
MH  - tryptophan
MH  - united states
MH  - universities
RP  - NOT IN FILE
NT  - UI - 99158601DA - 19990405IS - 0006-3495LA - engID - GM54052-02/GM/NIGMSPT - Journal ArticleCY - UNITED STATESRN - 0 (Pyrimidine Dimers)RN - 146-14-5 (Flavin-Adenine Dinucleotide)RN - 73-22-3 (Tryptophan)RN - EC 4.1.99.3 (Deoxyribodipyrimidine Photo-Lyase)SB - IM
UR  - PM:10049308
SO  - Biophys J 1999 Mar ;76(3):1241-1249

1574
UI  - 20
AU  - Dimroth P
AU  - Wang H
AU  - Grabe M
AU  - Oster G
AD  - Mikrobiologisches Institut, Eidgenossische Technische Hochschule, ETH- Zentrum, Schmelzbergstrasse 7, CH-8092 Zurich, Switzerland
TI  - Energy transduction in the sodium F-ATPase of Propionigenium modestum
AB  - The F-ATPase of the bacterium Propionigenium modestum is driven by an electrochemical sodium gradient between the cell interior and its environment. Here we present a mechanochemical model for the transduction of transmembrane sodium-motive force into rotary torque. The same mechanism is likely to operate in other F-ATPases, including the proton-driven F-ATPases of Escherichia coli
RP  - NOT IN FILE
NT  - UI - 99238459LA - engRN - 0 (Bacterial Proteins)RN - 0 (Molecular Motors)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19990610IS - 0027-8424SB - IMCY - UNITED STATESJC - PV3
UR  - PM:10220395
SO  - Proc Natl Acad Sci U S A 1999 Apr 27 ;96(9):4924-4929

1575
UI  - 103
AU  - Dmitriev O
AU  - Jones PC
AU  - Jiang W
AU  - Fillingame RH
AD  - Department of Biomolecular Chemistry, University of Wisconsin Medical School, Madison, Wisconsin 53706, USA
TI  - Structure of the membrane domain of subunit b of the Escherichia coli F0F1 ATP synthase
AB  - The structure of the N-terminal transmembrane domain (residues 1-34) of subunit b of the Escherichia coli F0F1-ATP synthase has been solved by two-dimensional 1H NMR in a membrane mimetic solvent mixture of chloroform/methanol/H2O (4:4:1). Residues 4-22 form an alpha-helix, which is likely to span the hydrophobic domain of the lipid bilayer to anchor the largely hydrophilic subunit b in the membrane. The helical structure is interrupted by a rigid bend in the region of residues 23- 26 with alpha-helical structure resuming at Pro-27 at an angle offset by 20 degrees from the transmembrane helix. In native subunit b, the hinge region and C-terminal alpha-helical segment would connect the transmembrane helix to the cytoplasmic domain. The transmembrane domains of the two subunit b in F0 were shown to be close to each other by cross-linking experiments in which single Cys were substituted for residues 2-21 of the native subunit and b-b dimer formation tested after oxidation with Cu(II)(phenanthroline)2. Cys residues that formed disulfide cross-links were found with a periodicity indicative of one face of an alpha-helix, over the span of residues 2-18, where Cys at positions 2, 6, and 10 formed dimers in highest yield. A model for the dimer is presented based upon the NMR structure and distance constraints from the cross-linking data. The transmembrane alpha- helices are positioned at a 23 degrees angle to each other with the side chains of Thr-6, Gln-10, Phe-14, and Phe-17 at the interface between subunits. The change in direction of helical packing at the hinge region may be important in the functional interaction of the cytoplasmic domains
RP  - NOT IN FILE
NT  - UI - 99269098LA - engRN - 0 (Membrane Proteins)RN - 0 (Peptide Fragments)RN - 0 (Solvents)RN - 52-90-4 (Cysteine)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM23105/GM/NIGMSID - R02301/PHSID - RR02301/RR/NCRRDA - 19990629IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:10336456
SO  - J Biol Chem 1999 May 28 ;274(22):15598-15604

1576
UI  - 10
AU  - Dmitriev OY
AU  - Jones PC
AU  - Fillingame RH
AD  - Department of Biomolecular Chemistry, University of Wisconsin Medical School, Madison, WI 53706, USA
TI  - Structure of the subunit c oligomer in the F1Fo ATP synthase: model derived from solution structure of the monomer and cross-linking in the native enzyme
AB  - The structure of the subunit c oligomer of the H+-transporting ATP synthase of Escherichia coli has been modeled by molecular dynamics and energy minimization calculations from the solution structure of monomeric subunit c and 21 intersubunit distance constraints derived from cross-linking of subunits. Subunit c folds in a hairpin-like structure with two transmembrane helices. In the c12 oligomer model, the subunits pack to form a compact hollow cylinder with an outer diameter of 55-60 A and an inner space with a minimal diameter of 11-12 A. Phospholipids are presumed to pack in the inner space in the native membrane. The transmembrane helices pack in two concentric rings with helix 1 inside and helix 2 outside. The calculations strongly favor this structure versus a model with helix 2 inside and helix 1 outside. Asp-61, the H+-transporting residue, packs toward the center of the four transmembrane helices of two interacting subunits. From this position at the front face of one subunit, the Asp-61 carboxylate lies proximal to side chains of Ala-24, Ile-28, and Ala-62, projecting from the back face of a second subunit. These interactions were predicted from previous mutational analyses. The packing supports the suggestion that a c-c dimer is the functional unit. The positioning of the Asp-61 carboxyl in the center of the interacting transmembrane helices, rather than at the periphery of the cylinder, has important implications regarding possible mechanisms of H+-transport-driven rotation of the c oligomer during ATP synthesis
RP  - NOT IN FILE
NT  - UI - 99324140LA - engRN - 0 (Cross-Linking Reagents)RN - 0 (Macromolecular Systems)RN - 0 (Phospholipids)RN - 0 (Solutions)RN - 56-84-8 (Aspartic Acid)RN - 6898-94-8 (Alanine)RN - 7004-09-3 (Isoleucine)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM23105/GM/NIGMSDA - 19990826IS - 0027-8424SB - IMCY - UNITED STATESJC - PV3
UR  - PM:10393899
SO  - Proc Natl Acad Sci U S A 1999 Jul 6 ;96(14):7785-7790

1577
UI  - 285
AU  - Feniouk BA
AU  - Cherepanov DA
AU  - Junge W
AU  - Mulkidjanian AY
AD  - Division of Biophysics, Faculty of Biology/Chemistry, University of Osnabruck, Germany
TI  - ATP-synthase of Rhodobacter capsulatus: coupling of proton flow through F0 to reactions in F1 under the ATP synthesis and slip conditions
AB  - A stepwise increasing membrane potential was generated in chromatophores of the phototrophic bacterium Rhodobacter capsulatus by illumination with short flashes of light. Proton transfer through ATP- synthase (measured by electrochromic carotenoid bandshift and by pH- indicators) and ATP release (measured by luminescence of luciferin- luciferase) were monitored. The ratio between the amount of protons translocated by F0F1 and the ATP yield decreased with the flash number from an apparent value of 13 after the first flash to about 5 when averaged over three flashes. In the absence of ADP, protons slipped through F0F1. The proton transfer through F0F1 after the first flash contained two kinetic components, of about 6 ms and 20 ms both under the ATP synthesis conditions and under slip. The slower component of proton transfer was substantially suppressed in the absence of ADP. We attribute our observations to the mechanism of energy storage in the ATP-synthase needed to couple the transfer of four protons with the synthesis of one molecule of ATP. Most probably, the transfer of initial protons of each tetrad creates a strain in the enzyme that slows the translocation of the following protons
RP  - NOT IN FILE
NT  - UI - 99192321LA - engRN - 0 (Dyes)RN - 0 (Phosphates)RN - 0 (Protons)RN - 553-24-2 (Neutral Red)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19990420IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:10094498
SO  - FEBS Lett 1999 Feb 26 ;445(2-3):409-414

1578
UI  - 7
AU  - Fillingame RH
AD  - Department of Biomolecular Chemistry, University of Wisconsin Medical School, Madison, WI 53706, USA rhfillin@facstaffwiscedu
TI  - Molecular rotary motors
RP  - NOT IN FILE
NT  - UI - 20064537LA - engRN - 0 (Actins)RN - 0 (Molecular Motors)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - CommentPT - Journal ArticleDA - 19991214IS - 0036-8075SB - IMCY - UNITED STATESJC - UJ7
UR  - PM:10610565
SO  - Science 1999 Nov 26 ;286(5445):1687-1688

1579
UI  - 13
AU  - Fillingame RH
AU  - Divall S
AD  - Department of Biomolecular Chemistry, University of Wisconsin Medical School, Madison 53706, USA
TI  - Proton ATPases in bacteria: comparison to Escherichia coli F1F0 as the prototype
AB  - The F1F0 ATP synthase complex of Escherichia coli functions reversibly in coupling proton translocation to ATP synthesis or hydrolysis. The structural organization and subunit composition corresponds to that seen in many other bacteria, i.e. a membrane extrinsic F1 sector with five subunits in an alpha 3 beta 3 gamma delta epsilon stoichiometry, and a membrane-traversing F0 sector with three subunits in an a1b2c12 stoichiometry. The structure of much of the F1 sector is known from a X- ray diffraction model. During function, The gamma subunit is known to rotate within a hexameric ring of alternating alpha and beta subunits to promote sequential substrate binding and product release from catalytic sites on the three beta subunits. Proton transport through F0 must be coupled to this rotation. Subunit c folds in the membrane as a hairpin to two alpha helices to generate the proton-binding site in F0. Its structure was determined by NMR, and the structure of the c oligomer was deduced by cross-linking experiments and molecular mechanics calculations. The implications of the oligomeric structure of subunit c will be considered and related to the H+/ATP pumping ratio, P/O ratios and the cation-binding site in other types of F0. The possible limits of the structure in changing the ion-binding specificity, stoichiometry and routes of proton entrance/exit to the binding site will be considered
RP  - NOT IN FILE
NT  - UI - 99224329LA - engRN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialID - GM-23105/GM/NIGMSDA - 19990611SB - IMCY - ENGLANDJC - C3Y
UR  - PM:10207922
SO  - Novartis Found Symp 1999  ;221():218-229

1580
UI  - 9886
AU  - Fischer S
AU  - Graber P
TI  - Comparison of Delta pH- and Delta phi-driven ATP synthesis catalyzed by the H+-ATPases from Escherichia coli or chloroplasts reconstituted into liposomes
AB  - The H+-ATPases from Escherichia coli, EF0FI1, and from chloroplasts, CF0F1, were reconstituted in liposomes from phosphatidylcholine/phosphatidic acid. The proteoliposomes mere energized by an acid-base transition and a K+/valinomycin diffusion potential and the initial rate of ATP synthesis was measured as a function of the transmembrane pH difference, Delta pH, and the electric potential difference, Delta phi With EF0F1, a rate of 80 s(-1) is observed at Delta pH = 4.1 and Delta phi approximate to 140 mV, The rate decreases sigmoidally with Delta phi and at Delta phi approximate to 0 mV, the rate is about 1 s(-1) although Delta pH is still 4,1, Under the same conditions with CF0F1, a rate of 280 s(-1) is observed which decreases to 190 s(-1) when Delta phi is abolished, i.e. ATP synthesis catalyzed by EF0F1 and CF0F1 depends in a different way on Delta pH and Delta phi EF0F1-catalyzed ATP synthesis was measured as a function of Delta pH at a constant Delta phi The rate depends sigmoidally on Delta pH reaching a maximal rate which cannot be further increased by increasing Delta pH. However, this maximal rate depends on Delta phi i.e. Delta pH and Delta phi are not kinetically equivalent in driving ATP synthesis. We assume that EF0F1 must be converted into a metastable, active state before it catalyzes proton transport- coupled ATP synthesis. For EF0F1, this activation step depends only on Delta phi whereas for CF0F1, the activation depends on Delta pH and Delta phi (C) 1999 Federation of European Biochemical Societies
MH  - ACTIVATION
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - CF0F1
MH  - chloroplast
MH  - Chloroplasts
MH  - COMPLEX
MH  - DELTA-PH
MH  - DEPENDENCE
MH  - Diffusion
MH  - diffusion potential
MH  - EF0F1
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0F1
MH  - FIELD
MH  - H+-ATPase
MH  - Kinetics
MH  - Liposomes
MH  - Phosphorylation
MH  - proteoliposome
MH  - PSI
MH  - RHODOBACTER-CAPSULATUS
MH  - SYNTHASE
MH  - transport
RP  - NOT IN FILE
NT  - JournalArticleELSEVIER SCIENCE BVSEP 3233YJAMSTERDAMFischer S Univ Freiburg, Inst Chem Phys, Albertstr 23A, D-79104 Freiburg, GermanyFEBS LETTPO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS
AV  - Univ Freiburg, Inst Chem Phys, Albertstr 23A, D-79104 Freiburg, Germany Univ Freiburg, Inst Chem Phys, D-79104 Freiburg, Germany
UR  - ISI:000082454500008
SO  - Febs Letters 1999  ;457(3):327-332

1581
UI  - 20990
AU  - Fischer S
AU  - Graber P
AD  - Institut fur Physikalische Chemie, Universitat Freiburg, Albertstr 23a, D-79104, Freiburg, Germany fischsus@rufuni-freiburgde
TI  - Comparison of DeltapH- and Delta***&phi;***-driven ATP synthesis catalyzed by the H(+)-ATPases from Escherichia coli or chloroplasts reconstituted into liposomes
AB  - The H(+)-ATPases from Escherichia coli, EF(0)F(1), and from chloroplasts, CF(0)F(1), were reconstituted in liposomes from phosphatidylcholine/phosphatidic acid. The proteoliposomes were energized by an acid-base transition and a K(+)/valinomycin diffusion potential and the initial rate of ATP synthesis was measured as a function of the transmembrane pH difference, DeltapH, and the electric potential difference, Delta&phi;. With EF(0)F(1), a rate of 80 s(-1) is observed at DeltapH=4.1 and Delta&phi; approximately 140 mV. The rate decreases sigmoidally with Delta&phi; and at Delta&phi; approximately 0 mV, the rate is about 1 s(-1) although DeltapH is still 4.1. Under the same conditions with CF(0)F(1), a rate of 280 s(-1) is observed which decreases to 190 s(-1) when Delta&phi; is abolished, i.e. ATP synthesis catalyzed by EF(0)F(1) and CF(0)F(1) depends in a different way on DeltapH and Delta&phi;. EF(0)F(1)-catalyzed ATP synthesis was measured as a function of DeltapH at a constant Delta&phi;. The rate depends sigmoidally on DeltapH reaching a maximal rate which cannot be further increased by increasing DeltapH. However, this maximal rate depends on Delta&phi;, i.e. DeltapH and Delta&phi; are not kinetically equivalent in driving ATP synthesis. We assume that EF(0)F(1) must be converted into a metastable, active state before it catalyzes proton transport- coupled ATP synthesis. For EF(0)F(1), this activation step depends only on Delta&phi;, whereas for CF(0)F(1), the activation depends on DeltapH and Delta&phi;
MH  - A
MH  - ACID
MH  - ACTIVATION
MH  - ACTIVE
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthesis
MH  - ATPase
MH  - chloroplast
MH  - Chloroplasts
MH  - CONSTANT
MH  - Diffusion
MH  - diffusion potential
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - function
MH  - H(+)ATPase
MH  - Liposomes
MH  - pH
MH  - Phosphatidic Acids
MH  - Phosphatidylcholines
MH  - Potassium
MH  - Potassium Chloride
MH  - proteoliposome
MH  - proton
MH  - Protons
MH  - synthesis
MH  - transport
RP  - NOT IN FILE
NT  - UI - 99402946LA - engRN - 0 (Liposomes)RN - 0 (Phosphatidic Acids)RN - 0 (Phosphatidylcholines)RN - 0 (Protons)RN - 56-65-5 (Adenosine Triphosphate)RN - 7447-40-7 (Potassium Chloride)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19991012IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:10471802
SO  - FEBS Lett 1999 Sep 3 ;457(3):327-332

1582
UI  - 385
AU  - Galkin MA
AU  - Vinogradov AD
AD  - Department of Biochemistry, School of Biology, Moscow State University, Russia
TI  - Energy-dependent transformation of the catalytic activities of the mitochondrial F0 x F1-ATP synthase
AB  - The ADP(Mg2+)-deactivated, azide-trapped F0 x F1-ATPase of coupled submitochondrial particles is capable of ATP synthesis being incapable of ATP hydrolysis and ATP-dependent delta muH+ generation [FEBS Lett. (1995) 366, 29-32]. This puzzling phenomenon was studied further. No ATPase activity of the submitochondrial particles catalyzing succinate- supported oxidative phosphorylation in the presence of azide was observed when ATP was added to the assay mixture after an uncoupler. Rapid ATP hydrolysis was detected in the same system when ATP followed by an uncoupler was added. Less than 5% of the original ATPase activity was seen when the reaction (assayed with ATP-regenerating system) was initiated by the addition of ATP to the azide-trapped coupled particles oxidizing succinate either in the presence or in the absence of the uncoupler. High ATP hydrolytic activity was revealed when the reaction was started by the simultaneous addition of the ATP plus uncoupler to the particles generating delta muH+. The energy-dependent conversion of the enzyme into latent uncoupler-activated ATPase was prevented by free ADP (Ki approximately 20 microM) and was greatly enhanced after multiple turnovers in oxidative phosphorylation. The results suggest that the catalytic properties of F0 x F1 are delta muH+-dependent which is in accord with our hypothesis on different conformational states of the enzyme participating in ATP synthesis or hydrolysis
RP  - NOT IN FILE
NT  - UI - 99231775LA - engRN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19990601IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:10217423
SO  - FEBS Lett 1999 Apr 1 ;448(1):123-126

1583
UI  - 6805
AU  - Gopta OA
AU  - Cherepanov DA
AU  - Junge W
AU  - Mulkidjanian AY
AD  - Division of Biophysics, University of Osnabruck, D-49069 Osnabruck, Germany
TI  - Proton transfer from the bulk to the bound ubiquinone QB of the reaction center in chromatophores of Rhodobacter sphaeroides: retarded conveyance by neutral water
AB  - The mechanism of proton transfer from the bulk into the membrane protein interior was studied. The light-induced reduction of a bound ubiquinone molecule Q(B) by the photosynthetic reaction center is accompanied by proton trapping. We used kinetic spectroscopy to measure (i) the electron transfer to Q(B) (at 450 nm), (ii) the electrogenic proton delivery from the surface to the Q(B) site (by electrochromic carotenoid response at 524 nm), and (iii) the disappearance of protons from the bulk solution (by pH indicators). The electron transfer to Q(B)(-) and the proton-related electrogenesis proceeded with the same time constant of approximately 100 microseconds (at pH 6.2), whereas the alkalinization in the bulk was distinctly delayed (tau approximately 400 microseconds). We investigated the latter reaction as a function of the pH indicator concentration, the added pH buffers, and the temperature. The results led us to the following conclusions: (i) proton transfer from the surface-located acidic groups into the Q(B) site followed the reduction of Q(B) without measurable delay; (ii) the reprotonation of these surface groups by pH indicators and hydronium ions was impeded, supposedly, because of their slow diffusion in the surface water layer; and (iii) as a result, the protons were slowly donated by neutral water to refill the proton vacancies at the surface. It is conceivable that the same mechanism accounts for the delayed relaxation of the surface pH changes into the bulk observed previously with bacteriorhodopsin membranes and thylakoids. Concerning the coupling between proton pumps in bioenergetic membranes, our results imply a tendency for the transient confinement of protons at the membrane surface
MH  - A
MH  - Bacteriorhodopsin
MH  - Biophysics
MH  - buffer
MH  - Buffers
MH  - carotenoid
MH  - chromatophore
MH  - chromatophores
MH  - CONSTANT
MH  - coupling
MH  - Diffusion
MH  - electrogenic
MH  - electron
MH  - electron transfer
MH  - Electron-transfer
MH  - function
MH  - indicator
MH  - ion
MH  - Ions
MH  - mechanism
MH  - membrane
MH  - Membranes
MH  - microsecond
MH  - pH
MH  - pH-indicator
MH  - protein
MH  - proton
MH  - Proton Pump
MH  - Proton transfer
MH  - Proton-transfer
MH  - Protons
MH  - reaction center
MH  - relaxation
MH  - Rhodobacter
MH  - Rhodobacter sphaeroides
MH  - Site
MH  - spectroscopy
MH  - sphaeroides
MH  - SURFACE
MH  - Temperature
MH  - thylakoid
MH  - thylakoids
MH  - Time
MH  - TRANSFER
MH  - trapping
MH  - Ubiquinone
MH  - Water
RP  - NOT IN FILE
NT  - UI - 20027517LA - engRN - 0 (Protons)RN - 1339-63-5 (Ubiquinone)RN - 7732-18-5 (Water)PT - Journal ArticleDA - 19991213IS - 0027-8424SB - IMCY - UNITED STATESJC - PV3
UR  - PM:10557290
SO  - Proc Natl Acad Sci U S A 1999 Nov 9 ;96(23):13159-13164

1584
UI  - 9946
AU  - Groth G
AU  - Schirwitz K
AD  - Heinrich-Heine-Universitat Dusseldorf, Biochemie der Pflanzen, Germany georggroth@uni-duesseldorfde
TI  - Rapid purification of membrane extrinsic F1-domain of chloroplast ATP synthase in monodisperse form suitable for 3D-crystallization
AB  - A new chromatographic procedure for purification of the membrane extrinsic F1-domain of chloroplast ATP synthase is presented. The purification is achieved by a single anion exchange chromatography step. Determination of the enzyme-bound nucleotides reveals only 1 mole of ADP per complex. The purified enzyme shows a latent Ca(2+)-dependent ATPase activity of 1.0 mumol.mg-1 min-1 and a Mg(2+)-dependent activity of 4.4 mumol.mg-1 .min-1. Both activities are increased up to 8-10-fold after dithiothreitol activation. Analysis of the purified F1-complex by SDS/PAGE, silver staining and immunoblotting revealed that the preparation is uncontaminated by fragmented subunits or ribulose-1,5- bisphosphate carboxylase/oxygenase. Gel filtration experiments indicate that the preparation is homogenous and monodisperse. In order to determine the solubility minimum of the purified F1-complex the isoelectric point of the preparation was calculated from pH mapping on ion exchange columns. In agreement with calculations based on the amino acid sequence, a slightly acidic pI of 5.7 was found. Using ammonium sulphate as a precipitant the purified CF1-complex could be crystallized by MicroBatch
MH  - ACTIVATION
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - ADP
MH  - Amino Acid Sequence
MH  - analysis
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Calcium
MH  - chloroplast
MH  - COMPLEX
MH  - Dithiothreitol
MH  - H(+)-Transporting ATP Synthase
MH  - Magnesium
MH  - Membrane Proteins
MH  - Nucleotides
MH  - Proteins
MH  - purification
MH  - Solubility
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 99191679LA - engRN - 0 (Membrane Proteins)RN - 0 (Nucleotides)RN - 0 (Plant Proteins)RN - 3483-12-3 (Dithiothreitol)RN - 58-64-0 (Adenosine Diphosphate)RN - 7439-95-4 (Magnesium)RN - 7440-70-2 (Calcium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19990422IS - 0014-2956SB - IMCY - GERMANYJC - EMZ
UR  - PM:10091579
SO  - Eur J Biochem 1999 Feb ;260(1):15-21

1585
UI  - 281
AU  - Hasler K
AU  - Panke O
AU  - Junge W
AD  - Department of Biology/Chemistry, Division of Biophysics, Universitat Osnabruck, Germany
TI  - On the stator of rotary ATP synthase: the binding strength of subunit delta to (alpha beta)3 as determined by fluorescence correlation spectroscopy
AB  - ATP synthase is conceived as a rotary enzyme. Proton flow drives the rotor (namely, subunits c12 epsilon gamma) relative to the stator (namely, subunits ab2 delta(alpha beta)3) and extrudes spontaneously formed ATP from three symmetrically arranged binding sites on (alpha beta)3 into the solution. We asked whether the binding of subunit delta to (alpha beta)3 is of sufficient strength to hold against the elastic strain, which is generated during the operation of this enzyme. According to current estimates, the elastically stored energy is about 50 kJ/mol. Subunit delta was specifically labeled without impairing its function. Its association with solubilized (alpha beta)3 gamma in detergent-free buffer was studied by fluorescence correlation spectroscopy (FCS). A very strong tendency of delta to dimerize in detergent-free buffer was apparent (K(d) </= 0.2 nM). Taking the upper limit of this figure into account, the dissociation constant between monomeric delta and (alpha beta)3 gamma was 0.8 nM if not smaller. It is equivalent to a free energy of binding of at least 52 kJ/mol and therewith is sufficient for the assumed hold-function of delta in the stator. Our data were compatible with a single binding site for delta on the hexagon of (alpha beta)3
RP  - NOT IN FILE
NT  - UI - 99452717LA - engRN - 0 (Detergents)RN - 0 (Fatty Acids)RN - 0 (Fluorescent Dyes)RN - 0 (Peptide Fragments)RN - 0 (Rhodamines)RN - 0 (Solutions)RN - 3416-24-8 (Glucosamine)RN - 62669-72-1 (tetramethylrhodamine)RN - 85261-19-4 (nonanoyl-N-methylglucamide)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19991117IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:10521283
SO  - Biochemistry 1999 Oct 12 ;38(41):13759-13765

1586
UI  - 138
AU  - Hausrath AC
AU  - Gruber G
AU  - Matthews BW
AU  - Capaldi RA
AD  - Institute of Molecular Biology, Howard Hughes Medical Institute, Department of Physics, 1229 University of Oregon, Eugene, OR 97403- 1229, USA
TI  - Structural features of the gamma subunit of the Escherichia coli F(1) ATPase revealed by a 4.4-A resolution map obtained by x-ray crystallography
AB  - The F(1) part of the F(1)F(O) ATP synthase from Escherichia coli has been crystallized and its structure determined to 4.4-A resolution by using molecular replacement based on the structure of the beef-heart mitochondrial enzyme. The bacterial F(1) consists of five subunits with stoichiometry alpha(3), beta(3), gamma, delta, and epsilon. delta was removed before crystallization. In agreement with the structure of the beef-heart mitochondrial enzyme, although not that from rat liver, the present study suggests that the alpha and beta subunits are arranged in a hexagonal barrel but depart from exact 3-fold symmetry. In the structures of both beef heart and rat-liver mitochondrial F(1), less than half of the structure of the gamma subunit was seen because of presumed disorder in the crystals. The present electron-density map includes a number of rod-shaped features which appear to correspond to additional alpha-helical regions within the gamma subunit. These suggest that the gamma subunit traverses the full length of the stalk that links the F(1) and F(O) parts and makes significant contacts with the c subunit ring of F(O)
RP  - NOT IN FILE
NT  - UI - 20040613LA - engRN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM20066/GM/NIGMSID - HL24526/HL/NHLBIDA - 20000106IS - 0027-8424SB - IMCY - UNITED STATESJC - PV3
UR  - PM:10570135
SO  - Proc Natl Acad Sci U S A 1999 Nov 23 ;96(24):13697-13702

1587
UI  - 21240
AU  - Heinz WF
AU  - Hoh JH
AD  - Department of Physiology, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205 USA
TI  - Relative surface charge density mapping with the atomic force microscope
AB  - An experimental approach for producing relative charge density maps of biological surfaces using the atomic force microscope is presented. This approach, called D minus D (D-D) mapping, uses isoforce surfaces collected at different salt concentrations to remove topography and isolate electrostatic contributions to the tip-sample interaction force. This approach is quantitative for surface potentials below 25 mV, and does not require prior knowledge of the cantilever spring constant, tip radius, or tip charge. In addition, D-D mapping does not require tip-sample contact. The performance of D-D mapping is demonstrated on surfaces of constant charge and varying topography (mechanically roughened mica and stacked bilayers of dipalmitolphosphatidylserine), a surface of varying charge and varying topography (patches of dipalmitolphosphatidylcholine on mica), and bacteriorhopsin membranes adsorbed to mica
MH  - A
MH  - Bacteriorhodopsin
MH  - CONSTANT
MH  - Lipid Bilayers
MH  - membrane
MH  - Membranes
MH  - physiology
MH  - Polystyrenes
MH  - SURFACE
RP  - NOT IN FILE
NT  - UI - 99093415LA - engRN - 0 (Aluminum Silicates)RN - 0 (Lipid Bilayers)RN - 0 (Phosphatidylserines)RN - 0 (Polystyrenes)RN - 12001-26-2 (mica)RN - 2644-64-6 (1,2-Dipalmitoylphosphatidylcholine)RN - 3036-82-6 (dipalmitoylphosphatidylserine)RN - 53026-44-1 (Bacteriorhodopsins)PT - Journal ArticleID - T32 GM-08043/GM/NIGMSDA - 19990222IS - 0006-3495SB - IMCY - UNITED STATES
UR  - PM:9876166
SO  - Biophys J 1999 Jan ;76(1 Pt 1):528-538

1588
UI  - 11
AU  - Hermolin J
AU  - Dmitriev OY
AU  - Zhang Y
AU  - Fillingame RH
AD  - Department of Biomolecular Chemistry, University of Wisconsin Medical School, Madison, Wisconsin 53706, USA
TI  - Defining the domain of binding of F1 subunit epsilon with the polar loop of F0 subunit c in the Escherichia coli ATP synthase
AB  - We have previously shown that the E31C-substituted epsilon subunit of F1 can be cross-linked by disulfide bond formation to the Q42C- substituted c subunit of F0 in the Escherichia coli F1F0-ATP synthase complex (Zhang, Y., and Fillingame, R. H. (1995) J. Biol. Chem. 270, 24609-24614). The interactions of subunits epsilon and c are thought to be central to the coupling of H+ transport through F0 to ATP synthesis in F1. To further define the domains of interaction, we have introduced additional Cys into subunit epsilon and subunit c and tested for cross- link formation following sulfhydryl oxidation. The results show that Cys, in a continuous stretch of residues 26-33 in subunit epsilon, can be cross-linked to Cys at positions 40, 42, and 44 in the polar loop region of subunit c. The results are interpreted, and the subunit interaction is modeled using the NMR and x-ray diffraction structures of the monomeric subunits together with information on the packing arrangement of subunit c in a ring of 12 subunits. In the model, residues 26-33 form a turn of antiparallel beta-sheet which packs between the polar loop regions of adjacent subunit c at the cytoplasmic surface of the c12 oligomer
RP  - NOT IN FILE
NT  - UI - 99287899LA - engRN - 0 (ATPase inhibitory protein)RN - 0 (Cross-Linking Reagents)RN - 0 (Disulfides)RN - 0 (Proteins)RN - 52-90-4 (Cysteine)RN - EC 3.6.1.- (ceroid lipofuscinosis protein)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM23105/GM/NIGMSDA - 19990706IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:10358051
SO  - J Biol Chem 1999 Jun 11 ;274(24):17011-17016

1589
UI  - 694
AU  - Hisabori T
AU  - Kondoh A
AU  - Yoshida M
AD  - Research Laboratory of Resources Utilization, Tokyo Institute of Technology, Nagatsuta 4259, Midori-ku, Yokohama, Japan thisabor@restitechacjp
TI  - The gamma subunit in chloroplast F(1)-ATPase can rotate in a unidirectional and counter-clockwise manner
AB  - Rotation of the gamma subunit in chloroplast F(1)-ATPase (CF(1)) was investigated by using a single molecule observation technique, which is developed by Noji et al. to observe the rotation of a central gamma subunit portion in the alpha(3)beta(3)gamma sub-complex of F(1)-ATPase from thermophilic Bacillus PS3 (TF(1)) during ATP hydrolysis [Noji, H. et al. (1997) Nature 386, 299-302]. We used two cysteines of the gamma subunit (Cys-199 and Cys-205) of CF(1)-ATPase, which are involved in the regulation of this enzyme, to fix the fluorochrome-labeled actin filament. Then we successfully observed a unidirectional, counter- clockwise rotation of the actin filament with the fluorescent microscope indicating the rotation of the gamma subunit in CF(1)- ATPase. We conclude that the rotation of the gamma subunit in the F(1)- motor is a ubiquitous phenomenon in all F(1)-ATPases in prokaryotes as well as in eukaryotes
RP  - NOT IN FILE
NT  - UI - 20069441LA - engRN - 0 (Actins)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 20000203IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:10601633
SO  - FEBS Lett 1999 Dec 10 ;463(1-2):35-38

1590
UI  - 981
AU  - Jungas C
AU  - Ranck JL
AU  - Rigaud JL
AU  - Joliot P
AU  - Vermeglio A
TI  - Supramolecular organization of the photosynthetic apparatus of Rhodobacter sphaeroides
AB  - Native tubular membranes were purified from the purple non-sulfur bacterium Rhodobacter sphaeroides. These tubular structures contain all the membrane components of the photosynthetic apparatus, in the relative ratio of one cytochrome bc1 complex to two reaction centers, and approximately 24 bacteriochlorophyll molecules per reaction center. Electron micrographs of negative-stained membranes diffract up to 25 A and allow the calculation of a projection map at 20 A. The unit cell (a = 198 A, b = 120 A and gamma = 103 degrees) contains an elongated S-shaped supercomplex presenting a pseudo-2-fold symmetry. Comparison with density maps of isolated reaction center and light-harvesting complexes allowed interpretation of the projection map. Each supercomplex is composed of light-harvesting 1 complexes that take the form of two C-shaped structures of approximately 112 A in external diameter, facing each other on the open side and enclosing the two reaction centers. The remaining positive density is tentatively attributed to one cytochrome bc1 complex. These features shed new light on the association of the reaction center and the light-harvesting complexes. In particular, the organization of the light-harvesting complexes in C-shaped structures ensures an efficient exchange of ubihydroquinone/ubiquinone between the reaction center and the cytochrome bc1 complex
MH  - Bacteria
MH  - Chemistry
MH  - Intracellular Membranes
MH  - Light
MH  - Macromolecular Systems
MH  - Membranes
MH  - metabolism
MH  - Microscopy,Electron
MH  - Models,Molecular
MH  - Photosynthesis
MH  - Photosynthetic Reaction Center,Bacterial
MH  - Rhodobacter sphaeroides
MH  - Ubiquinol-Cytochrome-c Reductase
MH  - ultrastructure
MH  - X-Ray Diffraction
RP  - NOT IN FILE
NT  - CEA/Cadarache-DSV-DEVM Laboratoire de Bioenergetique Cellulaire, 13108 St Paul-lez-Durance Cedex
SO  - EMBO J 1999 Feb 1 ;18(3):534-542

1591
UI  - 284
AU  - Junge W
AD  - Abteilung Biophysik, Fachbereich Biologie/Chemie, Universitat Osnabruck, D-49069 Germany junge@uosde
TI  - ATP synthase and other motor proteins
RP  - NOT IN FILE
NT  - UI - 99238422LA - engRN - 0 (Bacterial Proteins)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - CommentPT - Journal ArticleDA - 19990610IS - 0027-8424SB - IMCY - UNITED STATESJC - PV3
UR  - PM:10220358
SO  - Proc Natl Acad Sci U S A 1999 Apr 27 ;96(9):4735-4737

1592
UI  - 18
AU  - Kaim G
AU  - Dimroth P
AD  - Mikrobiologisches Institut, Eidgenossische Technische Hochschule, ETH- Zentrum, Schmelzbergstrasse 7, CH-8092 Zurich, Switzerland
TI  - ATP synthesis by F-type ATP synthase is obligatorily dependent on the transmembrane voltage
AB  - ATP synthase is the universal enzyme that manufactures cellular ATP using the energy stored in a transmembrane ion gradient. This energy gradient has two components: the concentration difference (DeltapH or DeltapNa(+)) and the electrical potential difference DeltaPsi, which are thermodynamically equivalent. However, they are not kinetically equivalent, as the mitochondrial and bacterial ATP synthases require a transmembrane potential, DeltaPsi, but the chloroplast enzyme has appeared to operate on DeltapH alone. Here we show that, contrary to the accepted wisdom, the 'acid bath' procedure used to study the chloroplast enzyme develops not only a DeltapH but also a membrane potential, and that this potential is essential for ATP synthesis. Thus, for the chloroplast and other ATP synthases, the membrane potential is the fundamental driving force for their normal operation. We discuss the biochemical reasons for this phenomenon and a model that is consistent with these new experimental facts
RP  - NOT IN FILE
NT  - UI - 99359340LA - engRN - 0 (Carboxylic Acids)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19990916IS - 0261-4189SB - IMCY - ENGLANDJC - EMB
UR  - PM:10428951
SO  - EMBO J 1999 Aug 2 ;18(15):4118-4127

1593
UI  - 21033
AU  - Kakinuma Y
AU  - Yamato I
AU  - Murata T
AD  - Faculty of Pharmaceutical Sciences, Chiba University, Japan
TI  - Structure and function of vacuolar Na+-translocating ATPase in Enterococcus hirae
AB  - A Na+-translocating ATPase was discovered in a gram-positive bacterium Enterococcus hirae. Our biochemical and molecular biological studies revealed that this Na+-ATPase belongs to the vacuolar-type enzyme. Purified Na+-ATPase consisted of nine subunits: NtpA, B, C, D, E, F, G, I, and K; reconstituted proteoliposomes showed ATP-driven electrogenic Na+ translocation. All these subunits were encoded by the ntp operon: ntpFIKECGABDHJ. The deduced amino acid sequences of the major subunits, A, B, and K (16 kDa proteolipid), were highly similar to those of A, B, and proteolipid subunits of vacuolar ATPases, although the similarities of other subunits were moderate. The ntpJ gene encoded a K+ transporter independent of the Na+-ATPase. Expression of this operon, encoding two transport systems for Na+ and K+ ions, was regulated at transcriptional level by intracellular Na+ as the signal. Two related cation pumps, vacuolar Na+-ATPase and F0F1, H+-ATPase, coexist in this bacterium
MH  - A
MH  - ACID
MH  - Adenosinetriphosphatase
MH  - Amino Acid Sequence
MH  - ATPase
MH  - Bacteria
MH  - Bacterial Proteins
MH  - electrogenic
MH  - F0F1
MH  - function
MH  - H+-ATPase
MH  - ion
MH  - Ions
MH  - membrane
MH  - Membrane Proteins
MH  - protein
MH  - Proteins
MH  - proteoliposome
MH  - review
MH  - structure
MH  - SUBUNIT
MH  - SYSTEM
MH  - SYSTEMS
MH  - translocation
MH  - transport
RP  - NOT IN FILE
NT  - UI - 99270692LA - engRN - 0 (Bacterial Proteins)RN - 0 (Membrane Proteins)RN - 0 (Sodium-Hydrogen Antiporter)RN - EC 3.6.1.- (NtpJ protein)RN - EC 3.6.1.- (sodium-translocating ATPase)RN - EC 3.6.1.- (vacuolar H+-ATPase)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 19990723IS - 0145-479XSB - IMCY - UNITED STATES
UR  - PM:10340844
SO  - J Bioenerg Biomembr 1999 Feb ;31(1):7-14

1594
UI  - 21177
AU  - Kalaidzidis IV
AU  - Belevich IN
AU  - Kalaidzidis YL
AU  - Kaulen AD
AD  - Department of Photobiochemistry, AN Belozersky Institute of Physico- Chemical Biology, Moscow State University, 119899, Moscow, Russia
TI  - Membrane potential stabilizes the O intermediate in liposomes containing bacteriorhodopsin
AB  - In the bacteriorhodopsin-containing proteoliposomes, a laser flash is found to induce formation of a bathointermediate decaying in several seconds, the difference spectrum being similar to the purple-blue transition. Different pH buffers do not affect the intermediate, whereas an uncoupler, gramicidin A, and lipophilic ions accelerate decay of the intermediate or inhibit its formation. In the liposomes containing E204Q bacteriorhodopsin mutant, formation of the intermediate is suppressed. In the wild-type bacteriorhodopsin liposomes, the bathointermediate formation is pH-independent within the pH 5-7 range. The efficiency of the long-lived O intermediate formation increases at a low pH. In the wild-type as well as in the E204Q mutant purple membrane, the O intermediate decay is slowed down at slightly higher pH values than that of the purple-blue transition. It is suggested that the membrane potential affects the equilibrium between the bacteriorhodopsin ground state (Glu-204 is protonated and Asp-85 is deprotonated) and the O intermediate (Asp-85 is protonated and Glu-204 is deprotonated), stabilizing the latter by changing the relative affinity of Asp-85 and Glu-204 to H(+). At a low pH, protonation of a proton-releasing group (possibly Glu-194) in the bacteriorhodopsin ground state seems to prevent deprotonation of the Glu-204 during the photocycle. Thus, all protonatable residues of the outward proton pathway should be protonated in the O intermediate. Under such conditions, membrane potential stabilization of the O intermediate in the liposomes can be attributed to the direct effect of the potential on the pK value of Asp-85
MH  - A
MH  - Asp-85
MH  - Bacteriorhodopsin
MH  - buffer
MH  - Buffers
MH  - flash
MH  - intermediate
MH  - ion
MH  - Ions
MH  - liposome
MH  - Liposomes
MH  - membrane
MH  - Membrane Potential
MH  - mutant
MH  - pH
MH  - proteoliposome
MH  - proton
MH  - Proton Pump
MH  - protonation
MH  - purple membrane
MH  - RESIDUE
MH  - spectra
RP  - NOT IN FILE
NT  - UI - 99439727LA - engRN - 0 (Liposomes)RN - 0 (Proton Pumps)RN - 53026-44-1 (Bacteriorhodopsins)PT - Journal ArticleDA - 19991101IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:10508934
SO  - FEBS Lett 1999 Oct 1 ;459(1):143-147

1595
UI  - 324
AU  - Karrasch S
AU  - Walker JE
AD  - Laboratory of Molecular Biology, MRC, Hills Road, Cambridge, CB2 2QH, United Kingdom karrsch@rufuni-freiburgde
TI  - Novel features in the structure of bovine ATP synthase
AB  - The F1F0-ATP synthase from bovine heart mitochondria catalyses the synthesis of ATP from ADP and inorganic phosphate by using the energy of an electrochemical proton gradient derived from electron transport. The enzyme consists of three major domains: the globular F1catalytic domain of known atomic structure lies outside the lipid bilayer and is attached by a central stalk to the intrinsic membrane domain, F0, which transports protons through the membrane. Proton transport through F0evokes structural changes that are probably transmitted by rotation of the stalk to the catalytic sites in F1. In an alpha3beta3gamma1subcomplex, the rotation of the central gamma subunit driven by ATP hydrolysis has been visualised by optical microscopy. In order to prevent the alpha3beta3structure from following the rotation of the central gamma subunit, it has been proposed that the enzyme might have a stator connecting static parts in F0to alpha3beta3,thereby keeping it fixed relative to the rotating parts. Here we present electron microscopy images that reveal three new features in bovine F1F0-ATPase, one of which could be a stator. The second feature is a collar structure above the membrane domain and the third feature is some additional density on top of the F1domain
RP  - NOT IN FILE
NT  - UI - 99321980LA - engRN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19990730IS - 0022-2836SB - IMCY - ENGLANDJC - J6V
UR  - PM:10390338
SO  - J Mol Biol 1999 Jul 9 ;290(2):379-384

1596
UI  - 695
AU  - Kato-Yamada Y
AU  - Bald D
AU  - Koike M
AU  - Motohashi K
AU  - Hisabori T
AU  - Yoshida M
AD  - Research Laboratory of Resources Utilization, Tokyo Institute of Technology, 4259 Nagatsuta, Yokohama, 226-8503, Japan
TI  - Epsilon subunit, an endogenous inhibitor of bacterial F(1)-ATPase, also inhibits F(0)F(1)-ATPase
AB  - Since the report by Sternweis and Smith (Sternweis, P. C., and Smith, J. B. (1980) Biochemistry 19, 526-531), the epsilon subunit, an endogenous inhibitor of bacterial F(1)-ATPase, has long been thought not to inhibit activity of the holo-enzyme, F(0)F(1)-ATPase. However, we report here that the epsilon subunit is exerting inhibition in F(0)F(1)-ATPase. We prepared a C-terminal half-truncated epsilon subunit (epsilon(DeltaC)) of the thermophilic Bacillus PS3 F(0)F(1)- ATPase and reconstituted F(1)- and F(0)F(1)-ATPase containing epsilon(DeltaC). Compared with F(1)- and F(0)F(1)-ATPase containing intact epsilon, those containing epsilon(DeltaC) showed uninhibited activity; severalfold higher rate of ATP hydrolysis at low ATP concentration and the start of ATP hydrolysis without an initial lag at high ATP concentration. The F(0)F(1)-ATPase containing epsilon(DeltaC) was capable of ATP-driven H(+) pumping. The time-course of pumping at low ATP concentration was faster than that by the F(0)F(1)-ATPase containing intact epsilon. Thus, the comparison with noninhibitory epsilon(DeltaC) mutant shed light on the inhibitory role of the intact epsilon subunit in F(0)F(1)-ATPase
RP  - NOT IN FILE
NT  - UI - 20036533LA - engRN - 0 (ATPase inhibitory protein)RN - 0 (Bacterial Proteins)RN - 0 (Liposomes)RN - 0 (Proteins)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19991229IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:10567363
SO  - J Biol Chem 1999 Nov 26 ;274(48):33991-33994

1597
UI  - 499
AU  - Ko YH
AU  - Hong S
AU  - Pedersen PL
AD  - Department of Biological Chemistry, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205-2185, USA
TI  - Chemical mechanism of ATP synthase. Magnesium plays a pivotal role in formation of the transition state where ATP is synthesized from ADP and inorganic phosphate
AB  - The chemical mechanism by which ATP synthases catalyze the synthesis of ATP remains unknown despite the recent elucidation of the three- dimensional structures of two forms of the F(1) catalytic sector (subunit stoichiometry, alpha(3)beta(3)gammadeltaepsilon). Lacking is critical information about the chemical events taking place at the catalytic site of each beta-subunit in the transition state. In an earlier report (Ko, Y. H., Bianchet, M. A., Amzel, L.M., and Pedersen, P. L. (1997) J. Biol. Chem. 272, 18875-18881), we provided evidence for transition state formation in the presence of Mg(2+), ADP, and orthovanadate (V(i)), a photoreactive phosphate analog with a trigonal bipyramidal geometry resembling that of the gamma-P of ATP in the transition state of enzymes like myosin. In the presence of ultraviolet light and O(2,) the MgADP.V(i)-F(1) complex was cleaved within the P- loop (GGAGVGKT) of a single beta-subunit at alanine 158, implicating this residue as within contact distance of the gamma-P of ATP in the transition state. Here, we report that ADP, although facilitating transition state formation, is not essential. In the presence of Mg(2+) and V(i) alone the catalytic activity of the resultant MgV(i)-F(1) complex is inhibited to nearly the same extent as that observed for the MgADP. V(i)-F(1) complex. Inhibition is not observed with ADP, Mg(2+), or V(i) alone. Significantly, in the presence of ultraviolet light and O(2,) the MgV(i)-F(1) complex is cleaved also within the P-loop of a single beta-subunit at alanine 158 as confirmed by Western blot analyses with two different antibodies, by N-terminal sequence analyses, and by quantification of the amount of unreacted beta- subunits. These novel findings indicate that Mg(2+) plays a pivotal role in transition state formation during ATP synthesis catalyzed by ATP synthases, a role that involves both its preferential coordination with P(i) and the repositioning of the P-loop to bring the nonpolar alanine 158 into the catalytic pocket. A reaction scheme for ATP synthases depicting a role for Mg(2+) in transition state formation is proposed here for the first time
RP  - NOT IN FILE
NT  - UI - 99436076LA - engRN - 0 (Multienzyme Complexes)RN - 0 (Peptide Fragments)RN - 0 (Phosphates)RN - 0 (Vanadates)RN - 58-64-0 (Adenosine Diphosphate)RN - 7439-95-4 (Magnesium)RN - 7782-44-7 (Oxygen)RN - EC 2.7.4 (Phosphotransferases (Phosphate Group Acceptor))RN - EC 2.7.4.- (ATP synthetase complex)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - CA 10951/CA/NCIDA - 19991109IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:10506126
SO  - J Biol Chem 1999 Oct 8 ;274(41):28853-28856

1598
UI  - 21153
AU  - Kramer DM
AU  - Sacksteder CA
AU  - Cruz JA
TI  - How acidic is the lumen?
AB  - Proton motive force (pmf), established across the thylakoid membrane by photosynthetic electron transfer, functions both to drive the synthesis of ATP and initiate processes that down-regulate photosynthesis. At the same time, excessively low lumen pH can lead to the destruction of some lumenal components and sensitization of the photosynthetic apparatus to photoinhibition. Therefore, in order to understand the energy budget of photosynthesis, its regulation and responses to environmental stresses, it is essential to know the magnitude of pmf, its distribution between "pH and the electric field ("φv;) as well as the relationships between these parameters and "GATP, and down-regulatory and inhibitory processes. We review past estimates of lumen pH and propose a model that can explain much of the divergent data in the literature. In this model, in intact plants under permissive conditions, photosynthesis is regulated so that lumen pH remains moderate (between 5.8 and 6.5), where it modulates the activity of the violaxanthin deepoxidase, does not significantly restrict the turnover of the cytochrome b6f complex, and does not destabilize the oxygen evolving complex. Only under stressed conditions, where light input exceeds the capacity of both photosynthesis and down-regulatory processes, does lumen pH decrease below 5, possibly contributing to photoinhibition. A value of n = 4 for the stoichiometry of protons pumped through the ATP synthase per ATP synthesized, and a minor contribution of "φv; to pmf, will allow moderate lumen pH to sustain the observed levels of "GATP.
MH  - proton
MH  - pmf
MH  - thylakoid
MH  - thylakoid membrane
MH  - membrane
MH  - electron
MH  - electron transfer
MH  - Electron-transfer
MH  - TRANSFER
MH  - function
MH  - synthesis
MH  - atp
MH  - Photosynthesis
MH  - Time
MH  - pH
MH  - regulation
MH  - FIELD
MH  - review
MH  - A
MH  - model
MH  - Plants
MH  - plant
MH  - cytochrome
MH  - b6f
MH  - COMPLEX
MH  - Oxygen
MH  - Light
MH  - Protons
MH  - ATP synthase
MH  - SYNTHASE
RP  - NOT IN FILE
SO  - Photosynth Research 1999  ;60():151-163

1599
UI  - 21073
AU  - Krulwich TA
AU  - Guffanti AA
AU  - Ito M
AD  - Department of Biochemistry, Mount Sinai School of Medicine, New York, NY 10029, USA
TI  - pH tolerance in Bacillus: alkaliphiles versus non-alkaliphiles
AB  - Monovalent cation/proton antiporters that catalyse electrogenic uptake of H+ in exchange for cytoplasmic K+ and/or Na+ are centrally involved in bacterial pH homeostasis under alkaline challenge. Systematic attempts have identified some, but not yet all, of the genes encoding such antiporters that participate in pH homeostasis in the neutrophilic Bacillus subtilis and the extremely alkaliphilic Bacillus firmus OF4. In each organism there are at least three distinct antiporters involved in pH homeostasis. They differ in cation requirement, with pH homeostasis specifically utilizing Na+/H+ antiport in the alkaliphile and using either Na+ or K+/H+ antiport in B. subtilis. Some of the antiporters involved in pH homeostasis are constitutive and are in place to respond to sudden pH shifts, but there is also an inducible component. At least two sets of homologous antiporters (NhaC and Mrp/Pha) function in both alkaliphiles and neutrophiles. An additional antiporter of a different transport protein family, the Gram-positive tetracycline-metal/H+ antiporter, is important in pH homeostasis in B. subtilis but has not yet been shown to be present in any alkaliphile. There are also differences outside of the antiporters themselves that contribute to the greater capacity of the alkaliphiles for pH homeostasis, including cation re-entry capacity and possible surface properties
MH  - A
MH  - Alkalies
MH  - Bacillus
MH  - Biochemistry
MH  - electrogenic
MH  - England
MH  - function
MH  - H+
MH  - Homeostasis
MH  - pH
MH  - protein
MH  - SURFACE
MH  - Surface Properties
MH  - transport
RP  - NOT IN FILE
NT  - UI - 99224326LA - engRN - 0 (Alkalies)RN - 0 (Antiporters)PT - Journal ArticleID - GM28454/GM/NIGMSID - GM52837/GM/NIGMSDA - 19990611SB - IMCY - ENGLAND
UR  - PM:10207919
SO  - Novartis Found Symp 1999  ;221():167-179

1600
UI  - 21244
AU  - Lancaster CR
AU  - Kroger A
AU  - Auer M
AU  - Michel H
AD  - Max-Planck-Institut fur Biophysik, Frankfurt am Main, Germany lancaster@mpibp-frankfurtmpgde
TI  - Structure of fumarate reductase from Wolinella succinogenes at 2.2 A resolution
AB  - Fumarate reductase couples the reduction of fumarate to succinate to the oxidation of quinol to quinone, in a reaction opposite to that catalysed by the related complex II of the respiratory chain (succinate dehydrogenase). Here we describe the crystal structure at 2.2 A resolution of the three protein subunits containing fumarate reductase from the anaerobic bacterium Wolinella succinogenes. Subunit A contains the site of fumarate reduction and a covalently bound flavin adenine dinucleotide prosthetic group. Subunit B contains three iron-sulphur centres. The menaquinol-oxidizing subunit C consists of five membrane- spanning, primarily helical segments and binds two haem b molecules. On the basis of the structure, we propose a pathway of electron transfer from the dihaem cytochrome b to the site of fumarate reduction and a mechanism of fumarate reduction. The relative orientations of the soluble and membrane-embedded subunits of succinate:quinone oxidoreductases appear to be unique
MH  - A
MH  - ACID
MH  - Bacteria
MH  - COMPLEX
MH  - cytochrome
MH  - Cytochrome b
MH  - electron
MH  - electron transfer
MH  - Electron-transfer
MH  - England
MH  - mechanism
MH  - membrane
MH  - Oxidoreductases
MH  - protein
MH  - Protein Subunits
MH  - Proteins
MH  - quinone
MH  - Quinones
MH  - resolution
MH  - Site
MH  - structure
MH  - SUBUNIT
MH  - succinate
MH  - TRANSFER
RP  - NOT IN FILE
NT  - UI - 20052158LA - engRN - 0 (Dicarboxylic Acids)RN - 0 (Iron-Sulfur Proteins)RN - 0 (Metals)RN - 0 (Quinones)RN - 146-14-5 (Flavin-Adenine Dinucleotide)RN - 14875-96-8 (Heme)RN - EC 1.3.99.1 (Succinate Dehydrogenase)PT - Journal ArticleDA - 19991210IS - 0028-0836SB - IMCY - ENGLAND
UR  - PM:10586875
SO  - Nature 1999 Nov 25 ;402(6760):377-385

1601
UI  - 325
AU  - Leslie AG
AU  - Abrahams JP
AU  - Braig K
AU  - Lutter R
AU  - Menz RI
AU  - Orriss GL
AU  - van Raaij MJ
AU  - Walker JE
AD  - MRC Laboratory of Molecular Biology, Cambridge, UK
TI  - The structure of bovine mitochondrial F1-ATPase: an example of rotary catalysis
AB  - There is now compelling evidence in support of a rotary catalytic mechanism in F1-ATPase, and, by extension, in the intact ATP synthase. Although models have been proposed to explain how protein translocation in F0 results in rotation of the gamma-subunit relative to the alpha 3/beta 3 assembly in F1 [22], these are still speculative. It seems likely that a satisfactory explanation of this mechanism will ultimately depend on structural information on the intact ATP synthase
RP  - NOT IN FILE
NT  - UI - 99193663LA - engRN - 0 (Enzyme Inhibitors)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 19990609IS - 0300-5127SB - IMCY - ENGLANDJC - E48
UR  - PM:10093703
SO  - Biochem Soc Trans 1999 Feb ;27(2):37-42

1602
UI  - 980
AU  - Levy D
AU  - Mosser G
AU  - Lambert O
AU  - Moeck GS
AU  - Bald D
AU  - Rigaud JL
TI  - Two-dimensional crystallization on lipid layer: A successful approach for membrane proteins
AB  - A considerable interest exists currently in designing innovative strategies to produce two-dimensional crystals of membrane proteins that are amenable to structural analysis by electron crystallography. We have developed a protocol for crystallizing membrane protein that is derived from the classical lipid-layer two-dimensional crystallization at the air/water interface used so far for soluble proteins. Lipid derivatized with a Ni(2+)-chelating head group provided a general approach to crystallizing histidine-tagged transmembrane proteins. The processes of protein binding and two-dimensional crystallization were analyzed by electron microscopy, using two prototypic membrane proteins: FhuA, a high-affinity receptor from the outer membrane of Escherichia coli, and the F(0)F(1)-ATP synthase from thermophilic Bacillus PS3. Conditions were found to avoid solubilization of the lipid layer by the detergent present with the purified membrane proteins and thus to allow binding of micellar proteins to the functionalized lipid head groups. After detergent removal using polystyrene beads, membrane sheets of several hundreds of square micrometers were reconstituted at the interface. High protein density in these membrane sheets allowed further formation of planar two-dimensional crystals. We believe that this strategy represents a new promising alternative to conventional dialysis methods for membrane protein 2D crystallization, with the additional advantage of necessitating little purified protein
MH  - Bacillus
MH  - Bacterial Outer Membrane Proteins
MH  - Chelating Agents
MH  - Chemistry
MH  - Crystallization
MH  - Detergents
MH  - Escherichia coli
MH  - H(+)-Transporting ATP Synthase
MH  - Histidine
MH  - Lipid Bilayers
MH  - Membrane Proteins
MH  - Methods
MH  - Micelles
MH  - Microscopy
MH  - Nickel
MH  - Protein Binding
MH  - Proteins
MH  - Receptors,Cell Surface
MH  - Receptors,Virus
MH  - Support,Non-U.S.Gov't
MH  - Surface Properties
RP  - NOT IN FILE
NT  - Section de Recherche, Institut Curie, UMR-CNRS 168 and LRC-CEA 8, 11 Rue P et M Curie, Paris, 75231, France daniellevy@curiefr
SO  - J Struct Biol 1999 Aug ;127(1):44-52

1603
UI  - 809
AU  - Malyan AN
AU  - Vitseva OI
AU  - Strotmann H
AD  - Institute of Basic Biological Problems, Russian Academy of Sciences, Pushchino, Moscow Region, 142292, Russia Malyan@isspserpukhovsu
TI  - Study of the structure of the thylakoid membrane-bound chloroplast coupling factor CF1
AB  - The structure of thylakoid membrane-bound chloroplast coupling factor CF1 was studied by limited proteolysis followed by sodium dodecylsulfate polyacrylamide gel electrophoresis and N-terminal sequence analysis. The N-terminal fragment of the alpha-subunit was shown to have an exposed area including the peptide bond R21-E22. The cleavage of this peptide bond caused the alphaK24-V25 bond to be exposed to the outside. In the N-terminal fragment of the beta-subunit, the L14-E15 bond was identified and found to be subject to trypsinolysis. Also, the alphaR140-S141, alphaG160-R161, and betaG102- G103 bonds were accessible to the proteolytic attack. In general, the beta-subunit of membrane-bound CF1 is more sensitive to proteolysis than that of solubilized CF1. The products of proteolysis of the alpha- subunit did not contain the polypeptides typical of the reaction of cleavage of the alphaE17-G18 and alphaE22-V23 bonds in isolated CF1. These results suggest a significant structural difference between soluble and membrane-bound CF1. A number of peptide bonds, alphaG160- R161 in particular, were shown to be shielded from proteolytic attack by papain in illuminated thylakoid membranes, probably as a result of membrane energization. In contrast, the light-induced reduction of the gamma-subunit caused an increase in the accessibility of some peptide bonds to this protease, including the alphaG160-R161 bond
RP  - NOT IN FILE
NT  - UI - 99453389LA - engRN - 0 (Macromolecular Systems)RN - 0 (Peptide Fragments)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19991108IS - 0006-2979SB - IMCY - RUSSIAJC - CSQ
UR  - PM:10521716
SO  - Biochemistry (Mosc ) 1999 Sep ;64(9):1000-1004

1604
UI  - 21
AU  - Matthey U
AU  - Kaim G
AU  - Braun D
AU  - Wuthrich K
AU  - Dimroth P
AD  - Mikrobiologisches Institut, Eidgenossische Technische Hochschule, Zurich, Switzerland
TI  - NMR studies of subunit c of the ATP synthase from Propionigenium modestum in dodecylsulphate micelles
AB  - The structure of the Na+, Li+ or H+-binding c subunit of the ATP synthase from Propionigenium modestum was studied by NMR. Subunit c in dodecylsulphate micelles consists of four alpha-helical segments, I-IV, that are connected by short linker peptides with non-regular secondary structures. We propose that helices I (V4-I26) and IV (I69-V85) are membrane-spanning structures, and that helices II and III and the intervening hydrophilic loop are located in the cytoplasm. The Na+- binding residues Q32, E65 and S66 are located in the I-->II and III-- >IV helix connections, probably near the membrane surface on the cytoplasmic side
RP  - NOT IN FILE
NT  - UI - 99234020LA - engRN - 0 (Bacterial Proteins)RN - 0 (Membrane Proteins)RN - 0 (Recombinant Proteins)RN - 151-21-3 (Sodium Dodecyl Sulfate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19990517IS - 0014-2956SB - IMCY - GERMANYJC - EMZ
UR  - PM:10215857
SO  - Eur J Biochem 1999 Apr ;261(2):459-467

1605
UI  - 21034
AU  - Murata T
AU  - Takase K
AU  - Yamato I
AU  - Igarashi K
AU  - Kakinuma Y
AD  - Department of Biological Science and Technology, Science University of Tokyo, Noda, Chiba, 278-8510, Japan
TI  - Properties of the V0V1 Na+-ATPase from Enterococcus hirae and its V0 moiety
AB  - We report here the large-scale purification of vacuolar (V0V1)-type Na+- ATPase from Enterococcus hirae achieved using column anion-exchange and gel filtration chromatographies; 32 mg of purified enzyme comprising nine subunits, A, B, C, D, E, F, G, I, and K, was obtained from 20 liter culture. This amount is 500-fold larger than that reported in the previous paper [Murata, T., Takase, K., Yamato, I., Igarashi, K., and Kakinuma, Y. (1997) J. Biol. Chem. 272, 24885-24890]. The purified enzyme shows a high specific activity of ATP hydrolysis (35.7 micromol Pi released/min/mg protein). ATP-driven 22Na+ uptake by reconstituted V0V1-proteoliposomes exhibited an apparent Kt value for Na+ of 40 microM, which is near the Km value (20 microM) for Na+ of the ATP hydrolytic activity. Denatured gel electrophoresis revealed that six subunits, A, B, C, D, E, and F, are releasable as the V1 subunit from the V0V1 complex by incubation with ethylenediaminetetraacetic acid; subunit G was not identified. The remaining V0-liposomes containing I and K subunits catalyzed Na+ uptake in response to potassium diffusion potential (Deltapsi, inside negative); the Kt value for Na+ of this reaction was estimated to be about 2 mM. Inhibition by N,N'- dicyclohexylcarbodiimide (DCCD) of the Na+-ATPase activity and Deltapsi- driven Na+ uptake by the V0-liposomes was prevented by the presence of Na+, suggesting that the Na+ binding site overlaps with the DCCD- reactive site
MH  - A
MH  - ACID
MH  - atp
MH  - ATPase
MH  - BINDING
MH  - COMPLEX
MH  - Dicyclohexylcarbodiimide
MH  - Diffusion
MH  - diffusion potential
MH  - Electrophoresis
MH  - H+-ATPase
MH  - Hydrolysis
MH  - Potassium
MH  - protein
MH  - Proteolipids
MH  - proteoliposome
MH  - purification
MH  - Site
MH  - Sodium
MH  - SUBUNIT
RP  - NOT IN FILE
NT  - UI - 99145498LA - engRN - 0 (Proteolipids)RN - 0 (proteoliposomes)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 7440-23-5 (Sodium)RN - EC 3.6.1.- (vacuolar H+-ATPase)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19990615IS - 0021-924XSB - IMCY - JAPAN
UR  - PM:9990142
SO  - J Biochem (Tokyo ) 1999 Feb ;125(2):414-421

1606
UI  - 409
AU  - Nadanaciva S
AU  - Weber J
AU  - Senior AE
AD  - Department of Biochemistry and Biophysics, University of Rochester Medical Center, Rochester, New York 14642, USA
TI  - Binding of the transition state analog MgADP-fluoroaluminate to F1- ATPase
AB  - Escherichia coli F1-ATPase from mutant betaY331W was potently inhibited by fluoroaluminate plus MgADP but not by MgADP alone. beta-Trp-331 fluorescence was used to measure MgADP binding to catalytic sites. Fluoroaluminate induced a very large increase in MgADP binding affinity at catalytic site one, a smaller increase at site two, and no effect at site three. Mutation of either of the critical catalytic site residues beta-Lys-155 or beta-Glu-181 to Gln abolished the effects of fluoroaluminate on MgADP binding. The results indicate that the MgADP- fluoroaluminate complex is a transition state analog and independently demonstrate that residues beta-Lys-155 and (particularly) beta-Glu-181 are important for generation and stabilization of the catalytic transition state. Dicyclohexylcarbodiimide-inhibited enzyme, with 1% residual steady-state ATPase, showed normal transition state formation as judged by fluoroaluminate-induced MgADP binding affinity changes, consistent with a proposed mechanism by which dicyclohexylcarbodiimide prevents a conformational interaction between catalytic sites but does not affect the catalytic step per se. The fluorescence technique should prove valuable for future transition state studies of F1-ATPase
RP  - NOT IN FILE
NT  - UI - 99167465LA - engRN - 0 (Aluminum Compounds)RN - 0 (Chlorides)RN - 0 (Enzyme Inhibitors)RN - 21330-18-7 (fluoroaluminum)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 7429-90-5 (Aluminum)RN - 7446-70-0 (aluminum chloride)RN - 7681-49-4 (Sodium Fluoride)RN - 7782-41-4 (Fluorine)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSDA - 19990413IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:10066761
SO  - J Biol Chem 1999 Mar 12 ;274(11):7052-7058

1607
UI  - 407
AU  - Nadanaciva S
AU  - Weber J
AU  - Senior AE
AD  - Department of Biochemistry and Biophysics, University of Rochester Medical Center, New York 14642, USA
TI  - The role of beta-Arg-182, an essential catalytic site residue in Escherichia coli F1-ATPase
AB  - Beta-Arg-182 in Escherichia coli F1-ATPase (beta-Arg-189 in bovine mitochondrial F1) is a residue which lies close to catalytic site bound nucleotide (Abrahams et al. (1994) Nature 370, 621-628). Here we investigated the role of this residue by characterizing two mutants, betaR182Q and betaR182K. Oxidative phosphorylation and steady-state ATPase activity of purified F1 were severely impaired by both mutations. Catalytic site nucleotide-binding parameters were measured using the fluorescence quench of beta-Trp-331 that occurred upon nucleotide binding to purified F1 from betaR182Q/betaY331W and betaR182K/betaY331W double mutants. It was found that (a) beta-Arg-182 interacts with the gamma-phosphate of MgATP, particularly at catalytic sites 1 and 2, (b) beta-Arg-182 has no functional interaction with the beta-phosphate of MgADP or with the magnesium of the magnesium- nucleotide complex in the catalytic sites, and (c) beta-Arg-182 is directly involved in the stabilization of the catalytic transition state. In these features the role of beta-Arg-182 resembles that of another positively charged residue in the catalytic site, the conserved lysine of the Walker A motif, beta-Lys-155. A further role of beta-Arg- 182 is suggested, namely involvement in conformational change at the catalytic site beta-alpha subunit interface that is required for multisite catalysis
RP  - NOT IN FILE
NT  - UI - 99316169LA - engRN - 21330-18-7 (fluoroaluminum)RN - 56-65-5 (Adenosine Triphosphate)RN - 56-85-9 (Glutamine)RN - 56-87-1 (Lysine)RN - 58-64-0 (Adenosine Diphosphate)RN - 7004-12-8 (Arginine)RN - 73-22-3 (Tryptophan)RN - 7429-90-5 (Aluminum)RN - 7782-41-4 (Fluorine)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSDA - 19990715IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:10387006
SO  - Biochemistry 1999 Jun 15 ;38(24):7670-7677

1608
UI  - 406
AU  - Nadanaciva S
AU  - Weber J
AU  - Wilke-Mounts S
AU  - Senior AE
AD  - Department of Biochemistry and Biophysics, University of Rochester Medical Center, New York 14642, USA
TI  - Importance of F1-ATPase residue alpha-Arg-376 for catalytic transition state stabilization
AB  - The functional role of essential residue alpha-Arg-376 in the catalytic site of F1-ATPase was studied. The mutants alpha R376C, alpha R376Q, and alpha R376K were constructed, and combined with the mutation beta Y331W, to investigate catalytic site nucleotide-binding parameters, and to assess catalytic transition state formation by measurement of MgADP- fluoroaluminate binding. Each mutation caused large impairment of ATP synthesis and hydrolysis. Despite the apparent proximity of alpha-Arg- 376 to bound nucleoside di- and triphosphate in published X-ray structures, the mutations had little effect on MgADP or MgATP binding affinities, particularly at the highest affinity catalytic site, site 1. Both Cys and Gln mutants abolished transition state formation, demonstrating that alpha-Arg-376 is normally involved at this step of catalysis. A model of the F1-ATPase catalytic transition state structure is presented and discussed. The Lys mutant, although severely impaired, supported transition state formation, suggesting that an additional essential role for the alpha-Arg-376 guanidinium group exists, likely in alpha/beta conformational signal transmission required for steady-state catalysis. Parallels between alpha-Arg-376 and GAP/G-protein "arginine finger" residues are evident
RP  - NOT IN FILE
NT  - UI - 20039872LA - engRN - 21330-18-7 (fluoroaluminum)RN - 52-90-4 (Cysteine)RN - 55520-40-6 (Tyrosine)RN - 56-65-5 (Adenosine Triphosphate)RN - 56-85-9 (Glutamine)RN - 56-87-1 (Lysine)RN - 58-64-0 (Adenosine Diphosphate)RN - 7004-12-8 (Arginine)RN - 73-22-3 (Tryptophan)RN - 7429-90-5 (Aluminum)RN - 7782-41-4 (Fluorine)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSDA - 19991216IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:10569931
SO  - Biochemistry 1999 Nov 23 ;38(47):15493-15499

1609
UI  - 20922
AU  - Nakamoto RK
AU  - Ketchum CJ
AU  - al Shawi MK
AD  - Department of Molecular Physiology and Biological Physics, University of Virginia, Charlottesville 22906, USA rkn3c@virginiaedu
TI  - Rotational coupling in the F0F1 ATP synthase
AB  - The F0F1 ATP synthase is a large multisubunit complex that couples translocation of protons down an electrochemical gradient to the synthesis of ATP. Recent advances in structural analyses have led to the demonstration that the enzyme utilizes a rotational catalytic mechanism. Kinetic and biochemical evidence is consistent with the expected equal participation of the three catalytic sites in the alpha 3 beta 3 hexamer, which operate in sequential, cooperative reaction pathways. The rotation of the core gamma subunit plays critical roles in establishing the conformation of the sites and the cooperative interactions. Mutational analyses have shown that the rotor subunits are responsible for coupling and in doing so transmit specific conformational information between transport and catalysis
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Catalysis
MH  - COMPLEX
MH  - conformation
MH  - coupling
MH  - F0F1
MH  - Macromolecular Systems
MH  - mechanism
MH  - physiology
MH  - proton
MH  - Protons
MH  - review
MH  - rotation
MH  - Site
MH  - SUBUNIT
MH  - SYNTHASE
MH  - synthesis
MH  - SYSTEM
MH  - SYSTEMS
MH  - translocation
MH  - transport
RP  - NOT IN FILE
NT  - UI - 99338990LA - engRN - 0 (Macromolecular Systems)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticlePT - ReviewPT - Review, AcademicID - R01-GM50957/GM/NIGMSID - R01-GM52502/GM/NIGMSDA - 19990908IS - 1056-8700SB - IMCY - UNITED STATES
UR  - PM:10410801
SO  - Annu Rev Biophys Biomol Struct 1999  ;28():205-234

1610
UI  - 699
AU  - Noji H
AU  - Yoshida M
AD  - CREST Genetic Programming Team 13, Teikyo University Biotechnology Research Center 3 F, Kawasaki
TI  - [F1-ATPase; the stepping rotary motor in ATP synthase]
RP  - NOT IN FILE
NT  - UI - 99166093LA - jpnRN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 19990428IS - 0037-1017SB - IMCY - JAPANJC - ILZ
UR  - PM:10067121
SO  - Seikagaku 1999 Jan ;71(1):34-50

1611
UI  - 282
AU  - Noji H
AU  - Hasler K
AU  - Junge W
AU  - Kinosita K
AU  - Yoshida M
AU  - Engelbrecht S
AD  - CREST team13 (Core Research for Evolutional Science and Technology) "Genetic Programming" Team 13, Teikyo University Biotechnology Research Center 3F, Nogawa 907, Kawasaki, Miyamae-ku, 216-0001, Japan
TI  - Rotation of Escherichia coli F(1)-ATPase
AB  - By applying the same method used for F(1)-ATPase (TF(1)) from thermophilic Bacillus PS3 (Noji, H., Yasuda, R., Yoshida, M., and Kinosita, K., Jr. (1997) Nature 386, 299-302), we observed ATP-driven rotation of a fluorescent actin filament attached to the gamma subunit in Escherichia coli F(1)-ATPase. The torque value and the direction of the rotation were the same as those observed for TF(1). F(1)-ATPases seem to share common properties of rotation irrespective of the sources
RP  - NOT IN FILE
NT  - UI - 99333374LA - engRN - 0 (Actins)RN - 0 (Recombinant Fusion Proteins)RN - 56-65-5 (Adenosine Triphosphate)RN - 9013-20-1 (Streptavidin)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19990816IS - 0006-291XSB - IMCY - UNITED STATESJC - 9Y8
UR  - PM:10403811
SO  - Biochem Biophys Res Commun 1999 Jul 14 ;260(3):597-599

1612
UI  - 141
AU  - Ogilvie I
AU  - Capaldi RA
AD  - Institute of Molecular Biology, University of Oregon, Eugene 97403- 1229, USA
TI  - Mutation of the mitochrondrially encoded ATPase 6 gene modeled in the ATP synthase of Escherichia coli
AB  - Defects of respiratory chain protein complexes and the ATP synthase are becoming increasingly implicated in human disease. Recently, mutations in the ATPase 6 gene have been shown to cause several different neurological disorders. The product of this gene is homologous to the a subunit of the ATP synthase of Escherichia coli. Here, mutations equivalent to those described in humans have been introduced into the a subunit of E. coli by site-directed mutagenesis, and the effects of these mutations on the ATPase activity, ATP synthesis and ability of the enzyme to pump protons studied in detail. The effects of the mutations varied considerably. The mutation L262P (9185 T-C equivalent) caused a 70% loss of ATP synthesis activity, reduced DCCD sensitivity, and lowered proton pumping activity. The L207P (8993 T-C equivalent) reduced ATP synthesis by 50%, affected DCCD sensitivity, while proton pumping was only marginally affected when measured by the standard AMCA quenching assay. The other mutations studied affected the functioning of the ATP synthase much less. The results confirm that modeling of these point mutations in the E. coli enzyme is a useful approach to determining how alterations in the ATPase 6 gene affect enzyme function and, therefore, how a pathogenic effect can be exerted
RP  - NOT IN FILE
NT  - UI - 99330192LA - engRN - 0 (Bacterial Proteins)RN - 0 (Proton Pump)RN - 0 (uncB protein, Escherichia coli)PT - Journal ArticleID - HL24526/HL/NHLBIDA - 19990802IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:10403398
SO  - FEBS Lett 1999 Jun 18 ;453(1-2):179-182

1613
UI  - 21286
AU  - Omelyan IP
TI  - Longitudinal wevevector- and frequency-dependent dielectric constant of the TIP4P water model
MH  - CONSTANT
MH  - Water
MH  - model
RP  - IN FILE
SO  - Mol Phys 1999  ;93():123-135

1614
UI  - 20840
AU  - Omote H
AU  - Sambonmatsu N
AU  - Saito K
AU  - Sambongi Y
AU  - Iwamoto-Kihara A
AU  - Yanagida T
AU  - Wada Y
AU  - Futai M
AD  - Division of Biological Sciences, Institute of Scientific and Industrial Research, Osaka University, CREST of Japan Science and Technology Corporation, Ibaraki, Osaka 567-0047, Japan
TI  - The gamma-subunit rotation and torque generation in F1-ATPase from wild- type or uncoupled mutant Escherichia coli
AB  - The rotation of the gamma-subunit has been included in the binding- change mechanism of ATP synthesis/hydrolysis by the proton ATP synthase (FOF1). The Escherichia coli ATP synthase was engineered for rotation studies such that its ATP hydrolysis and synthesis activity is similar to that of wild type. A fluorescently labeled actin filament connected to the gamma-subunit of the F1 sector rotated on addition of ATP. This progress enabled us to analyze the gammaM23K (the gamma-subunit Met-23 replaced by Lys) mutant, which is defective in energy coupling between catalysis and proton translocation. We found that the F1 sector produced essentially the same frictional torque, regardless of the mutation. These results suggest that the gammaM23K mutant is defective in the transformation of the mechanical work into proton translocation or vice versa
MH  - A
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BINDING
MH  - Catalysis
MH  - CHANGE MECHANISM
MH  - COLI ATP SYNTHASE
MH  - coupling
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - Hydrolysis
MH  - Macromolecular Systems
MH  - mechanism
MH  - mutant
MH  - protein
MH  - Proteins
MH  - proton
MH  - rotation
MH  - SYNTHASE
MH  - synthesis
MH  - SYSTEM
MH  - SYSTEMS
MH  - translocation
RP  - NOT IN FILE
NT  - UI - 99324139LA - engRN - 0 (Actins)RN - 0 (Macromolecular Systems)RN - 0 (Recombinant Proteins)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19990826IS - 0027-8424SB - IMCY - UNITED STATES
UR  - PM:10393898
SO  - Proc Natl Acad Sci U S A 1999 Jul 6 ;96(14):7780-7784

1615
UI  - 19906
AU  - Oster G
AU  - Wang H
AD  - Departments of Molecular & Cellular Biology, College of Natural Resources, University of California, Berkeley, California 94720-3112, USA goster@natureberkeleyedu
TI  - ATP synthase: two motors, two fuels
AB  - FoF1 ATPase is the universal protein responsible for ATP synthesis. The enzyme comprises two reversible rotary motors: Fo is either an ion 'turbine' or an ion pump, and F1 is either a hydrolysis motor or an ATP synthesizer. Recent biophysical and biochemical studies have helped to elucidate the operating principles for both motors
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - ATPase
MH  - England
MH  - F1
MH  - H(+)-Transporting ATP Synthase
MH  - Hydrolysis
MH  - ion
MH  - Macromolecular Systems
MH  - protein
MH  - review
MH  - SYNTHASE
MH  - synthesis
MH  - SYSTEM
MH  - SYSTEMS
RP  - NOT IN FILE
NT  - UI - 99216542LA - engRN - 0 (Macromolecular Systems)RN - 0 (Molecular Motors)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 19990610IS - 0969-2126SB - IMCY - ENGLANDJC - DEB
UR  - PM:10196130
SO  - Structure Fold Des 1999 Apr 15 ;7(4):R67-R72

1616
UI  - 798
AU  - Panke O
AU  - Rumberg B
AD  - Max-Volmer-Institut, Technische Universitat Berlin, D-10623, Berlin, Germany
TI  - Kinetic modeling of rotary CF0F1-ATP synthase: storage of elastic energy during energy transduction
AB  - F0F1-ATP synthase uses proton-motive force to produce ATP from ADP and Pi. With regard to its rotary mechanics, this energy transducing molecular machine assumes a unique position among all enzymes. In the work presented here we put forward a detailed functional model which is based on experimental results obtained with ATP synthase from spinach chloroplasts. We focus on the role of the elastic element, realized by the stalk-like subunit gamma, whose function is energy transduction between F0 and F1 taking into account the H+/ATP coupling ratio of four. Fitting parameters are the rate constants and the torsional rigidity of gamma, which have been adjusted according to the experimental results where the influence of transmembrane DeltapH on the rates of ATP synthesis/hydrolysis is put to the test. We show that the input and output of torsional energy are regulated by purely statistical principles, giving rise to the amount of transiently stored energy to be sliding, depending on DeltapH. During conditions of maximal turnover gamma turns out to be wound up towards 102 degrees which corresponds to a torque of 5.3. 10-20 N.m
RP  - NOT IN FILE
NT  - UI - 0LA - EngPT - JOURNAL ARTICLEDA - 19990728IS - 0006-3002SB - IMJC - A0W
UR  - PM:0010393255
SO  - Biochim Biophys Acta 1999 Jun 30 ;1412(2):118-128

1617
UI  - 21296
AU  - Pertsemlidis A
AU  - Soper AK
AU  - Sorenson JM
AU  - Head-Gordon T
AD  - Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720, USA
TI  - Evidence for microscopic, long-range hydration forces for a hydrophobic amino acid
AB  - We have combined neutron solution scattering experiments with molecular dynamics simulation to isolate an excess experimental signal that is caused solely by N-acetyl-leucine-amide (NALA) correlations in aqueous solution. This excess signal contains information about how NALA molecule centers are correlated in water, and we show how these solute- solute correlations might be determined at dilute concentrations in the small angle region. We have tested qualitatively different pair distribution functions for NALA molecule centers-gas, cluster, and aqueous forms of gc(r)-and have found that the excess experimental signal is adequate enough to rule out gas and cluster pair distribution functions. The aqueous form of gc(r) that exhibits a solvent-separated minimum, and possibly longer-ranged correlations as well, is not only physically sound but reproduces the experimental data reasonably well. This work demonstrates that important information in the small angle region can be mined to resolve solute-solute correlations, their lengthscales, and thermodynamic consequences even at dilute concentrations. The hydration forces that operate on the microscopic scale of individual amino acid side chains, implied by the small angle scattering data, could have significant effects on the early stages of protein folding, on ligand binding, and on other intermolecular interactions
MH  - A
MH  - ACID
MH  - Amino Acids
MH  - BINDING
MH  - biology
MH  - cell
MH  - data
MH  - function
MH  - INTERACTION
MH  - MOLECULAR-DYNAMICS
MH  - protein
MH  - SIMULATION
MH  - united states
MH  - universities
MH  - Water
RP  - NOT IN FILE
NT  - UI - 99110916LA - engRN - 0 (Amides)RN - 0 (Amino Acids)RN - 0 (N-acetylleucinamide)RN - 61-90-5 (Leucine)RN - 7732-18-5 (Water)PT - Journal ArticleDA - 19990316IS - 0027-8424SB - IMCY - UNITED STATES
UR  - PM:9892659
SO  - Proc Natl Acad Sci U S A 1999 Jan 19 ;96(2):481-486

1618
UI  - 9941
AU  - Poetsch A
AU  - Seelert H
AU  - Meyer zu TJ
AU  - Dencher NA
AD  - Abteilung Physikalische Biochemie, Technische Universitat Darmstadt, Petersenstrasse 22, Darmstadt, D-64287, Germany
TI  - Detergent effect on anion exchange perfusion chromatography and gel filtration of intact chloroplast H(+)-ATP synthase
AB  - To gain a pure enzyme preparation for functional and crystallization studies, an additional purification step in the isolation of the chloroplast ATP synthase (CF(0)F(1)) has been introduced. By applying gel filtration or anion exchange perfusion chromatography in presence of the detergents CHAPS and n-dodecyl-beta-d-maltoside, respectively, Rubisco and other contaminants were separated from CF(0)F(1). The purity and activity depended on the chromatographic method and the detergent employed. The highest purity and activity were achieved by anion exchange chromatography for the detergent dodecyl-maltoside and by gel filtration for the detergent CHAPS. The detergent Triton X-100, which is frequently used to solubilize CF(0)F(1), was found to be inadequate to stabilize the ATP synthase during chromatography
MH  - atp
MH  - ATP synthase
MH  - chloroplast
MH  - Cholic Acid
MH  - Cholic Acids
MH  - Crystallization
MH  - Detergents
MH  - Glucosides
MH  - H(+)-Transporting ATP Synthase
MH  - Octoxynol
MH  - purification
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 20025377LA - engRN - 0 (Cholic Acids)RN - 0 (Detergents)RN - 0 (Glucosides)RN - 69227-93-6 (dodecyl maltoside)RN - 75621-03-3 (3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate)RN - 9002-93-1 (Octoxynol)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19991220IS - 0006-291XSB - IMCY - UNITED STATESJC - 9Y8
UR  - PM:10558901
SO  - Biochem Biophys Res Commun 1999 Nov 19 ;265(2):520-524

1619
UI  - 810
AU  - Ponomarenko S
AU  - Volfson I
AU  - Strotmann H
AD  - Institut fur Biochemie der Pflanzen, Heinrich Heine Universitat Dusseldorf, Germany
TI  - Proton gradient-induced changes of the interaction between CF0 and CF1 related to activation of the chloroplast ATP synthase
AB  - Thylakoid energization by light causes destabilization of CF0CF1 so that the peripheral CF1 sector is more readily detached from the membrane by intermediate concentrations of the chaotropic salt NaSCN. Here we have investigated the correlation between the proton gradient- induced change of CF0CF1 interaction and CF0CF1 activation. The results indicate a close relationship between the two phenomena. The effect is most probably due to reduction of the electrostatic interaction between the two subcomplexes CF0 and CF1 as a consequence of protonations in the interface region
RP  - NOT IN FILE
NT  - UI - 99142697LA - engRN - 0 (Protons)RN - 0 (Sulfhydryl Compounds)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19990302IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:9989591
SO  - FEBS Lett 1999 Jan 25 ;443(2):136-138

1620
UI  - 21176
AU  - Radionov AN
AU  - Klyachko VA
AU  - Kaulen AD
AD  - Department of Photobiochemistry, Belozersky Institute of Physico- Chemical Biology, Lomonosov Moscow State University, Moscow, 119899, Russia
TI  - Formation of the M(N) (M(open)) intermediate in the wild-type bacteriorhodopsin photocycle is accompanied by an absorption spectrum shift to shorter wavelength, like that in the mutant D96N bacteriorhodopsin photocycle
AB  - Maximum of the M intermediate difference spectrum in the wild-type Halobacterium salinarium purple membrane is localized at 405-406 nm under conditions favoring accumulation of the M(N) intermediate (6 M guanidine chloride, pH 9.6), whereas immediately after laser flash the maximum is localized at 412 nm. The maximum is also localized at 412 nm 0.1 msec after the flash in the absence of guanidine chloride at pH 11.3. Within several milliseconds the maximum is shifted to short- wavelength region by 5-6 nm. This shift is similar to that in the D96N mutant which accompanies the M(N) (M(open)) intermediate formation. The main two differences are: 1) the rate of the shift is slower in the wild-type bacteriorhodopsin, and is similar to the rate of the M to N intermediate transition (t1/2 approximately 2 msec); 2) the shift in the wild-type bacteriorhodopsin is observed at alkaline pH values which are higher than pK of the Schiff base (approximately 10.8 at 1 M NaCl) in the N intermediate with the deprotonated Asp-96. Thus, the M(N) (M(open)) intermediate with open water-permeable inward proton channel is observed only at high pH, when the Schiff base and Asp-96 are deprotonated. The data confirmed our earlier conclusion that the M intermediate observed at lower pH has the closed inward proton channel
MH  - absorption
MH  - Bacteriorhodopsin
MH  - BASE
MH  - flash
MH  - Halobacterium
MH  - Halobacterium salinarium
MH  - intermediate
MH  - M
MH  - M-intermediate
MH  - membrane
MH  - mutant
MH  - pH
MH  - proton
MH  - purple membrane
MH  - Schiff base
MH  - Schiff-base
MH  - spectra
RP  - NOT IN FILE
NT  - UI - 20030184LA - engRN - 53026-44-1 (Bacteriorhodopsins)PT - Journal ArticleDA - 20000131IS - 0006-2979SB - IMCY - RUSSIA
UR  - PM:10561570
SO  - Biochemistry (Mosc ) 1999 Oct ;64(10):1210-1214

1621
UI  - 21178
AU  - Radionov AN
AU  - Kaulen AD
AD  - Department of Photobiochemistry, AN Belozersky Institute of Physico- Chemical Biology, Moscow State University, Russia
TI  - Two forms of N intermediate (N(open) and N(closed)) in the bacteriorhodopsin photocycle
AB  - Glutaraldehyde, aluminum ions and glycerol (that inhibit the M intermediate decay in the wild-type bacteriorhodopsin and azide-induced M decay in the D96N mutant by stabilization of the M(closed)) accelerate the N decay in the D96N mutant. The aluminum ions, the most potent activator of the N decay, induce a blue shift of the N difference spectrum by approximately 10 nm. Protonated azide as well as acetate and formate inhibit the N decay in both the D96N mutant and the wild-type protein. It is concluded that the N intermediate represents, in fact, an equilibrium mixture of the two ('open' and 'closed') forms. These two forms, like M(closed) and M(open), come to an equilibrium in the microseconds range. The absorption spectrum of the N(open) is slightly shifted to red in comparison to that of the N(closed). Again, this resembles the M forms. 13-cis-all-trans re-isomerization is assumed to occur in the N(closed) form only. Binding of 1-2 molecules of protonated azide stabilizes the N(open) form. Existence of the 'open' and 'closed' forms of the M and N intermediates provides the appropriate explanation of the cooperative phenomenon as well as some other effects on the bacteriorhodopsin photocycle. Summarizing the available data, we suggest that M(open) is identical to the M(N) form, whereas M1 and M2 are different substates of M(closed)
MH  - A
MH  - absorption
MH  - acetate
MH  - Bacteriorhodopsin
MH  - BINDING
MH  - intermediate
MH  - ion
MH  - Ions
MH  - M
MH  - M-intermediate
MH  - microsecond
MH  - mutant
MH  - protein
MH  - reaction center
MH  - spectra
MH  - Water
RP  - NOT IN FILE
NT  - UI - 99297565LA - engRN - 0 (Photosynthetic Reaction Center, Bacterial)RN - 53026-44-1 (Bacteriorhodopsins)RN - 7732-18-5 (Water)PT - Journal ArticleDA - 19990706IS - 0014-5793SB - IMCY - NETHERLANDS
UR  - PM:10371155
SO  - FEBS Lett 1999 May 21 ;451(2):147-151

1622
UI  - 9936
AU  - Rastogi VK
AU  - Girvin ME
AD  - Biochemistry Department, Albert Einstein College of Medicine, Bronx, New York 10461, USA
TI  - Structural changes linked to proton translocation by subunit c of the ATP synthase
AB  - F1F0 ATP synthases use a transmembrane proton gradient to drive the synthesis of cellular ATP. The structure of the cytosolic F1 portion of the enzyme and the basic mechanism of ATP hydrolysis by F1 are now well established, but how proton translocation through the transmembrane F0 portion drives these catalytic changes is less clear. Here we describe the structural changes in the proton-translocating F0 subunit c that are induced by deprotonating the specific aspartic acid involved in proton transport. Conformational changes between the protonated and deprotonated forms of subunit c provide the structural basis for an explicit mechanism to explain coupling of proton translocation by F0 to the rotation of subunits within the core of F1. Rotation of these subunits within F1 causes the catalytic conformational changes in the active sites of F1 that result in ATP synthesis
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - Biochemistry
MH  - conformational change
MH  - England
MH  - F0
MH  - F1
MH  - H(+)-Transporting ATP Synthase
MH  - Hydrolysis
MH  - mechanism
MH  - proton
MH  - Protons
MH  - SUBUNIT
MH  - SYNTHASE
MH  - transport
RP  - NOT IN FILE
NT  - UI - 20046441LA - engRN - 0 (Protons)RN - 56-65-5 (Adenosine Triphosphate)RN - 56-84-8 (Aspartic Acid)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19991210IS - 0028-0836SB - IMCY - ENGLANDJC - NSC
UR  - PM:10580496
SO  - Nature 1999 Nov 18 ;402(6759):263-268

1623
UI  - 9937
AU  - Rastogi VK
AU  - Girvin ME
TI  - 1H, 13C, and 15N assignments and secondary structure of the high pH form of subunit c of the F1F0 ATP synthase
MH  - atp
MH  - ATP synthase
MH  - Bacterial Proteins
MH  - H(+)-Transporting ATP Synthase
MH  - Hydrogen
MH  - Proteins
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 99169956LA - engRN - 0 (Bacterial Proteins)RN - 0 (Carbon Isotopes)RN - 0 (Nitrogen Isotopes)RN - 1333-74-0 (Hydrogen)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - LetterDA - 19990416IS - 0925-2738SB - IMCY - NETHERLANDSJC - BJM
UR  - PM:10070750
SO  - J Biomol NMR 1999 Jan ;13(1):91-92

1624
UI  - 20839
AU  - Sambongi Y
AU  - Iko Y
AU  - Tanabe M
AU  - Omote H
AU  - Iwamoto-Kihara A
AU  - Ueda I
AU  - Yanagida T
AU  - Wada Y
AU  - Futai M
AD  - Division of Biological Sciences, Institute of Scientific and Industrial Research, Osaka University, CREST (Core Research for Evolutional Science and Technology) of Japan Science and Technology Corporation, Ibaraki, Osaka 567-0047, Japan
TI  - Mechanical rotation of the c subunit oligomer in ATP synthase (F0F1): direct observation
AB  - F0F1, found in mitochondria or bacterial membranes, synthesizes adenosine 5'-triphosphate (ATP) coupling with an electrochemical proton gradient and also reversibly hydrolyzes ATP to form the gradient. An actin filament connected to a c subunit oligomer of F0 was able to rotate by using the energy of ATP hydrolysis. The rotary torque produced by the c subunit oligomer reached about 40 piconewton- nanometers, which is similar to that generated by the gamma subunit in the F1 motor. These results suggest that the gamma and c subunits rotate together during ATP hydrolysis and synthesis. Thus, coupled rotation may be essential for energy coupling between proton transport through F0 and ATP hydrolysis or synthesis in F1
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - coupling
MH  - Enzymes
MH  - F0
MH  - F0F1
MH  - F1
MH  - Hydrolysis
MH  - membrane
MH  - Membranes
MH  - Mitochondria
MH  - proton
MH  - rotation
MH  - SUBUNIT
MH  - SYNTHASE
MH  - synthesis
MH  - transport
MH  - Uncoupling Agents
RP  - NOT IN FILE
NT  - UI - 20044999LA - engRN - 0 (Actins)RN - 0 (Enzymes, Immobilized)RN - 0 (Molecular Motors)RN - 0 (Uncoupling Agents)RN - 0 (Venturicidins)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 19991214IS - 0036-8075SB - IMCY - UNITED STATES
UR  - PM:10576736
SO  - Science 1999 Nov 26 ;286(5445):1722-1724

1625
UI  - 140
AU  - Schulenberg B
AU  - Capaldi RA
AD  - Institute of Molecular Biology, University of Oregon, Eugene, Oregon 97403-1229, USA
TI  - The epsilon subunit of the F(1)F(0) complex of Escherichia coli. cross- linking studies show the same structure in situ as when isolated
AB  - Four double mutants in the epsilon subunit were generated, each containing two cysteines, which, based on the NMR structure of this subunit, should form internal disulfide bonds. Two of these were designed to generate interdomain cross-links that lock the C-terminal alpha-helical domain against the beta-sandwich (epsilonM49C/A126C and epsilonF61C/V130C). The second set should give cross-linking between the two C-terminal alpha-helices (epsilonA94C/L128C and epsilonA101C/L121C). All four mutants cross-linked with 90-100% efficiency upon CuCl(2) treatment in isolated Escherichia coli ATP synthase. This shows that the structure obtained for isolated epsilon is essentially the same as in the assembled complex. Functional studies revealed increased ATP hydrolysis after cross-linking between the two domains of the subunit but not after cross-linking between the C- terminal alpha-helices. None of the cross-links had any effect on proton pumping-coupled ATP hydrolysis, on DCCD sensitivity of this activity, or on ATP synthesis rates. Therefore, big conformational changes within epsilon can be ruled out as a part of the enzyme function. Protease digestion studies, however, showed that subtle changes do occur, since the epsilon subunit could be locked in an ADP or 5'-adenylyl-beta,gamma-imidodiphosphate conformation by the cross- linking with resulting differences in cleavage rates
RP  - NOT IN FILE
NT  - UI - 99428504LA - engRN - 0 (Adenine Nucleotides)RN - 0 (Cross-Linking Reagents)RN - EC 3.4.21.4 (Trypsin)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - HL 24526/HL/NHLBIDA - 19991102IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:10497194
SO  - J Biol Chem 1999 Oct 1 ;274(40):28351-28355

1626
UI  - 139
AU  - Schulenberg B
AU  - Aggeler R
AU  - Murray J
AU  - Capaldi RA
AD  - Institute of Molecular Biology, University of Oregon, Eugene, Oregon 97403-1229, USA
TI  - The gammaepsilon-c subunit interface in the ATP synthase of Escherichia coli. cross-linking of the epsilon subunit to the c subunit ring does not impair enzyme function, that of gamma to c subunits leads to uncoupling
AB  - Mutants with a cysteine residue in the gamma subunit at position 207 and the epsilon subunit at position 31 were expressed in combination with a c-dimer construct, which contains a single cysteine at position 42 of the second c subunit. These mutants are called gammaY207C/cc'Q42C and epsilonE31C/cc'Q42C, respectively. Cross-linking of epsilon to the c subunit ring was obtained almost to completion without significant effect on any enzyme function, i.e. ATP hydrolysis, ATP synthesis, and ATP hydrolysis-driven proton translocation were all close to that of wild type. The gamma subunit could also be linked to the c subunit ring in more than 90% yield, but this affected coupling. Thus, ATP hydrolysis was increased 2. 5-fold, ATP synthesis was dramatically decreased, and ATP hydrolysis-driven proton translocation was abolished, as measured by the 9-amino-6-chloro-2- methoxyacridinequenching method. These results for epsilonE31C/cc'Q42C indicate that the c subunit ring rotates with the central stalk element. That the gamma-epsilon cross-linked enzyme retains ATPase activity also argues for a gammaepsilon-c subunit rotor. However, the uncoupling induced by cross-linking of gamma to the c subunit ring points to important conformational changes taking place in the gammaepsilon-c subunit interface during this. Blocking these structural changes by cross-linking leads to a proton leak within the F(0)
RP  - NOT IN FILE
NT  - UI - 20036566LA - engRN - 0 (Cross-Linking Reagents)RN - 0 (Peptide Fragments)RN - 7440-50-8 (Copper)RN - 7758-89-6 (cuprous chloride)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - HL 24526/HL/NHLBIDA - 19991229IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:10567396
SO  - J Biol Chem 1999 Nov 26 ;274(48):34233-34237

1627
UI  - 21179
AU  - Siletsky S
AU  - Kaulen AD
AU  - Konstantinov AA
AD  - A N Belozersky Institute of Physico-Chemical Biology, Moscow State University, Moscow 119899, Russia
TI  - Resolution of electrogenic steps coupled to conversion of cytochrome c oxidase from the peroxy to the ferryl-oxo state
AB  - Charge translocation across the membrane coupled to transfer of the third electron in the reaction cycle of bovine cytochrome c oxidase (COX) has been studied. Flash-induced reduction of the peroxy intermediate (P) to the ferryl-oxo state (F) by tris-bipyridyl complex of Ru(II) in liposome-reconstituted COX is coupled to several phases of membrane potential generation that have been time-resolved with the use of an electrometric technique applied earlier in the studies of the ferryl-oxo-to-oxidized (F --> O) transition of the enzyme [Zaslavsky, D., et al. (1993) FEBS Lett. 336, 389-393]. As in the case of the F --> O transition, the electric response associated with photoreduction of P to F includes a rapid KCN-insensitive electrogenic phase with a tau of 40-50 microseconds (reduction of heme a by CuA) and a multiphasic slower part; this part is cyanide-sensitive and is assigned to vectorial transfer of protons coupled to reduction of oxygen intermediate in the binuclear center. The net KCN-sensitive phase of the response is approximately 4-fold more electrogenic than the rapid phase, which is similar to the characteristics of the F --> O electrogenic transition and is consistent with net transmembrane translocation of two protons per electron, including vectorial movement of both "chemical" and "pumped" protons. The protonic part of the P --> F electric response is faster than in the F --> O transition and can be deconvoluted into three exponential phases with tau values varying for different samples in the range of 0.25-0.33, 1-1.5, and 6-7.5 ms at pH 8. Of these three phases, the 1-1.5 ms component is the major one contributing 50-60%. The P --> F conversion induced by single electron photoreduction of the peroxy state as studied in this work is several times slower than the P --> F transition resolved during oxidation of the fully reduced oxidase by molecular oxygen. The role of the CuB redox state in controlling the rate of P --> F conversion of heme a3 is discussed
MH  - A
MH  - COMPLEX
MH  - cytochrome
MH  - Cytochrome c
MH  - Cytochrome-c Oxidase
MH  - electrogenic
MH  - electron
MH  - Hydrogen
MH  - Hydrogen Peroxide
MH  - intermediate
MH  - membrane
MH  - Membrane Potential
MH  - microsecond
MH  - Movement
MH  - Oxygen
MH  - P
MH  - pH
MH  - Potassium
MH  - proton
MH  - Protons
MH  - redox
MH  - resolution
MH  - Time
MH  - TRANSFER
MH  - translocation
RP  - NOT IN FILE
NT  - UI - 99218095LA - engRN - 0 (Ferrous Compounds)RN - 151-50-8 (Potassium Cyanide)RN - 7722-84-1 (Hydrogen Peroxide)RN - 7782-44-7 (Oxygen)RN - EC 1.9.3.1 (Cytochrome-c Oxidase)PT - Journal ArticleID - TW 00349/TW/FICDA - 19990517IS - 0006-2960SB - IMCY - UNITED STATES
UR  - PM:10200174
SO  - Biochemistry 1999 Apr 13 ;38(15):4853-4861

1628
UI  - 21254
AU  - Silver RB
AD  - Marine Biological Laboratory, Woods Hole, Massachusetts 02543, USA rsilver@mbledu
TI  - Imaging structured space-time patterns of Ca2+ signals: essential information for decisions in cell division
MH  - review
RP  - NOT IN FILE
NT  - UI - 20084147LA - engPT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 20000112IS - 0892-6638SB - IMCY - UNITED STATES
UR  - PM:10619129
SO  - FASEB J 1999 Dec ;13 Suppl 2():S209-S215

1629
UI  - 20810
AU  - Sorgen PL
AU  - Bubb MR
AU  - Cain BD
AD  - Department of Biochemistry and Molecular Biology, University of Florida, Gainesville, Florida 32610, USA
TI  - Lengthening the second stalk of F(1)F(0) ATP synthase in Escherichia coli
AB  - In Escherichia coli F(1)F(0) ATP synthase, the two b subunits dimerize forming the peripheral second stalk linking the membrane F(0) sector to F(1). Previously, we have demonstrated that the enzyme could accommodate relatively large deletions in the b subunits while retaining function (Sorgen, P. L., Caviston, T. L., Perry, R. C., and Cain, B. D. (1998) J. Biol. Chem. 273, 27873-27878). The manipulations of b subunit length have been extended by construction of insertion mutations into the uncF(b) gene adding amino acids to the second stalk. Mutants with insertions of seven amino acids were essentially identical to wild type strains, and mutants with insertions of up to 14 amino acids retained biologically significant levels of activity. Membranes prepared from these strains had readily detectable levels of F(1)F(0)- ATPase activity and proton pumping activity. However, the larger insertions resulted in decreasing levels of activity, and immunoblot analysis indicated that these reductions in activity correlated with reduced levels of b subunit in the membranes. Addition of 18 amino acids was sufficient to result in the loss of F(1)F(0) ATP synthase function. Assuming the predicted alpha-helical structure for this area of the b subunit, the 14-amino acid insertion would result in the addition of enough material to lengthen the b subunit by as much as 20 A. The results of both insertion and deletion experiments support a model in which the second stalk is a flexible feature of the enzyme rather than a rigid rod-like structure
MH  - A
MH  - ACID
MH  - Amino Acids
MH  - analysis
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Bacterial Proteins
MH  - Biochemistry
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - function
MH  - membrane
MH  - Membranes
MH  - model
MH  - mutant
MH  - P
MH  - protein
MH  - Proteins
MH  - proton
MH  - stalk
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 20062837LA - engRN - 0 (Bacterial Proteins)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleID - GM43495/GM/NIGMSDA - 20000127IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:10593914
SO  - J Biol Chem 1999 Dec 17 ;274(51):36261-36266

1630
UI  - 9885
AU  - Souza MO
AU  - Creczynski-Pasa TB
AU  - Graber P
AU  - Scofano HM
AU  - Mignaco JA
TI  - Hydrostatic pressure and heat modulate chloroplast ATP-synthase hydrolytic activities
MH  - chloroplast
MH  - Heat
RP  - NOT IN FILE
NT  - JournalMeeting AbstractFEDERATION AMER SOC EXP BIOLAPR 23S226QXBETHESDAUniv Fed Rio de Janeiro, Dept Bioquim Med, ICB, CCS, BR-21941 Rio De Janeiro, BrazilFASEB J9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 USA
AV  - Univ Fed Rio de Janeiro, Dept Bioquim Med, ICB, CCS, BR-21941 Rio De Janeiro, Brazil UFSC, Dept C Fisiol, CCB, Florianopolis, SC, Brazil Univ Freiburg, Inst Phys Chem, D-7800 Freiburg, Germany
UR  - ISI:000082033400686
SO  - Faseb Journal 1999  ;13(7):A1441-A1441

1631
UI  - 323
AU  - Stock D
AU  - Leslie AG
AU  - Walker JE
AD  - Medical Research Council Dunn Human Nutrition Unit, Hills Road, Cambridge CB2 2XY, UK
TI  - Molecular architecture of the rotary motor in ATP synthase
AB  - Adenosine triphosphate (ATP) synthase contains a rotary motor involved in biological energy conversion. Its membrane-embedded F0 sector has a rotation generator fueled by the proton-motive force, which provides the energy required for the synthesis of ATP by the F1 domain. An electron density map obtained from crystals of a subcomplex of yeast mitochondrial ATP synthase shows a ring of 10 c subunits. Each c subunit forms an alpha-helical hairpin. The interhelical loops of six to seven of the c subunits are in close contact with the gamma and delta subunits of the central stalk. The extensive contact between the c ring and the stalk suggests that they may rotate as an ensemble during catalysis
RP  - NOT IN FILE
NT  - UI - 20044992LA - engRN - 0 (Molecular Motors)RN - 0 (Protons)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19991214IS - 0036-8075SB - IMCY - UNITED STATESJC - UJ7
UR  - PM:10576729
SO  - Science 1999 Nov 26 ;286(5445):1700-1705

1632
UI  - 384
AU  - Syroeshkin AV
AU  - Galkin MA
AU  - Sedlov AV
AU  - Vinogradov AD
AD  - Department of Biochemistry, School of Biology, Lomonosov Moscow State University, Moscow, 119899, Russia
TI  - Kinetic mechanism of Fo x F1 mitochondrial ATPase: Mg2+ requirement for Mg x ATP hydrolysis
AB  - The initial rates of ATP hydrolysis catalyzed by Fo x F1 (bovine heart submitochondrial particles) preincubated in the presence of Pi for complete activation of the oligomycin-sensitive ATPase were measured as a function of ATP, Mg2+, and Mg x ATP concentrations. The results suggest the mechanism in which Mg x ATP complex is the true substrate of the ATPase and the second Mg2+ bound at a specific pH-dependent site is needed for the catalysis. Simple hyperbolic Michaelis--Menten dependences of the reaction rate on the substrate (Mg x ATP) and activating Mg2+ were found. In contrast to the generally accepted view, no inhibition of ATPase by free Mg2+ was found. Inhibition of the reaction by free ATP is due to a decrease of free Mg2+ needed for the catalysis. In the presence of both Ca2+ and Mg2+ the kinetics of ATP hydrolysis suggest that the Ca x ATP complex is neither hydrolyzed nor competes with Mg x ATP, and free Ca2+ does not affect the hydrolysis of Mg x ATP complex. A crucial role of free Mg2+ in the time-dependent inhibition of ATPase by azide is shown. The dependence of apparent Km for Mg x ATP on saturation of the Mg2+-specific site suggests the formal ping-pong mechanism in which bound Mg2+ participates in the overall reaction after dissociation of one product (most likely Pi) thus promoting either release of ADP (catalytic turnover) or slow isomerization of the enzyme--product complex (formation of the dead-end ADP(Mg2+)-inhibited enzyme). The rate of Mg x ATP hydrolysis only slightly depends on pH at saturating Mg2+. In the presence of limited amounts of free Mg2+ the pH dependence of the initial rate corresponds to the titration of a single group with pKa = 7.5. The simple competition between H+ and activating Mg2+ was observed. The specific role of Mg2+ as a coupling cation for energy transduction in Fo x F1- ATPase is discussed
RP  - NOT IN FILE
NT  - UI - 20030173LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - 7439-95-4 (Magnesium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 20000131IS - 0006-2979SB - IMCY - RUSSIAJC - CSQ
UR  - PM:10561559
SO  - Biochemistry (Mosc ) 1999 Oct ;64(10):1128-1137

1633
UI  - 21032
AU  - Takase K
AU  - Yamato I
AU  - Igarashi K
AU  - Kakinuma Y
AD  - Department of Biological Science and Technology, Science University of Tokyo, Chiba, Japan takase@y105hptbnodasutacjp
TI  - Indispensable glutamic acid residue-139 of NtpK proteolipid in the reaction of vacuolar Na(+)-translocating ATPase in Enterococcus hirae
AB  - Enterococcus hirae vacuolar ATPase catalyzes translocation of Na+ or Li+ coupled with ATP hydrolysis. It is suggested that the glutamic acid residue (Glu139) of NtpK proteolipid subunit of this multisubunit enzyme is the binding site of these ions for translocation. Here we established a complementation system for the ntpK gene with its deletion mutant, and found that the ATPase activity disappeared upon replacement of Glu139 by aspartic acid. The side-chain length of this acidic residue of NtpK is thus important for this ATPase reaction
MH  - A
MH  - ACID
MH  - Adenosinetriphosphatase
MH  - atp
MH  - ATPase
MH  - BINDING
MH  - glutamic acid
MH  - Hydrolysis
MH  - ion
MH  - Ions
MH  - mutant
MH  - RESIDUE
MH  - Site
MH  - Sodium
MH  - SUBUNIT
MH  - SYSTEM
MH  - translocation
RP  - NOT IN FILE
NT  - UI - 99356709LA - engRN - 0 (Plasmids)RN - 56-86-0 (Glutamic Acid)RN - 7440-23-5 (Sodium)RN - EC 3.6.1.- (sodium-translocating ATPase)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 19990928IS - 0916-8451SB - IMCY - JAPAN
UR  - PM:10427702
SO  - Biosci Biotechnol Biochem 1999 Jun ;63(6):1125-1129

1634
UI  - 700
AU  - Tsunoda SP
AU  - Muneyuki E
AU  - Amano T
AU  - Yoshida M
AU  - Noji H
AD  - Research Laboratory of Resources Utilization, Tokyo Institute of Technology, 4259 Nagatsuta, Yokohama 226, Japan
TI  - Cross-linking of two beta subunits in the closed conformation in F1- ATPase
AB  - In the crystal structure of mitochondrial F1-ATPase, two beta subunits with a bound Mg-nucleotide are in "closed" conformations, whereas the third beta subunit without bound nucleotide is in an "open" conformation. In this "CCO" (beta-closed beta-closed beta-open) conformational state, Ile-390s of the two closed beta subunits, even though they are separated by an intervening alpha subunit, have a direct contact. We replaced the equivalent Ile of the alpha3beta3gamma subcomplex of thermophilic F1-ATPase with Cys and observed the formation of the beta-beta cross-link through a disulfide bond. The analysis of conditions required for the cross-link formation indicates that: (i) F1-ATPase takes the CCO conformation when two catalytic sites are filled with Mg-nucleotide, (ii) intermediate(s) with the CCO conformation are generated during catalytic cycle, (iii) the Mg-ADP inhibited form is in the CCO conformation, and (iv) F1-ATPase dwells in conformational state(s) other than CCO when only one (or none) of catalytic sites is filled by Mg-nucleotide or when catalytic sites are filled by Mg2+-free nucleotide. The alpha3beta3gamma subcomplex containing the beta-beta cross-link retained the activity of uni-site catalysis but lost that of multiple catalytic turnover, suggesting that open-closed transition of beta subunits is required for the rotation of gamma subunit but not for hydrolysis of a single ATP
RP  - NOT IN FILE
NT  - UI - 99150357LA - engRN - 0 (Adenine Nucleotides)RN - 0 (Azides)RN - 0 (DNA Primers)RN - 7439-95-4 (Magnesium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19990318IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:10026189
SO  - J Biol Chem 1999 Feb 26 ;274(9):5701-5706

1635
UI  - 382
AU  - Vinogradov AD
AD  - Department of Biochemistry, School of Biology, Lomonosov Moscow State University, Moscow, 119899, Russia adv@biochembiomsusu
TI  - Mitochondrial ATP synthase: fifteen years later
AB  - Mitochondrial Fo.F1-H+-ATP synthase is the main enzyme responsible for the formation of ATP in aerobic cells. An alternating binding change mechanism is now generally accepted for the operation of the enzyme. This mechanism apparently leaves no room for the participation of nucleotides and Pi other than sequential binding to (release from) the catalytic sites. However, the kinetics of ATP hydrolysis by mitochondrial ATPase is very complex, and it is difficult to explain it in terms of the alternating binding change mechanism only. Fo.F1 catalyzes both delta muH+-dependent ATP synthesis and ATP-dependent delta muH+ generation. It is generally believed that this enzyme operates as the smallest molecular electromechanochemical reversible machine. This essay summarizes data which contradict this simple reversible mechanism and discusses a hypothesis in which different pathways are followed for ATP hydrolysis and ATP synthesis. A model for a reversible switch mechanism between ATP hydrolase and ATP synthase states of Fo. F1 is proposed
RP  - NOT IN FILE
NT  - UI - 20079351LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 20000128IS - 0006-2979SB - IMCY - RUSSIAJC - CSQ
UR  - PM:10611526
SO  - Biochemistry (Mosc ) 1999 Nov ;64(11):1219-1229

1636
UI  - 408
AU  - Weber J
AU  - Dunn SD
AU  - Senior AE
AD  - Department of Biochemistry and Biophysics, University of Rochester Medical Center, Rochester, New York 14642, USA
TI  - Effect of the epsilon-subunit on nucleotide binding to Escherichia coli F1-ATPase catalytic sites
AB  - The influence of the epsilon-subunit on the nucleotide binding affinities of the three catalytic sites of Escherichia coli F1-ATPase was investigated, using a genetically engineered Trp probe in the adenine-binding subdomain (beta-Trp-331). The interaction between epsilon and F1 was not affected by the mutation. Kd for binding of epsilon to betaY331W mutant F1 was approximately 1 nM, and epsilon inhibited ATPase activity by 90%. The only nucleotide binding affinities that showed significant differences in the epsilon-depleted and epsilon-replete forms of the enzyme were those for MgATP and MgADP at the high-affinity catalytic site 1. Kd1(MgATP) and Kd1(MgADP) were an order of magnitude higher in the absence of epsilon than in its presence. In contrast, the binding affinities for MgATP and MgADP at sites 2 and 3 were similar in the epsilon-depleted and epsilon-replete enzymes, as were the affinities at all three sites for free ATP and ADP. Comparison of MgATP binding and hydrolysis parameters showed that in the presence as well as the absence of epsilon, Km equals Kd3. Thus, in both cases, all three catalytic binding sites have to be occupied to obtain rapid (Vmax) MgATP hydrolysis rates
RP  - NOT IN FILE
NT  - UI - 99315853LA - engRN - 0 (ATPase inhibitory protein)RN - 0 (Bacterial Proteins)RN - 0 (Enzyme Inhibitors)RN - 0 (Ligands)RN - 0 (Proteins)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSDA - 19990727IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:10383416
SO  - J Biol Chem 1999 Jul 2 ;274(27):19124-19128

1637
UI  - 696
AU  - Yagi H
AU  - Tozawa K
AU  - Sekino N
AU  - Iwabuchi T
AU  - Yoshida M
AU  - Akutsu H
AD  - Department of Chemistry and Biotechnology, Faculty of Engineering, Yokohama National University, Yokohama 240-8501, Japan
TI  - Functional conformation changes in the TF(1)-ATPase beta subunit probed by 12 tyrosine residues
AB  - The effect of nucleotide binding on the structure of the F(1)-ATPase beta subunit from thermophilic bacillus PS-3 (TF(1)beta) was investigated by monitoring the NMR signals of the 12 tyrosine residues. The 3,5-proton resonances of 12 tyrosine residues could be observed for the specifically deuterated beta subunit. The assignment of 3,5-proton resonances of all of the tyrosine residues was accomplished using 14 mutant proteins, in each of which one or two tyrosine residues were replaced by phenylalanine. Binding of Mg. ATP induced an upfield shift of Tyr(341) resonance, suggesting that their aromatic rings are stacked to each other. Besides Tyr(341), the signal shift observed on Mg.ATP binding was restricted to the resonances of Tyr(148), Tyr(199), Tyr(238), and Tyr(307), suggesting that Mg.ATP induces a conformational change in the hinge region. This can be correlated to the change from the open to closed conformations as implicated in the crystal structure. Mg.ADP induced a similar but distinctly different conformational change. Therefore, the intrinsic conformational change in the beta subunit induced by the nucleotide binding is proposed to be one of the essential driving forces for the F(1) rotation. Reconstitution experiments showed that Tyr(277), one of the four conserved tyrosines, is essential to the formation of the alpha(3)beta(3)gamma complex
RP  - NOT IN FILE
NT  - UI - 99444145LA - engRN - 0 (Ligands)RN - 0 (Protons)RN - 25612-73-1 (Adenylyl Imidodiphosphate)RN - 55520-40-6 (Tyrosine)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 7782-39-0 (Deuterium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 19991122IS - 0006-3495SB - IMCY - UNITED STATESJC - A5S
UR  - PM:10512836
SO  - Biophys J 1999 Oct ;77(4):2175-2183

1638
UI  - 21234
AU  - Yeh IC
AU  - Berkowitz ML
TI  - Dielectric constant of water at high electric fields: Molecular dynamics study
AB  - Molecular dynamics computer simulations have been carried out for water enclosed between Pt(111) surfaces at high external electric fields. The dielectric constant of water as a function of electric fields has been calculated. Two-dimensional Ewald summation technique has been used for the calculation of long- range Coulombic forces. Simulations with a larger distance between walls, different surfaces, and bulk water have been done to confirm the macroscopic nature of the dielectric constant. Calculated dielectric constants have been compared with those obtained by a theoretical prediction and a recent simulation study. (C) 1999 American Institute of Physics. [S0021-9606(99)51815-1]
MH  - A
MH  - computer simulations
MH  - COMPUTER-SIMULATION
MH  - COMPUTER-SIMULATIONS
MH  - CONSTANT
MH  - FIELD
MH  - function
MH  - LIQUID WATER
MH  - MOLECULAR-DYNAMICS
MH  - SIMULATION
MH  - SIMULATIONS
MH  - SURFACE
MH  - Water
RP  - NOT IN FILE
NT  - JournalAPR 22186QCJ.Chem.Phys.
UR  - ISI:000079739600033
SO  - Journal of Chemical Physics 1999  ;110(16):7935-7942

1639
UI  - 688
AU  - Adachi K
AU  - Yasuda R
AU  - Noji H
AU  - Itoh H
AU  - Harada Y
AU  - Yoshida M
AU  - Kinosita K
AD  - Department of Physics, Faculty of Science, Kanazawa University, Kakuma- machi, Kanazawa 920-1192, Japan
TI  - Stepping rotation of F1-ATPase visualized through angle-resolved single- fluorophore imaging
AB  - Orientation dependence of single-fluorophore intensity was exploited in order to videotape conformational changes in a protein machine in real time. The fluorophore Cy3 attached to the central subunit of F(1)- ATPase revealed that the subunit rotates in the molecule in discrete 120 degrees steps and that each step is driven by the hydrolysis of one ATP molecule. These results, unlike those from the previous study under a frictional load, show that the 120 degrees stepping is a genuine property of this molecular motor. The data also show that the rate of ATP binding is insensitive to the load exerted on the rotor subunit
RP  - NOT IN FILE
NT  - UI - 20319011LA - engRN - 0 (Carbocyanines)RN - 0 (Fluorescent Dyes)RN - 0 (cyanine dye 3)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 20000731IS - 0027-8424SB - IMCY - UNITED STATESJC - PV3
UR  - PM:10840052
SO  - Proc Natl Acad Sci U S A 2000 Jun 20 ;97(13):7243-7247

1640
UI  - 20932
AU  - Altendorf K
AU  - Stalz W
AU  - Greie J
AU  - Deckers-Hebestreit G
AD  - Universitat Osnabruck, Fachbereich Biologie/Chemie, Abteilung Mikrobiologie, D-49069 Osnabruck, Germany
TI  - Structure and function of the F(o) complex of the ATP synthase from Escherichia coli
AB  - The membrane-bound ATP synthase (F(1)F(o)) from mitochondria, chloroplasts and bacteria plays a crucial role in energy-transducing reactions. In the case of Escherichia coli, the reversible, proton- translocating ATPase complex consists of two different entities, F(1) and F(o). The water-soluble F(1) part carries the catalytic sites for ATP synthesis and hydrolysis. It is associated with the membrane- embedded F(o) complex, which functions as a proton channel and consists of subunits a, b and c present in a stoichiometry of 1:2:12.Subunit b was isolated by preparative gel electrophoresis, acetone-precipitated and renatured in a cholate-containing buffer. Reconstituted subunit b together with purified ac subcomplex is active in proton translocation and F(1) binding, thereby demonstrating that subunit b had recovered its native conformation. Circular dichroism spectroscopy of subunit b reconstituted into liposomes revealed a rather high degree of alpha - helical conformation of 80%. After addition of a His(6)-tag to the N terminus of subunit a, a stable ab(2) subcomplex was purified instead of a single subunit a, arguing in favour of a direct interaction between these subunits. After addition of subunit c and reconstitution into phospholipid vesicles, an F(o) complex was obtained exhibiting rates of proton translocation and F(1) binding comparable with those of wild-type F(o).The epitopes of monoclonal antibodies against subunit c are located in the hydrophilic loop region (cL31-Q42) as mapped by enzyme-linked immunosorbent assay using overlapping synthetic heptapeptides. Binding studies revealed that all monoclonal antibodies (mAbs) bind to everted membrane vesicles irrespective of the presence or absence of F(1). Although the hydrophilic region of subunit c, and especially the highly conserved residues cA40, cR41, cQ42 and cP43, are known to interact with subunits gamma and epsilon of the F(1) part, the mAb molecules have no effect on the function of F(o), either in proton translocation or in F(1) binding. However, the F(1) part and the mAb molecule(s) are bound simultaneously to the F(o) complex, suggesting that not all c subunits are involved in the interaction with F(1)
MH  - A
MH  - ACTIVE
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - ATPase
MH  - Bacteria
MH  - BINDING
MH  - buffer
MH  - chloroplast
MH  - Chloroplasts
MH  - COMPLEX
MH  - conformation
MH  - Electrophoresis
MH  - England
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - function
MH  - Hydrolysis
MH  - Liposomes
MH  - membrane
MH  - membrane vesicles
MH  - Mitochondria
MH  - MONOCLONAL-ANTIBODIES
MH  - proton
MH  - reconstitution
MH  - RESIDUE
MH  - review
MH  - Site
MH  - spectroscopy
MH  - SUBUNIT
MH  - SYNTHASE
MH  - synthesis
MH  - translocation
RP  - NOT IN FILE
NT  - UI - 20123947LA - engRN - 0 (Antibodies, Monoclonal)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 20000217IS - 0022-0949SB - IMCY - ENGLAND
UR  - PM:10600669
SO  - J Exp Biol 2000 Jan ;203 Pt 1():19-28

1641
UI  - 318
AU  - Arechaga I
AU  - Miroux B
AU  - Karrasch S
AU  - Huijbregts R
AU  - de Kruijff B
AU  - Runswick MJ
AU  - Walker JE
AD  - The Medical Research Council Dunn Human Nutrition Unit, Cambridge, UK
TI  - Characterisation of new intracellular membranes in Escherichia coli accompanying large scale over-production of the b subunit of F(1)F(o) ATP synthase
AB  - Recombinant membrane proteins in Escherichia coli are either expressed at relatively low level in the cytoplasmic membrane or they accumulate as inclusion bodies. Here, we report that the abundant over-production of subunit b of E. coli F(1)F(o) ATP synthase in the mutant host strains E. coli C41(DE3) and C43(DE3) is accompanied by the proliferation of intracellular membranes without formation of inclusion bodies. Maximal levels of proliferation of intracellular membranes were observed in C43(DE3) cells over-producing subunit b. The new proliferated membranes contained all the over-expressed protein and could be recovered by a single centrifugation step. Recombinant subunit b represented up to 80% of the protein content of the membranes. The lipid:protein ratios and phospholipid compositions of the intracellular membranes differ from those of bacterial cytoplasmic membranes, and they are particularly rich in cardiolipin
RP  - NOT IN FILE
NT  - UI - 20480609LA - engRN - 0 (Lipids)RN - 0 (Peptide Fragments)RN - 0 (Phospholipids)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 20001023IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:11024463
SO  - FEBS Lett 2000 Oct 6 ;482(3):215-219

1642
UI  - 691
AU  - Bald D
AU  - Noji H
AU  - Stumpp MT
AU  - Yoshida M
AU  - Hisabori T
AD  - Chemical Resources Laboratory, Tokyo Institute of Technology, Nagatsuta 4259, Midori-ku, Yokohama 226-8503, Japan
TI  - ATPase activity of a highly stable alpha(3)beta(3)gamma subcomplex of thermophilic F(1) can be regulated by the introduced regulatory region of gamma subunit of chloroplast F(1)
AB  - A mutant F(1)-ATPase alpha(3)beta(3)gamma subcomplex from the thermophilic Bacillus PS3 was constructed, in which 111 amino acid residues (Val(92) to Phe(202)) from the central region of the gamma subunit were replaced by the 148 amino acid residues of the homologous region from spinach chloroplast F(1)-ATPase gamma subunit, including the regulatory stretch, and were designated as alpha(3)beta(3)gamma((TCT)) (Thermophilic-Chloroplast-Thermophilic). By the insertion of this regulatory region into the gamma subunit of thermophilic F(1), we could confer the thiol modulation property to the thermophilic alpha(3)beta(3)gamma subcomplex. The overexpressed alpha(3)beta(3)gamma((TCT)) was easily purified in large scale, and the ATP hydrolyzing activity of the obtained complex was shown to increase up to 3-fold upon treatment with chloroplast thioredoxin-f and dithiothreitol. No loss of thermostability compared with the wild type subcomplex was found, and activation by dithiothreitol was functional at temperatures up to 80 degrees C. alpha(3)beta(3)gamma((TCT)) was inhibited by the epsilon subunit from chloroplast F(1)-ATPase but not by the one from the thermophilic F(1)-ATPase, indicating that the introduced amino acid residues from chloroplast F(1)-gamma subunit are important for functional interaction with the epsilon subunit
RP  - NOT IN FILE
NT  - UI - 20239926LA - engRN - 0 (Amino Acids)RN - 0 (Plasmids)RN - 0 (Recombinant Proteins)RN - 3483-12-3 (Dithiothreitol)RN - 52500-60-4 (Thioredoxin)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 20000602IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:10777572
SO  - J Biol Chem 2000 Apr 28 ;275(17):12757-12762

1643
UI  - 9881
AU  - Bottcher B
AU  - Graber P
TI  - The structure of the H+-ATP synthase from chloroplasts and its subcomplexes as revealed by electron microscopy
AB  - The electron microscopic data available on CF0F1 and its subcomplexes, CF0, CF1, subunit III complex are collected and the CF1 data are compared with the high resolution structure of MF1. The data are based on electron microscopic investigation of negatively stained isolated CF1, CF0F1 and subunit III complex. In addition, two-dimensional crystals of CF0F1 and CF0F1 reconstituted liposomes were investigated by cryo- electron microscopy. Progress in the interpretation of electron microscopic data from biological samples has been made with the introduction of image analysis. Multi-reference alignment and classification of images have led to the differentiation between different conformational states and to the detection of a second stalk. Recently, the calculation of three-dimensional mars from the class averages led to the understanding of the spatial organisation of the enzyme. Such three-dimensional maps give evidence of the existence of a third connection between the F-0 part and F-1 part. (C) 2000 Elsevier Science B.V. All rights reserved
MH  - adenosine triphosphate synthase
MH  - analysis
MH  - ATOMIC-FORCE MICROSCOPY
MH  - CF0F1
MH  - chloroplast
MH  - Chloroplasts
MH  - COUPLING FACTOR-I
MH  - CROSS- LINKING
MH  - CRYOELECTRON MICROSCOPY
MH  - electron
MH  - electron microscopy
MH  - EPSILON-SUBUNIT
MH  - ESCHERICHIA-COLI F1-ATPASE
MH  - H+- translocating ATPase from chloroplast
MH  - H+-ATPase
MH  - IMMUNOELECTRON MICROSCOPY
MH  - Liposomes
MH  - Microscopy
MH  - MITOCHONDRIAL F1-ATPASE
MH  - MONOCLONAL-ANTIBODIES
MH  - review
MH  - SPINACH-CHLOROPLASTS
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - JournalReviewELSEVIER SCIENCE BVMAY 31327HXAMSTERDAMGraber P Inst Phys Chem, Albertstr 23A, D-79104 Freiburg, GermanyBBA-BIOENERGETICSPO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS
AV  - Inst Phys Chem, Albertstr 23A, D-79104 Freiburg, Germany Inst Phys Chem, D-79104 Freiburg, Germany European Mol Biol Lab, D-69012 Heidelberg, Germany
UR  - ISI:000087789400016
SO  - Biochimica et Biophysica Acta-Bioenergetics 2000  ;1458(2-3):404-416

1644
UI  - 20986
AU  - Bottcher B
AU  - Graber P
AD  - European Molecular Biology Laboratory, Heidelberg, Germany
TI  - The structure of the H(+)-ATP synthase from chloroplasts and its subcomplexes as revealed by electron microscopy
AB  - The electron microscopic data available on CF(0)F(1) and its subcomplexes, CF(0), CF(1), subunit III complex are collected and the CF(1) data are compared with the high resolution structure of MF(1). The data are based on electron microscopic investigation of negatively stained isolated CF(1), CF(0)F(1) and subunit III complex. In addition, two-dimensional crystals of CF(0)F(1) and CF(0)F(1) reconstituted liposomes were investigated by cryo-electron microscopy. Progress in the interpretation of electron microscopic data from biological samples has been made with the introduction of image analysis. Multi-reference alignment and classification of images have led to the differentiation between different conformational states and to the detection of a second stalk. Recently, the calculation of three-dimensional maps from the class averages led to the understanding of the spatial organisation of the enzyme. Such three-dimensional maps give evidence of the existence of a third connection between the F(0) part and F(1) part
MH  - A
MH  - analysis
MH  - ATPase
MH  - chloroplast
MH  - Chloroplasts
MH  - COMPLEX
MH  - CRYOELECTRON MICROSCOPY
MH  - electron
MH  - electron microscopy
MH  - ELECTRON-MICROSCOPY
MH  - Liposomes
MH  - Microscopy
MH  - Proteolipids
MH  - proteoliposome
MH  - resolution
MH  - review
MH  - stalk
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 20298319LA - engRN - 0 (Proteolipids)RN - 0 (proteoliposomes)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticlePT - ReviewPT - Review, AcademicDA - 20000727IS - 0006-3002SB - IMCY - NETHERLANDS
UR  - PM:10838054
SO  - Biochim Biophys Acta 2000 May 31 ;1458(2-3):404-416

1645
UI  - 20988
AU  - Bottcher B
AU  - Bertsche I
AU  - Reuter R
AU  - Graber P
AD  - Institut fur Physikalische Chemie, Albertstrasse 23a, Universitat Freiburg, D-79104, Germany bottcher@embl-heidelbergde
TI  - Direct visualisation of conformational changes in EF(0)F(1) by electron microscopy
AB  - The isolated H(+)-ATPase from Escherichia coli (EF(0)F(1)) was investigated by electron microscopy of samples of negatively stained monodisperse molecules, followed by single-particle image processing. The resulting three-dimensional maps showed that the F(1)-part is connected by a prominent stalk to a more peripheral part of F(0). The F(1)-part showed stain-accessible cavities inside. In three-dimensional maps from selected particles, a second stalk could be detected which was thinner than the main stalk and is thought to correspond to the stator.Three-dimensional maps of the enzyme in the absence and in the presence of the substrate analogue adenyl-beta, gamma-imidodiphosphate (AMP-PNP) were calculated. Upon binding of AMP-PNP the three- dimensional maps showed no significant changes in the F(0)-part of EF(0)F(1), whereas a major conformational change in the F(1)-part was observed. (1) The diameter of the F(1)-part decreased upon binding of AMP-PNP mainly in the upper half of F(1). (2) Enzyme particles prepared in the presence of AMP-PNP had a pointed cap at the top of the F(1)- part which was missing in its absence. (3) The stain-accessible cavity inside the F(1)-part altered its pattern significantly
MH  - A
MH  - ATPase
MH  - BINDING
MH  - conformational change
MH  - electron
MH  - electron microscopy
MH  - ELECTRON-MICROSCOPY
MH  - England
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - H(+)ATPase
MH  - image processing
MH  - Microscopy
MH  - stalk
RP  - NOT IN FILE
NT  - UI - 20135980LA - engRN - 25612-73-1 (Adenylyl Imidodiphosphate)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 20000323IS - 0022-2836SB - IMCY - ENGLAND
UR  - PM:10669600
SO  - J Mol Biol 2000 Feb 18 ;296(2):449-457

1646
UI  - 9883
AU  - Bottcher B
AU  - Bertsche I
AU  - Reuter R
AU  - Graber P
TI  - Direct visualisation of conformational changes in EF0F1 by electron microscopy
AB  - The isolated H+-ATPase from Escherichia coli (EF0F1) was investigated by electron microscopy of samples of negatively stained monodisperse molecules, followed by single-particle image processing. The resulting three-dimensional maps showed that the F-1-part is connected by a prominent stalk to a more peripheral part of F-0. The F-1-part showed stain-accessible cavities inside. In three-dimensional maps from selected particles, a second stalk could be detected which was thinner than the main stalk and is thought to correspond to the stator. Three-dimensional maps of the enzyme in the absence and in the presence of the substrate analogue adenyl-beta,gamma- imidodiphosphate (AMP-PNP) were calculated. Upon binding of AMP-PNP the three-dimensional maps showed no significant changes in the F-0-part of EF0F1, whereas a major conformational change in the F-1-part was observed. (1) The diameter of the F-1-part decreased upon binding of AMP-PNP mainly in the upper half of F-1. (2) Enzyme particles prepared in the presence of AMP-PNP had a pointed cap at the top of the F-1-part which was missing in its absence. (3) The stain- accessible cavity inside the F-1-part altered its pattern significantly. (C) 2000 Academic Press
MH  - 3D-map
MH  - ALPHA-SUBUNIT
MH  - COLI ATP SYNTHASE
MH  - conformational change
MH  - CROSS-LINKING
MH  - DELTA-SUBUNIT
MH  - electron
MH  - electron microscopy
MH  - England
MH  - EPSILON-SUBUNIT
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F-0 PARTS
MH  - F-ATPASE
MH  - F1
MH  - F1F0-ATP SYNTHASE
MH  - H+-ATPase
MH  - IMAGE DATA
MH  - image processing
MH  - Microscopy
RP  - NOT IN FILE
NT  - JournalArticleACADEMIC PRESS LTDFEB 18290VQLONDONBottcher B European Mol Biol Lab, Meyerhofstr 1,Postfach 102209, D-69012 Heidelberg, GermanyJ MOL BIOL24-28 OVAL RD, LONDON NW1 7DX, ENGLAND
AV  - European Mol Biol Lab, Meyerhofstr 1,Postfach 102209, D-69012 Heidelberg, Germany Univ Freiburg, Inst Phys Chem, D-79104 Freiburg, Germany
UR  - ISI:000085698900011
SO  - Journal of Molecular Biology 2000  ;296(2):449-457

1647
UI  - 19767
AU  - Boyer PD
AD  - Molecular Biology Institute, University of California at Los Angeles, Los Angeles, CA 90095-1570, USA pdboyer@uclaedu
TI  - Catalytic site forms and controls in ATP synthase catalysis
AB  - A suggested minimal scheme for substrate binding by and interconversion of three forms of the catalytic sites of the ATP synthase is presented. Each binding change, that drives simultaneous interchange of the three catalytic site forms, requires a 120 degrees rotation of the gamma with respect to the beta subunits. The binding of substrate(s) at two catalytic sites is regarded as sufficing for near maximal catalytic rates to be attained. Although three sites do not need to be filled for rapid catalysis, during rapid bisite catalysis some enzyme may be transiently present with three sites filled. Forms with preferential binding for ADP and P(i) or for ATP are considered to arise from the transition state and participate in other steps of the catalysis. Intermediate forms and steps that may be involved are evaluated. Experimental evidence for energy-dependent steps and for control of coupling to proton translocation and transition state forms are reviewed. Impact of relevant past data on present understanding of catalytic events is considered. In synthesis a key step is suggested in which proton translocation begins to deform an open site so as to increase the affinity for ADP and P(i), that then bind and pass through the transition state, and yield tightly bound ATP in one binding change. ADP binding appears to be a key parameter controlling rotation during synthesis. In hydrolysis ATP binding to a loose site likely precedes any proton translocation, with proton movement occurring as the tight site form develops. Aspects needing further study are noted. Characteristics of the related MgADP inhibition of the F(1) ATPases that have undermined many observations are summarized, and relations of three-site filling to catalysis are assessed
MH  - A
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - ADP
MH  - ADP BINDING
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BETA-SUBUNIT
MH  - BINDING
MH  - Catalysis
MH  - COMPLEX
MH  - H(+)-Transporting ATP Synthase
MH  - Hydrolysis
MH  - Movement
MH  - Multienzyme Complexes
MH  - Phosphates
MH  - Phosphotransferases
MH  - proton
MH  - Protons
MH  - review
MH  - rotation
MH  - SUBUNIT
MH  - SYNTHASE
MH  - synthesis
MH  - translocation
RP  - NOT IN FILE
NT  - UI - 20298306LA - engRN - 0 (Molecular Motors)RN - 0 (Multienzyme Complexes)RN - 0 (Phosphates)RN - 0 (Protons)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 2.7.4 (Phosphotransferases (Phosphate Group Acceptor))RN - EC 2.7.4.- (ATP synthetase complex)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 20000727IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:10838041
SO  - Biochim Biophys Acta 2000 May 31 ;1458(2-3):252-262

1648
UI  - 320
AU  - Braig K
AU  - Menz RI
AU  - Montgomery MG
AU  - Leslie AG
AU  - Walker JE
AD  - Institut fur Organische Chemie und Biochemie, Albert-Ludwigs Universitat Freiburg, Freiburg in Breisgau, D-79104, Germany
TI  - Structure of bovine mitochondrial F(1)-ATPase inhibited by Mg(2+) ADP and aluminium fluoride
AB  - BACKGROUND: The globular domain of the membrane-associated F(1)F(o)-ATP synthase complex can be detached intact as a water-soluble fragment known as F(1)-ATPase. It consists of five different subunits, alpha, beta, gamma, delta and epsilon, assembled with the stoichiometry 3:3:1:1:1. In the crystal structure of bovine F(1)-ATPase determined previously at 2.8 A resolution, the three catalytic beta subunits and the three noncatalytic alpha subunits are arranged alternately around a central alpha-helical coiled coil in the gamma subunit. In the crystals, the catalytic sites have different nucleotide occupancies. One contains the triphosphate form of the nucleotide, the second contains the diphosphate, and the third is unoccupied. Fluoroaluminate complexes have been shown to mimic the transition state in several ATP and GTP hydrolases. In order to understand more about its catalytic mechanism, F(1)-ATPase was inhibited with Mg(2+)ADP and aluminium fluoride and the structure of the inhibited complex was determined by X- ray crystallography. RESULTS: The structure of bovine F(1)-ATPase inhibited with Mg(2+)ADP and aluminium fluoride determined at 2.5 A resolution differs little from the original structure with bound AMP- PNP and ADP. The nucleotide occupancies of the alpha and beta subunits are unchanged except that both aluminium trifluoride and Mg(2+)ADP are bound in the nucleotide-binding site of the beta(DP) subunit. The presence of aluminium fluoride is accompanied by only minor adjustments in the surrounding protein. CONCLUSIONS: The structure appears to mimic a possible transition state. The coordination of the aluminofluoride group has many features in common with other aluminofluoride-NTP hydrolase complexes. Apparently, once nucleotide is bound to the catalytic beta subunit, no additional major structural changes are required for catalysis to occur
RP  - NOT IN FILE
NT  - UI - 20335018LA - engRN - 0 (Aluminum Compounds)RN - 0 (Enzyme Inhibitors)RN - 0 (Fluorides)RN - 58-64-0 (Adenosine Diphosphate)RN - 7784-18-1 (aluminum fluoride)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 20000830IS - 0969-2126SB - IMCY - ENGLANDJC - DEB
UR  - PM:10873854
SO  - Structure Fold Des 2000 Jun 15 ;8(6):567-573

1649
UI  - 21303
AU  - Brandsburg-Zabary S
AU  - Fried O
AU  - Marantz Y
AU  - Nachliel E
AU  - Gutman M
AD  - Laser Laboratory for Fast reactions in Biology, Biochemistry, Tel Aviv University, Tel Aviv, Israel
TI  - Biophysical aspects of intra-protein proton transfer
AB  - The passage of proton trough proteins is common to all membranal energy conserving enzymes. While the routes differ among the various proteins, the mechanism of proton propagation is based on the same chemical-physical principles. The proton progresses through a sequence of dissociation association steps where the protein and water molecules function as a solvent that lowers the energy penalty associated with the generation of ions in the protein. The propagation of the proton in the protein is a random walk, between the temporary proton binding sites that make the conducting path, that is biased by the intra-protein electrostatic potential. Kinetic measurements of proton transfer reactions, in the sub-ns up to micros time frame, allow to monitor the dynamics of the partial reactions of an overall proton transfer through a protein
MH  - A
MH  - Arylsulfonates
MH  - BINDING
MH  - BINDING SITE
MH  - Binding Sites
MH  - Biochemistry
MH  - biology
MH  - DYE
MH  - dyes
MH  - energy
MH  - enzyme
MH  - Enzymes
MH  - Fluorescent Dyes
MH  - function
MH  - ion
MH  - Ion Channels
MH  - Ions
MH  - mechanism
MH  - protein
MH  - Proteins
MH  - proton
MH  - Proton Pump
MH  - Proton transfer
MH  - Proton-transfer
MH  - Protons
MH  - review
MH  - Site
MH  - SOLVENT
MH  - Time
MH  - TRANSFER
MH  - universities
MH  - Water
RP  - NOT IN FILE
NT  - UI - 20273713DA - 20000630IS - 0006-3002LA - engPT - Journal ArticlePT - ReviewPT - Review, TutorialCY - NETHERLANDSRN - 0 (Arylsulfonates)RN - 0 (Fluorescent Dyes)RN - 0 (Ion Channels)RN - 0 (Porins)RN - 0 (Proteins)RN - 0 (Proton Pumps)RN - 0 (Protons)RN - 6358-69-6 (pyranine)RN - 7732-18-5 (Water)RN - 85130-31-0 (PhoE protein)SB - IM
UR  - PM:10812028
SO  - Biochim Biophys Acta 2000 May 12 ;1458(1):120-134

1650
UI  - 319
AU  - Cabezon E
AU  - Arechaga I
AU  - Jonathan P
AU  - Butler G
AU  - Walker JE
AD  - Medical Research Council Dunn Human Nutrition Unit, Cambridge CB2 2XY and The Medical Research Council Laboratory of Molecular Biology, Cambridge CB2 2QH, United Kingdom
TI  - Dimerization of bovine F1-ATPase by binding the inhibitor protein, IF1
AB  - In mitochondria, the hydrolytic activity of ATP synthase is regulated by a natural inhibitor protein, IF(1). The binding of IF(1) to ATP synthase depends on pH values, and below neutrality, IF(1) forms a stable complex with the enzyme. Bovine IF(1) has two oligomeric states, dimer and tetramer, depending on pH values. At pH 6.5, where it is active, IF(1) dimerizes by formation of an antiparallel alpha-helical coiled-coil in its C-terminal region. This arrangement places the inhibitory N-terminal regions in opposition, implying that active dimeric IF(1) can bind two F(1) domains simultaneously. Evidence of dimerization of F(1)-ATPase by binding to IF(1) is provided by gel filtration chromatography, analytical ultracentrifugation, and electron microscopy. At present, it is not known whether IF(1) can bring about the dimerization of the F(1)F(0)-ATPase complex
RP  - NOT IN FILE
NT  - UI - 20435783LA - engRN - 0 (Enzyme Inhibitors)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 20001013IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:10918052
SO  - J Biol Chem 2000 Sep 15 ;275(37):28353-28355

1651
UI  - 322
AU  - Cabezon E
AU  - Butler PJ
AU  - Runswick MJ
AU  - Walker JE
AD  - Medical Research Council Dunn Human Nutrition Unit, Cambridge, United Kingdom
TI  - Modulation of the oligomerization state of the bovine F1-ATPase inhibitor protein, IF1, by pH
AB  - Bovine IF(1), a basic protein of 84 amino acids, is involved in the regulation of the catalytic activity of the F(1) domain of ATP synthase. At pH 6.5, but not at basic pH values, it inhibits the ATP hydrolase activity of the enzyme. The oligomeric state of bovine IF(1) has been investigated at various pH values by sedimentation equilibrium analytical ultracentrifugation and by covalent cross-linking. Both techniques confirm that the protein forms a tetramer at pH 8, and below pH 6.5, the protein is predominantly dimeric. By covalent cross- linking, it has been found that at pH 8.0 the fragment of IF(1) consisting of residues 44-84 forms a dimer, whereas the fragment from residues 32-84 is tetrameric. Therefore, some or all of the residues between positions 32 and 43 are necessary for tetramer formation and are involved in the pH-sensitive interconversion between dimer and tetramer. One important residue in the interconversion is histidine 49. Mutation of this residue to lysine abolishes the pH-dependent activation-inactivation, and the mutant protein is active and dimeric at all pH values investigated. It is likely from NMR studies that the inhibitor protein dimerizes by forming an antiparallel alpha-helical coiled-coil over its C-terminal region and that at high pH values, where the protein is tetrameric, the inhibitory regions are masked. The mutation of histidine 49 to lysine is predicted to abolish coiled-coil formation over residues 32-43 preventing interaction between two dimers, forcing the equilibrium toward the dimeric state, thereby freeing the N-terminal inhibitory regions and allowing them to interact with F(1)
RP  - NOT IN FILE
NT  - UI - 20400477LA - engRN - 0 (ATPase inhibitory protein)RN - 0 (Amino Acids)RN - 0 (Cross-Linking Reagents)RN - 0 (Plasmids)RN - 0 (Proteins)RN - 0 (Recombinant Proteins)RN - 56-87-1 (Lysine)RN - 7006-35-1 (Histidine)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 20000921IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:10831597
SO  - J Biol Chem 2000 Aug 18 ;275(33):25460-25464

1652
UI  - 20808
AU  - Cain BD
AD  - Department of Biochemistry and Molecular Biology, University of Florida, Gainesville 32610, USA bcain@biochemmedufledu
TI  - Mutagenic analysis of the F0 stator subunits
AB  - The a and b subunits constitute the stator elements in the F0 sector of F1F0-ATP synthase. Both subunits have been difficult to study by physical means, so most of the information on structure and function relationships in the a and b subunits has been obtained using mutagenesis in combination with biochemical methods. These approaches were used to demonstrate that the a subunit in association with the ring of c subunits houses the proton channel through F1F0-ATP synthase. The map of the amino acids contributing to the proton channel is probably complete. The two b subunits dimerize, forming an extended flexible unit in the peripheral stalk linking the F1 and F0 sectors. The unique characteristics of specific amino acid substitutions affecting the a and b subunits suggested differential effects on rotation during F1F0-ATPase activity
MH  - A
MH  - ACID
MH  - Amino Acids
MH  - analysis
MH  - Biochemistry
MH  - F0
MH  - F1
MH  - F1F0-ATP SYNTHASE
MH  - function
MH  - method
MH  - Methods
MH  - mutagenesis
MH  - proton
MH  - rotation
MH  - stalk
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 21618402LA - engPT - Journal ArticleID - GM43495/GM/NIGMSDA - 20011220IS - 0145-479XSB - IMCY - United States
UR  - PM:11768298
SO  - J Bioenerg Biomembr 2000 Aug ;32(4):365-371

1653
UI  - 21414
AU  - Cain BD
TI  - Mutagenic analysis of the F0 stator subunits
AB  - The a and b subunits constitute the stator elements in the F0 sector of F1F0-ATP synthase. Both subunits have been difficult to study by physical means, so most of the information on structure and function relationships in the a and b subunits has been obtained using mutagenesis in combination with biochemical methods. These approaches were used to demonstrate that the a subunit in association with the ring of c subunits houses the proton channel through F1F0-ATP synthase. The map of the amino acids contributing to the proton channel is probably complete. The two b subunits dimerize, forming an extended flexible unit in the peripheral stalk linking the F1 and F0 sectors. The unique characteristics of specific amino acid substitutions affecting the a and b subunits suggested differential effects on rotation during F1F0-ATPase activity
MH  - A
MH  - ACID
MH  - Amino Acid Substitution
MH  - Amino Acids
MH  - analysis
MH  - Bacteria
MH  - Biochemistry
MH  - biology
MH  - Chemistry
MH  - enzymology
MH  - F0
MH  - F1
MH  - F1F0-ATP SYNTHASE
MH  - function
MH  - Human
MH  - metabolism
MH  - method
MH  - Methods
MH  - mutagenesis
MH  - Mutation
MH  - Protein Subunits
MH  - proton
MH  - Proton-Translocating ATPases
MH  - Recombinant Proteins
MH  - rotation
MH  - stalk
MH  - structure
MH  - SUBUNIT
MH  - Support,U.S.Gov't,P.H.S.
MH  - SYNTHASE
MH  - universities
RP  - NOT IN FILE
NT  - Department of Biochemistry and Molecular Biology, University of Florida, Gainesville 32610, USA bcain@biochemmedufleduFAU - Cain, B D
SO  - J Bioenerg Biomembr 2000 Aug ;32(4):365-371

1654
UI  - 137
AU  - Capaldi RA
AU  - Schulenberg B
AU  - Murray J
AU  - Aggeler R
AD  - Institute of Molecular Biology, University of Oregon, Eugene, OR 97403- 1229, USA rcapaldi@oregonuoregonedu
TI  - Cross-linking and electron microscopy studies of the structure and functioning of the Escherichia coli ATP synthase
AB  - ATP synthase, also called F(1)F(o)-ATPase, catalyzes the synthesis of ATP during oxidative phosphorylation. The enzyme is reversible and is able to use ATP to drive a proton gradient for transport purposes. Our work has focused on the enzyme from Escherichia coli (ECF(1)F(o)). We have used a combination of methods to study this enzyme, including electron microscopy and chemical cross-linking. The utility of these two approaches in particular, and the important insights they give into the structure and mechanism of the ATP synthase, are reviewed
RP  - NOT IN FILE
NT  - UI - 20123948LA - engRN - 0 (Cross-Linking Reagents)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialID - HL 24526/HL/NHLBIID - HL 58671/HL/NHLBIDA - 20000217IS - 0022-0949SB - IMCY - ENGLANDJC - I2F
UR  - PM:10600670
SO  - J Exp Biol 2000 Jan ;203 Pt 1():29-33

1655
UI  - 134
AU  - Capaldi RA
TI  - The changing face of mitochondrial research
RP  - NOT IN FILE
NT  - UI - 20245677LA - engRN - 0 (DNA, Mitochondrial)PT - EditorialDA - 20000628IS - 0968-0004SB - IMCY - ENGLANDJC - WEF
UR  - PM:10782087
SO  - Trends Biochem Sci 2000 May ;25(5):212-214

1656
UI  - 133
AU  - Capaldi RA
AU  - Schulenberg B
AD  - Institute of Molecular Biology, University of Oregon, Eugene, OR 97403, USA
TI  - The epsilon subunit of bacterial and chloroplast F(1)F(0) ATPases. Structure, arrangement, and role of the epsilon subunit in energy coupling within the complex
AB  - Recent studies show that the epsilon subunit of bacterial and chloroplast F(1)F(0) ATPases is a component of the central stalk that links the F(1) and F(0) parts. This subunit interacts with alpha, beta and gamma subunits of F(1) and the c subunit ring of F(0). Along with the gamma subunit, epsilon is a part of the rotor that couples events at the three catalytic sites sequentially with proton translocation through the F(0) part. Structural data on the epsilon subunit when separated from the complex and in situ are reviewed, and the functioning of this polypeptide in coupling within the ATP synthase is considered
RP  - NOT IN FILE
NT  - UI - 20298307LA - engRN - 0 (Bacterial Proteins)RN - 0 (Multienzyme Complexes)RN - 0 (Plant Proteins)RN - EC 2.7.4 (Phosphotransferases (Phosphate Group Acceptor))RN - EC 2.7.4.- (ATP synthetase complex)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialID - HL 24526/HL/NHLBIDA - 20000727IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:10838042
SO  - Biochim Biophys Acta 2000 May 31 ;1458(2-3):263-269

1657
UI  - 129
AU  - Carrozzo R
AU  - Murray J
AU  - Capuano O
AU  - Tessa A
AU  - Chichierchia G
AU  - Neglia MR
AU  - Capaldi RA
AU  - Santorelli FM
AD  - Department of Molecular Medicine and Neurology, IRCCS Bambino Gesu, I- 00165 Rome, Italy
TI  - A novel mtDNA mutation in the ATPase6 gene studied by E. coli modeling
AB  - This study aimed to understand the pathogenesis of a new mtDNA-related etiology of Leigh syndrome. We identified the T9176G mutation as the molecular basis of Leigh syndrome in a child and looked for alterations in cellular ATP production. We then modeled the new mtDNA mutation in E. coli and analyzed ATP synthesis, hydrolysis, and the ability of the mutated enzyme to pump protons. Our results suggest that the T9176G change results in a novel, fully assembled enzyme which inhibits the holoenzyme probably by blocking the proton pathway
RP  - NOT IN FILE
NT  - UI - 21274972LA - engPT - Journal ArticleDA - 20010530IS - 1590-1874SB - IMCY - ItalyJC - DRB
UR  - PM:11382202
SO  - Neurol Sci 2000  ;21(5 Suppl):S983-S984

1658
UI  - 132
AU  - Carrozzo R
AU  - Murray J
AU  - Santorelli FM
AU  - Capaldi RA
AD  - Unita di Medicina Moleculare, Ospedale Pediatrico, Rome, Italy
TI  - The T9176G mutation of human mtDNA gives a fully assembled but inactive ATP synthase when modeled in Escherichia coli
AB  - A new mutation in human F(1)F(0) ATPase6, T9176G, which changes Leu 217 to an Arg, has been described in two siblings with Leigh syndrome [Carrozzo et al. (2000) Neurology, in press]. This mutation was modeled in Escherichia coli by changing Leu 259 (the equivalent residue) to Arg and the properties of the altered ECF(1)F(0) were compared to those of previously characterized ATPase6 mutants also modeled in the E. coli enzyme. The L259R change produced a fully assembled ECF(1)F(0) which had no significant ATP hydrolysis, ATP synthesis or proton pumping functions. This is very different from previously described human ATPase6 mutations. The presence of Arg at position 259 in subunit a did not make membranes permeable to protons. We conclude that the mutation inhibits functioning by blocking the rotary motor action of the enzyme
RP  - NOT IN FILE
NT  - UI - 20570166LA - engRN - 0 (DNA, Mitochondrial)RN - 0 (Molecular Motors)RN - 53-84-9 (NAD)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.- (Atp6 protein (ATPase))RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 20010118IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:11119722
SO  - FEBS Lett 2000 Dec 15 ;486(3):297-299

1659
UI  - 21232
AU  - Cherepanov DA
AU  - Bibikov SI
AU  - Bibikova MV
AU  - Bloch DA
AU  - Drachev LA
AU  - Gopta OA
AU  - Oesterhelt D
AU  - Semenov AY
AU  - Mulkidjanian AY
AD  - Institute of Electrochemistry, Russian Academy of Sciences, Moscow
TI  - Reduction and protonation of the secondary quinone acceptor of Rhodobacter sphaeroides photosynthetic reaction center: kinetic model based on a comparison of wild-type chromatophores with mutants carrying Arg-->Ile substitution at sites 207 and 217 in the L-subunit
AB  - After the light-induced charge separation in the photosynthetic reaction center (RC) of Rhodobacter sphaeroides, the electron reaches, via the tightly bound ubiquinone QA, the loosely bound ubiquinone Q(B) After two subsequent flashes of light, Q(B) is reduced to ubiquinol Q(B)H2, with a semiquinone anion Q-(B) formed as an intermediate after the first flash. We studied Q(B)H2 formation in chromatophores from Rb. sphaeroides mutants that carried Arg-->Ile substitution at sites 207 and 217 in the L-subunit. While Arg-L207 is 17 A away from Q(B), Arg- L217 is closer (9 A) and contacts the Q(B)-binding pocket. From the pH dependence of the charge recombination in the RC after the first flash, we estimated deltaG(AB), the free energy difference between the Q- (A)Q(B) and Q(A)Q-(B) states, and pK212, the apparent pK of Glu-L212, a residue that is only 4 A away from Q(B). As expected, the replacement of positively charged arginines by neutral isoleucines destabilized the Q-(B) state in the L217RI mutant to a larger extent than in the L207RI one. Also as expected, pK212 increased by approximately 0.4 pH units in the L207RI mutant. The value of pK212 in the L217RI mutant decreased by 0.3 pH units, contrary to expectations. The rate of the Q-(A)Q-(B)-- >Q(A)Q(B)H2 transition upon the second flash, as monitored by electrometry via the accompanying changes in the membrane potential, was two times faster in the L207RI mutant than in the wild-type, but remained essentially unchanged in the L217RI mutant. To rationalize these findings, we developed and analyzed a kinetic model of the Q-(A)Q- (B)-->Q(A)Q(B)H2 transition. The model properly described the available experimental data and provided a set of quantitative kinetic and thermodynamic parameters of the Q(B) turnover. The non-electrostatic, 'chemical' affinity of the QB site to protons proved to be as important for the attracting protons from the bulk, as the appropriate electrostatic potential. The mutation-caused changes in the chemical proton affinity could be estimated from the difference between the experimentally established pK2J2 shifts and the expected changes in the electrostatic potential at Glu-L212, calculable from the X-ray structure of the RC. Based on functional studies, structural data and kinetic modeling, we suggest a mechanistic scheme of the QB turnover. The detachment of the formed ubiquinol from its proximal position next to Glu-L212 is considered as the rate-limiting step of the reaction cycle
MH  - A
MH  - acceptor
MH  - chromatophore
MH  - chromatophores
MH  - DEPENDENCE
MH  - Electrochemistry
MH  - electron
MH  - flash
MH  - intermediate
MH  - Light
MH  - membrane
MH  - Membrane Potential
MH  - model
MH  - mutant
MH  - pH
MH  - proton
MH  - protonation
MH  - Protons
MH  - quinone
MH  - Quinones
MH  - reaction center
MH  - RESIDUE
MH  - Rhodobacter
MH  - Rhodobacter sphaeroides
MH  - secondary
MH  - Site
MH  - sphaeroides
MH  - structure
MH  - Time
MH  - Ubiquinone
RP  - NOT IN FILE
NT  - UI - 20439551LA - engRN - 0 (Photosynthetic Reaction Center, Bacterial)RN - 0 (Protons)RN - 0 (Quinones)RN - 73-32-5 (Isoleucine)RN - 74-79-3 (Arginine)PT - Journal ArticleDA - 20000929IS - 0006-3002SB - IMCY - NETHERLANDS
UR  - PM:10924896
SO  - Biochim Biophys Acta 2000 Jul 20 ;1459(1):10-34

1660
UI  - 9882
AU  - Creczynski-Pasa TB
AU  - Possmayer FE
AU  - Scofano HM
AU  - Graber P
TI  - Characterization of nucleotide binding sites of the isolated H+-ATPase from spinach chloroplasts, CF0F1
AB  - Soluble purified CF0F1 from chloroplasts was either oxidized or reduced and then incubated with [alpha-P-32]ATP in the presence or in the absence of Mg2+. Depending on the conditions of incubation, the enzyme showed different tight-nucleotide binding sites. In the presence of EDTA, two sites bind [alpha- P-32]ATP from the reaction medium at different rates. Both sites promote ATP hydrolysis, since equimolar amounts of [alpha-P-32]ATP and [alpha-P-32]ADP are bound to the enzyme. In the presence of Mg2+, only one site appears during the first hour of incubation, with characteristics similar to those described in the absence of Mg2+. However, after this time a third site appears also permitting binding of ATP from the reaction medium, but in this case the bound ATP is not hydrolyzed, Covalent derivatization by 2-azido-[alpha-P-32]ATP was used to distinguish between catalytic and noncatalytic sites. In the presence of Mg2+, there are at least three distinct nucleotide binding sites that bind nucleotide tightly from the reaction medium: two of them are catalytic and one is noncatalytic, (C) 2000 Academic Press
MH  - ADP BINDING
MH  - atp
MH  - Binding Sites
MH  - Catalysis
MH  - CF0F1
MH  - chloroplast
MH  - chloroplast H+-ATPase
MH  - Chloroplasts
MH  - COUPLING FACTOR-I
MH  - F1
MH  - F1-ATPASE
MH  - H+-ATPase
MH  - Hydrolysis
MH  - mechanism
MH  - Mitochondria
MH  - NONCATALYTIC SITES
MH  - nucleotide binding
MH  - nucleotide binding sites
MH  - NUCLEOTIDE-BINDING-SITES
MH  - Spinach
MH  - SPINACH-CHLOROPLASTS
MH  - SUBUNIT
MH  - SYNTHASE
MH  - Time
RP  - NOT IN FILE
NT  - JournalArticleACADEMIC PRESS INCAPR 1299JJSAN DIEGOGraber P Univ Fed Rio de Janeiro, CCS, ICB, Dept Bioquim Med, Rio De Janeiro, BrazilARCH BIOCHEM BIOPHYS525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA
AV  - Univ Fed Rio de Janeiro, CCS, ICB, Dept Bioquim Med, Rio De Janeiro, Brazil Univ Fed Rio de Janeiro, CCS, ICB, Dept Bioquim Med, Rio De Janeiro, Brazil Univ Fed Santa Catarina, CCB, Dept Ciencias Fisiol, BR-88040900 Florianopolis, SC, Brazil Univ Freiburg, Inst Chem Phys, D-79014 Freiburg, Germany
UR  - ISI:000086195100015
SO  - Archives of Biochemistry and Biophysics 2000  ;376(1):141-148

1661
UI  - 19854
AU  - Cross RL
AD  - Department of Biochemistry and Molecular Biology, State University of New York, Health Science Center, Syracuse, NY 13210, USA crossr@hscsyredu
TI  - The rotary binding change mechanism of ATP synthases
AB  - The F(0)F(1) ATP synthase functions as a rotary motor where subunit rotation driven by a current of protons flowing through F(0) drives the binding changes in F(1) that are required for net ATP synthesis. Recent work that has led to the identification of components of the rotor and stator is reviewed. In addition, a model is proposed to describe the transmission of energy from four proton transport steps to the synthesis of one ATP. Finally, some of the requirements for efficient energy coupling by a rotary binding change mechanism are considered
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - BINDING
MH  - Biochemistry
MH  - CHANGE MECHANISM
MH  - COMPLEX
MH  - H(+)-Transporting ATP Synthase
MH  - mechanism
MH  - model
MH  - Multienzyme Complexes
MH  - Phosphotransferases
MH  - proton
MH  - Protons
MH  - review
MH  - rotation
MH  - SUBUNIT
MH  - SYNTHASE
MH  - synthesis
MH  - transport
RP  - NOT IN FILE
NT  - UI - 20298308LA - engRN - 0 (Molecular Motors)RN - 0 (Multienzyme Complexes)RN - 0 (Protons)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 2.7.4 (Phosphotransferases (Phosphate Group Acceptor))RN - EC 2.7.4.- (ATP synthetase complex)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialID - GM 23152/GM/NIGMSDA - 20000727IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:10838043
SO  - Biochim Biophys Acta 2000 May 31 ;1458(2-3):270-275

1662
UI  - 20930
AU  - Deckers-Hebestreit G
AU  - Greie J
AU  - Stalz W
AU  - Altendorf K
AD  - Abteilung Mikrobiologie, Fachbereich Biologie/Chemie, Universitat Osnabruck, D-49069, Osnabruck, Germany deckers-hebestreit@biologieuni- osnabrueckde
TI  - The ATP synthase of Escherichia coli: structure and function of F(0) subunits
AB  - In this review we discuss recent work from our laboratory concerning the structure and/or function of the F(0) subunits of the proton- translocating ATP synthase of Escherichia coli. For the topology of subunit a a brief discussion gives (i) a detailed picture of the C- terminal two-thirds of the protein with four transmembrane helices and the C terminus exposed to the cytoplasm and (ii) an evaluation of the controversial results obtained for the localization of the N-terminal region of subunit a including its consequences on the number of transmembrane helices. The structure of membrane-bound subunit b has been determined by circular dichroism spectroscopy to be at least 75% alpha-helical. For this purpose a method was developed, which allows the determination of the structure composition of membrane proteins in proteoliposomes. Subunit b was purified to homogeneity by preparative SDS gel electrophoresis, precipitated with acetone, and redissolved in cholate-containing buffer, thereby retaining its native conformation as shown by functional coreconstitution with an ac subcomplex. Monoclonal antibodies, which have their epitopes located within the hydrophilic loop region of subunit c, and the F(1) part are bound simultaneously to the F(0) complex without an effect on the function of F(0), indicating that not all c subunits are involved in F(1) interaction. Consequences on the coupling mechanism between ATP synthesis/hydrolysis and proton translocation are discussed
MH  - A
MH  - acceptor
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - buffer
MH  - COMPLEX
MH  - conformation
MH  - coupling
MH  - Cytoplasm
MH  - Electrophoresis
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - function
MH  - mechanism
MH  - membrane
MH  - Membrane Proteins
MH  - method
MH  - MONOCLONAL-ANTIBODIES
MH  - Multienzyme Complexes
MH  - Phosphotransferases
MH  - protein
MH  - Proteins
MH  - Proteolipids
MH  - proteoliposome
MH  - proton
MH  - review
MH  - spectroscopy
MH  - SUBUNIT
MH  - SYNTHASE
MH  - translocation
RP  - NOT IN FILE
NT  - UI - 20298316LA - engRN - 0 (Membrane Proteins)RN - 0 (Multienzyme Complexes)RN - 0 (Proteolipids)RN - 0 (proteoliposomes)RN - EC 2.7.4 (Phosphotransferases (Phosphate Group Acceptor))RN - EC 2.7.4.- (ATP Synthetase Complexes)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 20000727IS - 0006-3002SB - IMCY - NETHERLANDS
UR  - PM:10838051
SO  - Biochim Biophys Acta 2000 May 31 ;1458(2-3):364-373

1663
UI  - 9
AU  - Dimroth P
AU  - Matthey U
AU  - Kaim G
AD  - Institut fur Mikrobiologie, Eidgenossische Technische Hochschule, ETH- Zentrum, CH-8092, Zurich, Switzerland dimroth@microbioethzch
TI  - Critical evaluation of the one- versus the two-channel model for the operation of the ATP synthase's F(o) motor
AB  - The mechanism of converting an electrochemical gradient of protons or Na(+) ions across the membrane into rotational torque by the F(o) motor of the ATP synthase has been described by a two-channel model or by a one-channel model. Experimental evidence obtained with the F(o) motor from the Propionigenium modestum ATP synthase is described which is in accordance with the one-channel model, but not with the two-channel model. This evidence includes the ATP-dependent occlusion of one (22)Na(+) per ATP synthase with a mutated Na(+)-impermeable a subunit or the Na(+)(in)/(22)Na(+)(out) exchange which is not affected by modifying part of the c subunit sites with dicyclohexylcarbodiimide
RP  - NOT IN FILE
NT  - UI - 21061447LA - engRN - 0 (Cations)RN - 0 (Membrane Proteins)RN - 0 (Molecular Motors)RN - 0 (Protons)RN - 0 (Sodium Radioisotopes)RN - 56-65-5 (Adenosine Triphosphate)RN - 7440-23-5 (Sodium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 20010126IS - 0006-3002SB - IMCY - NetherlandsJC - A0W
UR  - PM:11004469
SO  - Biochim Biophys Acta 2000 Aug 15 ;1459(2-3):506-513

1664
UI  - 16
AU  - Dimroth P
AU  - Kaim G
AU  - Matthey U
AD  - Institut fur Mikrobiologie, Eidgenossische Technische Hochschule, ETH- Zentrum, Schmelzbergstrasse 7, CH-8092 Zurich, Switzerland dimroth@microbiolethzch
TI  - Crucial role of the membrane potential for ATP synthesis by F(1)F(o) ATP synthases
AB  - ATP, the universal carrier of cell energy, is manufactured from ADP and phosphate by the enzyme ATP synthase using the free energy of an electrochemical gradient of protons (or Na(+)). The proton-motive force consists of two components, the transmembrane proton concentration gradient (delta pH) and the membrane potential. The two components were considered to be not only thermodynamically but also kinetically equivalent, since the chloroplast ATP synthase appeared to operate on delta pH only. Recent experiments demonstrate, however, that the chloroplast ATP synthase, like those of mitochondria and bacteria, requires a membrane potential for ATP synthesis. Hence, the membrane potential and proton gradient are not equivalent under normal operating conditions far from equilibrium. These conclusions are corroborated by the finding that only the membrane potential induces a rotary torque that drives the counter-rotation of the a and c subunits in the F(o) motor of Propionigenium modestum ATP synthase
RP  - NOT IN FILE
NT  - UI - 20123951LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - 7440-23-5 (Sodium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 20000217IS - 0022-0949SB - IMCY - ENGLANDJC - I2F
UR  - PM:10600673
SO  - J Exp Biol 2000 Jan ;203 Pt 1():51-59

1665
UI  - 12
AU  - Dimroth P
AD  - Mikrobiologisches Institut, Eidgenossische Technische Hochschule, ETH- Zentrum, CH-8092, Zurich, Switzerland dimroth@microbiolethzch
TI  - Operation of the F(0) motor of the ATP synthase
AB  - ATP, the universal carrier of cell energy is manufactured from ADP and phosphate by the enzyme ATP synthase using the energy stored in a transmembrane ion gradient. The two components of the ion gradient (DeltapH or DeltapNa(+)) and the electrical potential difference Deltapsi are thermodynamically but not kinetically equivalent. In contrast to accepted wisdom, the electrical component is kinetically indispensable not only for bacterial ATP synthases but also for that from chloroplasts. Recent biochemical studies with the Na(+)- translocating ATP synthase of Propionigenium modestum have given a good idea of the ion translocation pathway in the F(0) motor. Taken together with biophysical data, the operating principles of the motor have been delineated
RP  - NOT IN FILE
NT  - UI - 20298317LA - engRN - 0 (Bacterial Proteins)RN - 0 (Chimeric Proteins)RN - 0 (Molecular Motors)RN - 0 (Multienzyme Complexes)RN - EC 2.7.4 (Phosphotransferases (Phosphate Group Acceptor))RN - EC 2.7.4.- (ATP synthetase complex)RN - EC 3.6.1.- (sodium ATPase)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 20000727IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:10838052
SO  - Biochim Biophys Acta 2000 May 31 ;1458(2-3):374-386

1666
UI  - 21416
AU  - Dunn SD
AU  - Revington M
AU  - Cipriano DJ
AU  - Shilton BH
TI  - The b subunit of Escherichia coli ATP synthase
AB  - The b subunit of ATP synthase is a major component of the second stalk connecting the F1 and F0 sectors of the enzyme and is essential for normal assembly and function. The 156-residue b subunit of the Escherichia coli ATP synthase has been investigated extensively through mutagenesis, deletion analysis, and biophysical characterization. The two copies of b exist as a highly extended, helical dimer extending from the membrane to near the top of F1, where they interact with the delta subunit. The sequence has been divided into four domains: the N-terminal membrane-spanning domain, the tether domain, the dimerization domain, and the C-terminal delta-binding domain. The dimerization domain, contained within residues 60-122, has many properties of a coiled-coil, while the delta-binding domain is more globular. Sites of crosslinking between b and the a, alpha, beta, and delta subunits of ATP synthase have been identified, and the functional significance of these interactions is under investigation. The b dimer may serve as an elastic element during rotational catalysis in the enzyme, but also directly influences the catalytic sites, suggesting a more active role in coupling
MH  - A
MH  - ACTIVE
MH  - alpha
MH  - Amino Acid Sequence
MH  - analysis
MH  - atp
MH  - ATP synthase
MH  - BETA
MH  - Biochemistry
MH  - Catalysis
MH  - catalytic
MH  - Chemistry
MH  - COLI ATP SYNTHASE
MH  - coupling
MH  - delta
MH  - DELTA-SUBUNIT
MH  - Dimerization
MH  - enzyme
MH  - enzymology
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - F1
MH  - function
MH  - INTERACTION
MH  - membrane
MH  - Models,Molecular
MH  - Molecular Sequence Data
MH  - mutagenesis
MH  - Protein Subunits
MH  - Proton-Translocating ATPases
MH  - RESIDUE
MH  - Sequence Deletion
MH  - Site
MH  - stalk
MH  - SUBUNIT
MH  - Support,Non-U.S.Gov't
MH  - SYNTHASE
MH  - universities
RP  - NOT IN FILE
NT  - Department of Biochemistry, University of Western Ontario, London, Canada sdunn@julianuwocaFAU - Dunn, S D
SO  - J Bioenerg Biomembr 2000 Aug ;32(4):347-355

1667
UI  - 21058
AU  - Dunn SD
AU  - Revington M
AU  - Cipriano DJ
AU  - Shilton BH
TI  - The b subunit of Escherichia coli ATP synthase
AB  - The b subunit of ATP synthase is a major component of the second stalk connecting the F1 and F0 sectors of the enzyme and is essential for normal assembly and function. The 156-residue b subunit of the Escherichia coli ATP synthase has been investigated extensively through mutagenesis, deletion analysis, and biophysical characterization. The two copies of b exist as a highly extended, helical dimer extending from the membrane to near the top of F1, where they interact with the delta subunit. The sequence has been divided into four domains: the N-terminal membrane-spanning domain, the tether domain, the dimerization domain, and the C-terminal delta-binding domain. The dimerization domain, contained within residues 60-122, has many properties of a coiled-coil, while the delta-binding domain is more globular. Sites of crosslinking between b and the a, alpha, beta, and delta subunits of ATP synthase have been identified, and the functional significance of these interactions is under investigation. The b dimer may serve as an elastic element during rotational catalysis in the enzyme, but also directly influences the catalytic sites, suggesting a more active role in coupling
MH  - A
MH  - ACTIVE
MH  - alpha
MH  - analysis
MH  - atp
MH  - ATP synthase
MH  - BETA
MH  - Biochemistry
MH  - Catalysis
MH  - COLI ATP SYNTHASE
MH  - coupling
MH  - delta
MH  - DELTA-SUBUNIT
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - F1
MH  - function
MH  - membrane
MH  - mutagenesis
MH  - RESIDUE
MH  - Site
MH  - stalk
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - Department of Biochemistry, University of Western Ontario, London, Canada sdunn@julianuwocaFAU - Dunn, S D
SO  - J Bioenerg Biomembr 2000 Aug ;32(4):347-355

1668
UI  - 19755
AU  - Feniouk BA
AU  - Cherepanov DA
AU  - Junge W
AU  - Mulkidjanian AY
TI  - FOF1-ATP-synthase of Rhodobacter capsulatus: Monitoring the ion flux through a single electrogenic transporter.
MH  - A
MH  - capsulatus
MH  - electrogenic
MH  - ion
MH  - Rhodobacter
MH  - rhodobacter capsulatus
MH  - RHODOBACTER-CAPSULATUS
RP  - NOT IN FILE
SO  - EBEC Short Reports 2000  ;11():190-190

1669
UI  - 19756
AU  - Feniouk BA
AU  - Cherepanov DA
AU  - Junge W
AU  - Mulkidjanian AY
TI  - ATP synthesis in chromatophores of Rhodobacter capsulatus: Evidence of a surface-confined proton transfer from the cytochrome-bc1 complex to the ATP synthase
MH  - A
MH  - atp
MH  - ATP synthesis
MH  - capsulatus
MH  - chromatophore
MH  - chromatophores
MH  - COMPLEX
MH  - electrogenic
MH  - ion
MH  - proton
MH  - Proton transfer
MH  - Rhodobacter
MH  - rhodobacter capsulatus
MH  - RHODOBACTER-CAPSULATUS
MH  - synthesis
MH  - TRANSFER
RP  - NOT IN FILE
SO  - EBEC Short Reports 2000  ;11():213-213

1670
UI  - 21106
AU  - Ferguson SJ
AD  - Department of Biochemistry, University of Oxford, South Parks Road, Oxford OX1 3QU, UK ferguson@biochoxacuk
TI  - Proton transfer: it's a stringent process
AB  - Proton transfer into and out of proteins is important, both for many enzyme reaction mechanisms and proton pumping across membranes. Recent work on several proteins has revealed stringent requirements for amino- acid side chains and subtle reorganisation of hydrogen-bond networks involving bound water molecules
MH  - A
MH  - ACID
MH  - Biochemistry
MH  - England
MH  - mechanism
MH  - MECHANISMS
MH  - membrane
MH  - Membranes
MH  - protein
MH  - Proteins
MH  - proton
MH  - Proton transfer
MH  - Proton-transfer
MH  - Protons
MH  - review
MH  - TRANSFER
MH  - Water
RP  - NOT IN FILE
NT  - UI - 20453778LA - engRN - 0 (Proteins)RN - 0 (Protons)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 20001107IS - 0960-9822SB - IMCY - ENGLAND
UR  - PM:10996087
SO  - Curr Biol 2000 Sep 7 ;10(17):R627-R630

1671
UI  - 21105
AU  - Ferguson SJ
AD  - Department of Biochemistry, University of Oxford, Oxford, UK ferguson@biochoxacuk
TI  - ATP synthase: what dictates the size of a ring?
AB  - Recent data suggest the source of F(0)F(1) ATP synthase determines a significant and surprising difference in the size of a putative rotating ring of integral membrane subunits of F(0); this can be correlated with biochemical data suggesting there is variation in the number of protons translocated per ATP synthesised
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Biochemistry
MH  - England
MH  - Macromolecular Systems
MH  - membrane
MH  - protein
MH  - Protein Subunits
MH  - proton
MH  - Proton-Translocating ATPases
MH  - Protons
MH  - SUBUNIT
MH  - SYNTHASE
MH  - SYSTEM
MH  - SYSTEMS
RP  - NOT IN FILE
NT  - UI - 20538942LA - engRN - 0 (Macromolecular Systems)RN - 0 (Protein Subunits)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 20001129IS - 0960-9822SB - IMCY - ENGLAND
UR  - PM:11084356
SO  - Curr Biol 2000 Nov 2 ;10(21):R804-R808

1672
UI  - 102
AU  - Fillingame RH
TI  - Getting to the bottom of the F1-ATPase
RP  - NOT IN FILE
NT  - UI - 20517589LA - engRN - 0 (Macromolecular Systems)RN - 0 (Protein Subunits)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - CommentPT - NewsDA - 20001117IS - 1072-8368SB - IMCY - UNITED STATESJC - B98
UR  - PM:11062550
SO  - Nat Struct Biol 2000 Nov ;7(11):1002-1004

1673
UI  - 8
AU  - Fillingame RH
AU  - Jiang W
AU  - Dmitriev OY
AD  - Department of Biomolecular Chemistry, University of Wisconsin Medical School, Madison, WI 53706, USA rhfillin@facstaffwiscedu
TI  - Coupling H(+) transport to rotary catalysis in F-type ATP synthases: structure and organization of the transmembrane rotary motor
AB  - H(+)-transporting F(1)F(o)-type ATP synthases utilize a transmembrane H(+) potential to drive ATP formation by a rotary catalytic mechanism. ATP is formed in alternating beta subunits of the extramembranous F(1) sector of the enzyme, synthesis being driven by rotation of the gamma subunit in the center of the F(1) molecule between the alternating catalytic sites. The H(+) electrochemical potential is thought to drive gamma subunit rotation by first coupling H(+) transport to rotation of an oligomeric rotor of c subunits within the transmembrane F(o) sector. The gamma subunit is forced to turn with the c(12) oligomeric rotor as a result of connections between subunit c and the gamma and epsilon subunits of F(1). In this essay, we will review recent studies on the Escherichia coli F(o) sector. The monomeric structure of subunit c, determined by nuclear magnetic resonance (NMR), is discussed first and used as a basis for the rest of the review. A model for the structural organization of the c(12) oligomer in F(o), deduced from extensive cross-linking studies and by molecular modeling, is then described. The interactions between the the a(1)b(2) 'stator' subcomplex of F(o) and the c(12) oligomer are then considered. A functional interaction between transmembrane helix 4 of subunit a (aTMH-4) and transmembrane helix 2 of subunit c (cTMH-2) during the proton-release step from Asp61 on cTMH-2 is suggested. Current a-c cross-linking data can only be explained by helix-helix swiveling or rotation during the proton transfer steps. A model that mechanically links helix rotation within a single subunit c to the incremental 30 degrees rotation of the c(12) oligomer is proposed. In the final section, the structural interactions between the surface residues of the c(12) oligomer and subunits epsilon and gamma are considered. A molecular model for the binding of subunit epsilon between the exposed, polar surfaces of two subunits c in the oligomer is proposed on the basis of cross-linking data and the NMR structures of the individual subunits
RP  - NOT IN FILE
NT  - UI - 20123946LA - engRN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialID - GM23105/GM/NIGMSDA - 20000217IS - 0022-0949SB - IMCY - ENGLANDJC - I2F
UR  - PM:10600668
SO  - J Exp Biol 2000 Jan ;203 Pt 1():9-17

1674
UI  - 6
AU  - Fillingame RH
AU  - Jiang W
AU  - Dmitriev OY
AU  - Jones PC
AD  - Department of Biomolecular Chemistry, University of Wisconsin Medical School, 1300 University Avenue, Madison, WI 53706, USA
TI  - Structural interpretations of F(0) rotary function in the Escherichia coli F(1)F(0) ATP synthase
AB  - F(1)F(0) ATP synthases are known to synthesize ATP by rotary catalysis in the F(1) sector of the enzyme. Proton translocation through the F(0) membrane sector is now proposed to drive rotation of an oligomer of c subunits, which in turn drives rotation of subunit gamma in F(1). The primary emphasis of this review will be on recent work from our laboratory on the structural organization of F(0), which proves to be consistent with the concept of a c(12) oligomeric rotor. From the NMR structure of subunit c and cross-linking studies, we can now suggest a detailed model for the organization of the c(12) oligomer in F(0) and some of the transmembrane interactions with subunits a and b. The structural model indicates that the H(+)-carrying carboxyl of subunit c is located between subunits of the c(12) oligomer and that two c subunits pack in a front-to-back manner to form the proton (cation) binding site. The proton carrying Asp61 side chain is occluded between subunits and access to it, for protonation and deprotonation via alternate entrance and exit half-channels, requires a swiveled opening of the packed c subunits and stepwise association with different transmembrane helices of subunit a. We suggest how some of the structural information can be incorporated into models of rotary movement of the c(12) oligomer during coupled synthesis of ATP in the F(1) portion of the molecule
RP  - NOT IN FILE
NT  - UI - 20298318LA - engRN - 0 (Bacterial Proteins)RN - 0 (Membrane Proteins)RN - 0 (Molecular Motors)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 20000727IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:10838053
SO  - Biochim Biophys Acta 2000 May 31 ;1458(2-3):387-403

1675
UI  - 9879
AU  - Fischer S
AU  - Graber P
AU  - Turina P
TI  - The activity of the ATP synthase from Escherichia coli is regulated by the transmembrane proton motive force
AB  - The ATP synthase from Escherichia coli was reconstituted into liposomes from phosphatidylcholine/phosphatidic acid. The proteoliposomes were energized by an acid-base transition and a K+/valinomycin diffusion potential, and one second after energization, the electrochemical proton gradient was dissipated by uncouplers, and the ATP hydrolysis measurement was started. In the presence of ADP and P-i, the initial rate of ATP hydrolysis was up to 9-fold higher with pre-energized proteoliposomes than with proteoliposomes that had not seen an electrochemical proton gradient. After dissipating the electrochemical proton gradient, the high rate of ATP hydrolysis decayed to the rate without preenergization within about 15 s. During this decay the enzyme carried out approximately 100 turnovers. In the absence of ADP and P-i, the rate of ATP hydrolysis was already high and could not be significantly increased by pre-energization. It is concluded that ATP hydrolysis is inhibited when ADP and P-i are bound to the enzyme and that a high Delta<(mu)over tilde>(H)+ is required to release ADP and P-i and to convert the enzyme into a high activity state. This high activity state is metastable and decays slowly when Delta<(mu)over tilde>(H)+ is abolished. Thus, the proton motive force does not only supply energy for ATP synthesis but also regulates the fraction of active enzymes
MH  - ADP
MH  - atp
MH  - Binding Sites
MH  - Biochemistry
MH  - CATALYTIC SITE NUCLEOTIDE
MH  - COUPLING FACTOR
MH  - DELTA-PH
MH  - Diffusion
MH  - Enzymes
MH  - Escherichia coli
MH  - H+-ATPase
MH  - Hydrolysis
MH  - INHIBITOR PROTEIN
MH  - INORGANIC-PHOSPHATE
MH  - Liposomes
MH  - MITOCHONDRIAL F1-ATPASE
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - JournalArticleAMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INCSEP 29358VMBETHESDATurina P Univ Bologna, Dept Biol, Biochem & Biophys Lab, Via Irnerio 42, I-40126 Bologna, ItalyJ BIOL CHEM9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA
AV  - Univ Bologna, Dept Biol, Biochem & Biophys Lab, Via Irnerio 42, I-40126 Bologna, Italy Univ Bologna, Dept Biol, Biochem & Biophys Lab, I-40126 Bologna, Italy Univ Freiburg, Inst Phys Chem, D-79104 Freiburg, Germany
UR  - ISI:000089577900035
SO  - Journal of Biological Chemistry 2000  ;275(39):30157-30162

1676
UI  - 20835
AU  - Futai M
AU  - Omote H
AU  - Sambongi Y
AU  - Wada Y
AD  - Division of Biological Sciences, Institute of Scientific and Industrial Research, Osaka University, CREST, Japan Science and Technology Corporation, Ibaraki, 567-0047, Osaka, Japan m-futai@sankenosaka- uacjp
TI  - Synthase (H(+) ATPase): coupling between catalysis, mechanical work, and proton translocation
AB  - Coupling with electrochemical proton gradient, ATP synthase (F(0)F(1)) synthesizes ATP from ADP and phosphate. Mutational studies on high- resolution structure have been useful in understanding this complicated membrane enzyme. We discuss mainly the mechanism of catalysis in the beta subunit of F(1) sector and roles of the gamma subunit in energy coupling. The gamma-subunit rotation during catalysis is also discussed
MH  - ADP
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BETA-SUBUNIT
MH  - Catalysis
MH  - coupling
MH  - mechanism
MH  - membrane
MH  - Nucleotides
MH  - proton
MH  - Protons
MH  - resolution
MH  - review
MH  - rotation
MH  - SUBUNIT
MH  - SYNTHASE
MH  - translocation
RP  - NOT IN FILE
NT  - UI - 20298309LA - engRN - 0 (Molecular Motors)RN - 0 (Nucleotides)RN - 0 (Protons)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 20000727IS - 0006-3002SB - IMCY - NETHERLANDS
UR  - PM:10838044
SO  - Biochim Biophys Acta 2000 May 31 ;1458(2-3):276-288

1677
UI  - 131
AU  - Garcia JJ
AU  - Ogilvie I
AU  - Robinson BH
AU  - Capaldi RA
AD  - Institute of Molecular Biology, University of Oregon, Eugene, Oregon 97403-1229, USA
TI  - Structure, functioning, and assembly of the ATP synthase in cells from patients with the T8993G mitochondrial DNA mutation. Comparison with the enzyme in Rho(0) cells completely lacking mtdna
AB  - The structure and functioning of the ATP synthase of human fibroblast cell lines with 91 and 100%, respectively, of the T8993G mutation have been studied, with MRC5 human fibroblasts and Rho(0) cells derived from this cell line as controls. ATP hydrolysis was normal but ATP synthesis was reduced by 60% in the 100% mutants. Both activities were highly oligomycin-sensitive. The levels of F(1)F(0) were close to normal, and the enzyme was stable. It is concluded that the loss of ATP synthesis is because of disruption of the proton translocation step within the F(0) part. This is supported by membrane potential measurements using the dye JC-1. Cells with a 91% mutation load grew well and showed only a 25% loss in ATP synthesis. This much reduced effect for only a 9% difference in mutation load mirrors the reduced pathogenicity in patients. F(1)F(0) has been purified for the first time from human cell lines. A partial complex was obtained from Rho(0) cells containing the F(1) subunits associated with several stalk, as well as F(0) subunits, including oligomycin sensitivity conferring protein, b, and c subunits. This partial complex no longer binds inhibitor protein
RP  - NOT IN FILE
NT  - UI - 20219152LA - engRN - 0 (DNA, Mitochondrial)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 20000505IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:10753912
SO  - J Biol Chem 2000 Apr 14 ;275(15):11075-11081

1678
UI  - 20809
AU  - Gardner JL
AU  - Cain BD
AD  - Department of Biochemistry and Molecular Biology, University of Florida, Gainesville, Florida 32610, USA
TI  - The a subunit ala-217 --> arg substitution affects catalytic activity of F(1)F(0) ATP synthase
AB  - A large number of mutations affecting the F(0) sector of Escherichia coli F(1)F(0) ATP synthase have been constructed and characterized. A subset of the missense mutations resulted in fully assembled enzyme complexes blocked in proton translocation and displaying marked decreases in ATP hydrolysis activity. The catalytic activities of one such mutant enzyme, a(ala-217-->arg), have been determined using both multisite and unisite catalysis conditions. As expected, the V(max) of the a(ala-217-->arg) enzyme was reduced under conditions of saturating substrate concentration. However, the F(0) sector amino acid substitution did not affect nucleotide occupancy of the noncatalytic sites. Moreover, the microscopic rate constants measured using unisite methods yielded no significant differences between the intact wild type F(1)F(0) ATP synthase and the a(ala-217-->arg) mutant enzyme. In general, the values for unisite activities in both preparations were very similar to numbers reported in the literature for E. coli F(1)- ATPase. The results suggest that the a(ala-217-->arg) substitution resulted in a defect in catalytic cooperativity and most likely altered the enzyme by inhibiting the rotational mechanism of F(1)F(0) ATP synthase
MH  - A
MH  - ACID
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Biochemistry
MH  - Catalysis
MH  - COMPLEX
MH  - CONSTANT
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - Hydrolysis
MH  - mechanism
MH  - method
MH  - Methods
MH  - mutant
MH  - NONCATALYTIC SITES
MH  - proton
MH  - rate constant
MH  - Site
MH  - SUBUNIT
MH  - SYNTHASE
MH  - translocation
MH  - UNISITE CATALYSIS
RP  - NOT IN FILE
NT  - UI - 20361781LA - engRN - 56-41-7 (Alanine)RN - 56-65-5 (Adenosine Triphosphate)RN - 74-79-3 (Arginine)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleID - GM43495/GM/NIGMSDA - 20000907IS - 0003-9861SB - IMCY - UNITED STATES
UR  - PM:10900150
SO  - Arch Biochem Biophys 2000 Aug 1 ;380(1):201-207

1679
UI  - 317
AU  - Gibbons C
AU  - Montgomery MG
AU  - Leslie AG
AU  - Walker JE
AD  - The Medical Research Council Dunn Human Nutrition Unit, Hills Road, Cambridge CB2 2XY, UK
TI  - The structure of the central stalk in bovine F(1)-ATPase at 2.4 A resolution
AB  - The central stalk in ATP synthase, made of gamma, delta and epsilon subunits in the mitochondrial enzyme, is the key rotary element in the enzyme's catalytic mechanism. The gamma subunit penetrates the catalytic (alpha beta)(3) domain and protrudes beneath it, interacting with a ring of c subunits in the membrane that drives rotation of the stalk during ATP synthesis. In other crystals of F(1)-ATPase, the protrusion was disordered, but with crystals of F(1)-ATPase inhibited with dicyclohexylcarbodiimide, the complete structure was revealed. The delta and epsilon subunits interact with a Rossmann fold in the gamma subunit, forming a foot. In ATP synthase, this foot interacts with the c-ring and couples the transmembrane proton motive force to catalysis in the (alpha beta)(3) domain
RP  - NOT IN FILE
NT  - UI - 20517602LA - engRN - 0 (Macromolecular Systems)RN - 0 (Protein Subunits)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 20001117IS - 1072-8368SB - IMCY - UNITED STATESJC - B98
UR  - PM:11062563
SO  - Nat Struct Biol 2000 Nov ;7(11):1055-1061

1680
UI  - 9877
AU  - Godbole R
AU  - Graber P
AU  - Hertel R
TI  - ATP production after pH shift in membrane vesicles from maize coleoptiles
AB  - Microsomal membrane vesicles and purified plasma membrane vesicles obtained from coleoptiles of maize (Zea mays L.) were subjected to pH shifts from pH 7.8 to 4.7. In the presence of ATPase inhibitors such as vanadate, net accumulation of radiolabelled butyric and indole-3-acetic acid (IAA) remained higher than in controls. When 2 min after the pH shift, at 4 degreesC, the microsomal vesicles were denatured the amount of ATP could be determined using the luciferin/luciferase assay. Significantly increased ATP production over control values - no pH-shift or ionophore treatment - was found. Therefore, such vesicles might produce ATP for in vitro transport processes such as auxin efflux
MH  - atp
MH  - ATP production
MH  - ATPase
MH  - auxin
MH  - AUXIN TRANSPORT
MH  - CUCURBITA
MH  - In Vitro
MH  - membrane vesicles
MH  - transport
MH  - Zea mays
RP  - NOT IN FILE
NT  - JournalArticleKLUWER ACADEMIC PUBLNOV417WMDORDRECHTHertel R Univ Freiburg, Inst Biol 3, Schanzlestr 1, D-79104 Freiburg, GermanyPLANT GROWTH REGULSPUIBOULEVARD 50, PO BOX 17, 3300 AA DORDRECHT, NETHERLANDS
AV  - Univ Freiburg, Inst Biol 3, Schanzlestr 1, D-79104 Freiburg, Germany Univ Freiburg, Inst Biol 3, D-79104 Freiburg, Germany Univ Freiburg, Inst Phys Chem, D-79104 Freiburg, Germany
UR  - ISI:000167858500007
SO  - Plant Growth Regulation 2000  ;32(2-3):151-155

1681
UI  - 19905
AU  - Grabe M
AU  - Wang H
AU  - Oster G
AD  - Departments of Physics, University of California, Berkeley 94720-3112, USA
TI  - The mechanochemistry of V-ATPase proton pumps
AB  - The vacuolar H(+)-ATPases (V-ATPases) are a universal class of proton pumps that are structurally similar to the F-ATPases. Both protein families are characterized by a membrane-bound segment (V(o), F(o)) responsible for the translocation of protons, and a soluble portion, (V(1), F(1)), which supplies the energy for translocation by hydrolyzing ATP. Here we present a mechanochemical model for the functioning of the V(o) ion pump that is consistent with the known structural features and biochemistry. The model reproduces a variety of experimental measurements of performance and provides a unified view of the many mechanisms of intracellular pH regulation
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Biochemistry
MH  - F-ATPASE
MH  - H(+)-Transporting ATP Synthase
MH  - H(+)ATPase
MH  - ion
MH  - Macromolecular Systems
MH  - mechanism
MH  - model
MH  - pH
MH  - protein
MH  - proton
MH  - Proton Pump
MH  - Protons
MH  - regulation
MH  - SYNTHASE
MH  - SYSTEM
MH  - SYSTEMS
MH  - translocation
RP  - NOT IN FILE
NT  - UI - 20287318LA - engRN - 0 (Macromolecular Systems)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.- (V-type ATPase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 20000710IS - 0006-3495SB - IMCY - UNITED STATESJC - A5S
UR  - PM:10827963
SO  - Biophys J 2000 Jun ;78(6):2798-2813

1682
UI  - 21415
AU  - Greie JC
AU  - Deckers-Hebestreit G
AU  - Altendorf K
TI  - Subunit organization of the stator part of the F0 complex from Escherichia coli ATP synthase
AB  - Membrane-bound ATP synthases (F1F0) catalyze the synthesis of ATP via a rotary catalytic mechanism utilizing the energy of an electrochemical ion gradient. The transmembrane potential is supposed to propel rotation of a subunit c ring of F0 together with subunits gamma and epsilon of F1, thereby forming the rotor part of the enzyme, whereas the remainder of the F1F0 complex functions as a stator for compensation of the torque generated during rotation. This review focuses on our recent work on the stator part of the F0 complex, e.g., subunits a and b. Using epitope insertion and antibody binding, subunit a was shown to comprise six transmembrane helixes with both the N- and C-terminus oriented toward the cytoplasm. By use of circular dichroism (CD) spectroscopy, the secondary structure of subunit b incorporated into proteoliposomes was determined to be 80% alpha-helical together with 14% beta turn conformation, providing flexibility to the second stalk. Reconstituted subunit b together with isolated ac subcomplex was shown to be active in proton translocation and functional F1 binding revealing the native conformation of the polypeptide chain. Chemical crosslinking in everted membrane vesicles led to the formation of subunit b homodimers around residues bQ37 to bL65, whereas bA32C could be crosslinked to subunit a, indicating a close proximity of subunits a and b near the membrane. Further evidence for the proposed direct interaction between subunits a and b was obtained by purification of a stable ab2 subcomplex via affinity chromatography using His tags fused to subunit a or b. This ab2 subcomplex was shown to be active in proton translocation and F1 binding, when coreconstituted with subunit c. Consequences of crosslink formation and subunit interaction within the F1F0 complex are discussed
MH  - A
MH  - ACTIVE
MH  - affinity
MH  - atp
MH  - ATP synthase
MH  - BETA
MH  - BINDING
MH  - catalytic
MH  - Chemistry
MH  - Circular Dichroism
MH  - COLI ATP SYNTHASE
MH  - COMPLEX
MH  - conformation
MH  - Cytoplasm
MH  - Dimerization
MH  - energy
MH  - enzyme
MH  - enzymology
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - F1
MH  - function
MH  - INTERACTION
MH  - ion
MH  - mechanism
MH  - membrane
MH  - membrane vesicles
MH  - Models,Molecular
MH  - Protein Structure,Secondary
MH  - Protein Subunits
MH  - proteoliposome
MH  - proton
MH  - Proton-Translocating ATPases
MH  - purification
MH  - RESIDUE
MH  - review
MH  - rotation
MH  - secondary
MH  - spectroscopy
MH  - stalk
MH  - structure
MH  - SUBUNIT
MH  - Support,Non-U.S.Gov't
MH  - SYNTHASE
MH  - synthesis
MH  - translocation
MH  - vesicles
RP  - NOT IN FILE
NT  - Universitat Osnabruck, Fachbereich Biologie/Chemie, Abteilung Mikrobiologie, Germany greie@biologieuni-osnabrueckdeFAU - Greie, J C
SO  - J Bioenerg Biomembr 2000 Aug ;32(4):357-364

1683
UI  - 20928
AU  - Greie JC
AU  - Deckers-Hebestreit G
AU  - Altendorf K
AD  - Universitat Osnabruck, Fachbereich Biologie/Chemie, Abteilung Mikrobiologie, Germany greie@biologieuni-osnabrueckde
TI  - Subunit organization of the stator part of the F0 complex from Escherichia coli ATP synthase
AB  - Membrane-bound ATP synthases (F1F0) catalyze the synthesis of ATP via a rotary catalytic mechanism utilizing the energy of an electrochemical ion gradient. The transmembrane potential is supposed to propel rotation of a subunit c ring of F0 together with subunits gamma and epsilon of F1, thereby forming the rotor part of the enzyme, whereas the remainder of the F1F0 complex functions as a stator for compensation of the torque generated during rotation. This review focuses on our recent work on the stator part of the F0 complex, e.g., subunits a and b. Using epitope insertion and antibody binding, subunit a was shown to comprise six transmembrane helixes with both the N- and C-terminus oriented toward the cytoplasm. By use of circular dichroism (CD) spectroscopy, the secondary structure of subunit b incorporated into proteoliposomes was determined to be 80% alpha-helical together with 14% beta turn conformation, providing flexibility to the second stalk. Reconstituted subunit b together with isolated ac subcomplex was shown to be active in proton translocation and functional F1 binding revealing the native conformation of the polypeptide chain. Chemical crosslinking in everted membrane vesicles led to the formation of subunit b homodimers around residues bQ37 to bL65, whereas bA32C could be crosslinked to subunit a, indicating a close proximity of subunits a and b near the membrane. Further evidence for the proposed direct interaction between subunits a and b was obtained by purification of a stable ab2 subcomplex via affinity chromatography using His tags fused to subunit a or b. This ab2 subcomplex was shown to be active in proton translocation and F1 binding, when coreconstituted with subunit c. Consequences of crosslink formation and subunit interaction within the F1F0 complex are discussed
MH  - A
MH  - ACTIVE
MH  - atp
MH  - ATP synthase
MH  - BINDING
MH  - COLI ATP SYNTHASE
MH  - COMPLEX
MH  - conformation
MH  - Cytoplasm
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - F1
MH  - function
MH  - ion
MH  - mechanism
MH  - membrane
MH  - membrane vesicles
MH  - proteoliposome
MH  - proton
MH  - purification
MH  - RESIDUE
MH  - review
MH  - rotation
MH  - secondary
MH  - spectroscopy
MH  - stalk
MH  - SUBUNIT
MH  - SYNTHASE
MH  - synthesis
MH  - translocation
RP  - NOT IN FILE
NT  - UI - 21618401LA - engPT - Journal ArticleDA - 20011220IS - 0145-479XSB - IMCY - United States
UR  - PM:11768297
SO  - J Bioenerg Biomembr 2000 Aug ;32(4):357-364

1684
UI  - 20931
AU  - Greie JC
AU  - Deckers-Hebestreit G
AU  - Altendorf K
AD  - Universitat Osnabruck, Fachbereich Biologie/Chemie, Abteilung Mikrobiologie, Germany greie@biologieuni-osnabrueckde
TI  - Secondary structure composition of reconstituted subunit b of the Escherichia coli ATP synthase
AB  - Subunit b of the Escherichia coli ATP synthase was isolated by preparative gel electrophoresis, acetone precipitated and after ion- pair extraction redissolved in a buffer either containing n-dodecyl- beta-D-maltoside or sodium cholate. The secondary structure of isolated subunit b was shown to be the same as within the FO complex, but was strongly dependent on the detergent used for replacement of the phospholipid environment. This was shown by an identical tryptic digestion pattern, which was strongly influenced by the detergent used for solubilization. An influence of the detergent n-dodecyl-beta-D- maltoside on the secondary structure of the hydrophilic part of subunit b was also shown for the soluble part of the polypeptide comprising residues Val25 to Leu156 (bsol) using CD spectroscopy. In order to determine the secondary structure of subunit b in its native conformation, isolated subunit b was reconstituted into E. coli lipid vesicles and analyzed with CD spectroscopy. The resulting spectrum revealed a secondary structure composition of 80% alpha helix together with 14% beta turn conformation. These results suggest that subunit b is not a rigid rod-like alpha helix simply linking F1 to FO, but rather provides an inherent flexibility for the storage of elastic energy within the second stalk generated by rotational movements within the F1FO complex
MH  - A
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - buffer
MH  - COLI ATP SYNTHASE
MH  - COMPLEX
MH  - conformation
MH  - Detergents
MH  - Electrophoresis
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1
MH  - Glucosides
MH  - ion
MH  - Movement
MH  - Proteolipids
MH  - proteoliposome
MH  - proton
MH  - Protons
MH  - RESIDUE
MH  - secondary
MH  - Sodium
MH  - spectra
MH  - spectroscopy
MH  - stalk
MH  - SUBUNIT
MH  - SYNTHASE
MH  - Trypsin
RP  - NOT IN FILE
NT  - UI - 20267835LA - engRN - 0 (Detergents)RN - 0 (Glucosides)RN - 0 (Proteolipids)RN - 0 (Protons)RN - 0 (proteoliposomes)RN - 69227-93-6 (dodecyl maltoside)RN - EC 3.4.21.4 (Trypsin)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 20000713IS - 0014-2956SB - IMCY - GERMANY
UR  - PM:10806404
SO  - Eur J Biochem 2000 May ;267(10):3040-3048

1685
UI  - 21057
AU  - Greie JC
AU  - Deckers-Hebestreit G
AU  - Altendorf K
TI  - Subunit organization of the stator part of the F0 complex from Escherichia coli ATP synthase
AB  - Membrane-bound ATP synthases (F1F0) catalyze the synthesis of ATP via a rotary catalytic mechanism utilizing the energy of an electrochemical ion gradient. The transmembrane potential is supposed to propel rotation of a subunit c ring of F0 together with subunits gamma and epsilon of F1, thereby forming the rotor part of the enzyme, whereas the remainder of the F1F0 complex functions as a stator for compensation of the torque generated during rotation. This review focuses on our recent work on the stator part of the F0 complex, e.g., subunits a and b. Using epitope insertion and antibody binding, subunit a was shown to comprise six transmembrane helixes with both the N- and C-terminus oriented toward the cytoplasm. By use of circular dichroism (CD) spectroscopy, the secondary structure of subunit b incorporated into proteoliposomes was determined to be 80% alpha-helical together with 14% beta turn conformation, providing flexibility to the second stalk. Reconstituted subunit b together with isolated ac subcomplex was shown to be active in proton translocation and functional F1 binding revealing the native conformation of the polypeptide chain. Chemical crosslinking in everted membrane vesicles led to the formation of subunit b homodimers around residues bQ37 to bL65, whereas bA32C could be crosslinked to subunit a, indicating a close proximity of subunits a and b near the membrane. Further evidence for the proposed direct interaction between subunits a and b was obtained by purification of a stable ab2 subcomplex via affinity chromatography using His tags fused to subunit a or b. This ab2 subcomplex was shown to be active in proton translocation and F1 binding, when coreconstituted with subunit c. Consequences of crosslink formation and subunit interaction within the F1F0 complex are discussed
MH  - A
MH  - ACTIVE
MH  - atp
MH  - ATP synthase
MH  - BETA
MH  - BINDING
MH  - COLI ATP SYNTHASE
MH  - COMPLEX
MH  - conformation
MH  - Cytoplasm
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - F1
MH  - function
MH  - ion
MH  - mechanism
MH  - membrane
MH  - membrane vesicles
MH  - proteoliposome
MH  - proton
MH  - purification
MH  - RESIDUE
MH  - review
MH  - rotation
MH  - secondary
MH  - spectroscopy
MH  - stalk
MH  - structure
MH  - SUBUNIT
MH  - SYNTHASE
MH  - synthesis
MH  - translocation
MH  - vesicles
RP  - NOT IN FILE
NT  - Universitat Osnabruck, Fachbereich Biologie/Chemie, Abteilung Mikrobiologie, Germany greie@biologieuni-osnabrueckdeFAU - Greie, J C
SO  - J Bioenerg Biomembr 2000 Aug ;32(4):357-364

1686
UI  - 9942
AU  - Groth G
AU  - Mills DA
AU  - Christiansen E
AU  - Richter ML
AU  - Huchzermeyer B
AD  - Department of Molecular Biosciences, University of Kansas, Lawrence, Kansas 66045, USA
TI  - Characterization of a phosphate binding domain on the alpha-subunit of chloroplast ATP synthase using the photoaffinity phosphate analogue 4- azido-2-nitrophenyl phosphate
AB  - The photoaffinity phosphate analogue 4-azido-2 nitrophenyl phosphate (ANPP) was shown previously (Pougeois, R., Lauquin, G. J.-M., and Vignais, P. V. (1983) Biochemistry 22, 1241-1245) to bind covalently and specifically to a single catalytic site on one of the three beta- subunits of the isolated chloroplast coupling factor 1 (CF(1)). Modification by ANPP strongly inhibited ATP hydrolysis activity. In this study, we examined labeling of membrane-bound CF(1) by ANPP by exposing thylakoid membranes to increasing concentrations of the reagent. ANPP exhibited saturable binding to two sites on CF(1), one on the beta-subunit and one on the alpha-subunit. Labeling by ANPP resulted in the complete inhibition of both ATP synthesis and ATP hydrolysis by the membrane-bound enzyme. Labeling of both sites by ANPP was reduced by more than 80% in the presence of P(i) (> or = 10 mM) and ATP (> or = 0.5 mM). ADP was less effective in competing with ANPP for binding, giving a maximum of approximately 35% inhibition at concentrations > or = 2 mM. ANPP-labeled tryptic peptides of the alpha- subunit were isolated and sequenced. The majority of the probe was contained in three peptides corresponding to residues Gln(173) to Arg(216), Gly(217) to Arg(253), and His(256) to Arg(272) of the alpha- subunit. In the mitochondrial F(1) (Abrahams, J. P., Leslie, A. G. W., Lutter, R., and Walker, J. E. (1994) Nature 370, 621-628), all three analogous peptides are located within the nucleotide binding pocket and within close proximity to the gamma-phosphate binding site. The data indicate, however, that the azidophenyl group of bound ANPP is oriented at approximately 180 degrees in the opposite direction to the adenine binding site with reference to the phosphate binding site on the alpha- subunit. The study has confirmed that ANPP is a bona fide phosphate analogue and suggests that it specifically targets the gamma-phosphate binding site within the nucleotide binding pockets on the alpha- and beta-subunits of CF(1). The study also indicates that in the resting state of the chloroplast F(1)-F(0) complex both the alpha- and beta- subunits are structurally asymmetric
MH  - ADP
MH  - ALPHA-SUBUNIT
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - Azides
MH  - BETA- SUBUNIT
MH  - BETA-SUBUNIT
MH  - BINDING
MH  - Biochemistry
MH  - chloroplast
MH  - COMPLEX
MH  - COUPLING FACTOR
MH  - H(+)-Transporting ATP Synthase
MH  - Hydrolysis
MH  - Membranes
MH  - nucleotide binding
MH  - Peptide Fragments
MH  - Phosphates
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 20530309LA - engRN - 0 (Azides)RN - 0 (Peptide Fragments)RN - 0 (Phosphates)RN - 0 (Photoaffinity Labels)RN - 74784-75-1 (4-azido-2-nitrophenyl phosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 20001130IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:11076517
SO  - Biochemistry 2000 Nov 14 ;39(45):13781-13787

1687
UI  - 9945
AU  - Groth G
AD  - Heinrich-Heine Universitat Dussseldorf, Biochemie der Pflanzen, Universitatsstr 1, 40225, Dusseldorf, Germany georggroth@uni- duesseldorfde
TI  - Molecular models of the structural arrangement of subunits and the mechanism of proton translocation in the membrane domain of F(1)F(0) ATP synthase
AB  - Subunit c of the proton-transporting ATP synthase of Escherichia coli forms an oligomeric complex in the membrane domain that functions in transmembrane proton conduction. The arrangement of subunit c monomers in this oligomeric complex was studied by scanning mutagenesis. On the basis of these studies and structural information on subunit c, different molecular models for the potential arrangement of monomers in the c-oligomer are discussed. Intersubunit contacts in the F(0) domain that have been analysed in the past by chemical modification and mutagenesis studies are summarised. Transient contacts of the c- oligomer with subunit a might play a crucial role in the mechanism of proton translocation. Schematic models presented by several authors that interpret proton transport in the F(0) domain by a relative rotation of the c-subunit oligomer against subunit a are reviewed against the background of the molecular models of the oligomer
MH  - atp
MH  - ATP synthase
MH  - COMPLEX
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - H(+)-Transporting ATP Synthase
MH  - mechanism
MH  - model
MH  - proton
MH  - Protons
MH  - review
MH  - SUBUNIT
MH  - SYNTHASE
MH  - transport
RP  - NOT IN FILE
NT  - UI - 20298320LA - engRN - 0 (3-trifluoromethyl-3-(3-iodophenyl)diazirineceramide)RN - 0 (Azirines)RN - 0 (Ceramides)RN - 0 (Disulfides)RN - 0 (Molecular Motors)RN - 0 (Photoaffinity Labels)RN - 0 (Protons)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 20000727IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:10838055
SO  - Biochim Biophys Acta 2000 May 31 ;1458(2-3):417-427

1688
UI  - 21417
AU  - Gruber G
TI  - Structural and functional features of the Escherichia coli F1-ATPase
AB  - The structural organization and overall dimensions of the Escherichia coli F1-ATPase in solution has been analyzed by synchroton X-ray scattering. Using an independent ab initio approach, the low-resolution shape of the hydrated enzyme was determined at 3.2 nm resolution. The shape permitted unequivocal identification of the volume occupied by the alpha3beta3gamma complex of the atomic model of the ECF1-ATPase. The position of the delta and epsilon subunits were found by interactive fitting of the solution scattering data and by cross-linking studies. Laser-induced covalent incorporation of 2-azido-ATP established a direct relationship between nucleotide binding affinity and the different interactions between the stalk subunits gamma and epsilon with the three catalytic subunits (beta) of the F1-ATPase. Mutants of the ECF1-ATPase with the introduction of Trp-for-Tyr replacement in the catalytic site of the complex made it possible to monitor the activated state for ATP synthesis (ATP conformation) in which the gamma and epsilon subunits are in close proximity to the alpha subunits and the ADP conformation, with the stalk subunits are linked to the beta subunit
MH  - 2-AZIDO-ATP
MH  - A
MH  - ADP
MH  - affinity
MH  - alpha
MH  - ALPHA-SUBUNIT
MH  - atp
MH  - ATP synthesis
MH  - BETA
MH  - BETA-SUBUNIT
MH  - BINDING
MH  - catalytic
MH  - Chemistry
MH  - COLI F1 ATPASE
MH  - COMPLEX
MH  - conformation
MH  - CROSS-LINKING
MH  - data
MH  - delta
MH  - enzyme
MH  - enzymology
MH  - EPSILON-SUBUNIT
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - INTERACTION
MH  - metabolism
MH  - model
MH  - mutant
MH  - nucleotide
MH  - nucleotide binding
MH  - Protein Conformation
MH  - Protein Subunits
MH  - Proton-Translocating ATPases
MH  - resolution
MH  - Site
MH  - stalk
MH  - SUBUNIT
MH  - Support,Non-U.S.Gov't
MH  - synthesis
MH  - universities
RP  - NOT IN FILE
NT  - University Osnabruck, Fachbereich Biologie/Chemie, Abteilung Zoophysiologie, Germany ggrueber@biologieuni-osnabrueckdeFAU - Gruber, G
SO  - J Bioenerg Biomembr 2000 Aug ;32(4):341-346

1689
UI  - 21408
AU  - Guerrieri F
AU  - Nicoletti C
AU  - Adorisio E
AU  - Caraccio G
AU  - Leonetti P
AU  - Zanotti F
AU  - Cantatore P
TI  - Correlation between decreased expression of mitochondrial F0F1-ATP synthase and low regenerating capability of the liver after partial hepatectomy in hypothyroid rats
AB  - In hypothyroid rats, partial hepatectomy does not induce liver regeneration until 120 h after surgical operation. when, instead, in normal rats a complete recovery of the liver mass, in this interval, is observed. In normal rats, a good efficiency of mitochondrial oxidative phosphorylation is needed as an energy source for liver regeneration (Guerrieri, F. et al., 1995); in hypothyroid rats the efficiency of mitochondrial oxidative phosphorylation is low in the 0-120 h interval after partial hepatectomy. This low efficiency of oxidative phosphorylation appears to be related to a low mitochondrial content of F0F1-ATP synthase, in liver of hypothyroid rats, which does not recover after partial hepatectomy. In the liver of hypothyroid rats, low levels of the nuclear-encoded mitochondrial catalytic betaF1 subunit and of its transcript are observed and they do not increase, as occurs in normal rats, after partial hepatectomy
MH  - A
MH  - Adenosine Triphosphate
MH  - adverse effects
MH  - Animal
MH  - Antithyroid Agents
MH  - Biochemistry
MH  - biology
MH  - catalytic
MH  - chemically induced
MH  - Disease Models,Animal
MH  - energy
MH  - Energy Metabolism
MH  - enzymology
MH  - Gene Expression
MH  - genetics
MH  - Hepatectomy
MH  - Hypothyroidism
MH  - Liver
MH  - Liver Regeneration
MH  - Male
MH  - metabolism
MH  - Mitochondria
MH  - Mitochondria,Liver
MH  - Oxidative Phosphorylation
MH  - Phosphorylation
MH  - physiology
MH  - Propylthiouracil
MH  - Proton-Translocating ATPases
MH  - Rats
MH  - Rats,Wistar
MH  - SUBUNIT
MH  - Support,Non-U.S.Gov't
MH  - SYNTHASE
MH  - Time Factors
MH  - universities
RP  - NOT IN FILE
NT  - Department of Medical Biochemistry and Biology and Centre for the Study of Mitochondria and Energy Metabolism (CNR), University of Bari, Italy ferguer@biochemunibaitFAU - Guerrieri, F
SO  - J Bioenerg Biomembr 2000 Apr ;32(2):183-191

1690
UI  - 21051
AU  - Guerrieri F
AU  - Nicoletti C
AU  - Adorisio E
AU  - Caraccio G
AU  - Leonetti P
AU  - Zanotti F
AU  - Cantatore P
TI  - Correlation between decreased expression of mitochondrial F0F1-ATP synthase and low regenerating capability of the liver after partial hepatectomy in hypothyroid rats
AB  - In hypothyroid rats, partial hepatectomy does not induce liver regeneration until 120 h after surgical operation. when, instead, in normal rats a complete recovery of the liver mass, in this interval, is observed. In normal rats, a good efficiency of mitochondrial oxidative phosphorylation is needed as an energy source for liver regeneration (Guerrieri, F. et al., 1995); in hypothyroid rats the efficiency of mitochondrial oxidative phosphorylation is low in the 0-120 h interval after partial hepatectomy. This low efficiency of oxidative phosphorylation appears to be related to a low mitochondrial content of F0F1-ATP synthase, in liver of hypothyroid rats, which does not recover after partial hepatectomy. In the liver of hypothyroid rats, low levels of the nuclear-encoded mitochondrial catalytic betaF1 subunit and of its transcript are observed and they do not increase, as occurs in normal rats, after partial hepatectomy
MH  - A
MH  - Biochemistry
MH  - Energy Metabolism
MH  - Liver
MH  - metabolism
MH  - Mitochondria
MH  - Oxidative Phosphorylation
MH  - Phosphorylation
MH  - Rats
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - Department of Medical Biochemistry and Biology and Centre for the Study of Mitochondria and Energy Metabolism (CNR), University of Bari, Italy ferguer@biochemunibaitFAU - Guerrieri, F
SO  - J Bioenerg Biomembr 2000 Apr ;32(2):183-191

1691
UI  - 690
AU  - Hara KY
AU  - Noji H
AU  - Bald D
AU  - Yasuda R
AU  - Kinosita K
AU  - Yoshida M
AD  - Chemical Resources Laboratory, R-1, Tokyo Institute of Technology, Nagatsuta 4259, Yokohama 226-8503, Japan
TI  - The role of the DELSEED motif of the beta subunit in rotation of F1- ATPase
AB  - F(1)-ATPase is a rotary motor protein, and ATP hydrolysis generates torque at the interface between the gamma subunit, a rotor shaft, and the alpha(3)beta(3) substructure, a stator ring. The region of conserved acidic "DELSEED" motif of the beta subunit has a contact with gamma subunit and has been assumed to be involved in torque generation. Using the thermophilic alpha(3)beta(3)gamma complex in which the corresponding sequence is DELSDED, we replaced each residue and all five acidic residues in this sequence with alanine. In addition, each of two conserved residues at the counterpart contact position of gamma subunit was also replaced. Surprisingly, all of these mutants rotated with as much torque as the wild-type. We conclude that side chains of the DELSEED motif of the beta subunit do not have a direct role in torque generation
RP  - NOT IN FILE
NT  - UI - 20261548LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 20000608IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:10799504
SO  - J Biol Chem 2000 May 12 ;275(19):14260-14263

1692
UI  - 21142
AU  - Heberle J
AD  - Research Centre Julich, IBI-2: Structural Biology, D-52425, Julich, Germany jheberle@fz-juelichde
TI  - Proton transfer reactions across bacteriorhodopsin and along the membrane
AB  - Bacteriorhodopsin is probably the best understood proton pump so far and is considered to be a model system for proton translocating membrane proteins. The basis of a molecular description of proton translocation is set by having the luxury of six highly resolved structural models at hand. Details of the mechanism and reaction dynamics were elucidated by a whole variety of biophysical techniques. The current molecular picture of catalysis by BR will be presented with examples from time-resolved spectroscopy. FT-IR spectroscopy monitors single proton transfer events within bacteriorhodopsin and judiciously positioned pH indicators detect proton migration at the membrane surface. Emerging properties are briefly outlined that underlie the efficient proton transfer across and along biological membranes
MH  - A
MH  - Bacteriorhodopsin
MH  - Catalysis
MH  - indicator
MH  - mechanism
MH  - membrane
MH  - Membrane Proteins
MH  - Membranes
MH  - model
MH  - pH
MH  - pH-indicator
MH  - protein
MH  - Proteins
MH  - proton
MH  - Proton Pump
MH  - Proton transfer
MH  - Proton-transfer
MH  - Protons
MH  - review
MH  - spectroscopy
MH  - SURFACE
MH  - SYSTEM
MH  - TRANSFER
MH  - translocation
RP  - NOT IN FILE
NT  - UI - 20273714LA - engRN - 0 (Membrane Proteins)RN - 0 (Proton Pumps)RN - 0 (Protons)RN - 53026-44-1 (Bacteriorhodopsins)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 20000630IS - 0006-3002SB - IMCY - NETHERLANDS
UR  - PM:10812029
SO  - Biochim Biophys Acta 2000 May 12 ;1458(1):135-147

1693
UI  - 20838
AU  - Hirata T
AU  - Nakamura N
AU  - Omote H
AU  - Wada Y
AU  - Futai M
AD  - Division of Biological Sciences, Institute of Scientific and Industrial Research, Osaka University, Osaka 567-0047, Japan
TI  - Regulation and reversibility of vacuolar H(+)-ATPase
AB  - Arabidopsis thaliana vacuolar H(+)-translocating pyrophosphatase (V- PPase) was expressed functionally in yeast vacuoles with endogenous vacuolar H(+)-ATPase (V-ATPase), and the regulation and reversibility of V-ATPase were studied using these vacuoles. Analysis of electrochemical proton gradient (DeltamuH) formation with ATP and pyrophosphate indicated that the proton transport by V-ATPase or V- PPase is not regulated strictly by the proton chemical gradient (DeltapH). On the other hand, vacuolar membranes may have a regulatory mechanism for maintaining a constant membrane potential (DeltaPsi). Chimeric vacuolar membranes showed ATP synthesis coupled with DeltamuH established by V-PPase. The ATP synthesis was sensitive to bafilomycin A(1) and exhibited two apparent K(m) values for ADP. These results indicate that V-ATPase is a reversible enzyme. The ATP synthesis was not observed in the presence of nigericin, which dissipates DeltapH but not DeltaPsi, suggesting that DeltapH is essential for ATP synthesis
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - ADP
MH  - analysis
MH  - Antibiotics
MH  - atp
MH  - ATP synthesis
MH  - ATPase
MH  - CONSTANT
MH  - H(+)ATPase
MH  - H+-ATPase
MH  - mechanism
MH  - membrane
MH  - Membrane Potential
MH  - Membranes
MH  - protein
MH  - Proteins
MH  - proton
MH  - Pyrophosphatases
MH  - regulation
MH  - synthesis
MH  - transport
RP  - NOT IN FILE
NT  - UI - 20085056LA - engRN - 0 (Antibiotics, Macrolide)RN - 0 (Recombinant Proteins)RN - 56-65-5 (Adenosine Triphosphate)RN - 88899-55-2 (bafilomycin A1)RN - EC 3.6.1.- (Pyrophosphatases)RN - EC 3.6.1.- (vacuolar H+-ATPase)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 20000131IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:10617629
SO  - J Biol Chem 2000 Jan 7 ;275(1):386-389

1694
UI  - 21277
AU  - Hunte C
AU  - Koepke J
AU  - Lange C
AU  - Rossmanith T
AU  - Michel H
AD  - Max-Planck-Institut fur Biophysik, Abt Molekulare Membranbiologie, Frankfurt, 60528, Germany hunte@mpibp-frankfurtmpgde
TI  - Structure at 2.3 A resolution of the cytochrome bc(1) complex from the yeast Saccharomyces cerevisiae co-crystallized with an antibody Fv fragment
AB  - BACKGROUND: The cytochrome bc(1) complex is part of the energy conversion machinery of the respiratory and photosynthetic electron transfer chains. This integral membrane protein complex catalyzes electron transfer from ubiquinol to cytochrome c. It couples the electron transfer to the electrogenic translocation of protons across the membrane via a so-called Q cycle mechanism. RESULTS: The cytochrome bc(1) complex from the yeast Saccharomyces cerevisiae was crystallized together with a bound antibody Fv fragment. The structure was determined at 2.3 A resolution using multiple isomorphous replacement, and refined to a crystallographic R factor of 22.2% (R(free) = 25.4%). The complex is present as a homodimer. Each 'monomer' of the refined model includes 2178 amino acid residues of subunits COR1, QCR2, COB, CYT1, RIP1, QCR6, QCR7, QCR8 and QCR9 of the cytochrome bc(1) complex and of the polypeptides V(H) and V(L) of the Fv fragment, the cofactors heme b(H), heme b(L), heme c(1), the [2Fe-2S] cluster and 346 water molecules. The Fv fragment binds to the extrinsic domain of the [2Fe- 2S] Rieske protein and is essential for formation of the crystal lattice. CONCLUSIONS: The approach to crystallize membrane proteins as complexes with specific antibody fragments appears to be of general importance. The structure of the yeast cytochrome bc(1) complex reveals in detail the binding sites of the natural substrate coenzyme Q6 and the inhibitor stigmatellin. Buried water molecules close to the binding sites suggest possible pathways for proton uptake and release. A comparison with other cytochrome bc(1) complexes shows features that are specific to yeast
MH  - A
MH  - ACID
MH  - BINDING
MH  - Binding Sites
MH  - COMPLEX
MH  - cytochrome
MH  - Cytochrome c
MH  - electrogenic
MH  - electron
MH  - electron transfer
MH  - Electron-transfer
MH  - England
MH  - inhibitor
MH  - mechanism
MH  - membrane
MH  - Membrane Proteins
MH  - model
MH  - protein
MH  - Proteins
MH  - proton
MH  - Protons
MH  - RESIDUE
MH  - resolution
MH  - Saccharomyces cerevisiae
MH  - Site
MH  - structure
MH  - SUBUNIT
MH  - TRANSFER
MH  - translocation
MH  - Ubiquinol-Cytochrome-c Reductase
MH  - Water
RP  - NOT IN FILE
NT  - UI - 20335027LA - engRN - 0 (Immunoglobulin Fragments)RN - 0 (immunoglobulin Fv)RN - EC 1.10.2.2 (Ubiquinol-Cytochrome-c Reductase)PT - Journal ArticleDA - 20000830SB - IMCY - ENGLAND
UR  - PM:10873857
SO  - Structure Fold Des 2000 Jun 15 ;8(6):669-684

1695
UI  - 5
AU  - Jones PC
AU  - Hermolin J
AU  - Jiang W
AU  - Fillingame RH
AD  - Department of Biomolecular Chemistry, University of Wisconsin Medical School, Madison, Wisconsin 53706, USA
TI  - Insights into the rotary catalytic mechanism of F0F1 ATP synthase from the cross-linking of subunits b and c in the Escherichia coli enzyme
AB  - The transmembrane sector of the F(0)F(1) rotary ATP synthase is proposed to organize with an oligomeric ring of c subunits, which function as a rotor, interacting with two b subunits at the periphery of the ring, the b subunits functioning as a stator. In this study, cysteines were introduced into the C-terminal region of subunit c and the N-terminal region of subunit b. Cys of N2C subunit b was cross- linked with Cys at positions 74, 75, and 78 of subunit c. In each case, a maximum of 50% of the b subunit could be cross-linked to subunit c, which suggests that either only one of the two b subunits lie adjacent to the c-ring or that both b subunits interact with a single subunit c. The results support a topological arrangement of these subunits, in which the respective N- and C-terminal ends of subunits b and c extend to the periplasmic surface of the membrane and cAsp-61 lies at the center of the membrane. The cross-linking of Cys between bN2C and cV78C was shown to inhibit ATP-driven proton pumping, as would be predicted from a rotary model for ATP synthase function, but unexpectedly, cross- linking did not lead to inhibition of ATPase activity. ATP hydrolysis and proton pumping are therefore uncoupled in the cross-linked enzyme. The c subunit lying adjacent to subunit b was shown to be mobile and to exchange with c subunits that initially occupied non-neighboring positions. The movement or exchange of subunits at the position adjacent to subunit b was blocked by dicyclohexylcarbodiimide. These experiments provide a biochemical verification that the oligomeric c- ring can move with respect to the b-stator and provide further support for a rotary catalytic mechanism in the ATP synthase
RP  - NOT IN FILE
NT  - UI - 20469499LA - engRN - 0 (Cross-Linking Reagents)RN - 0 (Lipid Bilayers)RN - 0 (Plasmids)RN - 0 (Protons)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM23105/GM/NIGMSDA - 20001023IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:10882728
SO  - J Biol Chem 2000 Oct 6 ;275(40):31340-31346

1696
UI  - 3
AU  - Jones PC
AU  - Hermolin J
AU  - Fillingame RH
AD  - Medical Research Council, Dunn Human Nutritional Unit, Cambridge CB2 2XY, United Kingdom
TI  - Mutations in single hairpin units of genetically fused subunit c provide support for a rotary catalytic mechanism in F(0)F(1) ATP synthase
AB  - Previously, we generated genetically fused dimers and trimers of subunit c of the Escherichia coli ATP synthase based upon the precedent of naturally occurring dimers in V-type H(+)-transporting ATPases. The c(2) and c(3) oligomers have proven useful in testing hypothesis regarding the mechanism of energy coupling. In the first part of this paper, the uncoupling Q42E substitution has been introduced into the second loop of the c(2) dimer or the third loop of the c(3) trimer. Both mutant proteins proved to be as functional as the wild type c(2) dimer or wild type c(3) trimer. The results argue against an obligatory movement of the epsilon subunit between loops of monomeric subunit c in the c(12) oligomer during rotary catalysis. Rather, the results support the hypothesis that the c-epsilon connection remains fixed as the c- oligomer rotates. In the second section of this paper, we report on the effect of substitution of the proton translocating Asp(61) in every second helical hairpin of the c(2) dimer, or in every third hairpin of the c(3) trimer. Based upon the precedent of V-type ATPases, where the c(2) dimer occurs naturally with a single proton translocating carboxyl in every second hairpin, these modified versions of the E. coli c(2) and c(3) fused proteins were predicted to have a functional H(+)- transporting ATPase activity, with a reduced H(+)/ATP stoichiometry, but to be inactive as ATP synthases. A variety of Asp(61)-substituted proteins proved to lack either activity indicating that the switch in function in V-type ATPases is a consequence of more than a single substitution
RP  - NOT IN FILE
NT  - UI - 20219189LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM23105/GM/NIGMSDA - 20000505IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:10753949
SO  - J Biol Chem 2000 Apr 14 ;275(15):11355-11360

1697
UI  - 968
AU  - Jucker BM
AU  - Dufour S
AU  - Ren J
AU  - Cao X
AU  - Previs SF
AU  - Underhill B
AU  - Cadman KS
AU  - Shulman GI
TI  - Assessment of mitochondrial energy coupling in vivo by 13C/31P NMR
AB  - The recently cloned uncoupling protein homolog UCP3 is expressed primarily in muscle and therefore may play a significant role in the regulation of energy expenditure and body weight. However, investigation into the regulation of uncoupling protein has been hampered by the inability to assess its activity in vivo. In this report, we demonstrate the use of a noninvasive NMR technique to assess mitochondrial energy uncoupling in skeletal muscle of awake rats by combining (13)C NMR to measure rates of mitochondrial substrate oxidation with (31)P NMR to assess unidirectional ATP synthesis flux. These combined (31)P/(13)C NMR measurements were performed in control, 10-day triiodo-l-thyronine (T(3))-treated (model of increased UCP3 expression), and acute 2,4-dinitrophenol (DNP)-treated (protonophore and mitochondrial uncoupler) rats. UCP3 mRNA and protein levels increased 8.1-fold (+/- 1.1) and 2.8-fold (+/- 0.8), respectively, in the T(3)-treated vs. control rat gastrocnemius muscle. (13)C NMR measurements of tricarboxylic acid cycle flux as an index of mitochondrial substrate oxidation were 61 +/- 21, 148 +/- 25, and 310 +/- 48 nmol/g per min in the control, T(3), and DNP groups, respectively. (31)P NMR saturation transfer measurements of unidirectional ATP synthesis flux were 83 +/- 14, 84 +/- 14, and 73 +/- 7 nmol/g per s in the control, T(3), and DNP groups, respectively. Together, these flux measurements, when normalized to the control group, suggest that acute administration of DNP (mitochondrial uncoupler) and chronic administration of T(3) decrease energy coupling by approximately 80% and approximately 60%, respectively, and that the latter treatment correlates with an increase in UCP3 mRNA and protein expression. This NMR approach could prove useful for exploring the regulation of uncoupling protein activity in vivo and elucidating its role in energy metabolism and obesity
MH  - Adenosine Triphosphate
MH  - Animal
MH  - biosynthesis
MH  - Citric Acid Cycle
MH  - Energy Metabolism
MH  - Magnetic Resonance Spectroscopy
MH  - metabolism
MH  - Mitochondria
MH  - Obesity
MH  - Oxidation-Reduction
MH  - physiology
MH  - Rats
MH  - Rats,Sprague-Dawley
MH  - Support,Non-U.S.Gov't
MH  - Support,U.S.Gov't,P.H.S.
RP  - NOT IN FILE
NT  - Departments of Internal Medicine and Cellular and Molecular Physiology, and Howard Hughes Medical Institute, Yale University School of Medicine, PO Box 9812, New Haven, CT 06510, USA
SO  - Proc Natl Acad Sci U S A 2000 Jun 6 ;97(12):6880-6884

1698
UI  - 687
AU  - Kato-Yamada Y
AU  - Yoshida M
AU  - Hisabori T
AD  - Chemical Resources Laboratory, Tokyo Institute of Technology, 4259 Nagatsuta, Yokohama 226-8503, Japan
TI  - Movement of the helical domain of the epsilon subunit is required for the activation of thermophilic F1-ATPase
AB  - The inhibitory effect of epsilon subunit in F(1)-ATPase from thermophilic Bacillus PS3 was examined focusing on the structure- function relationship. For this purpose, we designed a mutant for epsilon subunit similar to the one constructed by Schulenberg and Capaldi (Schulenberg, B., and Capaldi, R. A. (1999) J. Biol. Chem. 274, 28351-28355). We introduced two cysteine residues at the interface of N- terminal beta-sandwich domain (S48C) and C-terminal alpha-helical domain (N125C) of epsilon subunit. The alpha(3)beta(3)gammaepsilon complex containing the reduced form of this mutant epsilon subunit showed suppressed ATPase activity and gradual activation during the measurement. This activation pattern was similar to the complex with the wild type epsilon subunit. The conformation of the mutant epsilon subunit must be fixed and similar to the reported three-dimensional structure of the isolated epsilon subunit, when the intramolecular disulfide bridge was formed on this subunit by oxidation. This oxidized mutant epsilon subunit could form the alpha(3)beta(3)gammaepsilon complex but did not show any inhibitory effect. The complex was converted to the activated state, and the cross-link in the mutant epsilon subunit in the complex was efficiently formed in the presence of ATP-Mg, whereas no cross-link was observed without ATP-Mg, suggesting the conformation of the oxidized mutant epsilon subunit must be similar to that in the activated state complex. A non-hydrolyzable analog of ATP, 5'-adenylyl-beta,gamma-imidodiphosphate, could stimulate the formation of the cross-link on the epsilon subunit. Furthermore, the cross-link formation was stimulated by nucleotides even when this mutant epsilon subunit was assembled with a mutant alpha(3)beta(3)gamma complex lacking non-catalytic sites. These results indicate that binding of ATP to the catalytic sites induces a conformational change in the epsilon subunit and triggers transition of the complex from the suppressed state to the activated state
RP  - NOT IN FILE
NT  - UI - 20549575LA - engRN - 0 (Cross-Linking Reagents)RN - 0 (Disulfides)RN - 0 (Macromolecular Systems)RN - 0 (Protein Subunits)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 20001229IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:10958801
SO  - J Biol Chem 2000 Nov 17 ;275(46):35746-35750

1699
UI  - 21175
AU  - Kaulen AD
AD  - Department of Photobiochemistry, AN Belozersky Institute of Physico- Chemical Biology, Moscow State University, 119899, Moscow, Russia
TI  - Electrogenic processes and protein conformational changes accompanying the bacteriorhodopsin photocycle
AB  - The possible mechanisms of electrogenic processes accompanying proton transport in bacteriorhodopsin are discussed on the basis of recent structural data of the protein. Apparent inconsistencies between experimental data and their interpretation are considered. Special emphasis is placed on the protein conformational changes accompanying the reprotonation of chromophore and proton uptake stage in the bacteriorhodopsin photocycle
MH  - Bacteriorhodopsin
MH  - conformational change
MH  - conformational changes
MH  - electrogenic
MH  - mechanism
MH  - MECHANISMS
MH  - protein
MH  - proton
MH  - Protons
MH  - Retinaldehyde
MH  - review
MH  - transport
RP  - NOT IN FILE
NT  - UI - 20442121LA - engRN - 0 (Protons)RN - 116-31-4 (Retinaldehyde)RN - 53026-44-1 (Bacteriorhodopsins)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 20001103IS - 0006-3002SB - IMCY - NETHERLANDS
UR  - PM:10984601
SO  - Biochim Biophys Acta 2000 Aug 30 ;1460(1):204-219

1700
UI  - 498
AU  - Ko YH
AU  - Hullihen J
AU  - Hong S
AU  - Pedersen PL
AD  - Department of Biological Chemistry, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205-2185, USA
TI  - Mitochondrial F(0)F(1) ATP synthase. Subunit regions on the F1 motor shielded by F(0), Functional significance, and evidence for an involvement of the unique F(0) subunit F(6)
AB  - Studies reported here were undertaken to gain greater molecular insight into the complex structure of mitochondrial ATP synthase (F(0)F(1)) and its relationship to the enzyme's function and motor-related properties. Significantly, these studies, which employed N-terminal sequence, mass spectral, proteolytic, immunological, and functional analyses, led to the following novel findings. First, at the top of F(1) within F(0)F(1), all six N-terminal regions derived from alpha + beta subunits are shielded, indicating that one or more F(0) subunits forms a "cap." Second, at the bottom of F(1) within F(0)F(1), the N-terminal region of the single delta subunit and the C-terminal regions of all three alpha subunits are shielded also by F(0). Third, and in contrast, part of the gamma subunit located at the bottom of F(1) is already shielded in F(1), indicating that there is a preferential propensity for interaction with other F(1) subunits, most likely delta and epsilon. Fourth, and consistent with the first two conclusions above that specific regions at the top and bottom of F(1) are shielded by F(0), further proteolytic shaving of alpha and beta subunits at these locations eliminates the capacity of F(1) to couple a proton gradient to ATP synthesis. Finally, evidence was obtained that the F(0) subunit called "F(6)," unique to animal ATP synthases, is involved in shielding F(1). The significance of the studies reported here, in relation to current views about ATP synthase structure and function in animal mitochondria, is discussed
RP  - NOT IN FILE
NT  - UI - 20490733LA - engRN - 0 (Peptide Fragments)RN - 0 (Protein Subunits)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.4.19.3 (Pyroglutamyl-Peptidase I)RN - EC 3.4.21.4 (Trypsin)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - CA 10951/CA/NCIDA - 20001120IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:10887193
SO  - J Biol Chem 2000 Oct 20 ;275(42):32931-32939

1701
UI  - 808
AU  - Konno H
AU  - Yodogawa M
AU  - Stumpp MT
AU  - Kroth P
AU  - Strotmann H
AU  - Motohashi K
AU  - Amano T
AU  - Hisabori T
AD  - Chemical Resources Laboratory, Tokyo Institute of Technology, Nagatsuta 4259, Midori-Ku, Yokohama 226-8503, Japan
TI  - Inverse regulation of F1-ATPase activity by a mutation at the regulatory region on the gamma subunit of chloroplast ATP synthase
AB  - Chloroplast ATP synthase is a thiol-modulated enzyme whose DeltamuH(+)- linked activation is strongly influenced by reduction and the formation of a disulphide bridge between Cys(199) and Cys(205) on the gamma subunit. In solubilized chloroplast coupling factor 1 (CF(1)), reduction of the disulphide bond elicits the latent ATP-hydrolysing activity. To assess the regulatory importance of the amino acid residues around these cysteine residues, we focused on the three negatively charged residues Glu(210)-Asp-Glu(212) close to the two cysteine residues and also on the following region from Leu(213) to Ile(230), and investigated the modulation of ATPase activity by chloroplast thioredoxins. The mutant gamma subunits were reconstituted with the alpha and beta subunits from F(1) of the thermophilic bacterium Bacillus PS3; the active ATPase complexes obtained were purified by gel-filtration chromatography. The complex formed with a mutant gamma subunit in which Glu(210) to Glu(212) had been deleted was inactivated rather than activated by reduction of the disulphide bridge by reduced thioredoxin, indicating inverse regulation. This complex was insensitive to the inhibitory CF(1)-epsilon subunit when the mutant gamma subunit was oxidized. In contrast, the deletion of Glu(212) to Ile(230) converted the complex from a modulated state into a highly active state
RP  - NOT IN FILE
NT  - UI - 20558088LA - engRN - 0 (Chimeric Proteins)RN - 0 (Disulfides)RN - 0 (Plant Proteins)RN - 0 (Protein Subunits)RN - 0 (Reducing Agents)RN - 3483-12-3 (Dithiothreitol)RN - 52500-60-4 (Thioredoxin)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 20010117IS - 0264-6021SB - IMCY - ENGLANDJC - 9YO
UR  - PM:11104686
SO  - Biochem J 2000 Dec 15 ;352 Pt 3():783-788

1702
UI  - 20837
AU  - Le NP
AU  - Omote H
AU  - Wada Y
AU  - al Shawi MK
AU  - Nakamoto RK
AU  - Futai M
AD  - Division of Biological Sciences, The Institute of Scientific and Industrial Research, Osaka University, CREST (Core Research for Evolutional Science and Technology) of Japan Science and Technology Corporation, Ibaraki, Osaka 567-0047, Japan
TI  - Escherichia coli ATP synthase alpha subunit Arg-376: the catalytic site arginine does not participate in the hydrolysis/synthesis reaction but is required for promotion to the steady state
AB  - The three catalytic sites of the F(O)F(1) ATP synthase interact through a cooperative mechanism that is required for the promotion of catalysis. Replacement of the conserved alpha subunit Arg-376 in the Escherichia coli F(1) catalytic site with Ala or Lys resulted in turnover rates of ATP hydrolysis that were 2 x 10(3)-fold lower than that of the wild type. Mutant enzymes catalyzed hydrolysis at a single site with kinetics similar to that of the wild type; however, addition of excess ATP did not chase bound ATP, ADP, or Pi from the catalytic site, indicating that binding of ATP to the second and third sites failed to promote release of products from the first site. Direct monitoring of nucleotide binding in the alphaR376A and alphaR376K mutant F(1) by a tryptophan in place of betaTyr-331 (Weber et al. (1993) J. Biol. Chem. 268, 20126-20133) showed that the catalytic sites of the mutant enzymes, like the wild type, have different affinities and therefore, are structurally asymmetric. These results indicate that alphaArg-376, which is close to the beta- or gamma-phosphate group of bound ADP or ATP, respectively, does not make a significant contribution to the catalytic reaction, but coordination of the arginine to nucleotide filling the low-affinity sites is essential for promotion of rotational catalysis to steady-state turnover
MH  - A
MH  - ADP
MH  - ALPHA-SUBUNIT
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BINDING
MH  - Catalysis
MH  - COLI ATP SYNTHASE
MH  - Enzymes
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - Hydrolysis
MH  - Kinetics
MH  - mechanism
MH  - mutant
MH  - nucleotide binding
MH  - Phosphates
MH  - Site
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 20170906LA - engRN - 0 (Phosphates)RN - 74-79-3 (Arginine)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleID - GM-50957/GM/NIGMSDA - 20000419IS - 0006-2960SB - IMCY - UNITED STATES
UR  - PM:10704230
SO  - Biochemistry 2000 Mar 14 ;39(10):2778-2783

1703
UI  - 321
AU  - Leslie AG
AU  - Walker JE
AD  - MRC Laboratory of Molecular Biology, Cambridge, UK
TI  - Structural model of F1-ATPase and the implications for rotary catalysis
AB  - The crystal structure of bovine mitochondrial F1-ATPase is described. Several features of the structure are consistent with the binding change mechanism of catalysis, in which binding of substrates induces conformational changes that result in a high degree of cooperativity between the three catalytic sites. Furthermore, the structure also suggests that catalysis is accompanied by a physical rotation of the centrally placed gamma-subunit relative to the approximately spherical alpha3beta3 subassembly
RP  - NOT IN FILE
NT  - UI - 20294489LA - engRN - 0 (Macromolecular Systems)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 20000920IS - 0962-8436SB - IMCY - ENGLANDJC - P5Z
UR  - PM:10836500
SO  - Philos Trans R Soc Lond B Biol Sci 2000 Apr 29 ;355(1396):465-471

1704
UI  - 21247
AU  - Luecke H
AU  - Schobert B
AU  - Cartailler JP
AU  - Richter HT
AU  - Rosengarth A
AU  - Needleman R
AU  - Lanyi JK
AD  - Department of Molecular Biology and Biochemistry, University of California, Irvine, CA 92697, USA
TI  - Coupling photoisomerization of retinal to directional transport in bacteriorhodopsin
AB  - In order to understand how isomerization of the retinal drives unidirectional transmembrane ion transport in bacteriorhodopsin, we determined the atomic structures of the BR state and M photointermediate of the E204Q mutant, to 1.7 and 1.8 A resolution, respectively. Comparison of this M, in which proton release to the extracellular surface is blocked, with the previously determined M in the D96N mutant indicates that the changes in the extracellular region are initiated by changes in the electrostatic interactions of the retinal Schiff base with Asp85 and Asp212, but those on the cytoplasmic side originate from steric conflict of the 13-methyl retinal group with Trp182 and distortion of the pi-bulge of helix G. The structural changes suggest that protonation of Asp85 initiates a cascade of atomic displacements in the extracellular region that cause release of a proton to the surface. The progressive relaxation of the strained 13- cis retinal chain with deprotonated Schiff base, in turn, initiates atomic displacements in the cytoplasmic region that cause the intercalation of a hydrogen-bonded water molecule between Thr46 and Asp96. This accounts for the lowering of the pK(a) of Asp96, which then reprotonates the Schiff base via a newly formed chain of water molecules that is extending toward the Schiff base
MH  - A
MH  - Bacteriorhodopsin
MH  - BASE
MH  - Biochemistry
MH  - coupling
MH  - England
MH  - ion
MH  - Ion Transport
MH  - M
MH  - membrane
MH  - Membrane Proteins
MH  - mutant
MH  - photoisomerization
MH  - protein
MH  - Proteins
MH  - proton
MH  - proton release
MH  - protonation
MH  - Protons
MH  - relaxation
MH  - resolution
MH  - retinal
MH  - Retinaldehyde
MH  - Schiff base
MH  - Schiff Bases
MH  - Schiff-base
MH  - structure
MH  - SURFACE
MH  - transport
MH  - Water
RP  - NOT IN FILE
NT  - UI - 20366243LA - engRN - 0 (Membrane Proteins)RN - 0 (Protons)RN - 0 (Schiff Bases)RN - 116-31-4 (Retinaldehyde)RN - 53026-44-1 (Bacteriorhodopsins)RN - 7732-18-5 (Water)PT - Journal ArticleID - R01GM29498/GM/NIGMSID - R01GM59970/GM/NIGMSDA - 20000825IS - 0022-2836SB - IMSB - SCY - ENGLAND
UR  - PM:10903866
SO  - J Mol Biol 2000 Jul 28 ;300(5):1237-1255

1705
UI  - 19896
AU  - Marcus RA
TI  - On the theory of ion transfer rates across the interface of two immiscible liquids
AB  - Ion transfer across the interface of two immiscible liquids involves a mechanism for initiating desolvation from the first liquid, A, and concerted solvation by the second, B. In the present article a mechanism is considered in which this initiation is facilitated by the ion attaching itself to the tip of a solvent protrusion of B into A. (Protrusions have been observed in computer simulations and termed "fingers" or "cones.") It is presumed that the most effective protrusion represents a balance between two opposing effects: the more convex the protrusion the less probable the ion/protrusion formation but also the less the resistance to extrusion of the intervening liquid between the ion and the surface. An analogy of the latter to hydrodynamics is noted, namely, the more convex the surface the less the frictional force it exerts on the approaching ion. After diffusion in coordinate and solvation space across the interfacial region, the final detachment of the ion from solvent A is assumed to occur from a protrusion of A into B. Existing data on ion transfer rates are discussed, including the question of diffusion vs kinetic control. Computer simulations that correspond to the experimental conditions in realistic liquids for measurement of the electrochemical exchange current rate constant k(0) are suggested. They can be used to test specific theoretical features. With a suitable choice of systems the need (and a major barrier to the simulations) for having a base electrolyte in such simulations can be bypassed. An experiment for the real-time observation of an ion leaving the interface is also suggested. (C) 2000 American Institute of Physics. [S0021- 9606(00)50628-X]
MH  - A
MH  - BASE
MH  - CHARGE-TRANSFER PROCESSES
MH  - computer simulations
MH  - COMPUTER-SIMULATION
MH  - COMPUTER-SIMULATIONS
MH  - CONSTANT
MH  - Diffusion
MH  - DOUBLE-LAYER
MH  - ELECTRON-TRANSFER REACTIONS
MH  - FLUCTUATION ANALYSIS
MH  - ion
MH  - ion transfer rate
MH  - LASER-SCATTERING METHOD
MH  - LIQUID
MH  - LIQUID/LIQUID INTERFACE
MH  - mechanism
MH  - MOLECULAR-DYNAMICS
MH  - rate constant
MH  - SCANNING ELECTROCHEMICAL MICROSCOPY
MH  - SIMULATION
MH  - SIMULATIONS
MH  - SOLVATION
MH  - SOLVENT
MH  - SURFACE
MH  - SYSTEM
MH  - SYSTEMS
MH  - theory
MH  - TRANSFER
MH  - TRANSFER KINETICS
MH  - WATER/1;2-DICHLOROETHANE INTERFACE
RP  - NOT IN FILE
NT  - JournalJUL 22333RJMarcus RA CALTECH, Arthur Amos Noyes Lab Chem Phys, Pasadena, CA 91125 USAJ CHEM PHYS
AV  - CALTECH, Arthur Amos Noyes Lab Chem Phys, Pasadena, CA 91125 USA CALTECH, Arthur Amos Noyes Lab Chem Phys, Pasadena, CA 91125 USA
UR  - ISI:000088144000038
SO  - Journal of Chemical Physics 2000  ;113(4):1618-1629

1706
UI  - 693
AU  - Masaike T
AU  - Mitome N
AU  - Noji H
AU  - Muneyuki E
AU  - Yasuda R
AU  - Kinosita K
AU  - Yoshida M
AD  - The Research Laboratory of Resources Utilization, Tokyo Institute of Technology, Yokohama 226-8503, Japan
TI  - Rotation of F(1)-ATPase and the hinge residues of the beta subunit
AB  - Rotation of a motor protein, F(1)-ATPase, was demonstrated using a unique single-molecule observation system. This paper reviews what has been clarified by this system and then focuses on the role of residues at the hinge region of the beta subunit. We have visualised rotation of a single molecule of F(1)-ATPase by attaching a fluorescent actin filament to the top of the beta subunit in the immobilised F(1)-ATPase, thus settling a major controversy regarding the rotary catalysis. The rotation of the beta subunit was exclusively in one direction, as could be predicted by the crystal structure of bovine heart F(1)-ATPase. Rotation at low ATP concentrations revealed that one revolution consists of three 120 degrees steps, each fuelled by the binding of an ATP to the beta subunit. The mean work done by a 120 degrees step was approximately 80 pN nm, a value close to the free energy liberated by hydrolysis of one ATP molecule, implying nearly 100% efficiency of energy conversion. The torque is probably generated by the beta subunit, which undergoes large opening-closing domain motion upon binding of AT(D)P. We identified three hinge residues, betaHis179, betaGly180 and betaGly181, whose peptide bond dihedral angles are drastically changed during domain motion. Simultaneous substitution of these residues with alanine resulted in nearly complete loss (99%) of ATPase activity. Single or double substitution of the two Gly residues did not abolish the ATPase activity. However, reflecting the shift of the equilibrium between the open and closed forms of the beta subunit, single substitution caused changes in the propensity to generate the kinetically trapped Mg-ADP inhibited form: Gly180Ala enhanced the propensity and Gly181Ala abolished the propensity. In spite of these changes, the mean rotational torque was not changed significantly for any of the mutants
RP  - NOT IN FILE
NT  - UI - 20123945LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 20000217IS - 0022-0949SB - IMCY - ENGLANDJC - I2F
UR  - PM:10600667
SO  - J Exp Biol 2000 Jan ;203 Pt 1():1-8

1707
UI  - 21411
AU  - Mueller DM
TI  - Partial assembly of the yeast mitochondrial ATP synthase
AB  - The mitochondrial ATP synthase is a molecular motor that drives the phosphorylation of ADP to ATP. The yeast mitochondrial ATP synthase is composed of at least 19 different peptides, which comprise the F1 catalytic domain, the F0 proton pore, and two stalks, one of which is thought to act as a stator to link and hold F1 to F0, and the other as a rotor. Genetic studies using yeast Saccharomyces cerevisiae have suggested the hypothesis that the yeast mitochondrial ATP synthase can be assembled in the absence of 1, and even 2, of the polypeptides that are thought to comprise the rotor. However, the enzyme complex assembled in the absence of the rotor is thought to be uncoupled, allowing protons to freely flow through F0 into the mitochondrial matrix. Left uncontrolled, this is a lethal process and the cell must eliminate this leak if it is to survive. In yeast, the cell is thought to lose or delete its mitochondrial DNA (the petite mutation) thereby eliminating the genes encoding essential components of F0. Recent biochemical studies in yeast, and prior studies in E. coli, have provided support for the assembly of a partial ATP synthase in which the ATP synthase is no longer coupled to proton translocation
MH  - A
MH  - ADP
MH  - atp
MH  - ATP synthase
MH  - Biochemistry
MH  - biology
MH  - catalytic
MH  - catalytic domain
MH  - cell
MH  - Chemistry
MH  - COMPLEX
MH  - Dna
MH  - enzyme
MH  - enzymology
MH  - F0
MH  - F1
MH  - genetics
MH  - M
MH  - metabolism
MH  - Mitochondria
MH  - mutagenesis
MH  - Mutation
MH  - peptide
MH  - peptides
MH  - Phosphorylation
MH  - Protein Subunits
MH  - proton
MH  - Proton-Translocating ATPases
MH  - Protons
MH  - Recombinant Proteins
MH  - Saccharomyces cerevisiae
MH  - Sequence Deletion
MH  - stalk
MH  - Support,U.S.Gov't,P.H.S.
MH  - SYNTHASE
MH  - translocation
MH  - YEAST
RP  - NOT IN FILE
NT  - Department of Biochemistry and Molecular Biology, The Chicago Medical School, Illinois 60064, USA muellerd@misfinchcmseduFAU - Mueller, D M
SO  - J Bioenerg Biomembr 2000 Aug ;32(4):391-400

1708
UI  - 21054
AU  - Mueller DM
TI  - Partial assembly of the yeast mitochondrial ATP synthase
AB  - The mitochondrial ATP synthase is a molecular motor that drives the phosphorylation of ADP to ATP. The yeast mitochondrial ATP synthase is composed of at least 19 different peptides, which comprise the F1 catalytic domain, the F0 proton pore, and two stalks, one of which is thought to act as a stator to link and hold F1 to F0, and the other as a rotor. Genetic studies using yeast Saccharomyces cerevisiae have suggested the hypothesis that the yeast mitochondrial ATP synthase can be assembled in the absence of 1, and even 2, of the polypeptides that are thought to comprise the rotor. However, the enzyme complex assembled in the absence of the rotor is thought to be uncoupled, allowing protons to freely flow through F0 into the mitochondrial matrix. Left uncontrolled, this is a lethal process and the cell must eliminate this leak if it is to survive. In yeast, the cell is thought to lose or delete its mitochondrial DNA (the petite mutation) thereby eliminating the genes encoding essential components of F0. Recent biochemical studies in yeast, and prior studies in E. coli, have provided support for the assembly of a partial ATP synthase in which the ATP synthase is no longer coupled to proton translocation
MH  - A
MH  - ADP
MH  - atp
MH  - ATP synthase
MH  - Biochemistry
MH  - COMPLEX
MH  - F0
MH  - F1
MH  - M
MH  - Phosphorylation
MH  - proton
MH  - Protons
MH  - Saccharomyces cerevisiae
MH  - stalk
MH  - SYNTHASE
MH  - translocation
RP  - NOT IN FILE
NT  - Department of Biochemistry and Molecular Biology, The Chicago Medical School, Illinois 60064, USA muellerd@misfinchcmseduFAU - Mueller, D M
SO  - J Bioenerg Biomembr 2000 Aug ;32(4):391-400

1709
UI  - 689
AU  - Muneyuki E
AU  - Noji H
AU  - Amano T
AU  - Masaike T
AU  - Yoshida M
AD  - Research Laboratory of Resources Utilization, R-1, Tokyo Institute of Technology, Yokohama, Japan
TI  - F(0)F(1)-ATP synthase: general structural features of 'ATP-engine' and a problem on free energy transduction
RP  - NOT IN FILE
NT  - UI - 20298324LA - engRN - 0 (Bacterial Proteins)RN - 0 (Molecular Motors)RN - 0 (Protons)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 20000727IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:10838059
SO  - Biochim Biophys Acta 2000 May 31 ;1458(2-3):467-481

1710
UI  - 21031
AU  - Murata T
AU  - Igarashi K
AU  - Kakinuma Y
AU  - Yamato I
AD  - Department of Biological Science and Technology, Science University of Tokyo, 2641 Yamazaki, Noda-shi, Chiba 278-8510, Japan murata@y105hptbnodasutacjp
TI  - Na+ binding of V-type Na+-ATPase in Enterococcus hirae
AB  - Rotation catalysis theory has been successfully applied to the molecular mechanism of the ATP synthase (F(0)F(1)-ATPase) and probably of the vacuolar ATPase. We investigated the ion binding step to Enterococcus hirae Na(+)-translocating V-ATPase. The kinetics of Na(+) binding to purified V-ATPase suggested 6 +/- 1 Na(+) bound/enzyme molecule, with a single high affinity (K(d(Na(+()))) = 15 +/- 5 micrometer). The number of cation binding sites is consistent with the model that V-ATPase proteolipids form a rotor ring consisting of hexamers, each having one cation binding site. Release of the bound (22)Na(+) from purified molecules in a chasing experiment showed two phases: a fast component (about two-thirds of the total amount of bound Na(+); k(exchange) > 1.7 min(-1)) and a slow component (about one-third of the total; k(exchange) = 0.16 min(-1)), which changes to the fast component by adding ATP or ATPgammaS. This suggested that about two- thirds of the Na(+) binding sites of the Na(+)-ATPase are readily accessible from the aqueous phase and that the slow component is important for the transport reaction
MH  - A
MH  - Adenosinetriphosphatase
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BINDING
MH  - Binding Sites
MH  - Catalysis
MH  - ion
MH  - Kinetics
MH  - mechanism
MH  - model
MH  - Proteolipids
MH  - rotation
MH  - Site
MH  - Sodium
MH  - SYNTHASE
MH  - theory
MH  - transport
RP  - NOT IN FILE
NT  - UI - 20250894LA - engRN - 7440-23-5 (Sodium)RN - EC 3.6.1.- (sodium-translocating ATPase)RN - EC 3.6.1.3 (Adenosinetriphosphatase)PT - Journal ArticleDA - 20000601IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:10788452
SO  - J Biol Chem 2000 May 5 ;275(18):13415-13419

1711
UI  - 402
AU  - Nadanaciva S
AU  - Weber J
AU  - Senior AE
AD  - Department of Biochemistry and Biophysics, University of Rochester Medical Center, New York 14642, USA
TI  - New probes of the F1-ATPase catalytic transition state reveal that two of the three catalytic sites can assume a transition state conformation simultaneously
AB  - MgADP in combination with fluoroscandium (ScFx) is shown to form a potently inhibitory, tightly bound, noncovalent complex at the catalytic sites of F(1)-ATPase. The F(1).MgADP.ScFx complex mimics a catalytic transition state. Notably, ScFx caused large enhancement of MgADP binding affinity at both catalytic sites 1 and 2, with little effect at site 3. These results indicate that sites 1 and 2 may form a transition state conformation. A new direct optical probe of F(1)- ATPase catalytic transition state conformation is also reported, namely, substantial enhancement of fluorescence emission of residue beta-Trp-148 observed upon binding of MgADP.ScFx or MgIDP. ScFx. Using this fluorescence signal, titrations were performed with MgIDP.ScFx which demonstrated that catalytic sites 1 and 2 can both form a transition state conformation but site 3 cannot. Supporting data were obtained using MgIDP-fluoroaluminate. Current models of the MgATP hydrolysis mechanism uniformly make the assumption that only one catalytic site hydrolyzes MgATP at any one time. The fluorometal analogues demonstrate that two sites have the capability to form the transition state simultaneously
RP  - NOT IN FILE
NT  - UI - 20384248LA - engRN - 0 (Enzyme Inhibitors)RN - 55520-40-6 (Tyrosine)RN - 56-85-9 (Glutamine)RN - 56-87-1 (Lysine)RN - 58-64-0 (Adenosine Diphosphate)RN - 7004-12-8 (Arginine)RN - 73-22-3 (Tryptophan)RN - 7440-20-2 (Scandium)RN - 86-04-4 (Inosine Diphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSDA - 20000907IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:10924155
SO  - Biochemistry 2000 Aug 8 ;39(31):9583-9590

1712
UI  - 20921
AU  - Nakamoto RK
AU  - Ketchum CJ
AU  - Kuo PH
AU  - Peskova YB
AU  - al Shawi MK
AD  - Department of Molecular Physiology and Biological Physics, University of Virginia, PO Box 10011, Charlottesville, VA 22906-0011, USA rkn3c@virginiaedu
TI  - Molecular mechanisms of rotational catalysis in the F(0)F(1) ATP synthase
AB  - Rotation of the F(0)F(1) ATP synthase gamma subunit drives each of the three catalytic sites through their reaction pathways. The enzyme completes three cycles and synthesizes or hydrolyzes three ATP for each 360 degrees rotation of the gamma subunit. Mutagenesis studies have yielded considerable information on the roles of interactions between the rotor gamma subunit and the catalytic beta subunits. Amino acid substitutions, such as replacement of the conserved gammaMet-23 by Lys, cause altered interactions between gamma and beta subunits that have dramatic effects on the transition state of the steady state ATP synthesis and hydrolysis reactions. The mutations also perturb transmission of specific conformational information between subunits which is important for efficient conversion of energy between rotation and catalysis, and render the coupling between catalysis and transport inefficient. Amino acid replacements in the transport domain also affect the steady state catalytic transition state indicating that rotation is involved in coupling to transport
MH  - ACID
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - ATPase
MH  - BETA-SUBUNIT
MH  - Catalysis
MH  - coupling
MH  - Hydrolysis
MH  - mechanism
MH  - MECHANISMS
MH  - mutagenesis
MH  - physiology
MH  - review
MH  - rotation
MH  - Site
MH  - SUBUNIT
MH  - SYNTHASE
MH  - synthesis
MH  - transport
RP  - NOT IN FILE
NT  - UI - 20298310LA - engRN - 0 (Molecular Motors)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticlePT - ReviewPT - Review, TutorialID - R01-GM50957/GM/NIGMSID - R01-GM52502/GM/NIGMSDA - 20000727IS - 0006-3002SB - IMCY - NETHERLANDS
UR  - PM:10838045
SO  - Biochim Biophys Acta 2000 May 31 ;1458(2-3):289-299

1713
UI  - 21253
AU  - Nogi T
AU  - Fathir I
AU  - Kobayashi M
AU  - Nozawa T
AU  - Miki K
AD  - Department of Chemistry, Graduate School of Science, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan
TI  - Crystal structures of photosynthetic reaction center and high-potential iron-sulfur protein from Thermochromatium tepidum: thermostability and electron transfer
AB  - The reaction center (RC) of photosynthetic bacteria is a membrane protein complex that promotes a light-induced charge separation during the primary process of photosynthesis. In the photosynthetic electron transfer chain, the soluble electron carrier proteins transport electrons to the RC and reduce the photo-oxidized special-pair of bacteriochlorophyll. The high-potential iron-sulfur protein (HiPIP) is known to serve as an electron donor to the RC in some species, where the c-type cytochrome subunit, the peripheral subunit of the RC, directly accepts electrons from the HiPIP. Here we report the crystal structures of the RC and the HiPIP from Thermochromatium (Tch.) tepidum, at 2.2-A and 1.5-A resolution, respectively. Tch. tepidum can grow at the highest temperature of all known purple bacteria, and the Tch. tepidum RC shows some degree of stability to high temperature. Comparison with the RCs of mesophiles, such as Blastochloris viridis, has shown that the Tch. tepidum RC possesses more Arg residues at the membrane surface, which might contribute to the stability of this membrane protein. The RC and the HiPIP both possess hydrophobic patches on their respective surfaces, and the HiPIP is expected to interact with the cytochrome subunit by hydrophobic interactions near the heme- 1, the most distal heme to the special-pair
MH  - A
MH  - Bacteria
MH  - Carrier Proteins
MH  - Chemistry
MH  - COMPLEX
MH  - cytochrome
MH  - electron
MH  - electron transfer
MH  - Electron-transfer
MH  - Electrons
MH  - membrane
MH  - Photosynthesis
MH  - protein
MH  - Proteins
MH  - reaction center
MH  - RESIDUE
MH  - resolution
MH  - structure
MH  - SUBUNIT
MH  - SURFACE
MH  - Temperature
MH  - TRANSFER
MH  - transport
RP  - NOT IN FILE
NT  - UI - 20558546LA - engRN - 0 (Iron-Sulfur Proteins)RN - 0 (Photosynthetic Reaction Center, Bacterial)PT - Journal ArticleDA - 20001218IS - 0027-8424SB - IMCY - UNITED STATES
UR  - PM:11095707
SO  - Proc Natl Acad Sci U S A 2000 Dec 5 ;97(25):13561-13566

1714
UI  - 19904
AU  - Oster G
AU  - Wang H
AU  - Grabe M
AD  - University of California, Berkeley 94720-3112, USA
TI  - How Fo-ATPase generates rotary torque
AB  - The F-ATPases synthesize ATP using a transmembrane ionmotive force (IMF) established by the electron transport chain. This transduction involves first converting the IMF to a rotary torque in the transmembrane Fo portion. This torque is communicated from Fo to the F1 portion where the energy is used to release the newly synthesized ATP from the catalytic sites according to Boyer's binding change mechanism. Here we explain the principle by which an IMF generates this rotary torque in the Fo ion engine
MH  - A
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BINDING
MH  - CHANGE MECHANISM
MH  - electron
MH  - Electron Transport
MH  - England
MH  - F-ATPASE
MH  - F1
MH  - H(+)-Transporting ATP Synthase
MH  - ion
MH  - mechanism
MH  - review
MH  - Site
MH  - SYNTHASE
MH  - transport
RP  - NOT IN FILE
NT  - UI - 20294494LA - engRN - 0 (Molecular Motors)RN - EC 3.6.1.- (V-type ATPase)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 20000920IS - 0962-8436SB - IMCY - ENGLANDJC - P5Z
UR  - PM:10836505
SO  - Philos Trans R Soc Lond B Biol Sci 2000 Apr 29 ;355(1396):523-528

1715
UI  - 19903
AU  - Oster G
AU  - Wang H
AD  - Department of Molecular and Cellular Biology and College of Natural Resources, University of California, Berkeley, CA 94720-3112, USA goster@natureberkeleyedu
TI  - Reverse engineering a protein: the mechanochemistry of ATP synthase
AB  - ATP synthase comprises two rotary motors in one. The F(1) motor can generate a mechanical torque using the hydrolysis energy of ATP. The F(o) motor generates a rotary torque in the opposite direction, but it employs a transmembrane proton motive force. Each motor can be reversed: The F(o) motor can drive the F(1) motor in reverse to synthesize ATP, and the F(1) motor can drive the F(o) motor in reverse to pump protons. Thus ATP synthase exhibits two of the major energy transduction pathways employed by the cell to convert chemical energy into mechanical force. Here we show how a physical analysis of the F(1) and F(o) motors can provide a unified view of the mechanochemical principles underlying these energy transducers
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - analysis
MH  - atp
MH  - ATP synthase
MH  - H(+)-Transporting ATP Synthase
MH  - Hydrolysis
MH  - protein
MH  - proton
MH  - Protons
MH  - review
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 20298325LA - engRN - 0 (Molecular Motors)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 20000727IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:10838060
SO  - Biochim Biophys Acta 2000 May 31 ;1458(2-3):482-510

1716
UI  - 692
AU  - Ozawa K
AU  - Meikari T
AU  - Motohashi K
AU  - Yoshida M
AU  - Akutsu H
AD  - Department of Chemistry and Biotechnology, Faculty of Engineering, Yokohama National University, Hodogaya-ku, Yokohama 240-8501, Japan
TI  - Evidence for the presence of an F-type ATP synthase involved in sulfate respiration in Desulfovibrio vulgaris
AB  - Using a library of genomic DNA from Desulfovibrio vulgaris Miyazaki F, a strict anaerobe, and two synthetic deoxyoligonucleotide probes designed for F-type ATPases, the genes for open reading frames (ORFs) 1 to 5 were cloned and sequenced. The predicted protein sequences of the gene products indicate that they are composed of 172, 488, 294, 471, and 134 amino acids, respectively, and that they share considerable identity at the amino acid level with delta, alpha, gamma, beta, and epsilon subunits found in other F-type ATPases, respectively. Furthermore, a component carrying ATPase activity was partially purified from the cytoplasmic membrane fraction of the D. vulgaris Miyazaki F cells. The N-terminal amino acid sequences of three major polypeptides separated by sodium dodecyl sulfate-12% polyacrylamide gel electrophoresis were identical to those of the products predicted by the sequences of ORF-2, ORF-3, and ORF-4, suggesting that an F-type ATPase is functioning in the D. vulgaris Miyazaki F cytoplasmic membrane. The amount of the F-type ATPase produced in the D. vulgaris Miyazaki F cells is similar to that in the Escherichia coli cells cultured aerobically. It indicates that the enzyme works as an ATP synthase in the D. vulgaris Miyazaki F cells in connection with sulfate respiration
RP  - NOT IN FILE
NT  - UI - 20200358LA - engRN - 0 (Sulfates)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 20000418IS - 0021-9193SB - IMCY - UNITED STATESJC - HH3
UR  - PM:10735863
SO  - J Bacteriol 2000 Apr ;182(8):2200-2206

1717
UI  - 280
AU  - Panke O
AU  - Gumbiowski K
AU  - Junge W
AU  - Engelbrecht S
AD  - Universitat Osnabruck, Fachbereich Biologie, Abteilung Biophysik, Barbarastr 11, 49069, Osnabruck, Germany
TI  - F-ATPase: specific observation of the rotating c subunit oligomer of EF(o)EF(1)
AB  - The rotary motion in response to ATP hydrolysis of the ring of c subunits of the membrane portion, F(o), of ATP synthase, F(o)F(1), is still under contention. It was studied with EF(o)EF(1) (Escherichia coli) using microvideography with a fluorescent actin filament. To overcome the limited specificity of actin attachment through a Cys- maleimide couple which might have hampered the interpretation of previous work, we engineered a 'strep-tag' sequence into the C-terminal end of subunit c. It served (a) to purify the holoenzyme and (b) to monospecifically attach a fluorescent actin filament to subunit c. EF(o)EF(1) was immobilized on a Ni-NTA-coated glass slide by the engineered His-tag at the N-terminus of subunit beta. In the presence of MgATP we observed up to five counterclockwise rotating actin filaments per picture frame of 2000 microm(2) size, in some cases yielding a proportion of 5% rotating over total filaments. The rotation was unequivocally attributable to the ring of subunit c. The new, doubly engineered construct serves as a firmer basis for ongoing studies on torque and angular elastic distortions between F(1) and F(o)
RP  - NOT IN FILE
NT  - UI - 20245436LA - engRN - 0 (Actins)RN - 0 (Fluorescent Dyes)RN - 0 (Oligopeptides)RN - 58-85-5 (Biotin)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 20000602IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:10781800
SO  - FEBS Lett 2000 Apr 21 ;472(1):34-38

1718
UI  - 21053
AU  - Papa S
AU  - Zanotti F
AU  - Gaballo A
TI  - The structural and functional connection between the catalytic and proton translocating sectors of the mitochondrial F1F0-ATP synthase
AB  - The structural and functional connection between the peripheral catalytic F1 sector and the proton-translocating membrane sector F0 of the mitochondrial ATP synthase is reviewed. The observations examined show that the N-terminus of subunit gamma, the carboxy-terminal and central region of F0I-PVP(b), OSCP, and part of subunit d constitute a continuous structure, the lateral stalk, which connects the peripheries of F1 to F0 and surrounds the central element of the stalk, constituted by subunits gamma and delta. The ATPase inhibitor protein (IF1) binds at one side of the F1F0 connection. The carboxy-terminal segment of IF1 apparently binds to OSCP. The 42L-58K segment of IF1, which is per se the most active domain of the protein, binds at the surface of one of the three alpha/beta pairs of F1, thus preventing the cyclic interconversion of the catalytic sites required for ATP hydrolysis
MH  - A
MH  - ACTIVE
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Biochemistry
MH  - delta
MH  - F0
MH  - F1
MH  - F1F0-ATP SYNTHASE
MH  - Hydrolysis
MH  - inhibitor
MH  - INHIBITOR PROTEIN
MH  - membrane
MH  - protein
MH  - proton
MH  - Site
MH  - stalk
MH  - structure
MH  - SUBUNIT
MH  - SURFACE
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - Department of Medical Biochemistry and Biology, University of Bari, Italy papabehm@cimedocunibaitFAU - Papa, S
SO  - J Bioenerg Biomembr 2000 Aug ;32(4):401-411

1719
UI  - 21410
AU  - Papa S
AU  - Zanotti F
AU  - Gaballo A
TI  - The structural and functional connection between the catalytic and proton translocating sectors of the mitochondrial F1F0-ATP synthase
AB  - The structural and functional connection between the peripheral catalytic F1 sector and the proton-translocating membrane sector F0 of the mitochondrial ATP synthase is reviewed. The observations examined show that the N-terminus of subunit gamma, the carboxy-terminal and central region of F0I-PVP(b), OSCP, and part of subunit d constitute a continuous structure, the lateral stalk, which connects the peripheries of F1 to F0 and surrounds the central element of the stalk, constituted by subunits gamma and delta. The ATPase inhibitor protein (IF1) binds at one side of the F1F0 connection. The carboxy-terminal segment of IF1 apparently binds to OSCP. The 42L-58K segment of IF1, which is per se the most active domain of the protein, binds at the surface of one of the three alpha/beta pairs of F1, thus preventing the cyclic interconversion of the catalytic sites required for ATP hydrolysis
MH  - A
MH  - ACTIVE
MH  - Animal
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Biochemistry
MH  - biology
MH  - catalytic
MH  - catalytic domain
MH  - Chemistry
MH  - Crystallography,X-Ray
MH  - cyclic
MH  - delta
MH  - enzymology
MH  - Escherichia coli
MH  - F0
MH  - F1
MH  - F1F0-ATP SYNTHASE
MH  - Hydrolysis
MH  - inhibitor
MH  - INHIBITOR PROTEIN
MH  - Kinetics
MH  - membrane
MH  - metabolism
MH  - Mitochondria
MH  - Models,Molecular
MH  - Oligomycins
MH  - Peptide Fragments
MH  - pharmacology
MH  - protein
MH  - Protein Subunits
MH  - proton
MH  - Proton-Translocating ATPases
MH  - Site
MH  - stalk
MH  - structure
MH  - SUBUNIT
MH  - Support,Non-U.S.Gov't
MH  - SURFACE
MH  - SYNTHASE
MH  - universities
RP  - NOT IN FILE
NT  - Department of Medical Biochemistry and Biology, University of Bari, Italy papabehm@cimedocunibaitFAU - Papa, S
SO  - J Bioenerg Biomembr 2000 Aug ;32(4):401-411

1720
UI  - 21419
AU  - Pedersen PL
AU  - Ko YH
AU  - Hong S
TI  - ATP synthases in the year 2000: evolving views about the structures of these remarkable enzyme complexes
AB  - This introductory article briefly summarizes how our views about the structural features of ATP synthases (F0F1) have evolved over the past 30 years and also reviews some of our current views in the year 2000 about the structures of these remarkably unique enzyme complexes. Suffice it to say that as we approach the end of the first year of this new millinium, we can be conservatively confident that we have a reasonably good grasp of the overall "low-resolution" structural features of ATP synthases. Electron microscopy techniques, combined with the tools of biochemistry, molecular biology, and immunology, have played the leading role here by identifying the headpiece, basepiece, central stalk, side stalk, cap, and in the mitochondrial enzyme, the collar around the central stalk. We can be reasonably confident also that we have a fairly good grasp of much of the "high-resolution" structural features of both the F1 moiety comprised of fives subunit types (alpha, beta, gamma, delta, and epsilon) and parts of the F0 moiety comprised of either three (E. coli) or at least ten (mitochondria) subunit types. This information acquired in several different laboratories, either by X-ray crystallography or NMR spectroscopy, includes details about the active site and subunit relationships. Moreover, it is consistent with recently reported data that the F1 moiety may be an ATP driven motor, which, during ATP synthesis, is driven in reverse by the electrochemical proton gradient generated by the electron transport chain. The real structural challenges of the future are to acquire at high resolution "complete" ATP synthase complexes representative of different stages of the catalytic cycle during ATP synthesis and representative also of key regulatory states
MH  - A
MH  - ACTIVE
MH  - ACTIVE SITE
MH  - alpha
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - BETA
MH  - Biochemistry
MH  - biology
MH  - catalytic
MH  - Chemistry
MH  - COMPLEX
MH  - Crystallography,X-Ray
MH  - data
MH  - delta
MH  - electron
MH  - electron microscopy
MH  - Electron Transport
MH  - ELECTRON-MICROSCOPY
MH  - enzyme
MH  - enzymology
MH  - Escherichia coli
MH  - F0
MH  - F0F1
MH  - F1
MH  - Isoenzymes
MH  - Magnetic Resonance Spectroscopy
MH  - Microscopy
MH  - Microscopy,Electron
MH  - Mitochondria
MH  - Models,Molecular
MH  - NMR
MH  - P
MH  - Protein Conformation
MH  - Protein Subunits
MH  - proton
MH  - PROTON GRADIENT
MH  - Proton-Translocating ATPases
MH  - resolution
MH  - review
MH  - Site
MH  - spectroscopy
MH  - stalk
MH  - structure
MH  - SUBUNIT
MH  - Support,U.S.Gov't,P.H.S.
MH  - SYNTHASE
MH  - synthesis
MH  - transport
MH  - ultrastructure
MH  - universities
RP  - NOT IN FILE
NT  - Department of Biological Chemistry, Johns Hopkins University, School of Medicine, Baltimore, Maryland 21205-2185, USA ppederse@welchlinkwelchjhueduFAU - Pedersen, P L
SO  - J Bioenerg Biomembr 2000 Aug ;32(4):325-332

1721
UI  - 21059
AU  - Pedersen PL
AU  - Ko YH
AU  - Hong S
TI  - ATP synthases in the year 2000: evolving views about the structures of these remarkable enzyme complexes
AB  - This introductory article briefly summarizes how our views about the structural features of ATP synthases (F0F1) have evolved over the past 30 years and also reviews some of our current views in the year 2000 about the structures of these remarkably unique enzyme complexes. Suffice it to say that as we approach the end of the first year of this new millinium, we can be conservatively confident that we have a reasonably good grasp of the overall "low-resolution" structural features of ATP synthases. Electron microscopy techniques, combined with the tools of biochemistry, molecular biology, and immunology, have played the leading role here by identifying the headpiece, basepiece, central stalk, side stalk, cap, and in the mitochondrial enzyme, the collar around the central stalk. We can be reasonably confident also that we have a fairly good grasp of much of the "high-resolution" structural features of both the F1 moiety comprised of fives subunit types (alpha, beta, gamma, delta, and epsilon) and parts of the F0 moiety comprised of either three (E. coli) or at least ten (mitochondria) subunit types. This information acquired in several different laboratories, either by X-ray crystallography or NMR spectroscopy, includes details about the active site and subunit relationships. Moreover, it is consistent with recently reported data that the F1 moiety may be an ATP driven motor, which, during ATP synthesis, is driven in reverse by the electrochemical proton gradient generated by the electron transport chain. The real structural challenges of the future are to acquire at high resolution "complete" ATP synthase complexes representative of different stages of the catalytic cycle during ATP synthesis and representative also of key regulatory states
MH  - A
MH  - ACTIVE
MH  - alpha
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - BETA
MH  - Biochemistry
MH  - Chemistry
MH  - COMPLEX
MH  - delta
MH  - electron
MH  - electron microscopy
MH  - Electron Transport
MH  - ELECTRON-MICROSCOPY
MH  - F0
MH  - F0F1
MH  - F1
MH  - Microscopy
MH  - Mitochondria
MH  - NMR
MH  - P
MH  - proton
MH  - resolution
MH  - review
MH  - Site
MH  - spectroscopy
MH  - stalk
MH  - structure
MH  - SUBUNIT
MH  - SYNTHASE
MH  - synthesis
MH  - transport
RP  - NOT IN FILE
NT  - Department of Biological Chemistry, Johns Hopkins University, School of Medicine, Baltimore, Maryland 21205-2185, USA ppederse@welchlinkwelchjhueduFAU - Pedersen, P L
SO  - J Bioenerg Biomembr 2000 Aug ;32(4):325-332

1722
UI  - 21282
AU  - Petsev DN
AU  - Vekilov PG
TI  - Evidence for non-DLVO hydration interactions in solutions of the protein apoferritin
MH  - Solutions
MH  - protein
RP  - IN FILE
SO  - Phys Rev Lett 2000  ;84():1339-1342

1723
UI  - 9939
AU  - Poetsch A
AU  - Neff D
AU  - Seelert H
AU  - Schagger H
AU  - Dencher NA
AD  - Institut fur Biochemie, Abteilung Physikalische Biochemie, Technische Universitat Darmstadt, Germany
TI  - Dye removal, catalytic activity and 2D crystallization of chloroplast H(+)-ATP synthase purified by blue native electrophoresis
AB  - The proton-ATP synthase of thylakoid membranes from spinach chloroplasts (CF(O)F(1)) and its subcomplexes CF(O) and CF(1) were isolated by blue native electrophoresis (BN-PAGE) [Neff, D. and Dencher, N.A. (1999) Biochem. Biophys. Res. Commun. 259, 569-575] and subsequently electroeluted from the gel. A method was developed to remove most of the dye Coomassie G-250 (CBG) using gel filtration, a prerequisite for many biophysical investigations. The dye was removed from the electroeluted CF(O)F(1), CF(O) or CF(1) and exchanged with the detergent CHAPS. ATP hydrolysis activity of CF(1) and ATP synthesis activity of reconstituted CF(O)F(1) were determined before and after dye removal. The secondary structure of CF(O) was studied by CD spectroscopy in the presence and the absence of the dye. CBG neither abolishes the catalytic activity of the isolated CF(O)F(1) and CF(1) nor affects the subunit composition and the high alpha-helical content of CF(O). In crystallization attempts, 2D arrays of CF(O)F(1) and of CF(O) before and after dye removal were obtained. In the aggregates of CF(O), circular structures with a mean diameter of 6.7 nm were observed. Our results indicate that the combination of BN-PAGE and dye removal by gel filtration is a suitable approach to obtain catalytically active protein complexes for further functional and structural characterization
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - chloroplast
MH  - Chloroplasts
MH  - COMPLEX
MH  - Crystallization
MH  - Electrophoresis
MH  - H(+)-Transporting ATP Synthase
MH  - Hydrolysis
MH  - Indicators and Reagents
MH  - Membranes
MH  - Spinach
MH  - SPINACH-CHLOROPLASTS
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 20286312LA - engRN - 0 (Indicators and Reagents)RN - 0 (Rosaniline Dyes)RN - 6104-58-1 (coomassie Brilliant Blue)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 20000810IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:10825454
SO  - Biochim Biophys Acta 2000 Jun 1 ;1466(1-2):339-349

1724
UI  - 20984
AU  - Possmayer FE
AU  - Hartog AF
AU  - Berden JA
AU  - Graber P
TI  - Covalent modification of the catalytic sites of the H(+)-ATPase from chloroplasts with 2-nitreno-ADP. Modification of the catalytic site 1 (tight) and catalytic sites 1 and 2 together impairs both uni-site and multi-site catalysis of ATP synthesis and ATP hydrolysis
AB  - After isolation and purification, the H(+)-ATPase from chloroplasts, CF(0)F(1), contains one endogenous ADP at a catalytic site, and two endogenous ATP at non-catalytic sites. Incubation with 2-azido-[alpha- (32)P]ADP leads to tight binding of azido-nucleotides. Free nucleotides were removed by three consecutive passages through centrifugation columns, and upon UV-irradiation most of the label was covalently bound. The labelled enzyme was digested by trypsin, the peptides were separated by ion exchange chromatography into nitreno-AMP, nitreno-ADP and nitreno-ATP labelled peptides, and these were then separated by reversed phase chromatography. Amino acid sequence analysis was used to identify the type of the nucleotide binding site. After incubation with 2-azido-[alpha-(32)P]ADP, the covalently bound label was found exclusively at beta-Tyr-362. Incubation conditions with 2-azido-[alpha- (32)P]ADP were varied, and conditions were found which allow selective binding of the label to different catalytic sites, designated as 1, 2 and 3 in order of decreasing affinity for ADP, and either catalytic site 1 or catalytic sites 1 and 2 together were labelled. For measurements of the degree of inhibition by covalent modification, CF(0)F(1) was reconstituted into phosphatidylcholine liposomes, and the membranes were energised by an acid-base transition in the presence of a K(+)/valinomycin diffusion potential. The rate of ATP synthesis was 50-80 s(-1), and the rate of ATP hydrolysis was 15 s(-1) measured under multi-site conditions. Covalent modification of either catalytic site 1 or catalytic sites 1 and 2 together inhibited ATP synthesis and ATP hydrolysis equally, the degree of inhibition being proportional to the degree of modification. Extrapolation to complete inhibition indicates that derivatisation of catalytic site 1 leads to complete inhibition when 1 mol 2-nitreno-ADP is bound per mol CF(0)F(1). Derivatisation of catalytic sites 1 and 2 together extrapolates to complete inhibition when 2 mol 2-nitreno-ADP are bound per CF(0)F(1). The rate of ATP synthesis and the rate of ATP hydrolysis were measured as a function of the substrate concentration from multi-site to uni-site conditions with derivatised CF(0)F(1) and with non-derivatised CF(0)F(1). ATP synthesis and ATP hydrolysis under uni-site and under multi-site condition were inhibited by covalent modification of either catalytic site 1 or catalytic sites 1 and 2 together. The results indicate that derivatisation of site 1 inhibits activation of the enzyme and that cooperative interactions occur at least between the catalytic sites 2 and 3
MH  - A
MH  - ACID
MH  - ACTIVATION
MH  - ADP
MH  - Amino Acid Sequence
MH  - analysis
MH  - atp
MH  - ATP synthesis
MH  - BINDING
MH  - Catalysis
MH  - chloroplast
MH  - Chloroplasts
MH  - Diffusion
MH  - diffusion potential
MH  - function
MH  - H(+)ATPase
MH  - Hydrolysis
MH  - ion
MH  - ion exchange
MH  - Liposomes
MH  - membrane
MH  - Membranes
MH  - non-catalytic site
MH  - NONCATALYTIC SITES
MH  - nucleotide binding
MH  - Nucleotides
MH  - purification
MH  - Site
MH  - synthesis
MH  - Trypsin
RP  - NOT IN FILE
NT  - UI - 0LA - ENGPT - JOURNAL ARTICLEDA - 20000926IS - 0006-3002SB - IMJC - A0W
UR  - PM:11004435
SO  - Biochim Biophys Acta 2000 Jul 20 ;1459(1):202-217

1725
UI  - 9884
AU  - Possmayer FE
AU  - Hartog AF
AU  - Berden JA
AU  - Graber P
TI  - Covalent modification of the catalytic sites of the H+-ATPase from chloroplasts, CF0F1, with 2-azido-[alpha-P-32]ADP: modification of the catalytic site 2 (loose) and the catalytic site 3 (open) impairs multi-site, but not uni-site catalysis of both ATP synthesis and ATP hydrolysis
AB  - The H+-ATPase from chloroplasts, CF0F1, was isolated and purified. The enzyme contained one endogenous ADP at a catalytic site, and two endogenous ATP at non-catalytic sites. Incubation with 2-azido-[alpha-P-32]AD(T)P leads to a tight binding of the azido-nucleotides. Free nucleotides were removed by three consecutive passages through centrifugation columns, and after UV-irradiation, the label was covalently bound. The labelled enzyme was digested by trypsin, the peptides were separated by ion exchange chromatography into nitreno-AMP, nitreno-ADP and nitreno-ATP labelled peptides, and these were then separated by reversed phase chromatography. Amino acid sequence analysis was used to identify the type of the nucleotide binding site. After incubation with 2-azido-[alpha- P-32]ADP, the covalently bound label was found exclusively at beta-Tyr-362, i.e. binding occurs only to catalytic sites. Incubation conditions with 2-azido-[alpha-P-32]ADP were varied, and conditions were found which allow selective binding of the label to different catalytic sites, either to catalytic site 2 or to catalytic site 3. For measurements of the degree of inhibition by covalent modification, CF0F1 was reconstituted into phosphatidylcholine liposomes, and the membranes were energised by an acid-base transition in the presence of a K+/valinomycin diffusion potential. The rate of ATP synthesis was 120 s(-1), and the rate of ATP hydrolysis was 20 s(-1), both measured under multi-site conditions. Covalent modification of either catalytic site 2 or catalytic site 3 inhibited both ATP synthesis and ATP hydrolysis, the degree of inhibition being proportional to the degree of modification. Extrapolation to complete inhibition indicates that modification of one catalytic site, either site 2 or site 3, is sufficient to completely block multi-site ATP synthesis and ATP hydrolysis. The rate of ATP synthesis and the rate of ATP hydrolysis were measured as a function of the substrate concentration from multi-site to uni-site conditions with covalently modified CF0F1 and with nonmodified CF0F1. The result was that uni-site ATP synthesis and ATP hydrolysis were not inhibited by covalent modification of either catalytic site 2 or site 3. The results indicate cooperative interactions between catalytic nucleotide binding sites during multi-site catalysis, whereas neither uni-site ATP synthesis nor uni-site ATP hydrolysis require interaction with other sites. (C) 2000 Elsevier Science B.V. All rights reserved
MH  - 2- azido-nucleotide
MH  - ADENOSINE-TRIPHOSPHATASE
MH  - ADP
MH  - Amino Acid Sequence
MH  - analysis
MH  - atp
MH  - BETA-SUBUNIT
MH  - Binding Sites
MH  - Catalysis
MH  - CF0F1
MH  - CHANGE MECHANISM
MH  - chloroplast
MH  - Chloroplasts
MH  - COLI F1 ATPASE
MH  - COUPLING FACTOR-I
MH  - Diffusion
MH  - H(+)ATPase
MH  - H+-ATPase
MH  - HEART MITOCHONDRIAL ATPASE
MH  - Hydrolysis
MH  - Liposomes
MH  - Membranes
MH  - non-catalytic site
MH  - NONCATALYTIC SITES
MH  - nucleotide binding
MH  - nucleotide binding sites
MH  - NUCLEOTIDE-BINDING-SITES
MH  - Nucleotides
MH  - RECONSTITUTED CF0F1
MH  - Trypsin
MH  - uni-site catalysis
MH  - UNISITE CATALYSIS
RP  - NOT IN FILE
NT  - JournalArticleELSEVIER SCIENCE BVJAN 10276PLAMSTERDAMGraber P Univ Freiburg, Inst Phys Chem, Albertstr 23A, D-79104 Freiburg, GermanyBBA-BIOENERGETICSPO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS
AV  - Univ Freiburg, Inst Phys Chem, Albertstr 23A, D-79104 Freiburg, Germany Univ Freiburg, Inst Phys Chem, D-79104 Freiburg, Germany Univ Amsterdam, EC Slater Inst, NL-1018 TV Amsterdam, Netherlands
UR  - ISI:000084886500002
SO  - Biochimica et Biophysica Acta-Bioenergetics 2000  ;1456(2-3):77-98

1726
UI  - 21255
AU  - Sazanov LA
AU  - Walker JE
AD  - Medical Research Council Dunn Human Nutrition Unit, Wellcome Trust/MRC Building, Hills Road, Cambridge, CB2 2XY, UK
TI  - Cryo-electron crystallography of two sub-complexes of bovine complex I reveals the relationship between the membrane and peripheral arms
AB  - NADH:ubiquinone oxidoreductase (complex I) is the first and largest enzyme of the mitochondrial respiratory chain. The low-resolution structure of the complex is known from electron microscopy studies. The general shape of the complex is in the form of an L, with one arm in the membrane and the other peripheral. We have purified complex I from beef heart mitochondria and reconstituted the enzyme into lipid bilayers. Under different conditions, several two-dimensional crystal forms were obtained. Crystals belonging to space groups p222(1) and c12 (unit cell 488 Ax79 A) were obtained at 22 degrees C and contained only the membrane fragment of complex I similar to hydrophobic subcomplex Ibeta but lacking the ND5 subunit. A crystal form with larger unit cell (534 Ax81 A, space group c12) produced at 4 degrees C contained both the peripheral and membrane arms of the enzyme, except that ND5 was missing. Projection maps from frozen hydrated samples were calculated for all crystal forms. By comparing two different c12 crystal forms, extra electron density in the projection map of large crystal form was assigned to the peripheral arm of the enzyme. One of the features of the map is a deep, channel-like, cleft next to peripheral arm. Comparison with available structures of the intact enzyme indicates that large hydrophobic subunit ND5 is situated at the distal end of the membrane domain. Possible locations of subunit ND4 and of other subunits in the membrane domain are proposed. Implications of our findings for the mechanism of proton pumping by complex I are discussed
MH  - A
MH  - COMPLEX
MH  - Detergents
MH  - electron
MH  - electron microscopy
MH  - ELECTRON-MICROSCOPY
MH  - England
MH  - Human
MH  - Lipid Bilayers
MH  - mechanism
MH  - membrane
MH  - Membrane Proteins
MH  - Microscopy
MH  - Mitochondria
MH  - NADH Dehydrogenase
MH  - Nutrition
MH  - Oxidoreductases
MH  - Peptide Fragments
MH  - protein
MH  - Proteins
MH  - proton
MH  - structure
MH  - SUBUNIT
MH  - Ubiquinone
RP  - NOT IN FILE
NT  - UI - 20428906LA - engRN - 0 (Detergents)RN - 0 (Lipid Bilayers)RN - 0 (Membrane Proteins)RN - 0 (Peptide Fragments)RN - EC 1.6. (NADH, NADPH Oxidoreductases)RN - EC 1.6.5.3 (NADH dehydrogenase (ubiquinone))PT - Journal ArticleDA - 20001019IS - 0022-2836SB - IMCY - ENGLAND
UR  - PM:10970745
SO  - J Mol Biol 2000 Sep 15 ;302(2):455-464

1727
UI  - 9944
AU  - Schnick C
AU  - Forrest LR
AU  - Sansom MS
AU  - Groth G
AD  - Heinrich-Heine-Universitat Dusseldorf, Biochemie der Pflanzen, Germany
TI  - Molecular contacts in the transmembrane c-subunit oligomer of F-ATPases identified by tryptophan substitution mutagenesis
AB  - When isolated in its monomeric form, subunit c of the proton transporting ATP synthase of Escherichia coli was shown to fold in a hairpin-like structure consisting of two hydrophobic membrane spanning helices and a short connecting hydrophilic loop. In the plasma membrane of Escherichia coli, however, about 9-12 c-subunit monomers form an oligomeric complex that functions in transmembrane proton conduction and in energy transduction to the catalytic F1 domain. The arrangement of the monomers and the molecular architecture of the complex were studied by tryptophan scanning mutagenesis and restrained MD simulations. Residues 12-24 of the N-terminal transmembrane segment of subunit c were individually substituted by the large and moderately hydrophobic tryptophan side chain. Effects on the activity of the mutant proteins were studied in selective growth experiments and various ATP synthase specific activity assays. The results identify potential intersubunit contacts and structurally non-distorted, accessible residues in the c-oligomer and add constraints to the arrangement of monomers in the oligomeric complex. Results from our mutagenesis experiments were interpreted in structural models of the c- oligomer that have been obtained by restrained MD simulations. Different stoichiometries and monomer orientations were applied in these calculations. A cylindrical complex consisting of 10 monomers that are arranged in two concentric rings with the N-terminal helices of the monomers located at the periphery shows the best match with the experimental data
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthase
MH  - COMPLEX
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F-ATPASE
MH  - F1
MH  - H(+)-Transporting ATP Synthase
MH  - model
MH  - Proteins
MH  - proton
MH  - Protons
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 20439553LA - engRN - 0 (Culture Media)RN - 0 (Plasmids)RN - 0 (Protons)RN - 56-65-5 (Adenosine Triphosphate)RN - 73-22-3 (Tryptophan)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 20000929IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:10924898
SO  - Biochim Biophys Acta 2000 Jul 20 ;1459(1):49-60

1728
UI  - 9938
AU  - Seelert H
AU  - Poetsch A
AU  - Dencher NA
AU  - Engel A
AU  - Stahlberg H
AU  - Muller DJ
AD  - Department of Chemistry, Darmstadt University of Technology, Germany
TI  - Structural biology. Proton-powered turbine of a plant motor
MH  - atp
MH  - ATP synthase
MH  - Chemistry
MH  - England
MH  - H(+)-Transporting ATP Synthase
MH  - proton
MH  - Protons
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 20296339LA - engRN - 0 (Molecular Motors)RN - 0 (Protons)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 20000616IS - 0028-0836SB - IMCY - ENGLANDJC - NSC
UR  - PM:10839529
SO  - Nature 2000 May 25 ;405(6785):418-419

1729
UI  - 9940
AU  - Seelert H
AU  - Poetsch A
AU  - Rohlfs M
AU  - Dencher NA
AD  - Institut fur Biochemie, Abt Physikalische Biochemie, Technische Universitat Darmstadt, Petersenstr 22, D-64287 Darmstadt, Germany
TI  - Dye-ligand chromatographic purification of intact multisubunit membrane protein complexes: application to the chloroplast H+-FoF1-ATP synthase
AB  - n-Dodecyl-beta-D-maltoside was used as a detergent to solubilize the ammonium sulphate precipitate of chloroplast F(O)F(1)-ATP synthase, which was purified further by dye-ligand chromatography. Upon reconstitution of the purified protein complex into phosphatidylcholine/phosphatidic acid liposomes, ATP synthesis, driven by an artificial DeltapH/Deltapsi, was observed. The highest activity was achieved with ATP synthase solubilized in n-dodecyl-beta-D- maltoside followed by chromatography with Red 120 dye. The optimal dye for purification with CHAPS was Green 5. All known subunits were present in the monodisperse proton-translocating ATP synthase preparation obtained from chloroplasts
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - chloroplast
MH  - Chloroplasts
MH  - Cholic Acid
MH  - Cholic Acids
MH  - COMPLEX
MH  - England
MH  - Glucosides
MH  - H(+)-Transporting ATP Synthase
MH  - Ligands
MH  - Liposomes
MH  - Membrane Proteins
MH  - Proteins
MH  - purification
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 20125579LA - engRN - 0 (Cholic Acids)RN - 0 (Dyes)RN - 0 (Glucosides)RN - 0 (Ligands)RN - 0 (Liposomes)RN - 0 (Membrane Proteins)RN - 56-65-5 (Adenosine Triphosphate)RN - 69227-93-6 (dodecyl maltoside)RN - 75621-03-3 (3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 20000330IS - 0264-6021SB - IMCY - ENGLANDJC - 9YO
UR  - PM:10657237
SO  - Biochem J 2000 Feb 15 ;346 Pt 1():41-44

1730
UI  - 405
AU  - Senior AE
AU  - Nadanaciva S
AU  - Weber J
AD  - Department of Biochemistry, University of Rochester Medical Center, Rochester, NY 14642, USA alan_senior@urmcrochesteredu
TI  - Rate acceleration of ATP hydrolysis by F(1)F(o)-ATP synthase
AB  - The rate acceleration of ATP hydrolysis by F(1)F(o)-ATP synthase is of the order of 10(11)-fold. We present a cyclic enzyme mechanism for the reaction, relate it to known F(1) X-ray structure and speculate on the linkage between enzyme reaction intermediates and subunit rotation. Next, we describe five factors known to be important in the Escherichia coli enzyme for the rate acceleration. First, the provision of substrate binding energy by residues lining the catalytic site is substantial; beta-Lys155 and beta-Arg182 are specific examples, both of which differentially support substrate MgATP versus product MgADP binding. Second, octahedral coordination of the Mg(2+) in MgATP is crucial for both catalysis and catalytic site asymmetry. The residues involved are beta-Thr156, beta-Glu185 and beta-Asp242. Third, there is stabilization of a pentacoordinate phosphorus catalytic transition state by residues beta-Lys155, beta-Arg182 and alpha-Arg376. Fourth, residue beta-Glu181 binds the substrate water and stabilizes the catalytic transition state. Fifth, there is strong positive catalytic cooperativity, with binding of MgATP at all three sites yielding the maximum rate (V(max)); the molecular basis of this factor remains to be elucidated
RP  - NOT IN FILE
NT  - UI - 20123949LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialID - GM25349/GM/NIGMSDA - 20000217IS - 0022-0949SB - IMCY - ENGLANDJC - I2F
UR  - PM:10600671
SO  - J Exp Biol 2000 Jan ;203 Pt 1():35-40

1731
UI  - 21260
AU  - Shinkarev VP
AU  - Ugulava NB
AU  - Crofts AR
AU  - Wraight CA
AD  - Department of Plant Biology, University of Illinois at Urbana- Champaign, Illinois 61801, USA
TI  - DCCD inhibits the reactions of the iron-sulfur protein in Rhodobacter sphaeroides chromatophores
AB  - N,N'-dicyclohexylcarbodiimide (DCCD) has been reported to inhibit proton translocation by cytochrome bc(1) and b(6)f complexes without significantly altering the rate of electron transport, a process referred to as decoupling. To understand the possible role of DCCD in inhibiting the protonogenic reactions of cytochrome bc(1) complex, we investigated the effect of DCCD modification on flash-induced electron transport and electrochromic bandshift of carotenoids in Rb. sphaeroides chromatophores. DCCD has two distinct effects on phase III of the electrochromic bandshift of carotenoids reflecting the electrogenic reactions of the bc(1) complex. At low concentrations, DCCD increases the magnitude of the electrogenic process because of a decrease in the permeability of the membrane, probably through inhibition of F(o)F(1). At higher concentrations (>150 microM), DCCD slows the development of phase III of the electrochromic shift from about 3 ms in control preparations to about 23 ms at 1.2 mM DCCD, without significantly changing the amplitude. DCCD treatment of chromatophores also slows down the kinetics of flash-induced reduction of both cytochromes b and c, from 1.5-2 ms in control preparations to 8- 10 ms at 0.8 mM DCCD. Parallel slowing of the reduction of both cytochromes indicates that DCCD treatment modifies the reaction of QH(2) oxidation at the Q(o) site. Despite the similarity in the kinetics of both cytochromes, the onset of cytochrome c re-reduction is delayed 1-2 ms in comparison to cytochrome b reduction, indicating that DCCD inhibits the delivery of electrons from quinol to heme c(1). We conclude that DCCD treatment of chromatophores leads to modification of the rate of Q(o)H(2) oxidation by the iron-sulfur protein (ISP) as well as the donation of electrons from ISP to c(1), and we discuss the results in the context of the movement of ISP between the Q(o) site and cytochrome c(1)
MH  - A
MH  - carotenoid
MH  - Carotenoids
MH  - chromatophore
MH  - chromatophores
MH  - COMPLEX
MH  - cytochrome
MH  - Cytochrome b
MH  - Cytochrome c
MH  - Cytochromes
MH  - development
MH  - Dicyclohexylcarbodiimide
MH  - electrogenic
MH  - electron
MH  - Electron Transport
MH  - Electrons
MH  - Kinetics
MH  - membrane
MH  - Movement
MH  - Permeability
MH  - plant
MH  - protein
MH  - Proteins
MH  - proton
MH  - Rhodobacter
MH  - Rhodobacter sphaeroides
MH  - Site
MH  - sphaeroides
MH  - translocation
MH  - transport
MH  - Ubiquinol-Cytochrome-c Reductase
RP  - NOT IN FILE
NT  - UI - 20573618LA - engRN - 0 (Iron-Sulfur Proteins)RN - 36-88-4 (Carotenoids)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - EC 1.10.2.2 (Ubiquinol-Cytochrome-c Reductase)PT - Journal ArticleID - GM35438/GM/NIGMSID - GM53508/GM/NIGMSDA - 20010108IS - 0006-2960SB - IMCY - UNITED STATES
UR  - PM:11123950
SO  - Biochemistry 2000 Dec 26 ;39(51):16206-16212

1732
UI  - 21261
AU  - Shinkarev VP
AU  - Ugulava NB
AU  - Takahashi E
AU  - Crofts AR
AU  - Wraight CA
AD  - Department of Plant Biology, University of Illinois at Urbana- Champaign, 265 Morrill Hall, 505 South Goodwin Avenue, Urbana, Illinois 61801, USA
TI  - Aspartate-187 of cytochrome b is not needed for DCCD inhibition of ubiquinol: cytochrome c oxidoreductase in Rhodobacter sphaeroides chromatophores
AB  - N,N'-dicyclohexylcarbodiimide (DCCD) has been reported to inhibit steady-state proton translocation by cytochrome bc(1) and b(6)f complexes without significantly altering the rate of electron transport, a process referred to as decoupling. In chromatophores of the purple bacterium Rhodobacter sphaeroides, this has been associated with the specific labeling of a surface-exposed aspartate-187 of the cytochrome b subunit of the bc(1) complex [Wang et al. (1998) Arch. Biochem. Biophys. 352, 193-198]. To explore the possible role of this amino acid residue in the protonogenic reactions of cytochrome bc(1) complex, we investigated the effect of DCCD modification on flash- induced electron transport and the electrochromic bandshift of carotenoids in Rb. sphaeroides chromatophores from wild type (WT) and mutant cells, in which aspartate-187 of cytochrome b (Asp(B187)) has been changed to asparagine (mutant B187 DN). The kinetics and amplitude of phase III of the electrochromic shift of carotenoids, reflecting electrogenic reactions in the bc(1) complex, and of the redox changes of cytochromes and reaction center, were similar (+/- 15%) in both WT and B187DN chromatophores. DCCD effectively inhibited phase III of the carotenoid bandshift in both B187DN and WT chromatophores. The dependence of the kinetics and amplitude of phase III of the electrochromic shift on DCCD concentration was identical in WT and B187DN chromatophores, indicating that covalent modification of Asp(B187) is not specifically responsible for the effect of DCCD- induced effects of cytochrome bc(1) complex. Furthermore, no evidence for differential inhibition of electrogenesis and electron transport was found in either strain. We conclude that Asp(B187) plays no crucial role in the protonogenic reactions of bc(1) complex, since its replacement by asparagine does not lead to any significant effects on either the electrogenic reactions of bc(1) complex, as revealed by phase III of the electrochromic shift of carotenoids, or sensitivity of turnover to DCCD
MH  - A
MH  - ACID
MH  - Bacteria
MH  - carotenoid
MH  - Carotenoids
MH  - Cells
MH  - chromatophore
MH  - chromatophores
MH  - COMPLEX
MH  - cytochrome
MH  - Cytochrome b
MH  - Cytochrome c
MH  - Cytochrome-c Oxidase
MH  - Cytochromes
MH  - DEPENDENCE
MH  - Dicyclohexylcarbodiimide
MH  - electrogenic
MH  - electron
MH  - Electron Transport
MH  - flash
MH  - Kinetics
MH  - mutant
MH  - plant
MH  - proton
MH  - reaction center
MH  - redox
MH  - RESIDUE
MH  - Rhodobacter
MH  - Rhodobacter sphaeroides
MH  - sphaeroides
MH  - SUBUNIT
MH  - translocation
MH  - transport
MH  - Ubiquinone
RP  - NOT IN FILE
NT  - UI - 20541377LA - engRN - 1339-63-5 (Ubiquinone)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-84-8 (Aspartic Acid)RN - 56275-39-9 (ubiquinol)RN - 7006-34-0 (Asparagine)RN - 9035-37-4 (Cytochrome b)RN - EC 1.9.3.1 (Cytochrome-c Oxidase)PT - Journal ArticleID - GM 35438/GM/NIGMSID - GM53508/GM/NIGMSDA - 20001212IS - 0006-2960SB - IMCY - UNITED STATES
UR  - PM:11087372
SO  - Biochemistry 2000 Nov 21 ;39(46):14232-14237

1733
UI  - 19898
AU  - Slevin CJ
AU  - Unwin PR
TI  - Lateral proton diffusion rates along stearic acid monolayers
MH  - ACID
MH  - Diffusion
MH  - proton
RP  - NOT IN FILE
NT  - JournalMAR 22296ZTJ AMER CHEM SOC
UR  - ISI:000086057200024
SO  - Journal of the American Chemical Society 2000  ;122(11):2597-2602

1734
UI  - 21246
AU  - Soulimane T
AU  - Buse G
AU  - Bourenkov GP
AU  - Bartunik HD
AU  - Huber R
AU  - Than ME
AD  - Rheinisch-Westfalische Technische Hochschule Aachen, Institut fur Biochemie, Pauwelsstrasse 30, D-52057 Aachen, Germany tsoulimane@postklinikumrwth-aachende
TI  - Structure and mechanism of the aberrant ba(3)-cytochrome c oxidase from thermus thermophilus
AB  - Cytochrome c oxidase is a respiratory enzyme catalysing the energy- conserving reduction of molecular oxygen to water. The crystal structure of the ba(3)-cytochrome c oxidase from Thermus thermophilus has been determined to 2.4 A resolution using multiple anomalous dispersion (MAD) phasing and led to the discovery of a novel subunit IIa. A structure-based sequence alignment of this phylogenetically very distant oxidase with the other structurally known cytochrome oxidases leads to the identification of sequence motifs and residues that seem to be indispensable for the function of the haem copper oxidases, e.g. a new electron transfer pathway leading directly from Cu(A) to Cu(B). Specific features of the ba(3)-oxidase include an extended oxygen input channel, which leads directly to the active site, the presence of only one oxygen atom (O(2-), OH(-) or H(2)O) as bridging ligand at the active site and the mainly hydrophobic character of the interactions that stabilize the electron transfer complex between this oxidase and its substrate cytochrome c. New aspects of the proton pumping mechanism could be identified
MH  - A
MH  - ACTIVE
MH  - COMPLEX
MH  - Copper
MH  - cytochrome
MH  - Cytochrome b
MH  - Cytochrome c
MH  - Cytochrome-c Oxidase
MH  - electron
MH  - electron transfer
MH  - Electron-transfer
MH  - England
MH  - function
MH  - Ligands
MH  - mechanism
MH  - membrane
MH  - Membrane Proteins
MH  - Oxygen
MH  - protein
MH  - Proteins
MH  - proton
MH  - Protons
MH  - RESIDUE
MH  - resolution
MH  - Sequence Alignment
MH  - Site
MH  - structure
MH  - SUBUNIT
MH  - TRANSFER
MH  - Water
RP  - NOT IN FILE
NT  - UI - 20237613LA - engRN - 0 (Ligands)RN - 0 (Membrane Proteins)RN - 0 (Protons)RN - 7782-44-7 (Oxygen)RN - 9035-37-4 (Cytochrome b)RN - EC 1.- (cytochrome ba3)RN - EC 1.9.3.1 (Cytochrome-c Oxidase)PT - Journal ArticleDA - 20000613IS - 0261-4189SB - IMCY - ENGLAND
UR  - PM:10775261
SO  - EMBO J 2000 Apr 17 ;19(8):1766-1776

1735
UI  - 316
AU  - Stock D
AU  - Gibbons C
AU  - Arechaga I
AU  - Leslie AG
AU  - Walker JE
AD  - The Medical Research Council Dunn Human Nutrition Unit, Hills Road, CB2 2XY, Cambridge, UK
TI  - The rotary mechanism of ATP synthase
AB  - Since the chemiosmotic theory was proposed by Peter Mitchell in the 1960s, a major objective has been to elucidate the mechanism of coupling of the transmembrane proton motive force, created by respiration or photosynthesis, to the synthesis of ATP from ADP and inorganic phosphate. Recently, significant progress has been made towards establishing the complete structure of ATP synthase and revealing its mechanism. The X-ray structure of the F(1) catalytic domain has been completed and an electron density map of the F(1)-c(10) subcomplex has provided a glimpse of the motor in the membrane domain. Direct microscopic observation of rotation has been extended to F(1)- ATPase and F(1)F(o)-ATPase complexes
RP  - NOT IN FILE
NT  - UI - 20566955LA - engRN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 20010129IS - 0959-440XSB - IMCY - ENGLANDJC - B9V
UR  - PM:11114504
SO  - Curr Opin Struct Biol 2000 Dec ;10(6):672-679

1736
UI  - 686
AU  - Suzuki T
AU  - Suzuki J
AU  - Mitome N
AU  - Ueno H
AU  - Yoshida M
AD  - Chemical Resources Laboratory, Tokyo Institute of Technology, Nagatsuta 4259, Yokohama 226-8503, Japan
TI  - Second stalk of ATP synthase. Cross-linking of gamma subunit in F1 to truncated Fob subunit prevents ATP hydrolysis
AB  - ATP synthase consists of two portions, F(1) and F(o), connected by two stalks: a central rotor stalk containing gamma and epsilon subunits and a peripheral, second stalk formed by delta and two copies of F(o)b subunits. The second stalk is expected to keep the stator subunits from spinning along with the rotor. We isolated a TF(1)-b'(2) complex (alpha(3)beta(3)gammadeltaepsilonb'(2)) of a thermophilic Bacillus PS3, in which b' was a truncated cytoplasmic fragment of F(o)b subunit, and introduced a cysteine at its N terminus (bc'). Association of b'(2) or bc'(2) with TF(1) did not have significant effect on ATPase activity. A disulfide bond between the introduced cysteine of bc' and cysteine 109 of gamma subunit was readily formed, and this cross-link caused inactivation of ATPase. This implies that F(o)b subunit bound to stator subunits of F(1) with enough strength to resist rotation of gamma subunit and to prevent catalysis. Contrary to this apparent tight binding, some detergents such as lauryldodecylamine oxide tend to cause release of b'(2) from TF(1)
RP  - NOT IN FILE
NT  - UI - 20545540LA - engRN - 0 (DNA Primers)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 20001218IS - 0021-9258SB - IMCY - UNITED STATESJC - HIV
UR  - PM:10970900
SO  - J Biol Chem 2000 Dec 1 ;275(48):37902-37906

1737
UI  - 19761
AU  - Szpirer CY
AU  - Faelen M
AU  - Couturier M
AD  - Laboratoire de Genetique des Procaryotes, Institut de Biologie et de Medecine Moleculaires, Universite Libre de Bruxelles, 12, Rue Prof Jeener et Brachet, B-6041 Gosselies, Belgium ceszpir@dbmulbacbe
TI  - Interaction between the RP4 coupling protein TraG and the pBHR1 mobilization protein Mob
AB  - It is currently believed that interaction between the relaxosome of a mobilizable plasmid and the transfer machinery of the helper conjugative plasmid is mediated by a TraG family coupling protein. The coupling proteins appear as an essential determinant of mobilization specificity and efficiency. Using a two-hybrid system, we demonstrated for the first time the direct in vivo interaction between the coupling protein of a conjugative plasmid (the TraG protein of RP4) and the relaxase of a mobilizable plasmid (the Mob protein of pBHR1, a derivative of the broad host range plasmid pBBR1). This interaction was confirmed in vitro by an overlay assay and was shown to occur even in the absence of the transfer origin of pBHR1. We showed that, among 11 conjugative plasmids tested, pBHR1 is efficiently mobilized only by plasmids encoding an IncP-type transfer system. We also showed that the RP4 TraG coupling protein is essential for mobilization of a pBBR1 derivative and is the element that allows its mobilization by R388 plasmid (IncW) at a detectable frequency
MH  - A
MH  - Bacterial Proteins
MH  - England
MH  - In Vitro
MH  - Proteins
MH  - Time
MH  - TRANSFER
RP  - NOT IN FILE
NT  - UI - 20453448LA - engRN - 0 (Bacterial Proteins)RN - 0 (Mob protein)RN - 0 (Plasmids)RN - 0 (Recombinant Fusion Proteins)RN - 0 (TraGp protein)PT - Journal ArticleDA - 20001124IS - 0950-382XSB - IMCY - ENGLANDJC - MOM
UR  - PM:10998162
SO  - Mol Microbiol 2000 Sep ;37(6):1283-1292

1738
UI  - 20836
AU  - Toyomura T
AU  - Oka T
AU  - Yamaguchi C
AU  - Wada Y
AU  - Futai M
AD  - Division of Biological Sciences, Institute of Scientific and Industrial Research, Osaka University, Core Research for Evolutional Science and Technology (CREST) of the Japan Science and Technology Corporation, Osaka 567-0047, Japan
TI  - Three subunit a isoforms of mouse vacuolar H(+)-ATPase. Preferential expression of the a3 isoform during osteoclast differentiation
AB  - Vacuolar H(+)-ATPase (V-ATPase) is a multi-subunit enzyme with a membrane peripheral catalytic (V(1)) and an intrinsic (V(o)) sector. We have identified three cDNA clones coding for isoforms of mouse V(o) subunit a (a1, a2, and a3). They exhibit 48-52% identity with each other and high similarity to subunit a of other species. The a1 isoform was mainly expressed in brain and liver. The a2 isoform was observed in heart and kidney in addition to brain and liver. Transcripts for the a3 isoform were strongly expressed in heart and liver. The a3 isoform was induced during osteoclast differentiation, and localized in the plasma membrane and cytoplasmic filamentous structures. In contrast to a3, the a1 isoform was constitutively expressed and localized in the cytoplasmic endomembrane compartments of the same cells. These findings suggest that the a3 isoform is a component of the plasma membrane V- ATPase essential for bone resorption
MH  - A
MH  - ATPase
MH  - Brain
MH  - Cells
MH  - H(+)ATPase
MH  - H+-ATPase
MH  - Kidney
MH  - Liver
MH  - membrane
MH  - protein
MH  - SUBUNIT
RP  - NOT IN FILE
NT  - UI - 20187595LA - engRN - 0 (DNA, Complementary)RN - 0 (Protein Isoforms)RN - EC 3.6.1.- (vacuolar H+-ATPase)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 20000427IS - 0021-9258SB - IMCY - UNITED STATES
UR  - PM:10722719
SO  - J Biol Chem 2000 Mar 24 ;275(12):8760-8765

1739
UI  - 135
AU  - Tsunoda SP
AU  - Aggeler R
AU  - Noji H
AU  - Kinosita K
AU  - Yoshida M
AU  - Capaldi RA
AD  - Research Laboratory of Resources Utilization, Tokyo Institute of Technology, 4259 Nagatsuta, Yokohama, Japan
TI  - Observations of rotation within the F(o)F(1)-ATP synthase: deciding between rotation of the F(o)c subunit ring and artifact
AB  - F(o)F(1)-ATP synthase mediates coupling of proton flow in F(o) and ATP synthesis/hydrolysis in F(1) through rotation of central rotor subunits. A ring structure of F(o)c subunits is widely believed to be a part of the rotor. Using an attached actin filament as a probe, we have observed the rotation of the F(o)c subunit ring in detergent- solubilized F(o)F(1)-ATP synthase purified from Escherichia coli. Similar studies have been performed and reported recently [Sambongi et al. (1999) Science 286, 1722-1724]. However, in our hands this rotation has been observed only for the preparations which show poor sensitivity to dicyclohexylcarbodiimde, an F(o) inhibitor. We have found that detergents which adequately disperse the enzyme for the rotation assay also tend to transform F(o)F(1)-ATP synthase into an F(o) inhibitor- insensitive state in which F(1) can hydrolyze ATP regardless of the state of the F(o). Our results raise the important issue of whether rotation of the F(o)c ring in isolated F(o)F(1)-ATP synthase can be demonstrated unequivocally with the approach adopted here and also used by Sambongi et al
RP  - NOT IN FILE
NT  - UI - 20211418LA - engRN - 0 (Actins)RN - 0 (Biopolymers)RN - 0 (Detergents)RN - 0 (Molecular Probes)RN - 0 (Recombinant Fusion Proteins)RN - 0 (Uncoupling Agents)RN - 0 (Venturicidins)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 20000504IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:10745076
SO  - FEBS Lett 2000 Mar 31 ;470(3):244-248

1740
UI  - 21413
AU  - Turina P
TI  - Structural changes during ATP hydrolysis activity of the ATP synthase from Escherichia coli as revealed by fluorescent probes
AB  - F1F0-ATPase complexes undergo several changes in their tertiary and quaternary structure during their functioning. As a possible way to detect some of these different conformations during their activity, an environment-sensitive fluorescence probe was bound to cysteine residues, introduced by site-directed mutagenesis, in the gamma subunit of the Escherichia coli enzyme. Fluorescence changes and ATP hydrolysis rates were compared under various conditions in F1 and in reconstituted F1F0. The results are discussed in terms of possible modes of operation of the ATP synthases
MH  - A
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthase
MH  - biology
MH  - Chemistry
MH  - COMPLEX
MH  - conformation
MH  - Cysteine
MH  - enzyme
MH  - enzymology
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1
MH  - fluorescence
MH  - Fluorescent Dyes
MH  - Hydrolysis
MH  - Kinetics
MH  - metabolism
MH  - mutagenesis
MH  - P
MH  - Protein Conformation
MH  - Protein Structure,Quaternary
MH  - Protein Structure,Tertiary
MH  - Proton-Translocating ATPases
MH  - RESIDUE
MH  - site-directed
MH  - structure
MH  - SUBUNIT
MH  - SYNTHASE
MH  - universities
RP  - NOT IN FILE
NT  - Department of Biology, University of Bologna, Italy turina@almauniboitFAU - Turina, P
SO  - J Bioenerg Biomembr 2000 Aug ;32(4):373-381

1741
UI  - 21056
AU  - Turina P
TI  - Structural changes during ATP hydrolysis activity of the ATP synthase from Escherichia coli as revealed by fluorescent probes
AB  - F1F0-ATPase complexes undergo several changes in their tertiary and quaternary structure during their functioning. As a possible way to detect some of these different conformations during their activity, an environment-sensitive fluorescence probe was bound to cysteine residues, introduced by site-directed mutagenesis, in the gamma subunit of the Escherichia coli enzyme. Fluorescence changes and ATP hydrolysis rates were compared under various conditions in F1 and in reconstituted F1F0. The results are discussed in terms of possible modes of operation of the ATP synthases
MH  - A
MH  - atp
MH  - ATP synthase
MH  - COMPLEX
MH  - conformation
MH  - Cysteine
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1
MH  - fluorescence
MH  - Hydrolysis
MH  - mutagenesis
MH  - P
MH  - RESIDUE
MH  - site-directed
MH  - structure
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - Department of Biology, University of Bologna, Italy turina@almauniboitFAU - Turina, P
SO  - J Bioenerg Biomembr 2000 Aug ;32(4):373-381

1742
UI  - 20920
AU  - Turina P
AU  - Melandri BA
TI  - The mutation E210-->K in the a subunit of the Rhodobacter capsulatus ATP synthase
AB  -                       The photosynthetic bacterium Rb. capsulatus is a convenient system for investigating catalytic properties of                      the ATP synthase, since mutations con be easily introduced and functional studies can be carried out in the                      well-coupled internal membranes preparations (chromatophores). We will present functional measurements                      on the aGlu210-->Lys mutant. The mutant shows reduced ATP synthesis rates compared to wild-type but                      similar rates of ATP hydrolysis and proon pumping. Our working hypothesis is that the main role of Glu210                      (probably homologous to the Glu219 of E.coli) is to modulate the electrostatic environment of a                      proton-binding residue in the putative access channel of Fo.
MH  - A
MH  - atp
MH  - ATP synthesis
MH  - capsulatus
MH  - chromatophore
MH  - chromatophores
MH  - COMPLEX
MH  - electrogenic
MH  - ion
MH  - proton
MH  - Proton transfer
MH  - Rhodobacter
MH  - rhodobacter capsulatus
MH  - RHODOBACTER-CAPSULATUS
MH  - synthesis
MH  - TRANSFER
MH  - SUBUNIT
RP  - NOT IN FILE
SO  - EBEC Short Reports 2000  ;():

1743
UI  - 21055
AU  - Velours J
AU  - Arselin G
TI  - The Saccharomyces cerevisiae ATP synthase
AB  - The ATP synthase of the yeast Saccharomyces cerevisiae is composed of 20 different subunits whose primary structure is known. The organization of proteins that constitute the membranous domain is now under investigation. Cysteine insertions combined with the use of nonpermeant maleimide reagents and cross-linking reagents showing different lengths and specificity contribute to the knowledge of the location of the N- and C-termini of the subunits involved in the stator of the enzyme and their organization. This review summarizes data on yeast ATP synthase obtained in our laboratory since 1980
MH  - atp
MH  - ATP synthase
MH  - CROSS-LINKING
MH  - Cysteine
MH  - protein
MH  - Proteins
MH  - review
MH  - Saccharomyces cerevisiae
MH  - structure
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - Institut de Biochimie et Genetique Cellulaires du CNRS, Universite Victor Segalen, Bordeaux, France jeanvelours@ibgcu-bordeaux2frFAU - Velours, J
SO  - J Bioenerg Biomembr 2000 Aug ;32(4):383-390

1744
UI  - 21412
AU  - Velours J
AU  - Arselin G
TI  - The Saccharomyces cerevisiae ATP synthase
AB  - The ATP synthase of the yeast Saccharomyces cerevisiae is composed of 20 different subunits whose primary structure is known. The organization of proteins that constitute the membranous domain is now under investigation. Cysteine insertions combined with the use of nonpermeant maleimide reagents and cross-linking reagents showing different lengths and specificity contribute to the knowledge of the location of the N- and C-termini of the subunits involved in the stator of the enzyme and their organization. This review summarizes data on yeast ATP synthase obtained in our laboratory since 1980
MH  - Animal
MH  - atp
MH  - ATP synthase
MH  - Cell Nucleus
MH  - Chemistry
MH  - CROSS-LINKING
MH  - Cross-Linking Reagents
MH  - Cysteine
MH  - data
MH  - enzyme
MH  - enzymology
MH  - isolation & purification
MH  - Mammals
MH  - Mitochondria
MH  - Models,Molecular
MH  - primary
MH  - protein
MH  - Protein Subunits
MH  - Proteins
MH  - Proton-Translocating ATPases
MH  - review
MH  - Saccharomyces cerevisiae
MH  - structure
MH  - SUBUNIT
MH  - Support,Non-U.S.Gov't
MH  - SYNTHASE
MH  - YEAST
RP  - NOT IN FILE
NT  - Institut de Biochimie et Genetique Cellulaires du CNRS, Universite Victor Segalen, Bordeaux, France jeanvelours@ibgcu-bordeaux2frFAU - Velours, J
SO  - J Bioenerg Biomembr 2000 Aug ;32(4):383-390

1745
UI  - 383
AU  - Vinogradov AD
AD  - Department of Biochemistry, School of Biology, Moscow State University, Moscow 119899, Russian Federation adv@biochembiomsusu
TI  - Steady-state and pre-steady-state kinetics of the mitochondrial F(1)F(o) ATPase: is ATP synthase a reversible molecular machine?
AB  - H(+)-ATP synthase (F(1)F(o) ATPase) catalyzes the synthesis and/or hydrolysis of ATP, and the reactions are strongly affected by all the substrates (products) in a way clearly distinct from that expected of a simple reversibly operating enzyme. Recent studies have revealed the structure of F(1), which is ideally suited for the alternating binding change mechanism, with a rotating gamma-subunit as the energy-driven coupling device. According to this mechanism ATP, ADP, inorganic phosphate (P(i)) and Mg(2+) participate in the forward and reverse overall reactions exclusively as the substrates and products. However, both F(1) and F(1)F(o) demonstrate non-trivial steady-state and pre- steady-state kinetics as a function of variable substrate (product) concentrations. Several effectors cause unidirectional inhibition or activation of the enzyme. When considered separately, the unidirectional effects of ADP, P(i), Mg(2+) and energy supply on ATP synthesis or hydrolysis may possibly be explained by very complex kinetic schemes; taken together, the results suggest that different conformational states of the enzyme operate in the ATP hydrolase and ATP synthase reactions. A possible mechanism for an energy-dependent switch between the two states of F(1)F(o) ATPase is proposed
RP  - NOT IN FILE
NT  - UI - 20123950LA - engRN - 0 (Phosphates)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 7439-95-4 (Magnesium)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 20000217IS - 0022-0949SB - IMCY - ENGLANDJC - I2F
UR  - PM:10600672
SO  - J Exp Biol 2000 Jan ;203 Pt 1():41-49

1746
UI  - 21052
AU  - Vogel PD
TI  - Insights into ATP synthase structure and function using affinity and site-specific spin labeling
AB  - A variety of different approaches has been used during the last couple of decades to investigate structure and function relationships within the catalytic portion of the F0F1-ATP synthase and of its interactions with the proton-translocator F0. In our group, we employ ESR spectroscopy with the use of stable organic radicals, so-called spin labels, as reporter groups. The radicals are either attached to substrates/ligands or specifically inserted into the protein structure by "site-specific spin labeling." Both approaches bear intrinsic advantages for their special uses and result in the specific information that is available through ESR, e.g., structural changes due to binding of effector molecules (e.g., Mg2+ ions), conformational transitions during catalytic turnover, distance information on radicals bound at 20 A or less, and information on the binding characteristics of labeled substrates. This review summarizes the results of a variety of different approaches we have used during the last years to study, with the help of ESR spectroscopy, the structure of the nucleotide binding sites of F1-ATPases of different origins as well as interactions with F0 subunits
MH  - A
MH  - atp
MH  - ATP synthase
MH  - BINDING
MH  - Binding Sites
MH  - F0
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - function
MH  - ion
MH  - Ions
MH  - nucleotide binding
MH  - nucleotide binding sites
MH  - NUCLEOTIDE-BINDING-SITES
MH  - P
MH  - protein
MH  - review
MH  - Site
MH  - spectroscopy
MH  - structure
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - Fachbereich Chemie der Universitat Kaiserlautern, Germany vogel@-chemieuni-kldeFAU - Vogel, P D
SO  - J Bioenerg Biomembr 2000 Aug ;32(4):413-421

1747
UI  - 21409
AU  - Vogel PD
TI  - Insights into ATP synthase structure and function using affinity and site-specific spin labeling
AB  - A variety of different approaches has been used during the last couple of decades to investigate structure and function relationships within the catalytic portion of the F0F1-ATP synthase and of its interactions with the proton-translocator F0. In our group, we employ ESR spectroscopy with the use of stable organic radicals, so-called spin labels, as reporter groups. The radicals are either attached to substrates/ligands or specifically inserted into the protein structure by "site-specific spin labeling." Both approaches bear intrinsic advantages for their special uses and result in the specific information that is available through ESR, e.g., structural changes due to binding of effector molecules (e.g., Mg2+ ions), conformational transitions during catalytic turnover, distance information on radicals bound at 20 A or less, and information on the binding characteristics of labeled substrates. This review summarizes the results of a variety of different approaches we have used during the last years to study, with the help of ESR spectroscopy, the structure of the nucleotide binding sites of F1-ATPases of different origins as well as interactions with F0 subunits
MH  - A
MH  - affinity
MH  - Affinity Labels
MH  - atp
MH  - ATP synthase
MH  - BINDING
MH  - BINDING SITE
MH  - Binding Sites
MH  - catalytic
MH  - Chemistry
MH  - Chloroplasts
MH  - Crystallography,X-Ray
MH  - Electron Spin Resonance Spectroscopy
MH  - enzymology
MH  - Escherichia coli
MH  - F0
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - function
MH  - INTERACTION
MH  - ion
MH  - Ions
MH  - metabolism
MH  - nucleotide
MH  - nucleotide binding
MH  - nucleotide binding sites
MH  - NUCLEOTIDE-BINDING-SITES
MH  - P
MH  - protein
MH  - Protein Conformation
MH  - Proton-Translocating ATPases
MH  - review
MH  - Site
MH  - spectroscopy
MH  - Spin Labels
MH  - structure
MH  - SUBUNIT
MH  - Support,Non-U.S.Gov't
MH  - SYNTHASE
MH  - turnover
RP  - NOT IN FILE
NT  - Fachbereich Chemie der Universitat Kaiserlautern, Germany vogel@-chemieuni-kldeFAU - Vogel, P D
SO  - J Bioenerg Biomembr 2000 Aug ;32(4):413-421

1748
UI  - 20834
AU  - Wada Y
AU  - Sambongi Y
AU  - Futai M
AD  - Division of Biological Sciences, The Institute of Scientific and Industrial Research, Osaka University, CREST of Japan Science and Technology Corporation, Ibaraki, 567-0047, Japan
TI  - Biological nano motor, ATP synthase F(o)F(1): from catalysis to gammaepsilonc(10-12) subunit assembly rotation
AB  - Proton translocating ATPase (ATP synthase), a chemiosmotic enzyme, synthesizes ATP from ADP and phosphate coupling with the electrochemical ion gradient across the membrane. This enzyme has been studied extensively by combined genetic, biochemical and biophysical approaches. Such studies revealed a unique mechanism which transforms an electrochemical ion gradient into chemical energy through the rotation of a subunit assembly. Thus, this enzyme can be defined as a nano motor capable of coupling a chemical reaction and ion translocation, or more simply, as a protein complex carrying out rotational catalysis. In this article, we briefly discuss our recent work, emphasizing the rotation of subunit assembly (gammaepsilonc(10- 12)) which is formed from peripheral and intrinsic membrane subunits
MH  - A
MH  - ADP
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Catalysis
MH  - COMPLEX
MH  - coupling
MH  - ion
MH  - mechanism
MH  - membrane
MH  - Membrane Proteins
MH  - protein
MH  - Proteins
MH  - proton
MH  - Protons
MH  - review
MH  - rotation
MH  - SUBUNIT
MH  - SYNTHASE
MH  - translocation
RP  - NOT IN FILE
NT  - UI - 21061448LA - engRN - 0 (Membrane Proteins)RN - 0 (Molecular Motors)RN - 0 (Protons)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 20010126IS - 0006-3002SB - IMCY - Netherlands
UR  - PM:11004468
SO  - Biochim Biophys Acta 2000 Aug 15 ;1459(2-3):499-505

1749
UI  - 403
AU  - Weber J
AU  - Senior AE
AD  - Department of Biochemistry and Biophysics, Box 712, University of Rochester Medical Center, Rochester, NY 14642, USA
TI  - ATP synthase: what we know about ATP hydrolysis and what we do not know about ATP synthesis
AB  - In ATP synthase, X-ray structures, demonstration of ATP-driven gamma- subunit rotation, and tryptophan fluorescence techniques to determine catalytic site occupancy and nucleotide binding affinities have resulted in pronounced progress in understanding ATP hydrolysis, for which a mechanism is presented here. In contrast, ATP synthesis remains enigmatic. The molecular mechanism by which ADP is bound in presence of a high ATP/ADP concentration ratio is a fundamental unknown; similarly P(i) binding is not understood. Techniques to measure catalytic site occupancy and ligand binding affinity changes during net ATP synthesis are much needed. Relation of these parameters to gamma-rotation is a further goal. A speculative model for ATP synthesis is offered
RP  - NOT IN FILE
NT  - UI - 20298311LA - engRN - 0 (Molecular Motors)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 20000727IS - 0006-3002SB - IMCY - NETHERLANDSJC - A0W
UR  - PM:10838046
SO  - Biochim Biophys Acta 2000 May 31 ;1458(2-3):300-309

1750
UI  - 404
AU  - Weber J
AU  - Senior AE
AD  - Department of Biochemistry and Biophysics, Box 712, University of Rochester Medical Center, Rochester, New York 14642, USA joachim_weber@urmcrochesteredu
TI  - Features of F(1)-ATPase catalytic and noncatalytic sites revealed by fluorescence lifetimes and acrylamide quenching of specifically inserted tryptophan residues
AB  - Catalytic and noncatalytic nucleotide sites of the F(1) sector of ATP synthase were characterized by tryptophan fluorescence techniques. Seven Trp residues inserted in varied microenvironments in the catalytic sites, and one in the noncatalytic sites, were studied in mutant F(1) enzymes which were otherwise devoid of Trp. Parameters measured were fluorescence lifetimes and dynamic and static quenching by acrylamide in the absence or presence of nucleotide. The results indicated that the solution structures of the mutant enzymes were consistent with reported crystal structures. In enzyme with three empty noncatalytic sites, all sites were relatively inaccessible to acrylamide, indicating a closed conformation. In contrast, when the three catalytic sites were empty, they were relatively and equally accessible to acrylamide, indicating an open conformation. This was the case in the presence or absence of Mg(2+). Residue beta-Trp-331 has been extensively used previously to determine nucleotide binding parameters in F(1). Results here showed that in betaY331W mutant F(1), each of the three beta-Trp-331 residues has an unusually long fluorescence lifetime, confirming that each contributes equally to the overall fluorescence signal
RP  - NOT IN FILE
NT  - UI - 20281274LA - engRN - 0 (Nucleotides)RN - 73-22-3 (Tryptophan)RN - 7439-95-4 (Magnesium)RN - 79-06-1 (Acrylamide)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSDA - 20000706IS - 0006-2960SB - IMCY - UNITED STATESJC - A0G
UR  - PM:10819998
SO  - Biochemistry 2000 May 9 ;39(18):5287-5294

1751
UI  - 401
AU  - Weber J
AU  - Nadanaciva S
AU  - Senior AE
AD  - Department of Biochemistry and Biophysics, Box 712, University of Rochester Medical Center, 14642, Rochester, NY, USA
TI  - ATP-driven rotation of the gamma subunit in F(1)-ATPase
AB  - We present a mechanism for F(1)-ATPase in which hydrolysis of MgATP in the high-affinity catalytic site at the alpha/beta interface drives rotation of the gamma subunit via conformational changes in the alpha subunit. During hydrolysis, transition state formation and separation of P(i) from MgADP causes movement of portions of alpha, transmitted via two Arg residues which are hydrogen-bonded to the gamma-phosphate of MgATP, alphaArg376 and betaArg182; the latter is also hydrogen- bonded to interfacial alpha residues between alpha346 and alpha349. Changes in alpha conformation then push on gamma, resulting in rotation. Supporting evidence from the literature and from new data is discussed
RP  - NOT IN FILE
NT  - UI - 20490427LA - engRN - 0 (Protein Subunits)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM25349/GM/NIGMSDA - 20001107IS - 0014-5793SB - IMCY - NETHERLANDSJC - EUH
UR  - PM:11033345
SO  - FEBS Lett 2000 Oct 13 ;483(1):1-5

1752
UI  - 136
AU  - Wilkens S
AU  - Zhou J
AU  - Nakayama R
AU  - Dunn SD
AU  - Capaldi RA
AD  - Department of Biochemistry, University of California, Riverside, CA, 92521, USA stephanwilkens@ucredu
TI  - Localization of the delta subunit in the Escherichia coli F(1)F(0)- ATPsynthase by immuno electron microscopy: the delta subunit binds on top of the F(1)
AB  - The binding site of the delta subunit in the F(1)F(0)-ATPsynthase from Escherichia coli has been determined by electron microscopy of negatively stained, antibody-decorated enzyme molecules. The images show that the antibody is bound at the very top of the F(1) domain indicating that at least part of delta is bound in the dimple formed by the N termini of the alpha and beta subunits. The data may explain why there is only one binding site for delta on the F(1) despite there being three identical alphabeta pairs. The finding also implies that the b subunits of the F(0) have to extend all the way from the membrane surface to the very top of the F(1) domain to make contact with the delta subunit
RP  - NOT IN FILE
NT  - UI - 20090940LA - engRN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - RO1 HL58671/HL/NHLBIDA - 20000218IS - 0022-2836SB - IMCY - ENGLANDJC - J6V
UR  - PM:10623533
SO  - J Mol Biol 2000 Jan 21 ;295(3):387-391

1753
UI  - 21418
AU  - Wilkens S
TI  - F1F0-ATP synthase-stalking mind and imagination
AB  - Electron microscopy together with image analysis has been used to study the structure of the intact F1F0-ATPsynthase from Escherichia coli. A procedure has been developed which allows preparation of detergent-free enzyme. Aside from the well known two-domain structure, images of F1F0 prepared by this procedure show a number of additional features, including a second stalk, which can be seen extending all the way from the F0 to the top of the F1 in some images, and a small protein on the very top of the F1, which has been identified as the delta subunit by decoration with a monoclonal antibody. In light of these results, a refined model of the subunit arrangement of the complex is presented
MH  - A
MH  - analysis
MH  - Biochemistry
MH  - COMPLEX
MH  - CRYOELECTRON MICROSCOPY
MH  - delta
MH  - DELTA-SUBUNIT
MH  - electron
MH  - electron microscopy
MH  - ELECTRON-MICROSCOPY
MH  - enzyme
MH  - enzymology
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - F1
MH  - Light
MH  - Microscopy
MH  - Microscopy,Electron
MH  - model
MH  - MONOCLONAL-ANTIBODIES
MH  - protein
MH  - Protein Subunits
MH  - Proton-Translocating ATPases
MH  - stalk
MH  - structure
MH  - SUBUNIT
MH  - ultrastructure
MH  - universities
RP  - NOT IN FILE
NT  - Department of Biochemistry, University of California, Riverside, 92521, USA stephanwilkens@ucreduFAU - Wilkens, S
SO  - J Bioenerg Biomembr 2000 Aug ;32(4):333-339

1754
UI  - 896
AU  - Williams RJP
TI  - Mitochondria and chloroplasts: localized and delocalized bioenergetic transduction
MH  - Adenosine Triphosphate
MH  - Chloroplasts
MH  - Energy Metabolism
MH  - Energy Transfer
MH  - metabolism
MH  - Mitochondria
MH  - Models,Biological
RP  - NOT IN FILE
SO  - Trends Biochem Sci 2000 Oct ;25(10):479

1755
UI  - 21383
AU  - Ying H
AU  - Yu Y
AU  - Xu Y
TI  - Antisense of ATP synthase subunit e inhibits the growth of human hepatocellular carcinoma cells
AB  - Differentially expressed genes between normal and hepatocellular carcinoma tissues were investigated using differential display. We identified a cDNA fragment that was overexpressed in cancer tissue. Homology analysis showed that the sequence was identical to human ATP synthase subunit e (hAS-e). Moreover, Northern blot analysis demonstrated that hAS-e was overexpressed in 10 of 11 (91%) specimens of hepatocellular carcinoma compared with corresponding normal tissues. We introduced antisense hAS-e into a human hepatocellular carcinoma BEL-7404 cell and found that downregulation of the hAS-e led to cell growth inhibition. It was also found that the antisense transfection could decrease the serum-stimulated activation of mitogen-activated protein kinase (MAP kinase). Together, the results suggest that antisense of hAS-e can inhibit cell proliferation through the MAP kinase pathway. Our data indicate that hAS-e may become a new target in gene therapy
MH  - A
MH  - ACTIVATION
MH  - analysis
MH  - atp
MH  - ATP synthase
MH  - Base Sequence
MH  - Biochemistry
MH  - biology
MH  - Carcinoma,Hepatocellular
MH  - cell
MH  - Cell Division
MH  - Cells
MH  - Cloning,Molecular
MH  - data
MH  - DNA Primers
MH  - drug effects
MH  - genetics
MH  - Human
MH  - Liver Neoplasms
MH  - Mitochondrial Proton-Translocating ATPases
MH  - Oligodeoxyribonucleotides,Antisense
MH  - P
MH  - pharmacology
MH  - protein
MH  - Protein Subunits
MH  - SPECIMENS
MH  - SUBUNIT
MH  - Support,Non-U.S.Gov't
MH  - SYNTHASE
MH  - Tumor Cells,Cultured
RP  - NOT IN FILE
NT  - Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, P R ChinaFAU - Ying, H
SO  - Oncol Res 2000  ;12(11-12):485-490

1756
UI  - 19900
AU  - Yoshida T
AU  - Koga Y
AU  - Minowa H
AU  - Kamaya H
AU  - Ueda I
TI  - Interfacial lateral electrical conductance on lipid monolayer: Dose-dependent converse effect of alcohols
MH  - conductance
RP  - NOT IN FILE
NT  - JournalFEB 17285MYJ PHYS CHEM B
UR  - ISI:000085393900016
SO  - Journal of Physical Chemistry B 2000  ;104(6):1249-1252

1757
UI  - 21152
AU  - Adelroth P
AU  - Paddock ML
AU  - Tehrani A
AU  - Beatty JT
AU  - Feher G
AU  - Okamura MY
AD  - Department of Physics, University of California San Diego, La Jolla, California 92093, USA
TI  - Identification of the proton pathway in bacterial reaction centers: decrease of proton transfer rate by mutation of surface histidines at H126 and H128 and chemical rescue by imidazole identifies the initial proton donors
AB  - The pathway for proton transfer to Q(B) was studied in the reaction center (RC) from Rhodobacter sphaeroides. The binding of Zn(2+) or Cd(2+) to the RC surface at His-H126, His-H128, and Asp-H124 inhibits the rate of proton transfer to Q(B), suggesting that the His may be important for proton transfer [Paddock, M. L., Graige, M. S., Feher, G. and Okamura, M. Y. (1999) Proc. Natl. Acad. Sci. U.S.A. 96, 6183-6188]. To assess directly the role of the histidines, mutant RCs were constructed in which either one or both His were replaced with Ala. In the single His mutant RCs, no significant effects were observed. In contrast, in the double mutant RC at pH 8.5, the observed rates of proton uptake associated with both the first and the second proton- coupled electron-transfer reactions k(AB)(()(1)()) [Q(A)(-)(*)Q(B)-Glu(- ) + H(+) --> Q(A)(-)(*)Q(B)-GluH --> Q(A)Q(B)(-)(*)-GluH] and k(AB)(()(2)()) [Q(A)(-)(*)Q(B)(-)(*) + H(+) --> Q(A)(-)(*)(Q(B)H)(*) -- > Q(A)(Q(B)H)(-)], were found to be slowed by factors of approximately 10 and approximately 4, respectively. Evidence that the observed changes in the double mutant RC are due to a reduction in the proton- transfer rate constants are provided by the observations: (i) k(AB)(1) at pH approximately pK(a) of GluH became biphasic, indicating that proton transfer is slower than electron transfer and (ii) k(AB)(2) became independent of the driving force for electron transfer, indicating that proton transfer is the rate-limiting step. These changes were overcome by the addition of exogenous imidazole which acts as a proton donor in place of the imidazole groups of His that were removed in the double mutant RC. Thus, we conclude that His-H126 and His-H128 facilitate proton transfer into the RC, acting as RC-bound proton donors at the entrance of the proton-transfer pathways
MH  - A
MH  - BINDING
MH  - CONSTANT
MH  - electron
MH  - electron transfer
MH  - Electron-transfer
MH  - ELECTRON-TRANSFER REACTIONS
MH  - Histidine
MH  - M
MH  - mutant
MH  - pH
MH  - proton
MH  - Proton transfer
MH  - Proton-transfer
MH  - Protons
MH  - quinone
MH  - Quinones
MH  - rate constant
MH  - reaction center
MH  - Rhodobacter
MH  - Rhodobacter sphaeroides
MH  - sphaeroides
MH  - SURFACE
MH  - TRANSFER
RP  - NOT IN FILE
NT  - UI - 21581539LA - engRN - 0 (Imidazoles)RN - 0 (Photosynthetic Reaction Center, Bacterial)RN - 0 (Protons)RN - 0 (Quinones)RN - 56-41-7 (Alanine)RN - 71-00-1 (Histidine)PT - Journal ArticleID - GM-41637/GM/NIGMSDA - 20011128IS - 0006-2960SB - IMCY - United States
UR  - PM:11724567
SO  - Biochemistry 2001 Dec 4 ;40(48):14538-14546

1758
UI  - 9927
AU  - Arechaga I
AU  - Jones PC
AD  - The Medical Research Council Dunn Human Nutrition Unit, Wellcome Trust/MRC Building, Hills Road, CB2 2XY, Cambridge, UK ia@mrc- dunncamacuk
TI  - The rotor in the membrane of the ATP synthase and relatives
AB  - In recent years, structural information on the F(1) sector of the ATP synthase has provided an insight into the molecular mechanism of ATP catalysis. The structure strongly supports the proposal that the ATP synthase works as a rotary molecular motor. Insights into the membrane domain have just started to emerge but more detailed structural information is needed if the molecular mechanism of proton translocation coupled to ATP synthesis is to be understood. This review will focus mainly on the ion translocating rotor in the membrane domain of the F-type ATPase, and the related vacuolar and archaeal relatives
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - ATPase
MH  - Catalysis
MH  - H(+)-Transporting ATP Synthase
MH  - Human
MH  - Hydrogen
MH  - Ions
MH  - mechanism
MH  - Nutrition
MH  - proton
MH  - review
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 21195319LA - engRN - 0 (Ions)RN - 1333-74-0 (Hydrogen)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.- (V-type ATPase)RN - EC 3.6.1.- (ceroid lipofuscinosis protein)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 20010412IS - 0014-5793SB - IMCY - NetherlandsJC - EUH
UR  - PM:11297723
SO  - FEBS Lett 2001 Apr 6 ;494(1-2):1-5

1759
UI  - 9928
AU  - Arechaga I
AU  - Jones PC
AD  - The Medical Research Council, Dunn Human Nutrition Unit, Hills Road, CB2 2XY, Cambridge, UK
TI  - Quick guide: ATP synthase
MH  - atp
MH  - ATP synthase
MH  - England
MH  - H(+)-Transporting ATP Synthase
MH  - Human
MH  - Nutrition
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 21148092LA - engRN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - DirectoryDA - 20010316IS - 0960-9822SB - IMCY - EnglandJC - B44
UR  - PM:11250162
SO  - Curr Biol 2001 Feb 20 ;11(4):R117

1760
UI  - 681
AU  - Bald D
AU  - Noji H
AU  - Yoshida M
AU  - Hirono-Hara Y
AU  - Hisabori T
AD  - Chemical Resources Laboratory, Tokyo Institute of Technology, Yokohama, Kanagawa 226-8503
TI  - Redox regulation of the rotation of F1-ATP synthase
AB  - In F(1)-ATPase, the smallest known motor enzyme, unidirectional rotation of the central axis subunit gamma is coupled to ATP hydrolysis. In the present study, we report the redox-switching of the rotation of this enzyme. For the purpose, the switch region from the gamma subunit of the redox-sensitive chloroplast F(1)-ATPase was introduced into the bacterial F(1)-ATPase. The ATPase activity of the obtained complex was increased up to 3-fold upon reduction (Bald, D. et al., (2000) J. Biol. Chem. 275, 12757-12762). Here, we successfully observed the modulation of rotation of gamma in this chimeric complex by changes in the redox conditions. In addition we revealed that the suppressed enzymatic activity of the oxidized F(1)-ATPase complex was characterized by more frequent long pauses in the rotation of the gamma subunit. These findings obtained by the single molecule analysis therefore provide new insights into the mechanisms of enzyme regulation
RP  - NOT IN FILE
NT  - UI - 0LA - ENGPT - JOURNAL ARTICLEDA - 20010823IS - 0021-9258JC - HIV
UR  - PM:11518700
SO  - J Biol Chem 2001 Aug 22 ;():

1761
UI  - 21061
AU  - Belogrudov GI
AU  - Hatefi Y
TI  - Factor B and the mitochondrial ATP synthase complex
AB  - Factor B is a subunit of the mammalian ATP synthase complex, whose existence has been controversial. This paper describes the molecular and functional properties of a recombinant human factor B, which when added to bovine submitochondrial particles depleted of their factor B restores the energy coupling activity of the ATP synthase complexes. The mature human factor B has 175 amino acids and a molecular mass of 20,341 Da. The preparation is water-soluble, monomeric, and is inactivated by monothiol and especially dithiol modifying reagents, probably reacting at its cysteine residues C-92 and C-94. A likely factor B gene composed of 5 exons has been identified on chromosome 14q21.3, and the functional role of factor B in the mammalian ATP synthase complex has been discussed
MH  - A
MH  - ACID
MH  - Amino Acids
MH  - atp
MH  - ATP synthase
MH  - COMPLEX
MH  - coupling
MH  - Cysteine
MH  - Human
MH  - RESIDUE
MH  - Submitochondrial Particles
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, CA 92037
SO  - J Biol Chem 2001 Dec 14 ;.():

1762
UI  - 21436
AU  - Berggren KN
AU  - Chernokalskaya E
AU  - Lopez MF
AU  - Beechem JM
AU  - Patton WF
TI  - Comparison of three different fluorescent visualization strategies for detecting Escherichia coli ATP synthase subunits after sodium dodecyl sulfate-polyacrylamide gel electrophoresis
AB  - The correlation between protein molecular weight and the number of lysine or basic amino acid residues was found to be high for broad range molecular weight standards, subunits of Escherichia coli F1F0-ATP synthase and the translated open reading frame of E. coli. A relatively poor correlation between protein molecular weight and the number of cysteine residues was observed in all cases. The ability of amine-reactive, thiol-reactive and basic amino acid-binding fluorophores to detect the eight subunits of F1F0-ATP synthase complex was assessed using 2-methoxy-2,4-diphenyl-3(2H)-furanone (MDPF), monobromobimane (MBB) and SYPRO Ruby protein gel stain, respectively. Though experimentally none of the fluorophores provided accurate estimates of the subunit stoichiometry of this complex, MDPF and SYPRO Ruby protein gel stain were capable of semiquantitative detection of every subunit. MBB, however, failed to detect subunits a, b and c of the hydrophobic F0 complex, as well as subunit epsilon of the F1 complex. All three fluorescent detection procedures permitted subsequent identification of representative subunits by peptide mass profiling using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). The use of thiol-reactive fluorophores for the global analysis of protein expression profiles does not appear to be advisable as a significant number of proteins have few or no cysteine residues, thus escaping detection
MH  - A
MH  - ACID
MH  - Amino Acid Sequence
MH  - analysis
MH  - atp
MH  - ATP synthase
MH  - Bacterial Proton-Translocating ATPases
MH  - Bicyclo Compounds
MH  - Chemistry
MH  - COLI ATP SYNTHASE
MH  - Comparative Study
MH  - COMPLEX
MH  - Cysteine
MH  - Electrophoresis
MH  - enzymology
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - F1
MH  - F1F0-ATP SYNTHASE
MH  - Fluorescent Dyes
MH  - Furans
MH  - genetics
MH  - Genome,Bacterial
MH  - isolation & purification
MH  - molecular probes
MH  - Molecular Sequence Data
MH  - Molecular Weight
MH  - peptide
MH  - protein
MH  - Protein Subunits
MH  - Proteins
MH  - RESIDUE
MH  - Sodium
MH  - Spectrometry,Fluorescence
MH  - Spectrometry,Mass,Matrix-Assisted Laser Desorption-Ionization
MH  - stoichiometry
MH  - SUBUNIT
MH  - SUBUNIT-EPSILON
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - Proteomics Section, Molecular Probes, Inc, 4849 Pitchford Avenue, Eugene, Oregon 97402, USAFAU - Berggren, K N
SO  - Proteomics 2001 Jan ;1(1):54-65

1763
UI  - 497
AU  - Blum DJ
AU  - Ko YH
AU  - Hong S
AU  - Rini DA
AU  - Pedersen PL
AD  - Department of Biological Chemistry, Department of Art as Applied to Medicine, Johns Hopkins University, School of Medicine, 725 North Wolfe Street, Baltimore, Maryland, 21205-2185
TI  - Atp synthase motor components: proposal and animation of two dynamic models for stator function
AB  - Recent research indicates that ATP synthases (F(0)F(1)) contain two distinct nanomotors, one an electrochemically driven proton motor contained within F(0) that drives an ATP hydrolysis-driven motor (F(1)) in reverse during ATP synthesis. This is depicted in recent models as involving a series of events in which each of the three alphabeta pairs comprising F(1) is induced via a centrally rotating subunit (gamma) to undergo the sequential binding changes necessary to synthesize ATP (binding change mechanism). Stabilization of this rotary process (i.e., to minimize "wobble" of F(1)) is provided in current models by a peripheral stalk or "stator" that has recently been shown to extend from near the bottom of the ATP synthase molecule to the very top of F(1). Although quite elegant, these models envision the stator as fixed during ATP synthesis, i.e., bound to only a single alphabeta pair. This is despite the fact that the binding change mechanism views each alphabeta pair as going through the same sequential order of conformational changes which demonstrate a chemical equivalency among them. For this reason, we propose here two different dynamic models for stator function during ATP synthesis. Both models have been designed to maintain chemical equivalency among the three alphabeta pairs during ATP synthesis and both have been animated
RP  - NOT IN FILE
NT  - UI - 21458276LA - engPT - Journal ArticleDA - 20010927IS - 0006-291XSB - IMCY - United StatesJC - 9Y8
UR  - PM:11573932
SO  - Biochem Biophys Res Commun 2001 Oct 5 ;287(4):801-807

1764
UI  - 21426
AU  - Boyer PD
TI  - Toward an adequate scheme for the ATP synthase catalysis
AB  - The suggestions from the author's group over the past 25 years for how steps in catalysis by ATP synthase occur are reviewed. Whether rapid ATP hydrolysis requires the binding of ATP to a second site (bi-site activation) or to a second and third site (tri-site activation) is considered. Present evidence is regarded as strongly favoring bi-site activation. Presence of nucleotides at three sites during rapid ATP hydrolysis can be largely accounted for by the retention of ADP formed and/or by the rebinding of ADP formed. Menz, Leslie and Walker ((2001) FEBS Lett., 494, 11-14) recently attained an X-ray structure of a partially closed enzyme form that binds ADP better than ATP. This accomplishment and other considerations form the base for a revised reaction sequence. Three types of catalytic sites are suggested, similar to those proposed before the X-ray data became available. During net ATP synthesis a partially closed site readily binds ADP and Pi but not ATP. At a closed site, tightly bound ADP and Pi are reversibly converted to tightly bound ATP. ATP is released from a partially closed site that can readily bind ATP or ADP. ATP hydrolysis when protonmotive force is low or lacking occurs simply by reversal of all steps with the opposite rotation of the gamma subunit. Each type of site can exist in various conformations or forms as they are interconverted during a 120 degrees rotation. The conformational changes with the ATP synthase, including the vital change when bound ADP and Pi are converted to bound ATP, are correlated with those that occur in enzyme catalysis in general, as illustrated by recent studies of Rose with fumarase. The betaE structure of Walker's group is regarded as an unlikely, or only quite transient, intermediate. Other X-ray structures are regarded as closely resembling but not identical with certain forms participating in catalysis. Correlation of the suggested reaction scheme with other present information is considered
MH  - A
MH  - ACTIVATION
MH  - Adenosine Triphosphate
MH  - ADP
MH  - Animal
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - BASE
MH  - BINDING
MH  - Binding Sites
MH  - biology
MH  - biosynthesis
MH  - Catalysis
MH  - catalytic
MH  - Chemistry
MH  - Chloroplast Proton-Translocating ATPases
MH  - conformation
MH  - conformational change
MH  - conformational changes
MH  - Crystallography,X-Ray
MH  - data
MH  - enzyme
MH  - Enzyme Activation
MH  - Hydrolysis
MH  - intermediate
MH  - Kinetics
MH  - metabolism
MH  - Mitochondrial Proton-Translocating ATPases
MH  - nucleotide
MH  - Nucleotides
MH  - P
MH  - rotation
MH  - Site
MH  - structure
MH  - Substrate Specificity
MH  - SUBUNIT
MH  - SYNTHASE
MH  - synthesis
MH  - universities
RP  - NOT IN FILE
NT  - Molecular Biology Institute, Boyer Hall, University of California, Los Angeles, CA 90095-1570, USA pdboyer@uclaeduFAU - Boyer, P D
SO  - Biochemistry (Mosc ) 2001 Oct ;66(10):1058-1066

1765
UI  - 21065
AU  - Boyer PD
TI  - Toward an adequate scheme for the ATP synthase catalysis
AB  - The suggestions from the author's group over the past 25 years for how steps in catalysis by ATP synthase occur are reviewed. Whether rapid ATP hydrolysis requires the binding of ATP to a second site (bi-site activation) or to a second and third site (tri-site activation) is considered. Present evidence is regarded as strongly favoring bi-site activation. Presence of nucleotides at three sites during rapid ATP hydrolysis can be largely accounted for by the retention of ADP formed and/or by the rebinding of ADP formed. Menz, Leslie and Walker ((2001) FEBS Lett., 494, 11-14) recently attained an X-ray structure of a partially closed enzyme form that binds ADP better than ATP. This accomplishment and other considerations form the base for a revised reaction sequence. Three types of catalytic sites are suggested, similar to those proposed before the X-ray data became available. During net ATP synthesis a partially closed site readily binds ADP and Pi but not ATP. At a closed site, tightly bound ADP and Pi are reversibly converted to tightly bound ATP. ATP is released from a partially closed site that can readily bind ATP or ADP. ATP hydrolysis when protonmotive force is low or lacking occurs simply by reversal of all steps with the opposite rotation of the gamma subunit. Each type of site can exist in various conformations or forms as they are interconverted during a 120 degrees rotation. The conformational changes with the ATP synthase, including the vital change when bound ADP and Pi are converted to bound ATP, are correlated with those that occur in enzyme catalysis in general, as illustrated by recent studies of Rose with fumarase. The betaE structure of Walker's group is regarded as an unlikely, or only quite transient, intermediate. Other X-ray structures are regarded as closely resembling but not identical with certain forms participating in catalysis. Correlation of the suggested reaction scheme with other present information is considered
MH  - A
MH  - ACTIVATION
MH  - Adenosine Triphosphate
MH  - ADP
MH  - Animal
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - BASE
MH  - BINDING
MH  - Binding Sites
MH  - biosynthesis
MH  - Catalysis
MH  - Chemistry
MH  - Chloroplast Proton-Translocating ATPases
MH  - conformation
MH  - conformational change
MH  - conformational changes
MH  - Crystallography,X-Ray
MH  - Enzyme Activation
MH  - Hydrolysis
MH  - intermediate
MH  - Kinetics
MH  - metabolism
MH  - Mitochondrial Proton-Translocating ATPases
MH  - Nucleotides
MH  - P
MH  - rotation
MH  - Site
MH  - structure
MH  - Substrate Specificity
MH  - SUBUNIT
MH  - SYNTHASE
MH  - synthesis
RP  - NOT IN FILE
NT  - Molecular Biology Institute, Boyer Hall, University of California, Los Angeles, CA 90095-1570, USA pdboyer@uclaeduFAU - Boyer, P D
SO  - Biochemistry (Mosc ) 2001 Oct ;66(10):1058-1066

1766
UI  - 19766
AU  - Boyer PD
AD  - Molecular Biology Institute, University of California, Los Angeles, CA 90095, USA
TI  - New insights into one of nature's remarkable catalysts, the ATP synthase
AB  - In the August 10, 2001 issue of Cell, Menz et al. report the crystal structure of a novel inhibited form of the bovine mitochondrial F(1)- ATPase at 2 A resolution, with all three catalytic sites occupied by nucleotide, one of which binds ADP in preference to ATP
MH  - A
MH  - ADP
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - H(+)-Transporting ATP Synthase
MH  - Macromolecular Systems
MH  - resolution
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 21431971LA - engRN - 0 (Macromolecular Systems)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - CommentPT - Journal ArticleDA - 20010907IS - 1097-2765SB - IMCY - United StatesJC - C5E
UR  - PM:11545726
SO  - Mol Cell 2001 Aug ;8(2):246-247

1767
UI  - 21424
AU  - Cabezon E
AU  - Runswick MJ
AU  - Leslie AG
AU  - Walker JE
TI  - The structure of bovine IF(1), the regulatory subunit of mitochondrial F-ATPase
AB  - In mitochondria, the hydrolytic activity of ATP synthase is regulated by an inhibitor protein, IF(1). Its binding to ATP synthase depends on pH, and below neutrality, IF(1) is dimeric and forms a stable complex with the enzyme. At higher pH values, IF(1) forms tetramers and is inactive. In the 2.2 A structure of the bovine IF(1) described here, the four monomers in the asymmetric unit are arranged as a dimer of dimers. Monomers form dimers via an antiparallel alpha-helical coiled coil in the C-terminal region. Dimers are associated into oligomers and form long fibres in the crystal lattice, via coiled-coil interactions in the N-terminal and inhibitory regions (residues 14-47). Therefore, tetramer formation masks the inhibitory region, preventing IF(1) binding to ATP synthase
MH  - A
MH  - Animal
MH  - atp
MH  - ATP synthase
MH  - BINDING
MH  - Cattle
MH  - Chemistry
MH  - COMPLEX
MH  - Dimerization
MH  - enzyme
MH  - enzymology
MH  - F-ATPASE
MH  - Histidine
MH  - Human
MH  - Hydrogen-Ion Concentration
MH  - inhibitor
MH  - INHIBITOR PROTEIN
MH  - INTERACTION
MH  - metabolism
MH  - Mitochondria
MH  - Nutrition
MH  - pH
MH  - protein
MH  - Protein Binding
MH  - Protein Conformation
MH  - Proton-Translocating ATPases
MH  - RESIDUE
MH  - structure
MH  - SUBUNIT
MH  - Support,Non-U.S.Gov't
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - The Medical Research Council Dunn Human Nutrition Unit, Hills Road, Cambridge CB2 2XY, UKFAU - Cabezon, E
SO  - EMBO J 2001 Dec 17 ;20(24):6990-6996

1768
UI  - 21062
AU  - Cabezon E
AU  - Runswick MJ
AU  - Leslie AG
AU  - Walker JE
TI  - The structure of bovine IF(1), the regulatory subunit of mitochondrial F-ATPase
AB  - In mitochondria, the hydrolytic activity of ATP synthase is regulated by an inhibitor protein, IF(1). Its binding to ATP synthase depends on pH, and below neutrality, IF(1) is dimeric and forms a stable complex with the enzyme. At higher pH values, IF(1) forms tetramers and is inactive. In the 2.2 A structure of the bovine IF(1) described here, the four monomers in the asymmetric unit are arranged as a dimer of dimers. Monomers form dimers via an antiparallel alpha-helical coiled coil in the C-terminal region. Dimers are associated into oligomers and form long fibres in the crystal lattice, via coiled-coil interactions in the N-terminal and inhibitory regions (residues 14-47). Therefore, tetramer formation masks the inhibitory region, preventing IF(1) binding to ATP synthase
MH  - A
MH  - Animal
MH  - atp
MH  - ATP synthase
MH  - BINDING
MH  - Cattle
MH  - Chemistry
MH  - COMPLEX
MH  - Dimerization
MH  - enzymology
MH  - F-ATPASE
MH  - Histidine
MH  - Human
MH  - Hydrogen-Ion Concentration
MH  - inhibitor
MH  - INHIBITOR PROTEIN
MH  - metabolism
MH  - Mitochondria
MH  - Nutrition
MH  - pH
MH  - protein
MH  - Protein Binding
MH  - Protein Conformation
MH  - Proton-Translocating ATPases
MH  - RESIDUE
MH  - structure
MH  - SUBUNIT
MH  - Support,Non-U.S.Gov't
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - The Medical Research Council Dunn Human Nutrition Unit, Hills Road, Cambridge CB2 2XY, UKFAU - Cabezon, E
SO  - EMBO J 2001 Dec 17 ;20(24):6990-6996

1769
UI  - 21386
AU  - Cao NJ
AU  - Brusilow WS
AU  - Tomashek JJ
AU  - Woodbury DJ
TI  - Characterization of reconstituted Fo from wild-type Escherichia coli and identification of two other fluxes co-purifying with Fo
AB  - We purified the ATPase Fo sector from a nonoverexpressing strain of Escherichia coli, reconstituted it into lipid vesicles made of either asolectin or two different mixtures of purified lipids, and measured proton flux through the reconstituted proton channel. We measured single-channel conductances and found that Fo activity depends on both lipids and reconstitution methods. In asolectin vesicles, Fo has a single-channel conductance of about 0.2 fS. Additionally, the relatively impure Fo prepared from cells carrying single-copy ATPase genes allowed us to observe two other fluxes, a nonselective cation leak (C(L)) and a slow H+ flux (Hs). Unlike the Fo flux, these fluxes could not be blocked by the Fo inhibitor DCCD. The C, reduces the total apparent trapped volume inside vesicles and therefore must equilibrate both H+ and K+ in the vesicles that contain it. When reconstituted into bilayers, these Fo preparations displayed a 120 pS cation channel with characteristics consistent with C(L) flux. The Hs conducts only H+ but at a slower rate than the Fo. We were therefore able to: 1) quantitate the single-channel conductance of the Fo, 2) demonstrate that our Fo purification method co-purified other membrane proteins that have ion-conduction properties, and 3) show that certain lipids are necessary for functional reconstitution of Fo
MH  - A
MH  - Adenosinetriphosphatase
MH  - ATPase
MH  - Biological Transport
MH  - cell
MH  - Cell Membrane
MH  - Cells
MH  - Chemistry
MH  - conductance
MH  - Electrophoresis,Polyacrylamide Gel
MH  - enzymology
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - FO
MH  - H+
MH  - Hydrogen
MH  - inhibitor
MH  - Lipid Bilayers
MH  - Lipids
MH  - membrane
MH  - Membrane Proteins
MH  - metabolism
MH  - method
MH  - Methods
MH  - physiology
MH  - Poisson Distribution
MH  - protein
MH  - Proteins
MH  - proton
MH  - Proton-Translocating ATPases
MH  - Protons
MH  - purification
MH  - reconstitution
MH  - Support,U.S.Gov't,P.H.S.
MH  - Time Factors
MH  - universities
MH  - vesicles
RP  - NOT IN FILE
NT  - Department of Physiology, Wayne State University School of Medicine, Detroit, MI 48201, USAFAU - Cao, N J
SO  - Cell Biochem Biophys 2001  ;34(3):305-320

1770
UI  - 21173
AU  - Cao NJY
AU  - Brusilow WS
AU  - Tomashek JJ
AU  - Woodbury DJ
TI  - Characterization of reconstituted F-o from wild-type Escherichia coli and identification of two other fluxes co-purifying with F-o
AB  - We purified the ATPase F-o sector from a nonoverexpressing strain of Escherichia coli, reconstituted it intolipid vesicles made of either asolectin or two different mixtures of purified lipids, and measured proton fluxthrough the reconstituted proton channel. We measured single-channel conductances and found that F-o activitydepends on both lipids and reconstitution methods. In asolectin vesicles, F-o has a single-channel conductanceof about 0.2 fS. Additionally, the relatively impure F-o prepared from cells carrying single-copy ATPase genesallowed us to observe two other fluxes, a nonselective cation leak (C-L) and a slow H+ flux (H-s). Unlike theF-o flux, these fluxes could not be blocked by the F-o inhibitor DCCD. The C-L reduces the total apparenttrapped volume inside vesicles and therefore must equilibrate both H+ and K+ in the vesicles that contain it.When reconstituted into bilayers, these F-o preparations displayed a 120 pS cation channel with characteristicsconsistent with C-L flux. The H-s conducts only H+ but at a slower rate than the F-o. We were therefore ableto: 1) quantitate the single-channel conductance of the F-o, 2) demonstrate that our F-o purification methodco-purified other membrane proteins that have ion-conduction properties, and 3) show that certain lipids arenecessary for functional reconstitution of F-o.
MH  - ESCHERICHIA-COLI
MH  - Escherichia coli
MH  - ATPase
MH  - A
MH  - vesicles
MH  - Lipids
MH  - proton
MH  - conductance
MH  - reconstitution
MH  - Methods
MH  - method
MH  - Cells
MH  - H+
MH  - inhibitor
MH  - purification
MH  - membrane
MH  - Membrane Proteins
MH  - Proteins
MH  - protein
RP  - NOT IN FILE
SO  - Cell Biochemistry and Biophysics 2001  ;34():305-320

1771
UI  - 979
AU  - Chami M
AU  - Pehau-Arnaudet G
AU  - Lambert O
AU  - Ranck JL
AU  - Levy D
AU  - Rigaud JL
TI  - Use of octyl beta-thioglucopyranoside in two-dimensional crystallization of membrane proteins
AB  - A great interest exists in producing and/or improving two-dimensional (2D) crystals of membrane proteins amenable to structural analysis by electron crystallography. Here we report on the use of the detergent n-octyl beta-d-thioglucopyranoside in 2D crystallization trials of membrane proteins with radically different structures including FhuA from the outer membrane of Escherichia coli, light-harvesting complex II from Rubrivivax gelatinosus, and Photosystem I from cyanobacterium Synechococcus sp. We have analyzed by electron microscopy the structures reconstituted after detergent removal from lipid-detergent or lipid-protein-detergent micellar solutions containing either only n-octyl beta-d-thioglucopyranoside or n-octyl beta-d-thioglucopyranoside in combination with other detergents commonly used in membrane protein biochemistry. This allowed the definition of experimental conditions in which the use of n-octyl beta-d-thioglucopyranoside could induce a considerable increase in the size of reconstituted membrane structures, up to several micrometers. An other important feature was that, in addition to reconstitution of membrane proteins into large bilayered structures, this thioglycosylated detergent also was revealed to be efficient in crystallization trials, allowing the proteins to be analyzed in large coherent two-dimensional arrays. Thus, inclusion of n-octyl beta-d-thioglucopyranoside in 2D crystallization trials appears to be a promising method for the production of large and coherent 2D crystals that will be valuable for structural analysis by electron crystallography and atomic force microscopy. Copyright 2001 Academic Press
MH  - Bacterial Outer Membrane Proteins
MH  - Biochemistry
MH  - Chemistry
MH  - Crystallization
MH  - Cyanobacteria
MH  - Detergents
MH  - Escherichia coli
MH  - Liposomes
MH  - Membrane Proteins
MH  - metabolism
MH  - Micelles
MH  - Microscopy
MH  - Microscopy,Electron
MH  - Photosynthetic Reaction Center,Bacterial
MH  - Proteins
MH  - Proteobacteria
MH  - Receptors,Virus
MH  - Solutions
MH  - Support,Non-U.S.Gov't
MH  - Synechocystis Group
MH  - Thioglucosides
MH  - ultrastructure
RP  - NOT IN FILE
NT  - Section de Recherche, Institut Curie, UMR-CNRS 168 et LRC-CEA 8, 11 rue Pierre et Marie Curie, 75231 Paris, France
SO  - J Struct Biol 2001 Jan ;133(1):64-74

1772
UI  - 21063
AU  - Chang SY
AU  - Park SG
AU  - Kim S
AU  - Kang CY
TI  - Interaction of the C-terminal domain of p43 and the alpha subunit of ATP synthase: Its functional implication in endothelial cell proliferation
AB  - Human p43 is associated with macromolecular tRNA synthase complex and known as a precursor of EMAP II. Interestingly, p43 is also secreted to induce proinflammatory genes. Although p43 itself seems to be a cytokine working at physiological condition, most of the functional studies have been obtained with its C-terminal equivalent, EMAP II. To gain an insight into the working mechanism of p43/EMAP II, we used EMAP II and searched for an interacting cell surface molecule. The level of EMAP II-binding molecule(s) was significantly increased in serum starved tumor cells. Thus, the EMAP II-binding molecule was isolated from the membrane of the serum-starved CEM cell. The isolated protein was determined to be the alpha subunit of ATP synthase. The interaction of EMAP II and alpha-ATP synthase was confirmed by ELISA and in vitro pull down assays and blocked with the antibodies raised against EMAP II and alpha-ATP synthase. The binding of EMAP II to the surface of serum starved cells was inhibited in the presence of soluble alpha-ATP synthase. EMAP II inhibited the growth of endothelial cells and this effect was relieved by soluble alpha-ATP synthase. Anti-alpha-ATP synthase antibody also showed inhibitory effect on the proliferation of endothelial cells mimicking the activity of EMAP II. These results suggest the potential interaction of p43/EMAP II with alpha-ATP synthase and its role in the proliferation of endothelial cells
MH  - A
MH  - alpha
MH  - ALPHA-SUBUNIT
MH  - atp
MH  - ATP synthase
MH  - BINDING
MH  - Cells
MH  - COMPLEX
MH  - Human
MH  - In Vitro
MH  - mechanism
MH  - membrane
MH  - protein
MH  - SUBUNIT
MH  - SURFACE
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - Laboratory of Immunology, Seoul National University, Seoul 151-742
SO  - J Biol Chem 2001 Dec 11 ;.():

1773
UI  - 21304
AU  - Checover S
AU  - Marantz Y
AU  - Nachliel E
AU  - Gutman M
AU  - Pfeiffer M
AU  - Tittor J
AU  - Oesterhelt D
AU  - Dencher NA
AD  - Technische Universitat Darmstadt, Institut fur Biochemie, Darmstadt D-64287, Germany
TI  - Dynamics of the proton transfer reaction on the cytoplasmic surface of bacteriorhodopsin
AB  - The cytoplasmic surface of bacteriorhodopsin is characterized by a group of carboxylates that function as a proton attractive domain [Checover, S., Nachliel, E., Dencher, N. A., and Gutman, M. (1997) Biochemistry 36, 13919-13928]. To identify these carboxylates, we selectively mutated them into cysteine residues and monitored the effects of the dynamics of proton transfer between the bulk and the surface of the protein. The measurements were carried out without attachment of a pH-sensor to the cysteine residue, thus avoiding any structural perturbation and change in the surface charge caused by the attachment of a reporter group, and the protein was in its BR state. The purple membranes were suspended in an unbuffered solution of pyranine (8-hydroxypyrene-1,3,6-trisulfonate) and exposed to a train of 1000 laser pulses (2.1 mJ/pulse, lambda = 355 nm, at 10 Hz). The excitation of the dye ejected the hydroxyl's proton, and a few nanoseconds later, a pair of free protons and ground-state pyranine anion was formed. The experimental observation was the dynamics of the relaxation of the system to the prepulse state. The observed signals were reconstructed by a numeric method that replicates the chemical reactions proceeding in the perturbed space. The detailed reconstruction of the measured signal assigned the various proton-binding sites with rate constants for proton binding and proton exchange and the pK values. Comparison of the results obtained by the various mutants indicates that the dominant proton-binding cluster of the wild-type protein consists of D104, E161, and E234. The replacement of D104 or E161 with cysteine lowered the proton binding capacity of the cluster to approximately 60% of that of the native protein. The replacement of E234 with cysteine disrupted the structure of the cluster, causing the two remaining carboxylates to function as isolated residues that do not interact with each other. The possibility of proton transfer between monomers is discussed
MH  - A
MH  - ACID
MH  - Anions
MH  - Arylsulfonates
MH  - Bacteriorhodopsin
MH  - BINDING
MH  - Biochemistry
MH  - buffer
MH  - Buffers
MH  - CONSTANT
MH  - Cysteine
MH  - DYE
MH  - dyes
MH  - Fluorescent Dyes
MH  - function
MH  - glutamic acid
MH  - M
MH  - membrane
MH  - Membranes
MH  - method
MH  - mutant
MH  - peptide
MH  - Peptide Fragments
MH  - protein
MH  - proton
MH  - Proton transfer
MH  - Proton-transfer
MH  - Protons
MH  - purple membrane
MH  - rate constant
MH  - relaxation
MH  - RESIDUE
MH  - Site
MH  - structure
MH  - SURFACE
MH  - SYSTEM
MH  - TRANSFER
MH  - united states
RP  - NOT IN FILE
NT  - UI - 21181600DA - 20010404IS - 0006-2960LA - engPT - Journal ArticleCY - United StatesRN - 0 (Anions)RN - 0 (Arylsulfonates)RN - 0 (Buffers)RN - 0 (Carboxylic Acids)RN - 0 (Fluorescent Dyes)RN - 0 (Peptide Fragments)RN - 0 (Protons)RN - 53026-44-1 (Bacteriorhodopsins)RN - 56-84-8 (Aspartic Acid)RN - 56-86-0 (Glutamic Acid)RN - 6358-69-6 (pyranine)SB - IM
UR  - PM:11284684
SO  - Biochemistry 2001 Apr 10 ;40(14):4281-4292

1774
UI  - 21231
AU  - Cherepanov DA
AU  - Krishtalik LI
AU  - Mulkidjanian AY
AD  - Division of Biophysics, Faculty of Biology/Chemistry, University of Osnabruck, D-49069 Osnabruck, Germany
TI  - Photosynthetic electron transfer controlled by protein relaxation: analysis by Langevin stochastic approach
AB  - Relaxation processes in proteins range in time from picoseconds to seconds. Correspondingly, biological electron transfer (ET) could be controlled by slow protein relaxation. We used the Langevin stochastic approach to describe this type of ET dynamics. Two different types of kinetic behavior were revealed, namely: oscillating ET (that could occur at picoseconds) and monotonically relaxing ET. On a longer time scale, the ET dynamics can include two different kinetic components. The faster one reflects the initial, nonadiabatic ET, whereas the slower one is governed by the medium relaxation. We derived a simple relation between the relative extents of these components, the change in the free energy (DeltaG), and the energy of the slow reorganization Lambda. The rate of ET was found to be determined by slow relaxation at -DeltaG < or = Lambda. The application of the developed approach to experimental data on ET in the bacterial photosynthetic reaction centers allowed a quantitative description of the oscillating features in the primary charge separation and yielded values of Lambda for the slower low-exothermic ET reactions. In all cases but one, the obtained estimates of Lambda varied in the range of 70-100 meV. Because the vast majority of the biological ET reactions are only slightly exothermic (DeltaG > or = -100 meV), the relaxationally controlled ET is likely to prevail in proteins
MH  - A
MH  - analysis
MH  - Biophysics
MH  - electron
MH  - electron transfer
MH  - Electron-transfer
MH  - protein
MH  - Proteins
MH  - reaction center
MH  - relaxation
MH  - Time
MH  - TRANSFER
RP  - NOT IN FILE
NT  - UI - 21124256LA - engRN - 0 (Photosynthetic Reaction Center, Bacterial)PT - Journal ArticleDA - 20010306IS - 0006-3495SB - IMCY - United States
UR  - PM:11222272
SO  - Biophys J 2001 Mar ;80(3):1033-1049

1775
UI  - 21230
AU  - Cherepanov DA
AU  - Mulkidjanian AY
AD  - Abteilung Biophysik, Fachbereich Biologie/Chemie, Universitat Osnabruck, Germany
TI  - Proton transfer in Azotobacter vinelandii ferredoxin I: entatic Lys84 operates as elastic counterbalance for the proton-carrying Asp15
AB  - In ferredoxin I from Azotobacter vinelandii, the reduction of a [3Fe- 4S] iron-sulphur cluster is coupled with the protonation of the mu2S sulphur atom that is approx. 6 A away from the protein boundary. The recent study of the site-specific mutants of ferredoxin I led to the conclusion that a particular surface aspartic residue (Asp15) is solely responsible for the proton transfer to the mu2S atom by 'rapid penetrative excursions' (K. Chen, J. Hirst, R. Camba, C.A. Bonagura, C.D. Stout, B.K. Burgess, F.A. Armstrong, Nature 405 (2000) 814-817). In the same paper it has been reported that the replacement of Asp15 by glutamate led to the blockage of the enzyme, although glutamate, with its longer and more flexible side chain, should apparently do even better as a mobile proton carrier than aspartate. We tackled this puzzling incompetence of Glu15 by molecular dynamics simulations. It was revealed that the conformational alterations of Asp15 are energetically balanced by the straining of the nearby Lys84 side chain in wild-type ferredoxin I but not in the Asp15-->Glu mutant. Lys84 in ferredoxin I of A. vinelandii seems to represent the first case where the strained (entatic) conformation of a particular amino acid side chain could be directly identified in the ground state of an enzyme and assigned to a distinct mechanism of energy balance during the catalytic transition
MH  - A
MH  - ACID
MH  - conformation
MH  - mechanism
MH  - MOLECULAR-DYNAMICS
MH  - mutant
MH  - protein
MH  - proton
MH  - Proton Pump
MH  - Proton transfer
MH  - Proton-transfer
MH  - protonation
MH  - RESIDUE
MH  - SIMULATION
MH  - SIMULATIONS
MH  - SURFACE
MH  - TRANSFER
RP  - NOT IN FILE
NT  - UI - 21232880LA - engRN - 0 (Ferredoxins)RN - 0 (Proton Pumps)RN - 56-84-8 (Aspartic Acid)RN - 56-87-1 (Lysine)RN - 7006-34-0 (Asparagine)PT - Journal ArticleDA - 20010503IS - 0006-3002SB - IMCY - Netherlands
UR  - PM:11334783
SO  - Biochim Biophys Acta 2001 Jun 1 ;1505(2-3):179-184

1776
UI  - 278
AU  - Cherepanov DA
AU  - Junge W
AD  - Division of Biophysics, University of Osnabruck, D-49069 Osnabruck, Germany
TI  - Viscoelastic Dynamics of Actin Filaments Coupled to Rotary F-ATPase: Curvature as an Indicator of the Torque
AB  - ATP synthase (F-ATPase) operates as an electrochemical-to-mechanical-to- chemical energy transducer with an astounding 360 degrees rotary motion of subunits epsilongammac(10-14) (rotor) against delta(alphabeta)(3)ab(2) (stator). The enzyme's torque as a function of the angular reaction coordinate in relation to ATP- synthesis/hydrolysis, internal elasticity, and external load has remained an important issue. Fluorescent actin filaments of micrometer length have been used to detect the rotation as driven by ATP hydrolysis. We evaluated the viscoelastic dynamics of actin filaments under the influence of enzyme-generated torque, stochastic Langevin force, and viscous drag. Modeling with realistic parameters revealed the dominance of the lowest normal mode. Because of its slow relaxation (~100 ms), power strokes of the enzyme were expected to appear strongly damped in recordings of the angular velocity of the filament. This article describes the theoretical background for the alternative use of the filament as a spring balance. The enzyme's angular torque profile under load can be gauged by measuring the average curvature and the stochastic fluctuations of actin filaments. Pertinent experiments were analyzed in the companion paper
RP  - NOT IN FILE
NT  - UI - 21400327LA - engPT - Journal ArticleDA - 20010817IS - 0006-3495SB - IMCY - United StatesJC - A5S
UR  - PM:11509340
SO  - Biophys J 2001 Sep ;81(3):1234-1244

1777
UI  - 21257
AU  - Cotton NP
AU  - White SA
AU  - Peake SJ
AU  - McSweeney S
AU  - Jackson JB
AD  - School of Biosciences, University of Birmingham, Edgbaston, B15 2TT, Birmingham, United Kingdom
TI  - The crystal structure of an asymmetric complex of the two nucleotide binding components of proton-translocating transhydrogenase
AB  - BACKGROUND: Membrane-bound ion translocators have important functions in biology, but their mechanisms of action are often poorly understood. Transhydrogenase, found in animal mitochondria and bacteria, links the redox reaction between NAD(H) and NADP(H) to proton translocation across a membrane. Linkage is achieved through changes in protein conformation at the nucleotide binding sites. The redox reaction takes place between two protein components located on the membrane surface: dI, which binds NAD(H), and dIII, which binds NADP(H). A third component, dII, provides a proton channel through the membrane. Intact membrane-located transhydrogenase is probably a dimer (two copies each of dI, dII, and dIII). RESULTS: We have solved the high-resolution crystal structure of a dI:dIII complex of transhydrogenase from Rhodospirillum rubrum-the first from a transhydrogenase of any species. It is a heterotrimer, having two polypeptides of dI and one of dIII. The dI polypeptides fold into a dimer. The loop on dIII, which binds the nicotinamide ring of NADP(H), is inserted into the NAD(H) binding cleft of one of the dI polypeptides. The cleft of the other dI is not occupied by a corresponding dIII component. CONCLUSIONS: The redox step in the transhydrogenase reaction is readily visualized; the NC4 atoms of the nicotinamide rings of the bound nucleotides are brought together to facilitate direct hydride transfer with A-B stereochemistry. The asymmetry of the dI:dIII complex suggests that in the intact enzyme there is an alternation of conformation at the catalytic sites associated with changes in nucleotide binding during proton translocation
MH  - A
MH  - Animal
MH  - Bacteria
MH  - BINDING
MH  - Binding Sites
MH  - BOUND NUCLEOTIDES
MH  - COMPLEX
MH  - conformation
MH  - function
MH  - ion
MH  - mechanism
MH  - MECHANISMS
MH  - membrane
MH  - Mitochondria
MH  - Nad
MH  - Nadp
MH  - nucleotide binding
MH  - nucleotide binding sites
MH  - NUCLEOTIDE-BINDING-SITES
MH  - Nucleotides
MH  - protein
MH  - Protein Conformation
MH  - proton
MH  - Protons
MH  - redox
MH  - Rhodospirillum
MH  - Site
MH  - structure
MH  - SURFACE
MH  - TRANSFER
MH  - translocation
RP  - NOT IN FILE
NT  - UI - 21340778LA - engRN - 0 (Nucleotides)RN - 0 (Protons)RN - 53-59-8 (NADP)RN - 53-84-9 (NAD)RN - EC 1.6.1.1 (NADP Transhydrogenase)PT - Journal ArticleDA - 20010711IS - 0969-2126SB - IMCY - United States
UR  - PM:11250201
SO  - Structure (Camb ) 2001 Feb 7 ;9(2):165-176

1778
UI  - 100
AU  - Dmitriev OY
AU  - Fillingame RH
AD  - Department of Biomolecular Chemistry, University of Wisconsin Medical School, Madison, Wisconsin 53706, USA
TI  - Structure of Ala(20) --> Pro/Pro(64) --> Ala substituted subunit c of Escherichia coli ATP synthase in which the essential proline is switched between transmembrane helices
AB  - The structure of the A20P/P64A mutated subunit c of Escherichia coli ATP synthase, in which the essential proline has been switched from residue 64 of the second transmembrane helix (TMH) to residue 20 of the first TMH, has been solved by (15)N,(1)H NMR in a monophasic chloroform/methanol/water (4:4:1) solvent mixture. The cA20P/P64A mutant grows as well as wild type, and the F(0)F(1) complex is fully functional in ATPase-coupled H(+) pumping. Residues 20 and 64 lie directly opposite to each other in the hairpin-like structure of wild type subunit c, and the prolinyl 64 residue is thought to induce a slight bend in TMH-2 such that it wraps around a more straightened TMH- 1. In solution, the A20P/P64A substituted subunit c also forms a hairpin of two alpha-helices, with residues 41-45 forming a connecting loop as in the case of the wild type protein, but, in this case, Pro(20) induces a bend in TMH-1, which then packs against a more straightened TMH-2. The essential prolinyl residue, whether at position 64 or 20, lies close to the aspartyl 61 H(+) binding site. The prolinyl residue may introduce structural flexibility in this region of the protein, which may be necessary for the proposed movement of the alpha- helical segments during the course of the H(+) pumping catalytic cycle
RP  - NOT IN FILE
NT  - UI - 21347886LA - engRN - 0 (Membrane Proteins)RN - 147-85-3 (Proline)RN - 6898-94-8 (Alanine)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM23105/GM/NIGMSID - RR02301/RR/NCRRID - RR02781/RR/NCRRID - RR08438/RR/NCRRDA - 20010716IS - 0021-9258SB - IMCY - United StatesJC - HIV
UR  - PM:11331283
SO  - J Biol Chem 2001 Jul 20 ;276(29):27449-27454

1779
UI  - 21435
AU  - Dominguez-Ramirez L
AU  - Mendoza-Hernandez G
AU  - Carabez-Trejo A
AU  - Gomez-Puyou A
AU  - Tuena dG
TI  - Equilibrium between monomeric and dimeric mitochondrial F1-inhibitor protein complexes
AB  - Mg-ATP particles from bovine heart mitochondria have more than 95% of their F1 in complex with the inhibitor protein (IF1). The F1-IF1 complex was solubilized and purified. The question addressed was if this naturally occurring complex existed as monomers or dimers. Size exclusion chromatography and electron microscopy showed that most of the purified F1-IF1 complex was a dimer of two F1-IF1. As determined by the former method, the relative concentrations of dimeric and monomeric F1-IF1 depended on the concentration of protein that was applied to the column. Apparently, there is an equilibrium between the two forms of F1-IF1
MH  - A
MH  - Animal
MH  - antagonists & inhibitors
MH  - Ca(2+)-Transporting ATPase
MH  - Cattle
MH  - COMPLEX
MH  - Dimerization
MH  - electron
MH  - electron microscopy
MH  - ELECTRON-MICROSCOPY
MH  - equilibrium
MH  - F1
MH  - Hydrogen-Ion Concentration
MH  - inhibitor
MH  - INHIBITOR PROTEIN
MH  - metabolism
MH  - method
MH  - Microscopy
MH  - Mitochondria
MH  - Mitochondria,Heart
MH  - protein
MH  - Proteins
MH  - Proton-Translocating ATPases
RP  - NOT IN FILE
NT  - Departamento de Genetica Molecular, Instituto de Fisiologia Celular, Universidad Nacional Autonoma de MexicoFAU - Dominguez-Ramirez, L
SO  - FEBS Lett 2001 Oct 26 ;507(2):191-194

1780
UI  - 20929
AU  - Drose S
AU  - Boddien C
AU  - Gassel M
AU  - Ingenhorst G
AU  - Zeeck A
AU  - Altendorf K
AD  - Fachbereich Biologie/Chemie, Universitat Osnabruck, Barbarastrasse 11, Arbeitsgruppe Mikrobiologie, D-49076 Osnabruck, Germany
TI  - Semisynthetic derivatives of concanamycin A and C, as inhibitors of V- and P-type ATPases: structure-activity investigations and developments of photoaffinity probes
AB  - V-type ATPases are inhibited by the plecomacrolides bafilomycin and concanamycin, which exert their inhibitory potential at nanomolar concentrations. In addition, some P-type ATPases are inhibited at micromolar concentrations. We initiated intensive structure-activity investigations with semisynthetic concanamycin derivatives to approach the following two questions: (i) What is the pharmacophor, the structural key element, of the plecomacrolides that leads to their inhibitory potential against V- and P-type ATPases? (ii) Where is the binding site within these two different types of ATPases? In a first step, we examined where chemical modifications (O-acylations, substitutions, eliminations) could be placed without seriously affecting the inhibitory potential of the macrolides. In a second step, we used the knowledge of these structure-activity investigations to introduce traceable elements (fluorescent or radioactive) or nitrene- generating azido or carbene-generating diazirine-groups able to bind the inhibitors to their target covalently. These studies led finally to the synthesis of two photoaffinity probes that were used in labeling experiments with the purified plasma membrane V-type ATPase of Manduca sexta (described in a following paper, Huss, M., Gassel, M., Ingenhorst, G., Drose, S., Zeeck, A., Altendorf, K., Wieczorek, H., manuscript submitted)
MH  - A
MH  - Adenosinetriphosphatase
MH  - Antibiotics
MH  - ATPase
MH  - BINDING
MH  - Carrier Proteins
MH  - development
MH  - Enzyme Inhibitors
MH  - H+-ATPase
MH  - M
MH  - membrane
MH  - Potassium
MH  - protein
MH  - Proteins
MH  - Site
MH  - synthesis
RP  - NOT IN FILE
NT  - UI - 21159074LA - engRN - 0 (Antibiotics, Macrolide)RN - 0 (Carrier Proteins)RN - 0 (Enzyme Inhibitors)RN - 0 (Photoaffinity Labels)RN - 116764-51-3 (bafilomycin A)RN - 80890-47-7 (concanamycin A)RN - 81552-34-3 (concanamycin C)RN - 88899-56-3 (bafilomycin B1)RN - EC 3.6.1.- (potassium translocating Kdp-ATPase, E coli)RN - EC 3.6.1.- (vacuolar H+-ATPase)RN - EC 3.6.1.3 (Adenosinetriphosphatase)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 20010322IS - 0006-2960SB - IMCY - United States
UR  - PM:11258892
SO  - Biochemistry 2001 Mar 6 ;40(9):2816-2825

1781
UI  - 21443
AU  - Duby G
AU  - Degand H
AU  - Boutry M
TI  - Structure requirement and identification of a cryptic cleavage site in the mitochondrial processing of a plant F1-ATPase beta-subunit presequence
AB  - We sought to determine the structural features involved in the processing of the mitochondrial F1-ATPase beta-subunit (F1beta) presequence (54 residues) from Nicotiana plumbaginifolia. The cleavage efficiency of F1beta presequence mutants linked to the green fluorescent protein (GFP) was evaluated in vivo in tobacco by in situ microscopy and Western blotting. The residue at position -1 (Tyr) was required to be an aromatic residue and the residue at position +2 (Thr) was found to be important for F1beta processing, while, unexpectedly, changing the distal (Arg-15) and proximal (Arg-5) arginine residues did not strongly reduce processing. In addition, results also supported the requirement of a helical structure around the cleavage position. Sequencing of the mature form of a precursor containing the first 30 residues of the F1beta presequence linked to GFP revealed the presence of a cryptic cleavage site between residues 26 and 27, which showed the features of a classical mitochondrial processing site, suggesting dual processing of the F1beta presequence. In vitro processing confirmed these data and showed that processing was sensitive to o-phenanthroline, thus catalyzed by mitochondrial processing peptidase
MH  - A
MH  - Amino Acid Sequence
MH  - Arginine
MH  - BETA-SUBUNIT
MH  - Chemistry
MH  - data
MH  - enzymology
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - Hydrolysis
MH  - In Vitro
MH  - Luminescent Proteins
MH  - metabolism
MH  - Microscopy
MH  - Mitochondria
MH  - MITOCHONDRIAL F1-ATPASE
MH  - Molecular Sequence Data
MH  - mutant
MH  - plant
MH  - Plants,Toxic
MH  - protein
MH  - Protein Conformation
MH  - Protein Processing,Post-Translational
MH  - Proton-Translocating ATPases
MH  - Recombinant Fusion Proteins
MH  - RESIDUE
MH  - Site
MH  - structure
MH  - Support,Non-U.S.Gov't
MH  - Tobacco
RP  - NOT IN FILE
NT  - Unite de Biochimie Physiologique, Universite Catholique de Louvain, Croix du Sud 2-20, B-1348, Louvain-la-Neuve, BelgiumFAU - Duby, G
SO  - FEBS Lett 2001 Sep 21 ;505(3):409-413

1782
UI  - 20807
AU  - Eikerling M
AU  - Kornyshev AA
AU  - Kuznetsov AM
AU  - Ulstrup J
AU  - Walbran S
TI  - Mechanisms of proton conductance in polymer electrolyte membranes
MH  - conductance
MH  - mechanism
MH  - MECHANISMS
MH  - membrane
MH  - Membranes
MH  - proton
RP  - NOT IN FILE
NT  - JournalMAY 3428BWJ PHYS CHEM B
UR  - ISI:000168442100041
SO  - Journal of Physical Chemistry B 2001  ;105(17):3646-3662

1783
UI  - 20828
AU  - Feniouk BA
AU  - Cherepanov DA
AU  - Junge W
AU  - Mulkidjanian AY
TI  - Coupling of proton flow to ATP synthesis in Rhodobacter capsulatus: F(0)F(1)-ATP synthase is absent from about half of chromatophores
AB  - F(0)F(1)-ATP synthase (H(+)-ATP synthase, F(0)F(1)) utilizes the transmembrane protonmotive force to catalyze the formation of ATP from ADP and inorganic phosphate (P(i)). Structurally the enzyme consists of a membrane-embedded proton-translocating F(0) portion and a protruding hydrophilic F(1) part that catalyzes the synthesis of ATP. In photosynthetic purple bacteria a single turnover of the photosynthetic reaction centers (driven by a short saturating flash of light) generates protonmotive force that is sufficiently large to drive ATP synthesis. Using isolated chromatophore vesicles of Rhodobacter capsulatus, we monitored the flash induced ATP synthesis (by chemoluminescence of luciferin/luciferase) in parallel to the transmembrane charge transfer through F(0)F(1) (by following the decay of electrochromic bandshifts of intrinsic carotenoids). With the help of specific inhibitors of F(1) (efrapeptin) and of F(0) (venturicidin), we decomposed the kinetics of the total proton flow through F(0)F(1) into (i) those coupled to the ATP synthesis and (ii) the de-coupled proton escape through F(0). Taking the coupled proton flow, we calculated the H(+)/ATP ratio; it was found to be 3.3+/-0.6 at a large driving force (after one saturating flash of light) but to increase up to 5.1+/-0.9 at a smaller driving force (after a half-saturating flash). From the results obtained, we conclude that our routine chromatophore preparations contained three subsets of chromatophore vesicles. Chromatophores with coupled F(0)F(1) dominated in fresh material. Freezing/thawing or pre-illumination in the absence of ADP and P(i) led to an increase in the fraction of chromatophores with at least one de-coupled F(0)(F(1)). The disclosed fraction of chromatophores that lacked proton-conducting F(0)(F(1)) (approx. 40% of the total amount) remained constant upon these treatments
MH  - A
MH  - ADP
MH  - atp
MH  - ATP synthesis
MH  - Bacteria
MH  - capsulatus
MH  - carotenoid
MH  - Carotenoids
MH  - chromatophore
MH  - chromatophores
MH  - CONSTANT
MH  - coupling
MH  - flash
MH  - INORGANIC-PHOSPHATE
MH  - Kinetics
MH  - Light
MH  - proton
MH  - reaction center
MH  - Rhodobacter
MH  - rhodobacter capsulatus
MH  - RHODOBACTER-CAPSULATUS
MH  - SYNTHASE
MH  - synthesis
MH  - TRANSFER
RP  - NOT IN FILE
NT  - UI - 21638043LA - engPT - Journal ArticleDA - 20020107IS - 0006-3002SB - IMCY - Netherlands
UR  - PM:11779552
SO  - Biochim Biophys Acta 2001 Nov 1 ;1506(3):189-203

1784
UI  - 20831
AU  - Futai M
AU  - Sambongi Y
AU  - Wada Y
AD  - m-futai@sankenosaka-uacjp
TI  - [ATP synthase (nano machine that produces ATP) and proton pumping ATPase]
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATPase
MH  - H+-ATPase
MH  - proton
MH  - Proton Pump
MH  - Protons
MH  - review
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 21464031LA - jpnRN - 0 (Proton Pumps)RN - 0 (Protons)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.- (Vacuolar Proton-Translocating ATPases)RN - EC 3.6.1.- (vacuolar H+-ATPase)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 20011002IS - 0039-9450SB - IMCY - Japan
UR  - PM:11579576
SO  - Tanpakushitsu Kakusan Koso 2001 Aug ;46(11 Suppl):1754-1763

1785
UI  - 314
AU  - Gordon-Smith DJ
AU  - Carbajo RJ
AU  - Yang JC
AU  - Videler H
AU  - Runswick MJ
AU  - Walker JE
AU  - Neuhaus D
AD  - MRC Laboratory of Molecular Biology, Hills Road, Cambridge, CB2 2QH, UK
TI  - Solution structure of a C-terminal coiled-coil domain from bovine IF(1): the inhibitor protein of F(1) ATPase
AB  - Bovine IF(1) is a basic, 84 amino acid residue protein that inhibits the hydrolytic action of the F(1)F(0) ATP synthase in mitochondria under anaerobic conditions. Its oligomerization state is dependent on pH. At a pH value below 6.5 it forms an active dimer. At higher pH values, two dimers associate to form an inactive tetramer. Here, we present the solution structure of a C-terminal fragment of IF(1) (44- 84) containing all five of the histidine residues present in the sequence. Most unusually, the molecule forms an anti-parallel coiled- coil in which three of the five histidine residues occupy key positions at the dimer interface. Copyright 2001 Academic Press
RP  - NOT IN FILE
NT  - UI - 21226870LA - engRN - 0 (ATPase inhibitory protein)RN - 0 (Peptide Fragments)RN - 0 (Proteins)RN - 0 (Solutions)RN - 7006-35-1 (Histidine)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 20010430IS - 0022-2836SB - IMCY - EnglandJC - J6V
UR  - PM:11327770
SO  - J Mol Biol 2001 Apr 27 ;308(2):325-339

1786
UI  - 9943
AU  - Groth G
AU  - Pohl E
AD  - Heinrich-Heine-Universitat, Biochemie der Pflanzen, Universitatsstrasse 1, D-40225 Dusseldorf, Germany and EMBL Hamburg, Notkestrasse 85, D- 22603 Hamburg, Germany georggroth@uniduesseldorfde
TI  - The structure of the chloroplast F1-ATPase at 3.2 A resolution
AB  - The structure of the F(1)-ATPase from spinach chloroplasts was determined to 3.2 A resolution by molecular replacement based on the homologous structure of the bovine mitochondrial enzyme. The crystallized complex contains four different subunits in a stoichiometry of alpha(3)beta(3)gammaepsilon. Subunit delta was removed before crystallization to improve the diffraction of the crystals. The overall structure of the noncatalytic alpha-subunits and the catalytic beta-subunits is highly similar to those of the mitochondrial and thermophilic subunits. However, in the crystal structure of the chloroplast enzyme, all alpha- and beta-subunits adopt a closed conformation and appear to contain no bound adenine nucleotides. The superimposed crystallographic symmetry in the space group R32 impaired an exact tracing of the gamma- and epsilon-subunits in the complex. However, clear electron density was present at the core of the alpha(3)beta(3)-subcomplex, which probably represents the C-terminal domain of the gamma-subunit. The structure of the spinach chloroplast F(1) has a potential binding site for the phytotoxin, tentoxin, at the alphabeta-interface near betaAsp(83) and an insertion from betaGly(56)- Asn(60) in the N-terminal beta-barrel domain probably increases the thermal stability of the complex. The structure probably represents an inactive latent state of the ATPase, which is unique to chloroplast and cyanobacterial enzymes
MH  - Adenine Nucleotides
MH  - ADENINE-NUCLEOTIDES
MH  - ALPHA-SUBUNIT
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BETA-SUBUNIT
MH  - BINDING
MH  - chloroplast
MH  - Chloroplasts
MH  - COMPLEX
MH  - Crystallization
MH  - electron
MH  - Enzymes
MH  - EPSILON-SUBUNIT
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - H(+)-Transporting ATP Synthase
MH  - Nucleotides
MH  - Spinach
MH  - SPINACH-CHLOROPLASTS
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 21125671LA - engRN - 0 (Protein Subunits)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 20010306IS - 0021-9258SB - IMCY - United StatesJC - HIV
UR  - PM:11032839
SO  - J Biol Chem 2001 Jan 12 ;276(2):1345-1352

1787
UI  - 276
AU  - Gumbiowski K
AU  - Cherepanov D
AU  - Muller M
AU  - Panke O
AU  - Promto P
AU  - Winkler S
AU  - Junge W
AU  - Engelbrecht S
AD  - FB Biologie/Chemie, Universitt Osnabrck, Osnabrck 32130
TI  - F-ATPase: Forced full rotation of the rotor despite covalent cross-link with the stator
AB  - In ATP synthase (FOF1-ATPase) ion flow through the membrane-intrinsic portion, FO, drives the central "rotor", subunits c10epsilongamma, relative to the "stator" ab2delta(alphabeta)3. This converts ADP and Pi into ATP. Vice versa, ATP hydrolysis drives the rotation backwards. Covalent cross-links between rotor and stator subunits have been shown to inhibit these activities. Aiming at the rotary compliance of subunit gamma we introduced disulfide bridges between gamma (rotor) and alpha or beta (stator). We engineered cysteine residues into positions located roughly at the "top", "center", and "bottom" parts of the coiled coil portion of gamma and suitable residues on alpha or beta. This part of gamma is located at the center of the (alphabeta)3 domain with its C-terminal part at the "top" of F1 and the "bottom" part close to the FO complex. Disulfide bridge-formation under oxidizing conditions was quantitative as shown by SDS-PAGE and immunoblotting. As expected both the ATPase activities and the yield of rotating subunits gamma dropped to zero when the crosslink was formed at the "center" (gammaL262C <-> alphaA334C) and "bottom" (gammaC87 <-> betaD380C) positions. But much to our surprise disulfide-bridging neither impaired ATP hydrolysis activity nor the full rotation of gamma and the enzyme- generated torque of oxidized F1 which had been engineered at the "top" position (gammaA285C <-> alphaP280C). Apparently the high torque of this rotary engine uncoiled the alpha-helix and forced amino acids at the C-terminal portion of gamma into full rotation around their dihedral (Ramachandran) angles. This conclusion was supported by molecular dynamics simulations: If gammaC285-V286 are covalently attached to (alphabeta)3 and gammaA1-S281 is forced to rotate, gammaG282-A284 can serve as cardan shaft
RP  - NOT IN FILE
NT  - UI - 0LA - ENGPT - JOURNAL ARTICLEDA - 20010904IS - 0021-9258JC - HIV
UR  - PM:11533065
SO  - J Biol Chem 2001 Aug 31 ;():

1788
UI  - 21420
AU  - Hamilton CA
AU  - Good AG
AU  - Taylor GJ
TI  - Vacuolar H+-ATPase, but not mitochondrial F1F0-ATPase, is required for aluminum resistance in Saccharomyces cerevisiae
AB  - It was recently shown that vacuolar ATPase and mitochondrial F1F0-ATPase activities are induced by aluminum (Al) in an Al-resistant cultivar of wheat, suggesting that induction of these enzymes could be an adaptive trait involved in Al resistance. To test this hypothesis, we used the Saccharomyces cerevisiae model system. In yeast, unlike wheat, the activity, transcript and protein levels of mitochondrial F1F0-ATPase, but not vacuolar ATPase, are induced by Al, while plasma membrane P-ATPase activity is inhibited. However, yeast vacuolar ATPase mutant strains are hypersensitive to Al, while F1F0-ATPase mutant strains exhibit wild-type growth. These data suggest that vacuolar ATPase activity is involved in Al resistance, with ATP required for this activity supplied by mitochondrial F1F0-ATPase or fermentation
MH  - A
MH  - Aluminum
MH  - analysis
MH  - atp
MH  - ATPase
MH  - Blotting,Northern
MH  - data
MH  - Dose-Response Relationship,Drug
MH  - drug effects
MH  - Drug Resistance,Fungal
MH  - enzyme
MH  - Enzymes
MH  - enzymology
MH  - Fermentation
MH  - genetics
MH  - H+-ATPase
MH  - Immunoblotting
MH  - membrane
MH  - Mitochondrial Proton-Translocating ATPases
MH  - model
MH  - mutant
MH  - Mutation
MH  - pharmacology
MH  - physiology
MH  - protein
MH  - RESISTANCE
MH  - RNA,Fungal
MH  - RNA,Messenger
MH  - Saccharomyces cerevisiae
MH  - Support,Non-U.S.Gov't
MH  - SYSTEM
MH  - universities
MH  - Vacuolar Proton-Translocating ATPases
MH  - YEAST
RP  - NOT IN FILE
NT  - Department of Biological Sciences, University of Alberta, T6G 2E9, Edmonton, AB, Canada cah@ualbetacaFAU - Hamilton, C A
SO  - FEMS Microbiol Lett 2001 Dec 18 ;205(2):231-236

1789
UI  - 684
AU  - Hara KY
AU  - Kato-Yamada Y
AU  - Kikuchi Y
AU  - Hisabori T
AU  - Yoshida M
AD  - Chemical Resources Laboratory, R-1, Tokyo Institute of Technology, Nagatsuta 4259, Yokohama, 226-8503, Japan
TI  - The role of the betaDELSEED motif of F1-ATPase: propagation of the inhibitory effect of the epsilon subunit
AB  - In F(1)-ATPase, a rotary motor enzyme, the region of the conserved DELSEED motif in the beta subunit moves and contacts the rotor gamma subunit when the nucleotide fills the catalytic site, and the acidic nature of the motif was previously assumed to play a critical role in rotation. Our previous work, however, disproved the assumption (Hara, K. Y., Noji, H., Bald, D., Yasuda, R., Kinosita, K., Jr., and Yoshida, M. (2000) J. Biol. Chem. 275, 14260-14263), and the role of this motif remained unknown. Here, we found that the epsilon subunit, an intrinsic inhibitor, was unable to inhibit the ATPase activity of a mutant thermophilic F(1)-ATPase in which all of the five acidic residues in the DELSEED motif were replaced with alanines, although the epsilon subunit in the mutant F(1)-ATPase assumed the inhibitory form. In addition, the replacement of basic residues in the C-terminal region of the epsilon subunit by alanines caused a decrease of the inhibitory effect. Partial replacement of the acidic residues in the DELSEED motif of the beta subunit or of the basic residues in the C-terminal alpha- helix of the epsilon subunit induced a partial effect. We here conclude that the epsilon subunit exerts its inhibitory effect through the electrostatic interaction with the DELSEED motif of the beta subunit
RP  - NOT IN FILE
NT  - UI - 21316431LA - engRN - 0 (Protein Subunits)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 20010625IS - 0021-9258SB - IMCY - United StatesJC - HIV
UR  - PM:11279233
SO  - J Biol Chem 2001 Jun 29 ;276(26):23969-23973

1790
UI  - 127
AU  - Hausrath AC
AU  - Capaldi RA
AU  - Matthews BW
AD  - Institute of Molecular Biology, 1229 University of Oregon, Eugene, OR 97403-1229
TI  - The conformation of the epsilon and gamma subunits within the E. coli F1 ATPase
AB  - F1 is the water-soluble portion of the ubiquitous F1F0 ATP synthase. Its structure includes three alpha- and three beta-subunits, arranged as a hexameric disk, plus a gamma-subunit that penetrates the center of the disk akin to an axle. Recently Hausrath et al. obtained an electron density map of E. coli F1 at 4.4 resolution in which the coiled-coil alpha-helices of the gamma-subunit could be seen to extend 45 from the base of the alpha3beta3 hexamer [Hausrath, A.C., Grber, G., Matthews, B.W. & Capaldi, R.A. (1999) Proc. Natl. Acad. Sci. USA 96, 13697- 13702]. Subsequently the structure of a truncated form of the E. coli gamma-subunit in complex with epsilon has been described [Rodgers, A.J.W. & Wilce, M.C.J. (2000) Nature Struct. Biol. 7, 1051-1054]. In the present study the 4.4 resolution electron density map of E. coli F1 is re-evaluated in light of the newly-available data on the gamma- and epsilon-subunits. It is shown that the map of the F1 complex is consistent with the structure of the isolated subunits. When E. coli F1 is compared with that from beef heart, the structures of the E. coli gamma- and epsilon-subunits are seen to be generally similar to their counterparts in the bovine enzyme, but to undergo major shifts in position. In particular, the two, long, coiled-coil alpha-helices that lie along the axis of F1 both unwind and rotate. Also the epsilon- subunit rotates around the axis by 81 and undergoes a net translation of about 23. It is argued that these large-scale changes in conformation reflect distinct functional states that occur during the rotation of the gamma-subunit within the alpha3beta3 hexamer
RP  - NOT IN FILE
NT  - UI - 0LA - ENGPT - JOURNAL ARTICLEDA - 20011004IS - 0021-9258JC - HIV
UR  - PM:11585832
SO  - J Biol Chem 2001 Oct 3 ;():

1791
UI  - 21440
AU  - Hausrath AC
AU  - Capaldi RA
AU  - Matthews BW
TI  - The conformation of the epsilon- and gamma-subunits within the Escherichia coli F(1) ATPase
AB  - F(1) is the water-soluble portion of the ubiquitous F(1)F(0) ATP synthase. Its structure includes three alpha- and three beta-subunits, arranged as a hexameric disc, plus a gamma-subunit that penetrates the center of the disc akin to an axle. Recently Hausrath et al. (Hausrath, A. C., Gruber, G., Matthews, B. W., and Capaldi, R. A. (1999) Proc. Natl. Acad. Sci. U. S. A. 96, 13697-13702) obtained an electron density map of E. coli F(1) at 4.4-A resolution in which the coiled-coil alpha-helices of the gamma-subunit could be seen to extend 45 A from the base of the alpha(3)beta(3) hexamer. Subsequently the structure of a truncated form of the E. coli gamma-subunit in complex with epsilon has been described (Rodgers, A. J. W., and Wilce, M. C. J. (2000) Nat. Struct. Biol. 7, 1051-1054). In the present study the 4.4-A resolution electron density map of E. coli F(1) is re-evaluated in light of the newly available data on the gamma- and epsilon-subunits. It is shown that the map of the F(1) complex is consistent with the structure of the isolated subunits. When E. coli F(1) is compared with that from beef heart, the structures of the E. coli gamma- and epsilon-subunits are seen to be generally similar to their counterparts in the bovine enzyme but to undergo major shifts in position. In particular, the two long, coiled-coil alpha-helices that lie along the axis of F(1) both unwind and rotate. Also the epsilon-subunit rotates around the axis by 81 degrees and undergoes a net translation of about 23 A. It is argued that these large-scale changes in conformation reflect distinct functional states that occur during the rotation of the gamma-subunit within the alpha(3)beta(3) hexamer
MH  - A
MH  - alpha
MH  - Animal
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BASE
MH  - BETA-SUBUNIT
MH  - biology
MH  - Cattle
MH  - Chemistry
MH  - COMPLEX
MH  - conformation
MH  - Crystallography,X-Ray
MH  - data
MH  - electron
MH  - Electrons
MH  - enzyme
MH  - enzymology
MH  - EPSILON-SUBUNIT
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - Light
MH  - M
MH  - Models,Molecular
MH  - physiology
MH  - Protein Binding
MH  - Protein Conformation
MH  - Protein Structure,Secondary
MH  - Protein Structure,Tertiary
MH  - Proteins
MH  - Proton-Translocating ATPases
MH  - resolution
MH  - Ribonucleoproteins
MH  - RNA-Binding Proteins
MH  - rotation
MH  - structure
MH  - SUBUNIT
MH  - Support,U.S.Gov't,P.H.S.
MH  - SYNTHASE
MH  - universities
MH  - Yeasts
RP  - NOT IN FILE
NT  - Institute of Molecular Biology, Howard Hughes Medical Institute, University of Oregon, Eugene, OR 97403-1229, USAFAU - Hausrath, A C
SO  - J Biol Chem 2001 Dec 14 ;276(50):47227-47232

1792
UI  - 21068
AU  - Hirono-Hara Y
AU  - Noji H
AU  - Nishiura M
AU  - Muneyuki E
AU  - Hara KY
AU  - Yasuda R
AU  - Kinosita K
AU  - Yoshida M
TI  - Pause and rotation of F(1)-ATPase during catalysis
AB  - F(1)-ATPase is a rotary motor enzyme in which a single ATP molecule drives a 120 degrees rotation of the central gamma subunit relative to the surrounding alpha(3)beta(3) ring. Here, we show that the rotation of F(1)-ATPase spontaneously lapses into long (approximately 30 s) pauses during steady-state catalysis. The effects of ADP-Mg and mutation on the pauses, as well as kinetic comparison with bulk-phase catalysis, strongly indicate that the paused enzyme corresponds to the inactive state of F(1)-ATPase previously known as the ADP-Mg inhibited form in which F(1)-ATPase fails to release ADP-Mg from catalytic sites. The pausing position of the gamma subunit deviates from the ATP-waiting position and is most likely the recently found intermediate 90 degrees position
MH  - A
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - antagonists & inhibitors
MH  - atp
MH  - Catalysis
MH  - Chemistry
MH  - intermediate
MH  - Kinetics
MH  - Magnesium
MH  - Proton-Translocating ATPases
MH  - rotation
MH  - Site
MH  - SUBUNIT
MH  - Support,Non-U.S.Gov't
RP  - NOT IN FILE
NT  - Chemical Resources Laboratory, Tokyo Institute of Technology, 4259 Nagatsuta, Yokohama 226-8503, JapanFAU - Hirono-Hara, Y
SO  - Proc Natl Acad Sci U S A 2001 Nov ;%20;98(24):13649-13654

1793
UI  - 21434
AU  - Hirono-Hara Y
AU  - Noji H
AU  - Nishiura M
AU  - Muneyuki E
AU  - Hara KY
AU  - Yasuda R
AU  - Kinosita K
AU  - Yoshida M
TI  - Pause and rotation of F(1)-ATPase during catalysis
AB  - F(1)-ATPase is a rotary motor enzyme in which a single ATP molecule drives a 120 degrees rotation of the central gamma subunit relative to the surrounding alpha(3)beta(3) ring. Here, we show that the rotation of F(1)-ATPase spontaneously lapses into long (approximately 30 s) pauses during steady-state catalysis. The effects of ADP-Mg and mutation on the pauses, as well as kinetic comparison with bulk-phase catalysis, strongly indicate that the paused enzyme corresponds to the inactive state of F(1)-ATPase previously known as the ADP-Mg inhibited form in which F(1)-ATPase fails to release ADP-Mg from catalytic sites. The pausing position of the gamma subunit deviates from the ATP-waiting position and is most likely the recently found intermediate 90 degrees position
MH  - A
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - antagonists & inhibitors
MH  - atp
MH  - Catalysis
MH  - catalytic
MH  - Chemistry
MH  - enzyme
MH  - intermediate
MH  - Kinetics
MH  - Magnesium
MH  - Mutation
MH  - Proton-Translocating ATPases
MH  - rotation
MH  - Site
MH  - SUBUNIT
MH  - Support,Non-U.S.Gov't
RP  - NOT IN FILE
NT  - Chemical Resources Laboratory, Tokyo Institute of Technology, 4259 Nagatsuta, Yokohama 226-8503, JapanFAU - Hirono-Hara, Y
SO  - Proc Natl Acad Sci U S A 2001 Nov ;%20;98(24):13649-13654

1794
UI  - 19853
AU  - Hutcheon ML
AU  - Duncan TM
AU  - Ngai H
AU  - Cross RL
AD  - Department of Biochemistry and Molecular Biology, State University of New York Upstate Medical University, Syracuse, NY 13210, USA
TI  - Energy-driven subunit rotation at the interface between subunit a and the c oligomer in the F(O) sector of Escherichia coli ATP synthase
AB  - Subunit rotation within the F(1) catalytic sector of the ATP synthase has been well documented, identifying the synthase as the smallest known rotary motor. In the membrane-embedded F(O) sector, it is thought that proton transport occurs at a rotor/stator interface between the oligomeric ring of c subunits (rotor) and the single-copy a subunit (stator). Here we report evidence for an energy-dependent rotation at this interface. F(O)F(1) was expressed with a pair of substituted cysteines positioned to allow an intersubunit disulfide crosslink between subunit a and a c subunit [aN214C/cM65C; Jiang, W. & Fillingame, R. H. (1998) Proc. Natl. Acad. Sci. USA 95, 6607--6612]. Membranes were treated with N,N'-dicyclohexyl-[(14)C]carbodiimide to radiolabel the D61 residue on less than 20% of the c subunits. After oxidation to form an a--c crosslink, the c subunit properly aligned to crosslink to subunit a was found to contain very little (14)C label relative to other members of the c ring. However, exposure to MgATP before oxidation significantly increased the radiolabel in the a-c crosslink, indicating that a different c subunit was now aligned with subunit a. This increase was not induced by exposure to MgADP/P(i). Furthermore, preincubation with MgADP and azide to inhibit F(1) or with high concentrations of N,N'-dicyclohexylcarbodiimide to label most c subunits prevented the ATP effect. These results provide evidence for an energy-dependent rotation of the c ring relative to subunit a
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthase
MH  - Biochemistry
MH  - COLI ATP SYNTHASE
MH  - CROSS-LINKING
MH  - Dicyclohexylcarbodiimide
MH  - Enzyme Inhibitors
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - H(+)-Transporting ATP Synthase
MH  - membrane
MH  - Membranes
MH  - proton
MH  - rotation
MH  - SUBUNIT
MH  - SYNTHASE
MH  - transport
RP  - NOT IN FILE
NT  - UI - 21352981LA - engRN - 0 (Cross-Linking Reagents)RN - 0 (Enzyme Inhibitors)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM23152/GM/NIGMSDA - 20010718IS - 0027-8424SB - IMCY - United StatesJC - PV3
UR  - PM:11438702
SO  - Proc Natl Acad Sci U S A 2001 Jul 17 ;98(15):8519-8524

1795
UI  - 21438
AU  - Iko Y
AU  - Sambongi Y
AU  - Tanabe M
AU  - Iwamoto-Kihara A
AU  - Saito K
AU  - Ueda I
AU  - Wada Y
AU  - Futai M
TI  - ATP synthase F(1) sector rotation. Defective torque generation in the beta subunit Ser-174 to Phe mutant and its suppression by second mutations
AB  - Subunit gamma of the ATP synthase F(1) sector is located at the center of the alpha(3)beta(3) hexamer and rotates unidirectionally during ATP hydrolysis, generating the rotational torque of approximately 45 pN.nm. A mutant F(1) with the betaSer-174 to Phe substitution (betaS174F) in the beta subunit generated lower torque ( approximately 17 pN.nm), indicating that betaS174F is mechanically defective, the first such mutant reported. The defective rotation of betaS174F was suppressed by a second-site mutation, betaGly-149 to Ala, betaIle-163 to Ala, or betaIle-166 to Ala in the same subunit, but not by betaLeu-238 to Ala. These results suggest that the region between betaGly-149 and betaSer-174 plays an important role in the coupling between ATP hydrolysis and mechanical work
MH  - A
MH  - Actins
MH  - Adenosine Triphosphate
MH  - Alanine
MH  - atp
MH  - ATP synthase
MH  - BETA
MH  - BETA-SUBUNIT
MH  - Chemistry
MH  - coupling
MH  - enzymology
MH  - Escherichia coli
MH  - Glycine
MH  - Hydrolysis
MH  - Isoleucine
MH  - metabolism
MH  - Models,Molecular
MH  - mutant
MH  - Mutation
MH  - Phenylalanine
MH  - Protein Conformation
MH  - Protein Structure,Tertiary
MH  - Proton-Translocating ATPases
MH  - rotation
MH  - Serine
MH  - SUBUNIT
MH  - Support,Non-U.S.Gov't
MH  - SYNTHASE
MH  - Time Factors
MH  - universities
RP  - NOT IN FILE
NT  - Division of Biological Sciences, Institute of Scientific and Industrial Research, Osaka University, CREST (Core Research for Evolutional Science and Technology) of Japan Science and Technology Corporation, Ibaraki, Osaka 567-0047, JapanFAU - Iko, Y
SO  - J Biol Chem 2001 Dec 14 ;276(50):47508-47511

1796
UI  - 20830
AU  - Iko Y
AU  - Sambongi Y
AU  - Tanabe M
AU  - Iwamoto-Kihara A
AU  - Saito K
AU  - Ueda I
AU  - Wada Y
AU  - Futai M
AD  - Division of Biological Sciences, Institute of Scientific and Industrial Research, Osaka University, CREST (Core Research for Evolutional Science and Technology) of Japan Science and Technology Corporation, Ibaraki, Osaka 567-0047, Japan
TI  - ATP synthase F(1) sector rotation. Defective torque generation in the beta subunit Ser-174 to Phe mutant and its suppression by second mutations
AB  - Subunit gamma of the ATP synthase F(1) sector is located at the center of the alpha(3)beta(3) hexamer and rotates unidirectionally during ATP hydrolysis, generating the rotational torque of approximately 45 pN.nm. A mutant F(1) with the betaSer-174 to Phe substitution (betaS174F) in the beta subunit generated lower torque ( approximately 17 pN.nm), indicating that betaS174F is mechanically defective, the first such mutant reported. The defective rotation of betaS174F was suppressed by a second-site mutation, betaGly-149 to Ala, betaIle-163 to Ala, or betaIle-166 to Ala in the same subunit, but not by betaLeu-238 to Ala. These results suggest that the region between betaGly-149 and betaSer- 174 plays an important role in the coupling between ATP hydrolysis and mechanical work
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BETA-SUBUNIT
MH  - coupling
MH  - Hydrolysis
MH  - mutant
MH  - rotation
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 21601665LA - engRN - 0 (Actins)RN - 56-40-6 (Glycine)RN - 56-41-7 (Alanine)RN - 56-45-1 (Serine)RN - 56-65-5 (Adenosine Triphosphate)RN - 63-91-2 (Phenylalanine)RN - 73-32-5 (Isoleucine)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 20011212IS - 0021-9258SB - IMCY - United States
UR  - PM:11590180
SO  - J Biol Chem 2001 Dec 14 ;276(50):47508-47511

1797
UI  - 21064
AU  - Jernejc K
AU  - Legisa M
TI  - Activation of plasma membrane H+-ATPase by ammonium ions in Aspergillus niger
AB  - The addition of ammonium ions to Aspergillus niger cells originally growing on another nitrogen source resulted in rapid medium acidification. The addition of glucose or other fermentable sugars to the mycelium growing on glycerol did not have the same effect. The enzyme responsible for acidification seems to be plasma membrane H+-ATPase, which is most probably triggered by phosphorylation. Using specific activators and inhibitors, we tried to figure out which signalling pathway is involved in the process. No activation of H+-ATPase could be detected in the presence of diacylglycerol and other activators of protein kinase C, indicating that the stimulus is transmitted by another signalling chain. In the presence of inhibitors known to suppress the phosphatidyl-inositol signalling pathway, such as neomycin, compound 48/80 and calmidazolium, no increased H+-ATPase activity could be detected after the addition of ammonium ions. However, some tested inhibitors of the cAMP signalling pathway could not prevent activation of the enzyme by the stimulant. These results support the model in which ammonium-induced activation of proton extrusion in A. niger is mediated via the phosphatidyl-inositol signalling pathway, involving Ca2+/calmoduline-dependent protein kinase but not protein kinase C
MH  - A
MH  - ACTIVATION
MH  - Cells
MH  - Chemistry
MH  - Glucose
MH  - H+-ATPase
MH  - inhibitor
MH  - ion
MH  - Ions
MH  - membrane
MH  - model
MH  - Phosphorylation
MH  - protein
MH  - proton
RP  - NOT IN FILE
NT  - National Institute of Chemistry, Ljubljana, SloveniaFAU - Jernejc, K
SO  - Appl Microbiol Biotechnol 2001 Oct ;57(3):368-373

1798
UI  - 101
AU  - Jiang W
AU  - Hermolin J
AU  - Fillingame RH
AD  - Department of Biomolecular Chemistry, University of Wisconsin Medical School, 1300 University Avenue, Madison, WI 53706, USA
TI  - The preferred stoichiometry of c subunits in the rotary motor sector of Escherichia coli ATP synthase is 10
AB  - The stoichiometry of c subunits in the H(+)-transporting F(o) rotary motor of ATP synthase is uncertain, the most recent suggestions varying from 10 to 14. The stoichiometry will determine the number of H(+) transported per ATP synthesized and will directly relate to the P/O ratio of oxidative phosphorylation. The experiments described here show that the number of c subunits in functional complexes of F(o)F(1) ATP synthase from Escherichia coli can be manipulated, but that the preferred number is 10. Mixtures of genetically fused cysteine- substituted trimers (c(3)) and tetramers (c(4)) of subunit c were coexpressed and the c subunits crosslinked in the plasma membrane. Prominent products corresponding to oligomers of c(7) and c(10) were observed in the membrane and purified F(o)F(1) complex, indicating that the c(10) oligomer formed naturally. Oligomers larger than c(10) were also observed in the membrane fraction of cells expressing c(3) or c(4) individually, or in cells coexpressing c(3) and c(4) together, but these larger oligomers did not copurify with the functional F(o)F(1) complex and were concluded to be aberrant products of assembly in the membrane
RP  - NOT IN FILE
NT  - UI - 21221062LA - engRN - 0 (Cross-Linking Reagents)RN - 0 (Membrane Proteins)RN - 0 (Molecular Motors)RN - 0 (Protein Subunits)RN - 0 (Protons)RN - 0 (Recombinant Fusion Proteins)RN - 52-90-4 (Cysteine)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - GM23105/GM/NIGMSDA - 20010426IS - 0027-8424SB - IMCY - United StatesJC - PV3
UR  - PM:11320246
SO  - Proc Natl Acad Sci U S A 2001 Apr 24 ;98(9):4966-4971

1799
UI  - 9929
AU  - Jones PC
AD  - Dunn Human Nutrition Unit, Medical Research Council, Cambridge CB2 2XY, United Kingdom pcj@mrc-dunncamacuk
TI  - Introduction of a carboxyl group in the first transmembrane helix of Escherichia coli F1Fo ATPase subunit c and cytoplasmic pH regulation
AB  - The multicopy subunit c of the H(+)-transporting F1Fo ATP synthase of Escherichia coli folds across the membrane as a hairpin of two hydrophobic alpha helices. The subunits interact in a front-to-back fashion, forming an oligomeric ring with helix 1 packing in the interior and helix 2 at the periphery. A conserved carboxyl, Asp(61) in E. coli, centered in the second transmembrane helix is essential for H+ transport. A second carboxylic acid in the first transmembrane helix is found at a position equivalent to Ile28 in several bacteria, some the cause of serious infectious disease. This side chain has been predicted to pack proximal to the essential carboxyl in helix 2. It appears that in some of these bacteria the primary function of the enzyme is H+ pumping for cytoplasmic pH regulation. In this study, Ile28 was changed to Asp and Glu. Both mutants were functional. However, unlike the wild type, the mutants showed pH-dependent ATPase-coupled H+ pumping and passive H+ transport through Fo. The results indicate that the presence of a second carboxylate enables regulation of enzyme function in response to cytoplasmic pH and that the ion binding pocket is aqueous accessible. The presence of a single carboxyl at position 28, in mutants I28D/D61G and I28E/D61G, did not support growth on a succinate carbon source. However, I28E/D61G was functional in ATPase-coupled H+ transport. This result indicates that the side chain at position 28 is part of the ion binding pocket
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Bacteria
MH  - BINDING
MH  - Carbon Dioxide
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - H(+)-Transporting ATP Synthase
MH  - H+
MH  - Human
MH  - Nutrition
MH  - regulation
MH  - SUBUNIT
MH  - succinate
MH  - SYNTHASE
MH  - transport
RP  - NOT IN FILE
NT  - UI - 21101844LA - engRN - 124-38-9 (Carbon Dioxide)RN - 14485-07-5 (carboxyl radical)RN - 56-84-8 (Aspartic Acid)RN - 56-85-9 (Glutamine)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 20010222IS - 0021-9193SB - IMCY - United StatesJC - HH3
UR  - PM:11160082
SO  - J Bacteriol 2001 Mar ;183(5):1524-1530

1800
UI  - 21048
AU  - Jones RP
AU  - Hunt IE
AU  - Jaeger J
AU  - Ward A
AU  - O'Reilly J
AU  - Barratt EA
AU  - Findlay JB
AU  - Harrison MA
TI  - Expression, purification and secondary structure analysis of Saccharomyces cerevisiae vacuolar membrane H+-ATPase subunit F (Vma7p)
AB  - The vacuolar H+-ATPase is an acid pump found in virtually all eukaryotic cells. It shares a common macromolecular organization with the F1F0-ATPase, and some V-ATPase subunits are structural and functional homologues of F-ATPase components. However, the vacuolar complex contains several subunits which do not resemble F-ATPase subunits at the sequence level, and which currently have no specific function assigned. One example is subunit F, the Vma7p polypeptide of Saccharomyces cerevisiae. A recombinant form of Vma7p was expressed in Escherichia coli and purified to homogeneity. Mass spectroscopy confirmed a mass of 13460 Da for Vma7p, and dynamic light scattering showed that the polypeptide was globular and monodisperse even at high concentrations. Analysis of secondary structure by circular dichroism and FTIR showed that Vma7p comprises 30% alpha-helix and 32-42% beta-sheet. The protein fold recognition programme 'Threader 2' produced highly significant matches between Vma7p and five alpha-beta sandwich folds. Relative proportions of secondary structure elements within these folds were broadly consistent with the spectroscopic data. Although Vma7p does not share sequence similarity with the F-ATPase epsilon subunit, the analysis suggests that the polypeptides not only have similar masses and assemble into homologous core complexes, but also share similar secondary structures. It is possible that the two polypeptides are homologous and perform similar functions within their respective ATPases. The production of high yields of homogeneous, folded, monodisperse protein will facilitate high resolution crystallography and NMR spectroscopy studies
MH  - A
MH  - ACID
MH  - analysis
MH  - ATPase
MH  - Biochemistry
MH  - Cells
MH  - COMPLEX
MH  - EPSILON-SUBUNIT
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F-ATPASE
MH  - function
MH  - H+-ATPase
MH  - Light
MH  - membrane
MH  - NMR
MH  - P
MH  - protein
MH  - purification
MH  - resolution
MH  - Saccharomyces cerevisiae
MH  - secondary
MH  - spectroscopy
MH  - structure
MH  - SUBUNIT
RP  - NOT IN FILE
NT  - School of Biochemistry and Molecular Biology, University of Leeds, UKFAU - Jones, R P
SO  - Mol Membr Biol 2001 Oct ;18(4):283-290

1801
UI  - 277
AU  - Junge W
AU  - Panke O
AU  - Cherepanov DA
AU  - Gumbiowski K
AU  - Muller M
AU  - Engelbrecht S
AD  - Division of Biophysics, University of Osnabruck, D-49069, Osnabruck, Germany junge@uosde
TI  - Inter-subunit rotation and elastic power transmission in F0F1-ATPase
AB  - ATP synthase (F-ATPase) produces ATP at the expense of ion-motive force or vice versa. It is composed from two motor/generators, the ATPase (F1) and the ion translocator (F0), which both are rotary steppers. They are mechanically coupled by 360 degrees rotary motion of subunits against each other. The rotor, subunits gamma(epsilon)C10-14, moves against the stator, (alphabeta)3delta(ab2). The enzyme copes with symmetry mismatch (C3 versus C10-14) between its two motors, and it operates robustly in chimeric constructs or with drastically modified subunits. We scrutinized whether an elastic power transmission accounts for these properties. We used the curvature of fluorescent actin filaments, attached to the rotating c ring, as a spring balance (flexural rigidity of 8.10(-26) N x m2) to gauge the angular profile of the output torque at F0 during ATP hydrolysis by F1. The large average output torque (56 pN nm) proved the absence of any slip. Angular variations of the torque were small, so that the output free energy of the loaded enzyme decayed almost linearly over the angular reaction coordinate. Considering the three-fold stepping and high activation barrier (>40 kJ/mol) of the driving motor (F1) itself, the rather constant output torque seen by F0 implied a soft elastic power transmission between F1 and F0. It is considered as essential, not only for the robust operation of this ubiquitous enzyme under symmetry mismatch, but also for a high turnover rate under load of the two counteracting and stepping motors/generators
RP  - NOT IN FILE
NT  - UI - 21423569LA - engRN - 0 (Actins)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 20010904IS - 0014-5793SB - IMCY - NetherlandsJC - EUH
UR  - PM:11532447
SO  - FEBS Lett 2001 Aug 31 ;504(3):152-160

1802
UI  - 21030
AU  - Kakinuma Y
AU  - Miyauchi T
AU  - Yuki K
AU  - Murakoshi N
AU  - Goto K
AU  - Yamaguchi I
AD  - Cardiovascular Division, Department of Internal Medicine, Institute of Clinical Medicine, Institute of Basic Medical Sciences, University of Tsukuba, Ibaraki, Japan
TI  - Novel molecular mechanism of increased myocardial endothelin-1 expression in the failing heart involving the transcriptional factor hypoxia-inducible factor-1alpha induced for impaired myocardial energy metabolism
AB  - BACKGROUND:Hypoxia-inducible factor (HIF)-1alpha is an important transcriptional factor that activates the gene expression of glycolytic enzymes, which are activated as compensation for impaired beta- oxidation of fatty acid in the failing heart. We reported that cardiac endothelin (ET)-1 expression is markedly increased in heart failure. The mechanism, however, is unknown. Because we found an HIF-1alpha binding site in the 5'-promoter region of the ET-1 gene, we hypothesized that HIF-1alpha is involved in this mechanism. Methods and RESULTS:In rat cardiomyocytes, luciferase assay and electrophoretic mobility shift assay showed that HIF-1alpha transcriptionally activates ET-1 gene expression by direct interaction with the predicted DNA binding site in the 5'-promoter region. HIF-1alpha mRNA and ET-1 mRNA in the failing heart increased during the aggravation of heart failure in vivo in animal models, ie, rats with myocardial infarction and hamsters with cardiomyopathy. In cultured cardiomyocytes treated with a mitochondrial inhibitor, HIF-1alpha mRNA and ET-1 mRNA were markedly increased with activated glycolysis, and antisense oligonucleotide for HIF-1alpha largely inhibited the increased gene expression of ET-1. CONCLUSIONS:The present study revealed a novel molecular mechanism of upregulation of myocardial ET-1 in heart failure, indicating that induction of HIF-1alpha to stimulate glycolysis as an adaptation in heart failure against impaired energy metabolism alternatively causes an elevation of cardiac ET-1 gene expression as a maladaptation
MH  - A
MH  - ACID
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - Animal
MH  - BETA
MH  - BINDING
MH  - Energy Metabolism
MH  - Enzymes
MH  - Glycolysis
MH  - Hamsters
MH  - inhibitor
MH  - luciferase
MH  - mechanism
MH  - metabolism
MH  - method
MH  - Methods
MH  - model
MH  - protein
MH  - Proteins
MH  - Rats
MH  - Site
RP  - NOT IN FILE
NT  - UI - 21266772LA - engRN - 0 (DNA, Antisense)RN - 0 (DNA-Binding Proteins)RN - 0 (Endothelin-1)RN - 0 (HIF-1 protein)RN - 0 (HIF1alpha protein)RN - 0 (Nuclear Proteins)RN - 0 (RNA, Messenger)RN - 0 (Recombinant Fusion Proteins)RN - 56-65-5 (Adenosine Triphosphate)RN - 7440-48-4 (Cobalt)RN - 7646-79-9 (cobaltous chloride)RN - EC 1.13.12.- (Luciferase)PT - Journal ArticleDA - 20010524IS - 1524-4539SB - AIMSB - IMCY - United States
UR  - PM:11352889
SO  - Circulation 2001 May 15 ;103(19):2387-2394

1803
UI  - 21174
AU  - Kalaidzidis IV
AU  - Kaulen AD
AU  - Radionov AN
AU  - Khitrina LV
AD  - Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Moscow, 119899, Russia innak@phtbiogenebeemsusu
TI  - Photoelectrochemical cycle of bacteriorhodopsin
AB  - The scheme of the bacteriorhodopsin photocycle associated with a transmembrane proton transfer and electrogenesis is considered. The role of conformational changes in the polypeptide chain during the proton transport is discussed
MH  - A
MH  - Bacteriorhodopsin
MH  - conformational change
MH  - conformational changes
MH  - proton
MH  - Proton transfer
MH  - Proton-transfer
MH  - Protons
MH  - review
MH  - TRANSFER
MH  - transport
RP  - NOT IN FILE
NT  - UI - 21609810LA - engRN - 0 (Protons)RN - 53026-44-1 (Bacteriorhodopsins)PT - Journal ArticlePT - ReviewPT - Review, AcademicDA - 20011217IS - 0006-2979SB - IMCY - United States
UR  - PM:11743867
SO  - Biochemistry (Mosc ) 2001 Nov ;66(11):1220-1233

1804
UI  - 21432
AU  - Kipp JL
AU  - Ramirez VD
TI  - Effect of estradiol, diethylstilbestrol, and resveratrol on F0F1-ATPase activity from mitochondrial preparations of rat heart, liver, and brain
AB  - The question of whether estrogens or estrogen-like compounds would alter differentially the enzymatic activity of the FOF1-ATPase was addressed. Mitochondrial fractions of the liver, brain, and heart were obtained from adult male rats and solubilized by digitonin. About 85% of the adenosine triphosphate hydrolysis by these three preparations come from the mitochondrial FOF1-ATPase. The enzymatic activity differed in the following order: liver < brain < heart. A concentration of 13 nM estradiol stimulated the FOF1-ATPase activity in heart by 10% (p < 0.01), but not in liver or brain. 17beta-estradiol competed off the binding of estradiol-17beta-17-(O-carboxymethyl)oxime:125I-labeled bovine serium albumin to mitochondrial preparations of the heart, revealing two binding sites. Resveratrol inhibited the F0F1-ATPase activity in both heart and liver with an IC50 of 13-15 microM, which confirmed our previous report in preparations of brain. Lower doses (picomolar to nanomolar) of resveratrol stimulated the FOF1-ATPase activity in liver by 10% but not in heart. At 6.7 microM, diethylstilbestrol (DES) inhibited the FOF1-ATPase activity in the three preparations by 61-67%. This study demonstrates that estradiol activates rat heart mitochondrial FOF1-ATPase at physiologic concentrations and that the FOF1-ATPase activity is markedly different in rat liver, brain, and heart. In addition, estradiol, DES, and resveratrol alter the FOF1-ATPase activity selectively, probably via different mechanisms
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - Adult
MH  - Animal
MH  - antagonists & inhibitors
MH  - BINDING
MH  - BINDING SITE
MH  - Binding Sites
MH  - Binding,Competitive
MH  - Brain
MH  - Comparative Study
MH  - Diethylstilbestrol
MH  - Digitonin
MH  - drug effects
MH  - Enzyme Activation
MH  - Enzyme Inhibitors
MH  - enzymology
MH  - Estradiol
MH  - Hydrolysis
MH  - Iodine Radioisotopes
MH  - Lactate Dehydrogenase
MH  - Liver
MH  - Male
MH  - mechanism
MH  - MECHANISMS
MH  - metabolism
MH  - Mitochondria
MH  - Mitochondria,Heart
MH  - Mitochondria,Liver
MH  - P
MH  - pharmacology
MH  - physiology
MH  - Proton-Translocating ATPases
MH  - Pyruvate Kinase
MH  - Rats
MH  - Rats,Sprague-Dawley
MH  - Serum Albumin,Bovine
MH  - Site
MH  - Solubility
MH  - Stilbenes
MH  - Support,U.S.Gov't,P.H.S.
MH  - ultrastructure
MH  - universities
RP  - NOT IN FILE
NT  - Department of Molecular and Integrative Physiology, University of Illinois at Urbana-Champaign, Urbana 61801, USA liu4@uiuceduFAU - Kipp, J L
SO  - Endocrine 2001 Jul ;15(2):165-175

1805
UI  - 21067
AU  - Kipp JL
AU  - Ramirez VD
TI  - Effect of estradiol, diethylstilbestrol, and resveratrol on F0F1-ATPase activity from mitochondrial preparations of rat heart, liver, and brain
AB  - The question of whether estrogens or estrogen-like compounds would alter differentially the enzymatic activity of the FOF1-ATPase was addressed. Mitochondrial fractions of the liver, brain, and heart were obtained from adult male rats and solubilized by digitonin. About 85% of the adenosine triphosphate hydrolysis by these three preparations come from the mitochondrial FOF1-ATPase. The enzymatic activity differed in the following order: liver < brain < heart. A concentration of 13 nM estradiol stimulated the FOF1-ATPase activity in heart by 10% (p < 0.01), but not in liver or brain. 17beta-estradiol competed off the binding of estradiol-17beta-17-(O-carboxymethyl)oxime:125I-labeled bovine serium albumin to mitochondrial preparations of the heart, revealing two binding sites. Resveratrol inhibited the F0F1-ATPase activity in both heart and liver with an IC50 of 13-15 microM, which confirmed our previous report in preparations of brain. Lower doses (picomolar to nanomolar) of resveratrol stimulated the FOF1-ATPase activity in liver by 10% but not in heart. At 6.7 microM, diethylstilbestrol (DES) inhibited the FOF1-ATPase activity in the three preparations by 61-67%. This study demonstrates that estradiol activates rat heart mitochondrial FOF1-ATPase at physiologic concentrations and that the FOF1-ATPase activity is markedly different in rat liver, brain, and heart. In addition, estradiol, DES, and resveratrol alter the FOF1-ATPase activity selectively, probably via different mechanisms
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - Adult
MH  - BINDING
MH  - Binding Sites
MH  - Brain
MH  - Hydrolysis
MH  - Liver
MH  - Male
MH  - mechanism
MH  - MECHANISMS
MH  - P
MH  - physiology
MH  - Rats
MH  - Site
RP  - NOT IN FILE
NT  - Department of Molecular and Integrative Physiology, University of Illinois at Urbana-Champaign, Urbana 61801, USA liu4@uiuceduFAU - Kipp, J L
SO  - Endocrine 2001 Jul ;15(2):165-175

1806
UI  - 21072
AU  - Krulwich TA
AU  - Ito M
AU  - Guffanti AA
AD  - Department of Biochemistry and Molecular Biology, Mount Sinai School of Medicine, New York, NY 10029, USA terrykrulwich@mssmedu
TI  - The Na(+)-dependence of alkaliphily in Bacillus
AB  - A Na(+) cycle plays a central role in the remarkable capacity of aerobic, extremely alkaliphilic Bacillus species for pH homeostasis. The capacity for pH homeostasis, in turn, appears to set the upper pH limit for growth. One limb of the alkaliphile Na(+) cycle consists of Na(+)/H(+) antiporters that achieve net H(+) accumulation that is coupled to Na(+) efflux. The major antiporter on which pH homeostasis depends is thought to be the Mrp(Sha)-encoded antiporter, first identified from a partial clone in Bacillus halodurans C-125. Mrp(Sha) may function as a complex. While this antiporter is capable of secondary antiport energized by an imposed or respiration-generated protonmotive force, the possibility of a primary mode has not been excluded. In Bacillus pseudofirmus OF4, at least two additional antiporters, including NhaC, have supporting roles in pH homeostasis. Some of these additional antiporters may be especially important for antiport at low [Na(+)] or at near-neutral pH. The second limb of the Na(+) cycle facilitates Na(+) re-entry via Na(+)/solute symporters and, perhaps, the ion channel associated with the Na(+)-dependent flagellar motor. The process of pH homeostasis is also enhanced, perhaps especially during transitions to high pH, by different arrays of secondary cell wall polymers in the two alkaliphilic Bacillus species studied most intensively. The mechanisms whereby alkaliphiles handle the challenge of Na(+) stress at very elevated [Na(+)] are just beginning to be identified, and a hypothesis has been advanced to explain the finding that B. pseudofirmus OF4 requires a higher [Na(+)] for growth at near-neutral pH than at very alkaline pH values
MH  - A
MH  - Bacillus
MH  - Biochemistry
MH  - Cations
MH  - COMPLEX
MH  - function
MH  - Homeostasis
MH  - ion
MH  - Lipids
MH  - mechanism
MH  - MECHANISMS
MH  - membrane
MH  - pH
MH  - review
MH  - secondary
MH  - Sodium
RP  - NOT IN FILE
NT  - UI - 21145127LA - engRN - 0 (Cations, Monovalent)RN - 0 (Membrane Lipids)RN - 7440-23-5 (Sodium)PT - Journal ArticlePT - ReviewPT - Review, TutorialID - GM28454/GM/NIGMSDA - 20010315IS - 0006-3002SB - IMCY - Netherlands
UR  - PM:11248197
SO  - Biochim Biophys Acta 2001 May 1 ;1505(1):158-168

1807
UI  - 978
AU  - Levy D
AU  - Chami M
AU  - Rigaud JL
TI  - Two-dimensional crystallization of membrane proteins: the lipid layer strategy
AB  - Due to the difficulty to crystallize membrane proteins, there is a considerable interest to intensify research topics aimed at developing new methods of crystallization. In this context, the lipid layer crystallization at the air/water interface, used so far for soluble proteins, has been recently adapted successfully to produce two-dimensional (2D) crystals of membrane proteins, amenable to structural analysis by electron crystallography. Besides to represent a new alternative strategy, this approach gains the advantage to decrease significantly the amount of material needed in incubation trials, thus opening the field of crystallization to those membrane proteins difficult to surexpress and/or purify. The systematic studies that have been performed on different classes of membrane proteins are reviewed and the physico-chemical processes that lead to the production of 2D crystals are addressed. The different drawbacks, advantages and perspectives of this new strategy for providing structural information on membrane proteins are discussed
MH  - Air
MH  - Animal
MH  - Cell Membrane
MH  - Chemistry
MH  - Crystallography,X-Ray
MH  - Lipid Bilayers
MH  - Lipids
MH  - Membrane Proteins
MH  - metabolism
MH  - Methods
MH  - Micelles
MH  - Microscopy
MH  - Models,Biological
MH  - Protein Conformation
MH  - Proteins
MH  - Water
RP  - NOT IN FILE
NT  - Institut Curie, Section de Recherche, UMR-CNRS 168 and LRC-CEA 8, 11 Rue Pierre et Marie Curie, 75231 Cedex 05, Paris, France
SO  - FEBS Lett 2001 Aug 31 ;504(3):187-193

1808
UI  - 21049
AU  - Lodeyro AF
AU  - Calcaterra NB
AU  - Roveri OA
TI  - Inhibition of steady-state mitochondrial ATP synthesis by bicarbonate, an activating anion of ATP hydrolysis
AB  - Bicarbonate, an activating anion of ATP hydrolysis, inhibited ATP synthesis coupled to succinate oxidation in beef heart submitochondrial particles but diminished the lag time and increased the steady-state velocity of the (32)Pi-ATP exchange reaction. The latter effects exclude the possibility that bicarbonate is inducing an intrinsic uncoupling between ATP hydrolysis and proton translocation at the level of F(1)F(o) ATPase. The inhibition of ATP synthesis was competitive with respect to ADP at low fixed [Pi], mixed at high [Pi] and non-competitive towards Pi at any fixed [ADP]. From these results we can conclude that (i) bicarbonate does not bind to a Pi site in the mitochondrial F(1); (ii) it competes with the binding of ADP to a low-affinity site, likely the low-affinity non-catalytic nucleotide binding site. It is postulated that bicarbonate stimulates ATP hydrolysis and inhibits ATP synthesis by modulating the relative affinities of the catalytic site for ATP and ADP
MH  - A
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - ADP
MH  - Animal
MH  - atp
MH  - ATP synthesis
MH  - ATPase
MH  - BINDING
MH  - Binding Sites
MH  - Binding,Competitive
MH  - biosynthesis
MH  - Cattle
MH  - Chemistry
MH  - drug effects
MH  - Hydrolysis
MH  - metabolism
MH  - Mitochondria,Heart
MH  - nucleotide binding
MH  - pharmacology
MH  - Phosphates
MH  - proton
MH  - Proton-Translocating ATPases
MH  - Site
MH  - Sodium Bicarbonate
MH  - Submitochondrial Particles
MH  - succinate
MH  - Support,Non-U.S.Gov't
MH  - synthesis
MH  - Time
MH  - translocation
RP  - NOT IN FILE
NT  - Departamento de Quimica Biologica, Area Biofisica, Facultad de Ciencias Bioquimicas y Farmaceuticas, Universidad Nacional de Rosario, Suipacha 531, (S2002LRK) Rosario, ArgentinaFAU - Lodeyro, A F
SO  - Biochim Biophys Acta 2001 Nov 1 ;1506(3):236-243

1809
UI  - 21407
AU  - Lodeyro AF
AU  - Calcaterra NB
AU  - Roveri OA
TI  - Inhibition of steady-state mitochondrial ATP synthesis by bicarbonate, an activating anion of ATP hydrolysis
AB  - Bicarbonate, an activating anion of ATP hydrolysis, inhibited ATP synthesis coupled to succinate oxidation in beef heart submitochondrial particles but diminished the lag time and increased the steady-state velocity of the (32)Pi-ATP exchange reaction. The latter effects exclude the possibility that bicarbonate is inducing an intrinsic uncoupling between ATP hydrolysis and proton translocation at the level of F(1)F(o) ATPase. The inhibition of ATP synthesis was competitive with respect to ADP at low fixed [Pi], mixed at high [Pi] and non-competitive towards Pi at any fixed [ADP]. From these results we can conclude that (i) bicarbonate does not bind to a Pi site in the mitochondrial F(1); (ii) it competes with the binding of ADP to a low-affinity site, likely the low-affinity non-catalytic nucleotide binding site. It is postulated that bicarbonate stimulates ATP hydrolysis and inhibits ATP synthesis by modulating the relative affinities of the catalytic site for ATP and ADP
MH  - A
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - ADP
MH  - affinity
MH  - Animal
MH  - atp
MH  - ATP synthesis
MH  - ATPase
MH  - BINDING
MH  - BINDING SITE
MH  - Binding Sites
MH  - Binding,Competitive
MH  - biosynthesis
MH  - catalytic
MH  - Cattle
MH  - Chemistry
MH  - drug effects
MH  - Hydrolysis
MH  - metabolism
MH  - Mitochondria,Heart
MH  - nucleotide
MH  - nucleotide binding
MH  - pharmacology
MH  - Phosphates
MH  - proton
MH  - Proton-Translocating ATPases
MH  - Site
MH  - Sodium Bicarbonate
MH  - Submitochondrial Particles
MH  - succinate
MH  - Support,Non-U.S.Gov't
MH  - synthesis
MH  - Time
MH  - translocation
RP  - NOT IN FILE
NT  - Departamento de Quimica Biologica, Area Biofisica, Facultad de Ciencias Bioquimicas y Farmaceuticas, Universidad Nacional de Rosario, Suipacha 531, (S2002LRK) Rosario, ArgentinaFAU - Lodeyro, A F
SO  - Biochim Biophys Acta 2001 Nov 1 ;1506(3):236-243

1810
UI  - 21429
AU  - Lu YM
AU  - Miyazawa K
AU  - Yamaguchi K
AU  - Nowaki K
AU  - Iwatsuki H
AU  - Wakamatsu Y
AU  - Ichikawa N
AU  - Hashimoto T
TI  - Deletion of mitochondrial ATPase inhibitor in the yeast Saccharomyces cerevisiae decreased cellular and mitochondrial ATP levels under non-nutritional conditions and induced a respiration-deficient cell-type
AB  - T(1), a mutant yeast lacking three regulatory proteins of F(1)F(o)ATPase, namely ATPase inhibitor, 9K protein and 15K protein, grew on non-fermentable carbon source at the same rate as normal cells but was less viable when incubated in water. During the incubation, the cellular ATP content decreased rapidly in the T(1) cells but not in normal cells, and respiration-deficient cells appeared among the T(1) cells. The same mutation was also induced in D26 cells lacking only the ATPase inhibitor. Overexpression of the ATPase inhibitor in YC63 cells, which were derived from the D26 strain harboring an expression vector containing the gene of the ATPase inhibitor, prevented the decrease of cellular ATP level and the mutation. Isolated T(1) mitochondria exhibited ATP hydrolysis for maintenance of membrane potential when antimycin A was added to the mitochondrial suspension, while normal and YC63 mitochondria continued to show low hydrolytic activity and low membrane potential. Thus, it is likely that deletion of the ATPase inhibitor induces ATPase activity of F(1)F(o)ATPase to create a dispensable membrane potential under the non-nutritional conditions and that this depletes mitochondrial and cellular ATP. The depletion of mitochondrial ATP in turn leads to occurrence of aberrant DNA in mitochondria
MH  - A
MH  - Adenosine Triphosphate
MH  - antagonists & inhibitors
MH  - Antimycin A
MH  - atp
MH  - ATPase
MH  - cell
MH  - Cell Division
MH  - Cell Respiration
MH  - Cells
MH  - Chemistry
MH  - cytology
MH  - Dna
MH  - DNA,Mitochondrial
MH  - Gene Deletion
MH  - genetics
MH  - Hydrolysis
MH  - inhibitor
MH  - M
MH  - membrane
MH  - Membrane Potential
MH  - Membrane Potentials
MH  - metabolism
MH  - Mitochondria
MH  - mutant
MH  - Mutation
MH  - physiology
MH  - protein
MH  - Proteins
MH  - Proton-Translocating ATPases
MH  - Saccharomyces cerevisiae
MH  - Support,Non-U.S.Gov't
MH  - Water
MH  - YEAST
RP  - NOT IN FILE
NT  - Department of Applied Chemistry, Muroran Institute of Technology, Muroran, Hokkaido 050-8585, JapanFAU - Lu, Y M
SO  - J Biochem (Tokyo ) 2001 Dec ;130(6):873-878

1811
UI  - 21442
AU  - Martin-Galiano AJ
AU  - Ferrandiz MJ
AU  - de la Campa AG
TI  - The promoter of the operon encoding the F0F1 ATPase of Streptococcus pneumoniae is inducible by pH
AB  - The genes encoding the subunits of the F0F1 membrane ATPase of Streptococcus pneumoniae were cloned and sequenced. The eight genes, transcribed to one mRNA, are organized in an operon encoding the c, a, b, delta, alpha, gamma, beta and epsilon subunits of 66, 238, 165, 178, 501, 292, 471 and 139 amino acid residues, respectively, that were expressed in an Escherichia coli system. To investigate the role of the ATPase in the regulation of the intracellular pH, the expression of the operon between pH 5.7 and 7.5 was studied. An increase in both the ATPase activity and the amount of the alpha and beta F1 subunits as shown by Western blot analysis was observed as the pH decreased. These increases were accompanied by an increase in the atp-specific mRNA, as shown by Northern blot and slot-blot analysis. Primer extension experiments and transcriptional fusions between the atp promoter and the reporter cat gene demonstrated that this pH-dependent increase in the mRNA was regulated at the level of initiation of transcription. Transcription of the operon occurs from a promoter with a consensus -35 box (TTGACA) and a -10 box (TACACT) that differs from the consensus (TATAAT). A point mutation at the -10 box of the promoter (change to TGCACT) avoided this increase, suggesting a role for this sequence in the pH-inducible regulation
MH  - A
MH  - ACID
MH  - alpha
MH  - Amino Acid Sequence
MH  - analysis
MH  - atp
MH  - ATPase
MH  - Base Sequence
MH  - BETA
MH  - Chemistry
MH  - Cloning,Molecular
MH  - delta
MH  - DNA,Bacterial
MH  - enzymology
MH  - EPSILON-SUBUNIT
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0F1
MH  - F1
MH  - Gene Expression Regulation,Bacterial
MH  - Gene Expression Regulation,Enzymologic
MH  - Genes,Bacterial
MH  - genetics
MH  - Hydrogen-Ion Concentration
MH  - membrane
MH  - metabolism
MH  - Molecular Sequence Data
MH  - Mutation
MH  - Operon
MH  - pH
MH  - point mutation
MH  - Promoter Regions (Genetics)
MH  - Protein Subunits
MH  - Proton-Translocating ATPases
MH  - regulation
MH  - RESIDUE
MH  - Restriction Mapping
MH  - RNA,Bacterial
MH  - RNA,Messenger
MH  - Sequence Homology,Nucleic Acid
MH  - Streptococcus
MH  - Streptococcus pneumoniae
MH  - SUBUNIT
MH  - Support,Non-U.S.Gov't
MH  - SYSTEM
RP  - NOT IN FILE
NT  - Unidad de Genetica Bacteriana (CSIC), Centro Nacional de Biologia Fundamental, Instituto de Salud Carlos III, 28220 Majadahonda, Madrid, SpainFAU - Martin-Galiano, A J
SO  - Mol Microbiol 2001 Sep ;41(6):1327-1338

1812
UI  - 21439
AU  - McGeoch JE
AU  - Guidotti G
TI  - Batten disease and the control of the Fo subunit c pore by cGMP and calcium
AB  - Subunit c of ATP synthase functions as a high conductance ion channel, tightly regulated by calcium. We have suggested that the pathogenesis of Batten syndromes involving overaccumulation of subunit c are linked to the protein's ion channel function. In normal electrically excitable tissue the channel could act as a pacer setting nodal voltage via control of cation entry. The channel conductance is controlled by voltage, calcium, cyclic nucleotides and polyamines. We discuss the pathogenic role that subunit c could play in the electrically excitable tissues of retina, brain and heart where Batten neurodegeneration is seen. Focus is given to potential links between subunit c and the known mutant gene products in the Batten diseases, the process of apoptosis, and the requirement of the growing brain for gradients of cGMP, a ligand of the subunit c channel
MH  - A
MH  - Animal
MH  - Apoptosis
MH  - atp
MH  - ATP synthase
MH  - biology
MH  - Brain
MH  - Calcium
MH  - Cattle
MH  - conductance
MH  - cyclic
MH  - Cyclic GMP
MH  - FO
MH  - function
MH  - ion
MH  - Ion Channel Gating
MH  - metabolism
MH  - Microscopy,Electron
MH  - mutant
MH  - Nerve Degeneration
MH  - Neuronal Ceroid-Lipofuscinosis
MH  - nucleotide
MH  - Nucleotides
MH  - physiology
MH  - protein
MH  - Proton-Translocating ATPases
MH  - Rats
MH  - Sheep
MH  - SUBUNIT
MH  - Support,U.S.Gov't,Non-P.H.S.
MH  - SYNTHASE
MH  - ultrastructure
MH  - universities
MH  - voltage
RP  - NOT IN FILE
NT  - Department of Molecular and Cellular Biology, Harvard University, 7 Divinity Avenue, Cambridge, MA 02138, USA mcgeoch@fasharvardeduFAU - McGeoch, J E
SO  - Europ J Paediatr Neurol 2001  ;5 Suppl A:147-50.():147-150

1813
UI  - 21250
AU  - McLuskey K
AU  - Prince SM
AU  - Cogdell RJ
AU  - Isaacs NW
AD  - Department of Chemistry, Division of Biochemistry and Molecular Biology IBLS, University of Glasgow, Glasgow, Scotland G12 8QQ, UK
TI  - The crystallographic structure of the B800-820 LH3 light-harvesting complex from the purple bacteria Rhodopseudomonas acidophila strain 7050
AB  - The B800-820, or LH3, complex is a spectroscopic variant of the B800- 850 LH2 peripheral light-harvesting complex. LH3 is synthesized by some species and strains of purple bacteria when growing under what are generally classed as "stressed" conditions, such as low intensity illumination and/or low temperature (<30 degrees C). The apoproteins in these complexes modify the absorption properties of the chromophores to ensure that the photosynthetic process is highly efficient. The crystal structure of the B800-820 light-harvesting complex, an integral membrane pigment-protein complex, from the purple bacteria Rhodopseudomonas (Rps.) acidophila strain 7050 has been determined to a resolution of 3.0 A by molecular replacement. The overall structure of the LH3 complex is analogous to that of the LH2 complex from Rps. acidophila strain 10050. LH3 has a nonameric quaternary structure where two concentric cylinders of alpha-helices enclose the pigment molecules bacteriochlorophyll a and carotenoid. The observed spectroscopic differences between LH2 and LH3 can be attributed to differences in the primary structure of the apoproteins. There are changes in hydrogen bonding patterns between the coupled Bchla molecules and the protein that have an effect on the conformation of the C3-acetyl groups of the B820 molecules. The structure of LH3 shows the important role that the protein plays in modulating the characteristics of the light-harvesting system and indicates the mechanisms by which the absorption properties of the complex are altered to produce a more efficient light-harvesting component
MH  - A
MH  - absorption
MH  - Bacteria
MH  - Biochemistry
MH  - carotenoid
MH  - Carotenoids
MH  - Chemistry
MH  - COMPLEX
MH  - conformation
MH  - Hydrogen
MH  - Hydrogen Bonding
MH  - low temperature
MH  - mechanism
MH  - MECHANISMS
MH  - membrane
MH  - protein
MH  - reaction center
MH  - resolution
MH  - rhodopseudomonas
MH  - structure
MH  - SYSTEM
MH  - Temperature
RP  - NOT IN FILE
NT  - UI - 21361108LA - engRN - 0 (B800-820 light-harvesting complex)RN - 0 (Bacteriochlorophylls)RN - 0 (Photosynthetic Reaction Center, Bacterial)RN - 0 (Porphyrins)RN - 103428-20-2 (bacteriochlorin a)RN - 36-88-4 (Carotenoids)PT - Journal ArticleDA - 20010724IS - 0006-2960SB - IMCY - United States
UR  - PM:11467938
SO  - Biochemistry 2001 Jul 31 ;40(30):8783-8789

1814
UI  - 21302
AU  - Medvedev D
AU  - Stuchebrukhov AA
AD  - Department of Chemistry, University of California, Davis, CA 95616, USA
TI  - DNA repair mechanism by photolyase: electron transfer path from the photolyase catalytic cofactor FADH(-) to DNA thymine dimer
AB  - Photolyase is an enzyme that catalyses photorepair of thymine dimers in UV damaged DNA by electron transfer reaction. The structure of the photolyase/DNA complex is unknown at present. Using crystal structure coordinates of the substrate-free enzyme from E. coli, we have recently built a computer molecular model of a thymine dimer docked to photolyase catalytic site and studied molecular dynamics of the system. In this paper, we present analysis of the electronic coupling and electron transfer pathway between the catalytic cofactor FADH(-) and the pyrimidine dimer by the method of interatomic tunneling currents. Electronic structure is treated in the extended Huckel approximation. The root mean square transfer matrix element is about 6 cm(-1), which is consistent with the experimentally determined rate of transfer. We find that electron transfer mechanism responsible for the repair utilizes an unusual folded conformation of FADH(-) in photolyases, in which the isoalloxazine ring of the flavin and the adenine are in close proximity, and the peculiar features of the docked orientation of the dimer. The tunneling currents show explicitly that despite of the close proximity between the donor and acceptor complexes, the electron transfer mechanism between the flavin and the thymine bases is not direct, but indirect, with the adenine acting as an intermediate. These calculations confirm the previously made conclusion based on an indirect evidence for such mechanism
MH  - A
MH  - acceptor
MH  - analysis
MH  - BASE
MH  - catalytic
MH  - Chemistry
MH  - COMPLEX
MH  - conformation
MH  - coupling
MH  - electron
MH  - electron transfer
MH  - Electron-transfer
MH  - England
MH  - enzyme
MH  - intermediate
MH  - mechanism
MH  - method
MH  - model
MH  - MOLECULAR-DYNAMICS
MH  - Site
MH  - structure
MH  - SYSTEM
MH  - TRANSFER
MH  - universities
RP  - NOT IN FILE
NT  - UI - 21265198DA - 20010523IS - 0022-5193LA - engID - GM54052-02/GM/NIGMSPT - Journal ArticleCY - EnglandRN - 0 (Pyrimidine Dimers)RN - 146-14-5 (Flavin-Adenine Dinucleotide)RN - EC 4.1.99.3 (Deoxyribodipyrimidine Photo-Lyase)SB - IM
UR  - PM:11371177
SO  - J Theor Biol 2001 May 21 ;210(2):237-248

1815
UI  - 21444
AU  - Meier T
AU  - Matthey U
AU  - Henzen F
AU  - Dimroth P
AU  - Muller DJ
TI  - The central plug in the reconstituted undecameric c cylinder of a bacterial ATP synthase consists of phospholipids
AB  - The isolated rotor cylinder of the ATP synthase from Ilyobacter tartaricus was reconstituted into two-dimensional crystalline arrays. Atomic force microscopy imaging indicated a central cavity on one side of the rotor and a central plug protruding from the other side. Upon incubation with phospholipase C, the plug disappeared, but the appearance of the surrounding c subunit oligomer was not affected. This indicates that the plug consists of phospholipids. As the detergent-purified c cylinder is completely devoid of phospholipids, these are incorporated into the central hole from one side of the cylinder during the reconstitution procedure
MH  - A
MH  - atp
MH  - ATP synthase
MH  - Chemistry
MH  - enzymology
MH  - Fusobacterium
MH  - Microscopy
MH  - Microscopy,Atomic Force
MH  - Phospholipids
MH  - Proton-Translocating ATPases
MH  - reconstitution
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - Institut fur Mikrobiologie, Eidgenossische Technische Hochschule, ETH-Zentrum, Zurich, SwitzerlandFAU - Meier, T
SO  - FEBS Lett 2001 Sep 21 ;505(3):353-356

1816
UI  - 313
AU  - Menz RI
AU  - Walker JE
AU  - Leslie AG
AD  - Medical Research Council Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, United Kingdom
TI  - Structure of bovine mitochondrial F(1)-ATPase with nucleotide bound to all three catalytic sites: implications for the mechanism of rotary catalysis
AB  - The crystal structure of a novel aluminium fluoride inhibited form of bovine mitochondrial F(1)-ATPase has been determined at 2 A resolution. In contrast to all previously determined structures of the bovine enzyme, all three catalytic sites are occupied by nucleotide. The subunit that did not bind nucleotide in previous structures binds ADP and sulfate (mimicking phosphate), and adopts a "half-closed" conformation. This structure probably represents the posthydrolysis, pre-product release step on the catalytic pathway. A catalytic scheme for hydrolysis (and synthesis) at physiological rates and a mechanism for the ATP-driven rotation of the gamma subunit are proposed based on the crystal structures of the bovine enzyme
RP  - NOT IN FILE
NT  - UI - 21400444LA - engRN - 0 (Aluminum Compounds)RN - 0 (Enzyme Inhibitors)RN - 0 (Fluorides)RN - 0 (Nucleotides)RN - 0 (Protein Subunits)RN - 0 (Sulfates)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 7784-18-1 (aluminum fluoride)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 20010817IS - 0092-8674SB - IMCY - United StatesJC - CQ4
UR  - PM:11509182
SO  - Cell 2001 Aug 10 ;106(3):331-341

1817
UI  - 315
AU  - Menz RI
AU  - Leslie AG
AU  - Walker JE
AD  - The Medical Research Council Laboratory of Molecular Biology, Cambridge, UK
TI  - The structure and nucleotide occupancy of bovine mitochondrial F(1)- ATPase are not influenced by crystallisation at high concentrations of nucleotide
AB  - Analysis of tryptophan mutants of F(1)-ATPase from Escherichia coli [Lobau et al. (1997) FEBS Lett. 404, 15-18] suggested that nucleotide concentrations used to grow crystals for the determination of the structure of bovine F(1)-ATPase [Abrahams et al. (1994) Nature 370, 621- 628] would be sufficient to occupy only two catalytic sites, and that higher concentrations of nucleotide would result in all three sites being occupied. We have determined the structure of bovine F(1)-ATPase at 2.9 A resolution with crystals grown in the presence of 5 mM AMPPNP and 5 microM ADP. Similar to previous structures of bovine F(1)-ATPase determined with crystals grown in the presence of lower nucleotide concentrations, only two beta-subunits have bound nucleotide and the third subunit remains empty
RP  - NOT IN FILE
NT  - UI - 21195321LA - engRN - 0 (Nucleotides)RN - 25612-73-1 (Adenylyl Imidodiphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 20010412IS - 0014-5793SB - IMCY - NetherlandsJC - EUH
UR  - PM:11297725
SO  - FEBS Lett 2001 Apr 6 ;494(1-2):11-14

1818
UI  - 2
AU  - Muller DJ
AU  - Dencher NA
AU  - Meier T
AU  - Dimroth P
AU  - Suda K
AU  - Stahlberg H
AU  - Engel A
AU  - Seelert H
AU  - Matthey U
AD  - Max-Planck Institute of Molecular Cell Biology and Genetics, Dresden, Germany danielmueller@mpi-cbgde
TI  - ATP synthase: constrained stoichiometry of the transmembrane rotor
AB  - Recent structural data suggest that the number of identical subunits (c or III) assembled into the cation-powered rotor of F1F0 ATP synthase depends on the biological origin. Atomic force microscopy allowed individual subunits of the cylindrical transmembrane rotors from spinach chloroplast and from Ilyobacter tartaricus ATP synthase to be directly visualized in their native-like environment. Occasionally, individual rotors exhibit structural gaps of the size of one or more subunits. Complete rotors and arch-shaped fragments of incomplete rotors revealed the same diameter within one ATP synthase species. These results suggest the rotor diameter and stoichiometry to be determined by the shape of the subunits and their nearest neighbor interactions
RP  - NOT IN FILE
NT  - UI - 21423579LA - engRN - 0 (Multienzyme Complexes)RN - EC 2.7.4 (Phosphotransferases (Phosphate Group Acceptor))RN - EC 2.7.4.- (ATP synthetase complex)PT - Journal ArticleDA - 20010904IS - 0014-5793SB - IMCY - NetherlandsJC - EUH
UR  - PM:11532457
SO  - FEBS Lett 2001 Aug 31 ;504(3):219-222

1819
UI  - 21029
AU  - Murata T
AU  - Kakinuma Y
AU  - Yamato I
AD  - Department of Biological Science and Technology, Science University of Tokyo, 2641 Yamazaki, Noda-shi, Chiba 278-8510, Japan tm@mrc- dunncamacuk
TI  - ATP-dependent affinity change of Na+-binding sites of V-ATPase
AB  - V-type Na(+)-ATPase of Enterococcus hirae binds about six (6 +/- 1) Na(+) ions/enzyme molecule with a high affinity (Murata, T., Igarashi, K., Kakinuma, Y., and Yamato, I. (2000) J. Biol. Chem. 275, 13415- 13419). After the addition of 5 mm ATP, the binding capacity dropped to about 2 (1.8 +/- 0.3) Na(+) ions/enzyme molecule, returning to the initial value concomitant with the decrease of ATP hydrolysis rate. These findings suggest that the affinity of four of six Na(+)-binding sites of the enzyme changes (lowers) in enzyme reaction. The ATP analogs (adenosine 5'-O-(3-thiotriphosphate) or 5'-adenylylimido- diphosphate), ADP, or aluminum fluoride that is postulated to trap ATPases at their transition state did not inhibit the Na(+) binding capacity significantly. Therefore, the affinity decrease of Na(+)- binding sites was unlikely to be due to ATP binding alone or at the transition state of ATP hydrolysis. In the presence of 5 mm ATP, the ATPase showed strong negative cooperativity (n(H) = 0.16 +/- 0.03) for Na(+) stimulation of ATPase activity. The Hill coefficient (n(H)) increased to 1 in parallel to the decrease of ATP concentration in the reaction mixture. Thus, the ATP-dependent affinity change cooperatively occurs in continuous enzyme reaction
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - ADP
MH  - ANALOGS
MH  - atp
MH  - ATPase
MH  - BINDING
MH  - Binding Sites
MH  - H+-ATPase
MH  - Hydrolysis
MH  - Site
MH  - Sodium
RP  - NOT IN FILE
NT  - UI - 21611172LA - engRN - 56-65-5 (Adenosine Triphosphate)RN - 7440-23-5 (Sodium)RN - EC 3.6.1.- (Vacuolar Proton-Translocating ATPases)RN - EC 3.6.1.- (vacuolar H+-ATPase)PT - Journal ArticleDA - 20011217IS - 0021-9258SB - IMCY - United States
UR  - PM:11557766
SO  - J Biol Chem 2001 Dec 21 ;276(51):48337-48340

1820
UI  - 685
AU  - Noji H
AU  - Yoshida M
AD  - CREST (Core Research for Evolutional Science and Technology) "Genetic Programming" Team 13, Teikyo University Biotechnology Research Center 3F, Nogawa 907, Miyamae-ku, Kawasaki 216-0001, Japan
TI  - The rotary machine in the cell, ATP synthase
RP  - NOT IN FILE
NT  - UI - 21125712LA - engRN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticlePT - ReviewPT - Review, TutorialDA - 20010306IS - 0021-9258SB - IMCY - United StatesJC - HIV
UR  - PM:11080505
SO  - J Biol Chem 2001 Jan 19 ;276(3):1665-1668

1821
UI  - 683
AU  - Noji H
AU  - Bald D
AU  - Yasuda R
AU  - Itoh H
AU  - Yoshida M
AU  - Kinosita K
AD  - CREST "Genetic Programming" Team 13, Teikyo University Biotechnology Research Center 3F, Nogawa 907, Miyamae-ku, Kawasaki 216-0001, Japan
TI  - Purine but not pyrimidine nucleotides support rotation of F(1)-ATPase
AB  - The binding change model for the F(1)-ATPase predicts that its rotation is intimately correlated with the changes in the affinities of the three catalytic sites for nucleotides. If so, subtle differences in the nucleotide structure may have pronounced effects on rotation. Here we show by single-molecule imaging that purine nucleotides ATP, GTP, and ITP support rotation but pyrimidine nucleotides UTP and CTP do not, suggesting that the extra ring in purine is indispensable for proper operation of this molecular motor. Although the three purine nucleotides were bound to the enzyme at different rates, all showed similar rotational characteristics: counterclockwise rotation, 120 degrees steps each driven by hydrolysis of one nucleotide molecule, occasional back steps, rotary torque of approximately 40 piconewtons (pN).nm, and mechanical work done in a step of approximately 80 pN.nm. These latter characteristics are likely to be determined by the rotational mechanism built in the protein structure, which purine nucleotides can energize. With ATP and GTP, rotation was observed even when the free energy of hydrolysis was -80 pN.nm/molecule, indicating approximately 100% efficiency. Reconstituted F(o)F(1)-ATPase actively translocated protons by hydrolyzing ATP, GTP, and ITP, but CTP and UTP were not even hydrolyzed. Isolated F(1) very slowly hydrolyzed UTP (but not CTP), suggesting possible uncoupling from rotation
RP  - NOT IN FILE
NT  - UI - 21326046LA - engRN - 0 (Purine Nucleotides)RN - 0 (Pyrimidine Nucleotides)RN - 132-06-9 (Inosine Triphosphate)RN - 56-65-5 (Adenosine Triphosphate)RN - 63-39-8 (Uridine Triphosphate)RN - 86-01-1 (Guanosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 20010702IS - 0021-9258SB - IMCY - United StatesJC - HIV
UR  - PM:11279248
SO  - J Biol Chem 2001 Jul 6 ;276(27):25480-25486

1822
UI  - 20832
AU  - Oka T
AU  - Toyomura T
AU  - Honjo K
AU  - Wada Y
AU  - Futai M
AD  - Division of Biological Sciences, Institute of Scientific and Industrial Research, Osaka University, Japan
TI  - Four subunit a isoforms of Caenorhabditis elegans vacuolar H+-ATPase. Cell-specific expression during development
AB  - We have identified four genes (vha-5, vha-6, vha-7, and unc-32) coding for vacuolar-type proton-translocating ATPase (V-ATPase) subunit a in Caenorhabditis elegans, the first example of four distinct isoforms in eukaryotes. Their products had nine putative transmembrane regions, exhibited 43-60% identity and 62-84% similarity with the bovine subunit a1 isoform, and retained 11 amino acid residues essential for yeast V- ATPase activity (Leng, X. H., Manolson, M. F., and Forgac, M. (1998) J. Biol. Chem. 273, 6717-6723). The similarities, together with the results of immunoprecipitation, suggest that these isoforms are components of V-ATPase. Transgenic and immunofluorescence analyses revealed that these genes were strongly expressed in distinct cells; vha-5 was strongly expressed in an H-shaped excretory cell, vha-6 was strongly expressed in intestine, vha-7 was strongly expressed in hypodermis, and unc-32 was strongly expressed in nerve cells. Furthermore, the vha-7 and unc-32 genes were also expressed in the uteri of hermaphrodites. RNA interference analysis showed that the double-stranded RNA for unc-32 caused embryonic lethality similar to that seen with other subunit genes (vha-1, vha-4, and vha-11) (Oka, T., and Futai, M. (2000) J. Biol. Chem. 275, 29556-29561). The progenies of worms injected with the vha-5 or vha-6 double-stranded RNA became died at a specific larval stage, whereas the vha-7 double-stranded RNA showed no effect on development. These results suggest that V-ATPases with these isoforms generate acidic compartments essential for worm development in a cell-specific manner
MH  - A
MH  - ACID
MH  - analysis
MH  - ATPase
MH  - Cells
MH  - development
MH  - H+-ATPase
MH  - M
MH  - protein
MH  - Proteins
MH  - proton
MH  - Proton Pump
MH  - RESIDUE
MH  - SUBUNIT
RP  - NOT IN FILE
NT  - UI - 21413939LA - engRN - 0 (Isoenzymes)RN - 0 (Luminescent Proteins)RN - 0 (Protein Subunits)RN - 0 (Proton Pumps)RN - 0 (RNA, Double-Stranded)RN - 0 (Recombinant Fusion Proteins)RN - 147336-22-9 (green fluorescent protein)RN - EC 3.6.1.- (vacuolar H+-ATPase)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 20010827IS - 0021-9258SB - IMCY - United States
UR  - PM:11441002
SO  - J Biol Chem 2001 Aug 31 ;276(35):33079-33085

1823
UI  - 21441
AU  - Osanai T
AU  - Tanaka M
AU  - Kamada T
AU  - Nakano T
AU  - Takahashi K
AU  - Okada S
AU  - Sirato K
AU  - Magota K
AU  - Kodama S
AU  - Okumura K
TI  - Mitochondrial coupling factor 6 as a potent endogenous vasoconstrictor
AB  - We demonstrated recently that coupling factor 6, an essential component of the energy-transducing stalk of mitochondrial ATP synthase, suppresses the synthesis of prostacyclin in vascular endothelial cells. Here, we tested the hypothesis that coupling factor 6 is present on the cell surface and is involved in the regulation of systemic circulation. This peptide is present on the surface of CRL-2222 vascular endothelial cells and is released by these cells into the medium. In vivo, the peptide circulates in the vascular system of the rat, and its gene expression and plasma concentration are higher in spontaneously hypertensive rats (SHRs) than in normotensive controls. Elevation of blood pressure with norepinephrine did not affect the plasma concentration of coupling factor 6. Intravenous injection of recombinant peptide increased blood pressure, apparently by suppressing prostacyclin synthesis, whereas a specific Ab to coupling factor 6 decreased systemic blood pressure concomitantly with an increase in plasma prostacyclin. Interestingly, the antibody's hypotensive effect could be abolished by treating with the cyclooxygenase inhibitor indomethacin. These findings indicate that mitochondrial coupling factor 6 functions as a potent endogenous vasoconstrictor in the fashion of a circulating hormone and may suggest a new mechanism for hypertension
MH  - A
MH  - Amino Acid Sequence
MH  - Animal
MH  - atp
MH  - ATP synthase
MH  - biosynthesis
MH  - blood
MH  - Bradykinin
MH  - cell
MH  - Cells
MH  - Cells,Cultured
MH  - coupling
MH  - COUPLING FACTOR
MH  - cytology
MH  - Endothelium,Vascular
MH  - Epoprostenol
MH  - function
MH  - Gene Expression
MH  - genetics
MH  - inhibitor
MH  - Male
MH  - mechanism
MH  - metabolism
MH  - Mitochondria
MH  - Mitochondrial Proton-Translocating ATPases
MH  - Molecular Sequence Data
MH  - Oxidative Phosphorylation Coupling Factors
MH  - peptide
MH  - pharmacology
MH  - Rats
MH  - Rats,Inbred SHR
MH  - Rats,Inbred WKY
MH  - Recombinant Fusion Proteins
MH  - regulation
MH  - stalk
MH  - SURFACE
MH  - SYNTHASE
MH  - synthesis
MH  - SYSTEM
MH  - universities
MH  - Vasoconstrictor Agents
RP  - NOT IN FILE
NT  - The Second Department of Internal Medicine, Hirosaki University School of Medicine, Hirosaki, Japan osanait@cchirosaki-uacjpFAU - Osanai, T
SO  - J Clin Invest 2001 Oct ;108(7):1023-1030

1824
UI  - 21421
AU  - Osanai T
AU  - Okada S
AU  - Sirato K
AU  - Nakano T
AU  - Saitoh M
AU  - Magota K
AU  - Okumura K
TI  - Mitochondrial coupling factor 6 is present on the surface of human vascular endothelial cells and is released by shear stress
AB  - BACKGROUND: We showed that mitochondrial coupling factor 6 (CF6), an endogenous inhibitor of prostacyclin synthesis, is present in the systemic circulation as a pressor substance in rats. We investigated the possibility of vascular endothelial cells as a source of circulating CF6. METHODS AND RESULTS: We used 2 cultured endothelial cell lines, human umbilical vein endothelial cells (HUVECs) and ECV 304 cells (transformed HUVECs), for this study. Immunofluorescence microscopy of both ECV 304 and HUVECs confirmed the surface-associated immunoreactivity of anti-CF6 antibody on the plasma membrane. The concentration of CF6 in the medium increased gradually with time in both ECV 304 and HUVECs in static conditions. Exposure of ECV 304 and HUVECs to a fluid shear stress enhanced the release of CF6: In ECV 304, the concentration of CF6 in the medium (ng. well(-1). 6 hours(-1)) was 2.1+/-0.8 at baseline, 4.3+/-0.8 after shear at 15 dynes/cm(2), and 57.7+/-8.4 after shear at 25 dynes/cm(2). CF6 contents in the cell homogenate and mitochondria were both significantly increased after exposure of ECV 304 to 6-hour shear at 15 dynes/cm(2), whereas they were unchanged after shear stress at 25 dynes/cm(2). The ratio of CF6 to GAPDH mRNA was enhanced significantly, by 1.8+/-0.2-fold, after 6-hour shear stress at 25 dynes/cm(2). Flow cytometry analysis revealed that the surface-associated CF6 was significantly increased in a 3-hour static condition after the previous exposure of the cells to shear stress for 3 hours. CONCLUSIONS: Vascular endothelial cells are a source of CF6, and shear stress regulates the release of the surface-associated CF6
MH  - A
MH  - analysis
MH  - cell
MH  - Cell Line
MH  - Cell Membrane
MH  - Cells
MH  - Cells,Cultured
MH  - coupling
MH  - COUPLING FACTOR
MH  - cytology
MH  - Endothelium,Vascular
MH  - enzymology
MH  - Flow Cytometry
MH  - Gene Expression Regulation,Enzymologic
MH  - genetics
MH  - Human
MH  - inhibitor
MH  - membrane
MH  - metabolism
MH  - method
MH  - Methods
MH  - Microscopy
MH  - Microscopy,Fluorescence
MH  - Mitochondria
MH  - Mitochondrial Proton-Translocating ATPases
MH  - Oxidative Phosphorylation Coupling Factors
MH  - Radioimmunoassay
MH  - Rats
MH  - RNA,Messenger
MH  - secretion
MH  - Stress,Mechanical
MH  - Support,Non-U.S.Gov't
MH  - SURFACE
MH  - synthesis
MH  - Time
MH  - Time Factors
MH  - universities
RP  - NOT IN FILE
NT  - Second Department of Internal Medicine, Hirosaki University School of Medicine, Hirosaki, Japan osanait@cchirosaki-uacjpFAU - Osanai, T
SO  - Circulation 2001 Dec 18 ;104(25):3132-3136

1825
UI  - 21060
AU  - Osanai T
AU  - Okada S
AU  - Sirato K
AU  - Nakano T
AU  - Saitoh M
AU  - Magota K
AU  - Okumura K
TI  - Mitochondrial coupling factor 6 is present on the surface of human vascular endothelial cells and is released by shear stress
AB  - BACKGROUND: We showed that mitochondrial coupling factor 6 (CF6), an endogenous inhibitor of prostacyclin synthesis, is present in the systemic circulation as a pressor substance in rats. We investigated the possibility of vascular endothelial cells as a source of circulating CF6. METHODS AND RESULTS: We used 2 cultured endothelial cell lines, human umbilical vein endothelial cells (HUVECs) and ECV 304 cells (transformed HUVECs), for this study. Immunofluorescence microscopy of both ECV 304 and HUVECs confirmed the surface-associated immunoreactivity of anti-CF6 antibody on the plasma membrane. The concentration of CF6 in the medium increased gradually with time in both ECV 304 and HUVECs in static conditions. Exposure of ECV 304 and HUVECs to a fluid shear stress enhanced the release of CF6: In ECV 304, the concentration of CF6 in the medium (ng. well(-1). 6 hours(-1)) was 2.1+/-0.8 at baseline, 4.3+/-0.8 after shear at 15 dynes/cm(2), and 57.7+/-8.4 after shear at 25 dynes/cm(2). CF6 contents in the cell homogenate and mitochondria were both significantly increased after exposure of ECV 304 to 6-hour shear at 15 dynes/cm(2), whereas they were unchanged after shear stress at 25 dynes/cm(2). The ratio of CF6 to GAPDH mRNA was enhanced significantly, by 1.8+/-0.2-fold, after 6-hour shear stress at 25 dynes/cm(2). Flow cytometry analysis revealed that the surface-associated CF6 was significantly increased in a 3-hour static condition after the previous exposure of the cells to shear stress for 3 hours. CONCLUSIONS: Vascular endothelial cells are a source of CF6, and shear stress regulates the release of the surface-associated CF6
MH  - A
MH  - analysis
MH  - Cell Line
MH  - Cell Membrane
MH  - Cells
MH  - Cells,Cultured
MH  - coupling
MH  - COUPLING FACTOR
MH  - cytology
MH  - Endothelium,Vascular
MH  - enzymology
MH  - Flow Cytometry
MH  - Gene Expression Regulation,Enzymologic
MH  - genetics
MH  - Human
MH  - inhibitor
MH  - membrane
MH  - metabolism
MH  - method
MH  - Methods
MH  - Microscopy
MH  - Microscopy,Fluorescence
MH  - Mitochondria
MH  - Mitochondrial Proton-Translocating ATPases
MH  - Oxidative Phosphorylation Coupling Factors
MH  - Radioimmunoassay
MH  - Rats
MH  - RNA,Messenger
MH  - secretion
MH  - Stress,Mechanical
MH  - Support,Non-U.S.Gov't
MH  - SURFACE
MH  - synthesis
MH  - Time
MH  - Time Factors
RP  - NOT IN FILE
NT  - Second Department of Internal Medicine, Hirosaki University School of Medicine, Hirosaki, Japan osanait@cchirosaki-uacjpFAU - Osanai, T
SO  - Circulation 2001 Dec 18 ;104(25):3132-3136

1826
UI  - 19886
AU  - Paddison SJ
AU  - Paul R
AU  - Zawodzinski TA
TI  - Proton friction and diffusion coefficients in hydrated polymer electrolyte membranes: Computations with a non-equilibrium statistical mechanical model
MH  - A
MH  - Diffusion
MH  - membrane
MH  - Membranes
MH  - model
MH  - proton
RP  - NOT IN FILE
NT  - JournalOCT 22484YTJ CHEM PHYS
UR  - ISI:000171719200047
SO  - Journal of Chemical Physics 2001  ;115(16):7753-7761

1827
UI  - 279
AU  - Panke O
AU  - Cherepanov DA
AU  - Gumbiowski K
AU  - Engelbrecht S
AU  - Junge W
AD  - Division of Biophysics, University of Osnabruck, D-49069 Osnabruck, Germany
TI  - Viscoelastic dynamics of actin filaments coupled to rotary f-atpase: angular torque profile of the enzyme
AB  - ATP synthase (F(O)F(1)) operates as two rotary motor/generators coupled by a common shaft. Both portions, F(1) and F(O), are rotary steppers. Their symmetries are mismatched (C(3) versus C(10-14)). We used the curvature of fluorescent actin filaments, attached to the rotating c- ring, as a spring balance (flexural rigidity of 8. 10(-26) Nm(2)) to gauge the angular profile of the output torque at F(O) during ATP hydrolysis by F(1) (see theoretical companion article (. Biophys. J. 81:1234-1244.)). The large average output torque (50 +/- 6 pN. nm) proved the absence of any slip. Variations of the torque were small, and the output free energy of the loaded enzyme decayed almost linearly over the angular reaction coordinate. Considering the threefold stepping and high activation barrier of the driving motor proper, the rather constant output torque implied a soft elastic power transmission between F(1) and F(O). It is considered as essential, not only for the robust operation of this ubiquitous enzyme under symmetry mismatch, but also for a high turnover rate of the two counteracting and stepping motor/generators
RP  - NOT IN FILE
NT  - UI - 21400326LA - engPT - Journal ArticleDA - 20010817IS - 0006-3495SB - IMCY - United StatesJC - A5S
UR  - PM:11509339
SO  - Biophys J 2001 Sep ;81(3):1220-1233

1828
UI  - 19887
AU  - Paul R
AU  - Paddison SJ
TI  - A statistical mechanical model for the calculation of the permittivity of water in hydrated polymer electrolyte membrane pores
MH  - A
MH  - membrane
MH  - model
MH  - Water
RP  - NOT IN FILE
NT  - JournalOCT 22484YTJ CHEM PHYS
UR  - ISI:000171719200048
SO  - Journal of Chemical Physics 2001  ;115(16):7762-7771

1829
UI  - 966
AU  - Pfeiffer T
AU  - Schuster S
AU  - Bonhoeffer S
TI  - Cooperation and competition in the evolution of ATP-producing pathways
AB  - Heterotrophic organisms generally face a trade-off between rate and yield of adenosine triphosphate (ATP) production. This trade-off may result in an evolutionary dilemma, because cells with a higher rate but lower yield of ATP production may gain a selective advantage when competing for shared energy resources. Using an analysis of model simulations and biochemical observations, we show that ATP production with a low rate and high yield can be viewed as a form of cooperative resource use and may evolve in spatially structured environments. Furthermore, we argue that the high ATP yield of respiration may have facilitated the evolutionary transition from unicellular to undifferentiated multicellular organisms
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - Animal
MH  - Bacteria
MH  - Carbohydrates
MH  - Cells
MH  - development
MH  - Dictyostelium
MH  - Energy Metabolism
MH  - Evolution
MH  - Fermentation
MH  - Fungi
MH  - growth &amp
MH  - Mathematics
MH  - metabolism
MH  - Models,Biological
MH  - Oxidation-Reduction
MH  - Oxidative Phosphorylation
MH  - Oxygen Consumption
MH  - Respiration
MH  - Support,Non-U.S.Gov't
MH  - Thermodynamics
RP  - NOT IN FILE
NT  - Friedrich Miescher Institute, Post Office Box 2543, CH-4002 Basel, Switzerland, Max Delbruck Center for Molecular Medicine, D-13092 Berlin, Germany
SO  - Science 2001 Apr ;%20;292(5516):504-507

1830
UI  - 20982
AU  - Possmayer FE
AU  - Hartog AF
AU  - Berden JA
AU  - Graber P
AD  - Institut fur Physikalische Chemie, Universitat Freiburg, Germany
TI  - Covalent modification of the non-catalytic sites of the H(+)-ATPase from chloroplasts with 2-azido-[alpha-(32)P]ATP and its effect on ATP synthesis and ATP hydrolysis
AB  - Incubation of the isolated H(+)-ATPase from chloroplasts, CF(0)F(1), with 2-azido-[alpha-(32)P]ATP leads to the binding of this nucleotide to different sites. These sites were identified after removal of free nucleotides, UV-irradiation and trypsin treatment by separation of the tryptic peptides by ion exchange chromatography. The nitreno-AMP, nitreno-ADP and nitreno-ATP peptides were further separated on a reversed phase column, the main fractions were subjected to amino acid sequence analysis and the derivatized tyrosines were used to distinguish between catalytic (beta-Tyr362) and non-catalytic (beta- Tyr385) sites. Several incubation procedures were developed which allow a selective occupation of each of the three non-catalytic sites. The non-catalytic site with the highest dissociation constant (site 6) becomes half maximally filled at 50 microM 2-azido-[alpha-(32)P]ATP, that with the intermediate dissociation constant (site 5) at 2 microM. The ATP at the site with the lowest dissociation constant had to be hydrolyzed first to ADP before a replacement by 2-azido-[alpha- (32)P]ATP was possible. CF(0)F(1) with non-covalently bound 2-azido- [alpha-(32)P]ATP and after covalent derivatization was reconstituted into liposomes and the rates of ATP synthesis as well as ATP hydrolysis were measured after energization of the proteoliposomes by Delta pH/Delta phi. Non-covalent binding of 2-azido-ATP to any of the three non-catalytic sites does not influence ATP synthesis and ATP hydrolysis, whereas covalent derivatization of any of the three sites inhibits both, the degree being proportional to the degree of derivatization. Extrapolation to complete inhibition indicates that derivatization of one site (either 4 or 5 or 6) is sufficient to block completely multi-site catalysis. The rates of ATP synthesis and ATP hydrolysis were measured as a function of the ADP and ATP concentration from uni-site to multi-site conditions with covalently derivatized and non-derivatized CF(0)F(1). Uni-site ATP synthesis and ATP hydrolysis were not inhibited by covalent derivatization of any of the non- catalytic sites, whereas multi-site catalysis is inhibited. These results indicate that multi-site catalysis requires some flexibility between beta- and alpha-subunits which is abolished by covalent derivatization of beta-Tyr385 with a 2-nitreno-adenine nucleotide. Conformational changes connected with energy transduction between the F(0)-part and the F(1)-part are either not required for uni-site ATP synthesis or they are not impaired by the derivatization of any of the three beta-Tyr385
MH  - 2-AZIDO-ATP
MH  - A
MH  - ACID
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - ADP
MH  - Affinity Labels
MH  - ALPHA-SUBUNIT
MH  - Amino Acid Sequence
MH  - analysis
MH  - atp
MH  - ATP synthesis
MH  - ATPase
MH  - Azides
MH  - BINDING
MH  - Catalysis
MH  - chloroplast
MH  - Chloroplasts
MH  - conformational change
MH  - CONSTANT
MH  - function
MH  - H(+)ATPase
MH  - Hydrolysis
MH  - intermediate
MH  - ion
MH  - ion exchange
MH  - Liposomes
MH  - non-catalytic site
MH  - NONCATALYTIC SITES
MH  - Nucleotides
MH  - proteoliposome
MH  - Site
MH  - synthesis
MH  - Trypsin
RP  - NOT IN FILE
NT  - UI - 21240264LA - engRN - 0 (Affinity Labels)RN - 0 (Azides)RN - 56-65-5 (Adenosine Triphosphate)RN - 58-64-0 (Adenosine Diphosphate)RN - 72884-75-4 (2-azidoadenosine 5'-triphosphate)RN - EC 3.4.21.4 (Trypsin)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 20010508IS - 0006-3002SB - IMCY - Netherlands
UR  - PM:11342174
SO  - Biochim Biophys Acta 2001 Feb 9 ;1510(1-2):378-400

1831
UI  - 9878
AU  - Possmayer FE
AU  - Hartog AF
AU  - Berden JA
AU  - Graber P
TI  - Covalent modification of the non-catalytic sites of the H+- ATPase from chloroplasts with 2-azido-[alpha-P-32]ATP and its effect on ATP synthesis and ATP hydrolysis
AB  - Incubation of the isolated H+-ATPase from chloroplasts, CF0F1, with 2-azido-[alpha-P-32]ATP leads to the binding of this nucleotide to different sites. These sites were identified after removal of free nucleotides, UV-irradiation and trypsin treatment by separation of the tryptic peptides by ion exchange chromatography. The nitreno-AMP, nitreno-ADP and nitreno-ATP peptides were further separated on a reversed phase column, the main fractions were subjected to amino acid sequence analysis and the derivatized tyrosines were used to distinguish between catalytic (beta -Tyr362) and non-catalytic (beta -Tyr385) sites. Several incubation procedures were developed which allow a selective occupation of each of the three noncatalytic sites. The non-catalytic site with the highest dissociation constant (site 6) becomes half maximally filled at 50 muM 2-azido- [alpha-P-32]ATP, that with the intermediate dissociation constant (site 5) at 2 muM. The ATP at the site with the lowest dissociation constant had to be hydrolyzed first to ADP before a replacement by 2-azido-[alpha-P-32]ATP was possible. CF0F1 with non-covalently bound 2-azido-[alpha-P-32]ATP and after covalent derivatization was reconstituted into liposomes and the rates of ATP synthesis as well as ATP hydrolysis were measured after energization of the proteoliposomes by Delta pH/Delta . Noncovalent binding of 2-azido-ATP to any of the three non-catalytic sites does not influence ATP synthesis and ATP hydrolysis, whereas covalent derivatization of any of the three sites inhibits both, the degree being proportional to the degree of derivatization. Extrapolation to complete inhibition indicates that derivatization of one site (either 4 or 5 or 6) is sufficient to block completely multi-site catalysis. The rates of ATP synthesis and ATP hydrolysis were measured as a function of the ADP and ATP concentration from uni-site to multi-site conditions with covalently derivatized and non- derivatized CF0F1. Uni-site ATP synthesis and ATP hydrolysis were not inhibited by covalent derivatization of any of the non-catalytic sites, whereas multi-site catalysis is inhibited. These results indicate that multi-site catalysis requires some flexibility between beta- and a-subunits which is abolished by covalent derivatization of beta -Tyr385 with a 2-nitreno- adenine nucleotide. Conformational changes connected with energy transduction between the F-0-part and the F-1-part are either not required for uni-site ATP synthesis or they are not impaired by the derivatization of any of the three beta - Tyr385. (C) 2001 Elsevier Science B.V. All rights reserved
MH  - 2- azido-nucleotide
MH  - 2-AZIDO-ATP
MH  - Amino Acid Sequence
MH  - analysis
MH  - atp
MH  - ATPase
MH  - BETA-SUBUNIT
MH  - Catalysis
MH  - CF0F1
MH  - Chloroplasts
MH  - COUPLING FACTOR-I
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - H+-ATPase
MH  - Hydrolysis
MH  - Liposomes
MH  - MITOCHONDRIAL ADENOSINE- TRIPHOSPHATASE
MH  - non-catalytic site
MH  - NONCATALYTIC SITES
MH  - nucleotide binding
MH  - NUCLEOTIDE BINDING-SITES
MH  - Nucleotides
MH  - SYNTHASE
MH  - THERMOPHILIC BACILLUS PS3
MH  - Trypsin
MH  - uni-site catalysis
RP  - NOT IN FILE
NT  - JournalArticleELSEVIER SCIENCE BVFEB 9404DKAMSTERDAMGraber P Univ Freiburg, Inst Chem Phys, Albertstr 23A, D-79104 Freiburg, GermanyBBA-BIOMEMBRANESPO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS
AV  - Univ Freiburg, Inst Chem Phys, Albertstr 23A, D-79104 Freiburg, Germany Univ Freiburg, Inst Chem Phys, D-79104 Freiburg, Germany Univ Amsterdam, EC Slater Inst Biochem Res, Bioctr, NL-1018 TV Amsterdam, Netherlands
UR  - ISI:000167082000033
SO  - Biochimica et Biophysica Acta-Biomembranes 2001  ;1510(1-2):378-400

1832
UI  - 9948
AU  - Sali A
TI  - Modeller 4 software
MH  - modeller
MH  - Software
PB  - http://guitar.rockefeller.edu/modeller/modeller.htmlRP  - NOT IN FILE
UR  - http://guitar.rockefeller.edu/modeller/modeller.html
SO  -  2001  ;():

1833
UI  - 21402
AU  - Schagger H
TI  - Respiratory chain supercomplexes
AB  - Respiratory chain supercomplexes have been isolated from mammalian and yeast mitochondria, and bacterial membranes. Functional roles of respiratory chain supercomplexes are catalytic enhancement, substrate channelling, and stabilization of complex I by complex III in mammalian cells. Bacterial supercomplexes are characterized by their relatively high detergent-stability compared to yeast or mammalian supercomplexes that are stable to sonication. The mobility of substrate cytochrome c increases in the order bacterial, yeast, and mammalian respiratory chain. In bacterial supercomplexes, the electron transfer between complexes III and IV involves movement of the mobile head of a tightly bound cytochrome c, whereas the yeast S. cerevisiae seems to use substrate channelling of a mobile cytochrome c, and mammalian respiratory chains have been described to use a cytochrome c pool. Dimeric ATP synthase seems to be specific for mitochondrial OXPHOS systems. Monomeric complex V was found in Acetobacterium woodii and Paracoccus denitrificans
MH  - A
MH  - Animal
MH  - atp
MH  - ATP synthase
MH  - catalytic
MH  - cell
MH  - Cell Respiration
MH  - Cells
MH  - Chemistry
MH  - COMPLEX
MH  - cytochrome
MH  - Cytochrome c
MH  - electron
MH  - electron transfer
MH  - Electron Transport
MH  - Electron-transfer
MH  - Macromolecular Systems
MH  - membrane
MH  - Membranes
MH  - metabolism
MH  - Mitochondria
MH  - Mitochondrial Proton-Translocating ATPases
MH  - Movement
MH  - Oxidative Phosphorylation
MH  - Paracoccus denitrificans
MH  - Support,Non-U.S.Gov't
MH  - SYNTHASE
MH  - SYSTEM
MH  - SYSTEMS
MH  - TRANSFER
MH  - YEAST
MH  - Yeasts
RP  - NOT IN FILE
NT  - Zentrum der Biologischen Chemie, Universitatsklinikum Frankfurt, Frankfurt am Main, Germany schagger@zbcklinikuni-frankfurtdeFAU - Schagger, H
SO  - IUBMB Life 2001 Sep ;52(3-5):119-128

1834
UI  - 21046
AU  - Schagger H
TI  - Respiratory chain supercomplexes
AB  - Respiratory chain supercomplexes have been isolated from mammalian and yeast mitochondria, and bacterial membranes. Functional roles of respiratory chain supercomplexes are catalytic enhancement, substrate channelling, and stabilization of complex I by complex III in mammalian cells. Bacterial supercomplexes are characterized by their relatively high detergent-stability compared to yeast or mammalian supercomplexes that are stable to sonication. The mobility of substrate cytochrome c increases in the order bacterial, yeast, and mammalian respiratory chain. In bacterial supercomplexes, the electron transfer between complexes III and IV involves movement of the mobile head of a tightly bound cytochrome c, whereas the yeast S. cerevisiae seems to use substrate channelling of a mobile cytochrome c, and mammalian respiratory chains have been described to use a cytochrome c pool. Dimeric ATP synthase seems to be specific for mitochondrial OXPHOS systems. Monomeric complex V was found in Acetobacterium woodii and Paracoccus denitrificans
MH  - A
MH  - atp
MH  - ATP synthase
MH  - Cells
MH  - COMPLEX
MH  - cytochrome
MH  - Cytochrome c
MH  - electron
MH  - electron transfer
MH  - Electron-transfer
MH  - membrane
MH  - Membranes
MH  - Mitochondria
MH  - Movement
MH  - SYNTHASE
MH  - SYSTEM
MH  - SYSTEMS
MH  - TRANSFER
RP  - NOT IN FILE
NT  - Zentrum der Biologischen Chemie, Universitatsklinikum Frankfurt, Frankfurt am Main, Germany schagger@zbcklinikuni-frankfurtdeFAU - Schagger, H
SO  - IUBMB Life 2001 Sep ;52(3-5):119-128

1835
UI  - 21427
AU  - Schon EA
AU  - Santra S
AU  - Pallotti F
AU  - Girvin ME
TI  - Pathogenesis of primary defects in mitochondrial ATP synthesis
AB  - Maternally inherited mutations in the mtDNA-encoded ATPase 6 subunit of complex V (ATP synthase) of the respiratory chain/oxidative phosphorylation system are responsible for a subgroup of severe and often-fatal disorders characterized predominantly by lesions in the brain, particularly in the striatum. These include NARP (neuropathy, ataxia, and retinitis pigmentosa), MILS (maternally inherited Leigh syndrome), and FBSN (familial bilateral striatal necrosis). Of the five known pathogenic mutations causing these disorders, four are located at two codons (156 and 217), each of which can suffer mutations converting a conserved leucine to either an arginine or a proline. Based on the accumulating data on both the structure of ATP synthase and the mechanism by which rotary catalysis couples proton flow to ATP synthesis, we propose a model that may help explain why mutations at codons 156 and 217 are pathogenic
MH  - A
MH  - Adenosine Triphosphate
MH  - Amino Acid Sequence
MH  - Arginine
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - ATPase
MH  - biosynthesis
MH  - Brain
MH  - Catalysis
MH  - COMPLEX
MH  - data
MH  - enzymology
MH  - genetics
MH  - Human
MH  - Leucine
MH  - mechanism
MH  - Mitochondria
MH  - Mitochondrial Diseases
MH  - Mitochondrial Proton-Translocating ATPases
MH  - model
MH  - Mutation
MH  - Phosphorylation
MH  - primary
MH  - proton
MH  - structure
MH  - SUBUNIT
MH  - Support,Non-U.S.Gov't
MH  - Support,U.S.Gov't,P.H.S.
MH  - SYNTHASE
MH  - synthesis
MH  - SYSTEM
MH  - universities
RP  - NOT IN FILE
NT  - Columbia University College of Physicians and Surgeons, New York, NY, USA pas3@columbiaeduFAU - Schon, E A
SO  - Semin Cell Dev Biol 2001 Dec ;12(6):441-448

1836
UI  - 21066
AU  - Schon EA
AU  - Santra S
AU  - Pallotti F
AU  - Girvin ME
TI  - Pathogenesis of primary defects in mitochondrial ATP synthesis
AB  - Maternally inherited mutations in the mtDNA-encoded ATPase 6 subunit of complex V (ATP synthase) of the respiratory chain/oxidative phosphorylation system are responsible for a subgroup of severe and often-fatal disorders characterized predominantly by lesions in the brain, particularly in the striatum. These include NARP (neuropathy, ataxia, and retinitis pigmentosa), MILS (maternally inherited Leigh syndrome), and FBSN (familial bilateral striatal necrosis). Of the five known pathogenic mutations causing these disorders, four are located at two codons (156 and 217), each of which can suffer mutations converting a conserved leucine to either an arginine or a proline. Based on the accumulating data on both the structure of ATP synthase and the mechanism by which rotary catalysis couples proton flow to ATP synthesis, we propose a model that may help explain why mutations at codons 156 and 217 are pathogenic
MH  - A
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - ATPase
MH  - Brain
MH  - Catalysis
MH  - COMPLEX
MH  - mechanism
MH  - model
MH  - Phosphorylation
MH  - proton
MH  - structure
MH  - SUBUNIT
MH  - SYNTHASE
MH  - synthesis
MH  - SYSTEM
RP  - NOT IN FILE
NT  - Columbia University College of Physicians and Surgeons, New York, NY, USAFAU - Schon, E A
SO  - Semin Cell Dev Biol 2001 Dec ;12(6):441-448

1837
UI  - 21259
AU  - Shinkarev VP
AU  - Crofts AR
AU  - Wraight CA
AD  - Department of Biochemistry, University of Illinois at Urbana-Champaign, 156 Davenport Hall, 607 South Mathews Avenue, Urbana, Illinois 6l80l, USA vshinkar@uiucedu
TI  - The electric field generated by photosynthetic reaction center induces rapid reversed electron transfer in the bc1 complex
AB  - The cytochrome bc(1) complex is the central enzyme of respiratory and photosynthetic electron-transfer chains. It couples the redox work of quinol oxidation and cytochrome reduction to the generation of a proton gradient needed for ATP synthesis. When the quinone processing Q(i)- and Q(o)-sites of the complex are inhibited by both antimycin and myxothiazol, the flash-induced kinetics of the b-heme chain, which transfers electrons between these sites, are also expected to be inhibited. However, we have observed in Rhodobacter sphaeroides chromatophores, that when a fraction of heme b(H) is reduced, flash excitation induces fast (half-time approximately 0.1 ms) oxidation of heme b(H), even in the presence of antimycin and myxothiazol. The sensitivity of this oxidation to ionophores and uncouplers, and the absence of any delay in the onset of this reaction, indicates that it is due to a reversal of electron transfer between b(L) and b(H) hemes, driven by the electrical field generated by the photosynthetic reaction center. In the presence of antimycin A, but absence of myxothiazol, the second and following flashes induce a similar ( approximately 0.1 ms) transient oxidation of approximately 10% of the cytochrome b(H) reduced on the first flash. From the observed amplitude of the field-induced oxidation of heme b(H), we estimate that the equilibrium constant for sharing one electron between hemes b(L) and b(H) is 10-15 at pH 7. The small value of this equilibrium constant modifies our understanding of the thermodynamics of the Q-cycle, especially in the context of a dimeric structure of bc(1) complex
MH  - A
MH  - Antimycin A
MH  - atp
MH  - ATP synthesis
MH  - Biochemistry
MH  - chromatophore
MH  - chromatophores
MH  - COMPLEX
MH  - CONSTANT
MH  - cytochrome
MH  - electron
MH  - electron transfer
MH  - Electron-transfer
MH  - Electrons
MH  - FIELD
MH  - flash
MH  - Ionophores
MH  - Kinetics
MH  - pH
MH  - proton
MH  - quinone
MH  - reaction center
MH  - redox
MH  - Rhodobacter
MH  - Rhodobacter sphaeroides
MH  - Site
MH  - sphaeroides
MH  - structure
MH  - synthesis
MH  - Thermodynamics
MH  - Thiazoles
MH  - TRANSFER
MH  - Ubiquinol-Cytochrome-c Reductase
RP  - NOT IN FILE
NT  - UI - 21488063LA - engRN - 0 (Photosynthetic Reaction Center, Bacterial)RN - 0 (Thiazoles)RN - 11118-72-2 (antimycin)RN - 14875-96-8 (Heme)RN - 642-15-9 (Antimycin A)RN - 76706-55-3 (myxothiazol)RN - EC 1.10.2.2 (Ubiquinol-Cytochrome-c Reductase)PT - Journal ArticleID - GM 53508/GM/NIGMSDA - 20011016IS - 0006-2960SB - IMCY - United States
UR  - PM:11601982
SO  - Biochemistry 2001 Oct 23 ;40(42):12584-12590

1838
UI  - 21404
AU  - Sigyarto C
AU  - Hugosson M
AU  - Moberg P
AU  - Andreu D
AU  - Glaser E
TI  - L and D presequence peptides derived from the precursor of F1beta subunit of the ATP synthase inhibit mitochondrial protein import by interaction with import machinery
AB  - We investigated the effect of L and D enantiomers of a 25-residue peptide derived from the N-terminal region of the presequence of Nicotiana plumbaginifolia F1beta subunit of the ATP synthase, pF1beta(1, 25), on import into spinach leaf mitochondria. Three in vitro synthesized precursor proteins using different import pathways were used. Import of the precursor proteins of F1beta subunit of the ATP synthase, pre-F1beta, and the alternative oxidase, pre-AOX, required addition of external ATP. whereas the chimeric precursor containing the N-terminal 84 amino acids of the cytochrome b2 precursor protein linked to dihydrofolate reductase, pre-b2(1, 84)-DHFR was not dependent on ATP. Import of pre-F1beta, and pre-AOX was inhibited already at 1 microM and 3 microM concentration of the L and D enantiomers, whereas inhibition of import of pre-b2(1, 84)-DHFR, occurred at concentrations >10 microM of both enantiomers. Binding efficiency of the precursor proteins was not affected by addition of the L and D enantiomers. There was no correlation between inhibition of import of pre-F1beta and pre-AOX and dissipation of membrane potential measured as a decrease of Rhodamine 123 fluorescence quenching. The inhibitory effect of the L and D presequence enantiomers on import of pre-F1beta and pre-AOX was concluded to occur within the outer membrane translocase machinery beyond the initial precursor receptor interaction. Furthermore, the fact that the D enantiomer had the same effect as the natural peptide showed that interaction of the presequence with the import machinery was not dependent on chiral properties of the presequence
MH  - A
MH  - ACID
MH  - Amino Acids
MH  - atp
MH  - ATP synthase
MH  - BINDING
MH  - Biochemistry
MH  - Biological Transport
MH  - Biophysics
MH  - Chemistry
MH  - cytochrome
MH  - Enzyme Precursors
MH  - fluorescence
MH  - genetics
MH  - In Vitro
MH  - INTERACTION
MH  - membrane
MH  - Membrane Potential
MH  - Membrane Potentials
MH  - metabolism
MH  - Mitochondria
MH  - Oxidoreductases
MH  - peptide
MH  - peptides
MH  - physiology
MH  - Plant Proteins
MH  - protein
MH  - Protein Binding
MH  - Protein Sorting Signals
MH  - Protein Subunits
MH  - Proteins
MH  - Proton-Translocating ATPases
MH  - Spinach
MH  - Stereoisomerism
MH  - SUBUNIT
MH  - Support,Non-U.S.Gov't
MH  - SYNTHASE
MH  - universities
RP  - NOT IN FILE
NT  - Department of Biochemistry and Biophysics, Stockholm University, SwedenFAU - Sigyarto, C
SO  - Plant Mol Biol 2001 Dec ;47(6):815-826

1839
UI  - 21047
AU  - Sigyarto C
AU  - Hugosson M
AU  - Moberg P
AU  - Andreu D
AU  - Glaser E
TI  - L and D presequence peptides derived from the precursor of F1beta subunit of the ATP synthase inhibit mitochondrial protein import by interaction with import machinery
AB  - We investigated the effect of L and D enantiomers of a 25-residue peptide derived from the N-terminal region of the presequence of Nicotiana plumbaginifolia F1beta subunit of the ATP synthase, pF1beta(1, 25), on import into spinach leaf mitochondria. Three in vitro synthesized precursor proteins using different import pathways were used. Import of the precursor proteins of F1beta subunit of the ATP synthase, pre-F1beta, and the alternative oxidase, pre-AOX, required addition of external ATP. whereas the chimeric precursor containing the N-terminal 84 amino acids of the cytochrome b2 precursor protein linked to dihydrofolate reductase, pre-b2(1, 84)-DHFR was not dependent on ATP. Import of pre-F1beta, and pre-AOX was inhibited already at 1 microM and 3 microM concentration of the L and D enantiomers, whereas inhibition of import of pre-b2(1, 84)-DHFR, occurred at concentrations >10 microM of both enantiomers. Binding efficiency of the precursor proteins was not affected by addition of the L and D enantiomers. There was no correlation between inhibition of import of pre-F1beta and pre-AOX and dissipation of membrane potential measured as a decrease of Rhodamine 123 fluorescence quenching. The inhibitory effect of the L and D presequence enantiomers on import of pre-F1beta and pre-AOX was concluded to occur within the outer membrane translocase machinery beyond the initial precursor receptor interaction. Furthermore, the fact that the D enantiomer had the same effect as the natural peptide showed that interaction of the presequence with the import machinery was not dependent on chiral properties of the presequence
MH  - A
MH  - ACID
MH  - Amino Acids
MH  - atp
MH  - ATP synthase
MH  - BINDING
MH  - Biochemistry
MH  - Biological Transport
MH  - Biophysics
MH  - Chemistry
MH  - cytochrome
MH  - Enzyme Precursors
MH  - fluorescence
MH  - genetics
MH  - In Vitro
MH  - membrane
MH  - Membrane Potential
MH  - Membrane Potentials
MH  - metabolism
MH  - Mitochondria
MH  - Oxidoreductases
MH  - physiology
MH  - Plant Proteins
MH  - protein
MH  - Protein Binding
MH  - Protein Sorting Signals
MH  - Protein Subunits
MH  - Proteins
MH  - Proton-Translocating ATPases
MH  - Spinach
MH  - Stereoisomerism
MH  - SUBUNIT
MH  - Support,Non-U.S.Gov't
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - Department of Biochemistry and Biophysics, Stockholm University, SwedenFAU - Sigyarto, C
SO  - Plant Mol Biol 2001 Dec ;47(6):815-826

1840
UI  - 4
AU  - Stahlberg H
AU  - Muller DJ
AU  - Suda K
AU  - Fotiadis D
AU  - Engel A
AU  - Meier T
AU  - Matthey U
AU  - Dimroth P
AD  - ME Muller Institute for Structural Biology, Biozentrum, University of Basel, Klingelbergstrasse 70, CH-4056 Basel, Switzerland
TI  - Bacterial Na(+)-ATP synthase has an undecameric rotor
AB  - Synthesis of adenosine triphosphate (ATP) by the F(1)F(0) ATP synthase involves a membrane-embedded rotary engine, the F(0) domain, which drives the extra-membranous catalytic F(1) domain. The F(0) domain consists of subunits a(1)b(2) and a cylindrical rotor assembled from 9- 14 alpha-helical hairpin-shaped c-subunits. According to structural analyses, rotors contain 10 c-subunits in yeast and 14 in chloroplast ATP synthases. We determined the rotor stoichiometry of Ilyobacter tartaricus ATP synthase by atomic force microscopy and cryo-electron microscopy, and show the cylindrical sodium-driven rotor to comprise 11 c-subunits
RP  - NOT IN FILE
NT  - UI - 21174503LA - engRN - 0 (Protein Subunits)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 20010329IS - 1469-221XSB - IMCY - EnglandJC - D0T
UR  - PM:11266365
SO  - EMBO Rep 2001 Mar ;2(3):229-233

1841
UI  - 678
AU  - Suzuki T
AU  - Hisabori T
AU  - Yoshida M
TI  - [ATPase and ATP synthase]
RP  - NOT IN FILE
NT  - UI - 21464019LA - jpnPT - Journal ArticleDA - 20011002IS - 0039-9450SB - IMCY - JapanJC - Q7D
UR  - PM:11579564
SO  - Tanpakushitsu Kakusan Koso 2001 Aug ;46(11 Suppl):1668-1678

1842
UI  - 20833
AU  - Tanabe M
AU  - Nishio K
AU  - Iko Y
AU  - Sambongi Y
AU  - Iwamoto-Kihara A
AU  - Wada Y
AU  - Futai M
AD  - Division of Biological Sciences, Institute of Scientific and Industrial Research, Osaka University, Ibaraki, Osaka 567-0047, Japan
TI  - Rotation of a complex of the gamma subunit and c ring of Escherichia coli ATP synthase. The rotor and stator are interchangeable
AB  - ATP synthase (F0F1) transforms an electrochemical proton gradient into chemical energy (ATP) through the rotation of a subunit assembly. It has been suggested that a complex of the gamma subunit and c ring (c(10- 14)) of F0F1 could rotate together during ATP hydrolysis and synthesis (Sambongi, Y., Iko, Y., Tanabe, M., Omote, H., Iwamoto-Kihara, A., Ueda, I., Yanagida, T., Wada, Y., and Futai, M. (1999) Science 286, 1722-1724). We observed that the rotation of the c ring with the cI28T mutation (c subunit cIle-28 replaced by Thr) was less sensitive to venturicidin than that of the wild type, consistent with the antibiotic effect on the cI28T mutant and wild-type ATPase activities (Fillingame, R. H., Oldenburg, M., and Fraga, D. (1991) J. Biol. Chem. 266, 20934- 20939). Furthermore, we engineered F0F1 to see the alpha(3)beta(3) hexamer rotation; a biotin tag was introduced into the alpha or beta subunit, and a His tag was introduced into the c subunit. The engineered enzymes could be purified by metal affinity chromatography and density gradient centrifugation. They were immobilized on a glass surface through the c subunit, and an actin filament was connected to the alpha or beta subunit. The filament rotated upon the addition of ATP and generated essentially the same frictional torque as one connected to the c ring. These results indicate that the gammaepsilonc(10-14) complex is a mechanical unit of the enzyme and that it can be used as a rotor or a stator experimentally, depending on the subunit immobilized
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BETA-SUBUNIT
MH  - COLI ATP SYNTHASE
MH  - COMPLEX
MH  - Dicyclohexylcarbodiimide
MH  - Enzyme Inhibitors
MH  - Enzymes
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0F1
MH  - Hydrolysis
MH  - M
MH  - mutant
MH  - proton
MH  - rotation
MH  - SUBUNIT
MH  - SURFACE
MH  - SYNTHASE
MH  - synthesis
RP  - NOT IN FILE
NT  - UI - 21226775LA - engRN - 0 (Actins)RN - 0 (Enzyme Inhibitors)RN - 0 (Plasmids)RN - 0 (Venturicidins)RN - 11021-88-8 (venturicidin)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleDA - 20010430IS - 0021-9258SB - IMCY - United States
UR  - PM:11279047
SO  - J Biol Chem 2001 May 4 ;276(18):15269-15274

1843
UI  - 21069
AU  - Tavakoli N
AU  - Kluge C
AU  - Golldack D
AU  - Mimura T
AU  - Dietz KJ
TI  - Reversible redox control of plant vacuolar H+-ATPase activity is related to disulfide bridge formation in subunit E as well as subunit A
AB  - The plant vacuolar proton pump can be subjected to reversible redox regulation in vitro. The redox-dependent activity change involves disulfide bridge formation not only in Vatp A, as reported for bovine V-ATPase, but also in the stalk subunit Vatp E. Microsomal membranes isolated from barley leaves were analysed for their activity of bafilomycin-sensitive ATP hydrolysis and proton pumping using quinacrine fluorescence quenching in vesicle preparations. ATP hydrolysis and proton pumping activity were inhibited by H2O2. H2O2-deactivated ATPase was reactivated by cysteine and glutathione. The glutathione concentration needed for half maximal reactivation was 1 mmol l-1. The activity loss was accompanied by shifts in electrophoretic mobility of Vatp A and E which were reversed upon reductive reactivation. The redox-dependent shift was also seen with recombinant Vatp E, and was absent following site-directed mutagenesis of either of the two cys residues conserved throughout all plant Vatp E sequences. V-ATPase was also inhibited by oxidized thioredoxin. These results support the hypothesis that tuning of vacuolar ATPase activity can be mediated by redox control depending on the metabolic requirements
MH  - A
MH  - atp
MH  - ATPase
MH  - Cysteine
MH  - fluorescence
MH  - H+-ATPase
MH  - Hydrolysis
MH  - In Vitro
MH  - membrane
MH  - Membranes
MH  - mutagenesis
MH  - plant
MH  - proton
MH  - Proton Pump
MH  - redox
MH  - regulation
MH  - RESIDUE
MH  - site-directed
MH  - stalk
MH  - SUBUNIT
RP  - NOT IN FILE
NT  - Lehrstuhl fur Stoffwechselphysiologie und Biochemie der Pflanzen, W5, Universitat Bielefeld, 33501 Bielefeld, GermanyFAU - Tavakoli, N
SO  - Plant J 2001 Oct ;28(1):51-59

1844
UI  - 19889
AU  - Teschke O
AU  - Ceotto G
AU  - de Souza EF
TI  - Interfacial water dielectric-permittivity-profile measurements using atomic force microscopy - art. no. 011605
MH  - Microscopy
MH  - Water
RP  - NOT IN FILE
NT  - JournalJUL1453GAPHYS REV E
UR  - ISI:000169907100061
SO  - Physical Review e 2001  ;6401(1):011605-1-011605-10

1845
UI  - 19888
AU  - Teschke O
AU  - Ceotto G
AU  - de Souza EF
TI  - Rupture force of adsorbed self-assembled surfactant layers - Effect of the dielectric exchange force
RP  - NOT IN FILE
NT  - JournalAUG 31470FYCHEM PHYS LETT
UR  - ISI:000170860900001
SO  - Chemical Physics Letters 2001  ;344(5-6):429-433

1846
UI  - 21070
AU  - Thelly TM
AU  - Mohankumar C
TI  - H+-ATPase as a biochemical marker for early detection of root (wilt) disease in coconut palms (Cocos nucifera L)
AB  - H+-ATPase activity in leaves and roots of coconut palms growing in 'root wilt disease-prevalent areas' was compared with that of coconut palms growing in 'disease-free areas'. The activity was found to be significantly less in the leaves and roots of palms in the disease-prevalent zone as compared to that in disease-free zone. Histochemical examination of the leaves showed results that corroborated the biochemical findings. The possible application of H+-ATPase activity as a marker for the early detection of wilt disease in coconut palms is suggested
MH  - A
MH  - Biochemistry
MH  - Chemistry
MH  - Coconut
MH  - enzymology
MH  - H+-ATPase
MH  - isolation & purification
MH  - M
MH  - metabolism
MH  - Microscopy,Electron,Scanning
MH  - plant
MH  - Plant Diseases
MH  - Plant Leaves
MH  - Proton-Translocating ATPases
MH  - ultrastructure
RP  - NOT IN FILE
NT  - Plant Biochemistry Unit, Department of Botany, University College, Thiruvananthapuram, KeralaFAU - Thelly, T M
SO  - Indian J Biochem Biophys 2001 Jun ;38(3):199-202

1847
UI  - 21155
AU  - Tomashek JJ
AU  - Poposki JA
AU  - Brusilow WS
AD  - Wayne State University School of Medicine, Department of Biochemistry, Detroit, Michigan 48201, USA
TI  - A functional His-tagged c subunit of the Escherichia coli F-type ATPase/Synthase
AB  - The c subunit of the Escherichia coli F0 has been tagged with a hexahistidine motif at its C-terminus. The tagged subunit is capable of forming functional F0 complexes that translocate protons in the absence of the F1 complex. In the presence of F1, the two sectors associate and display all biochemical activities of the wildtype enzyme: DCCD- inhibitable ATPase activity, ATP synthase activity, and ATP-dependent proton pumping. The enzyme can be solubilized and purified as an intact complex under native conditions on immobilized-metal affinity chromatography (IMAC) resin. The purified complex can be reincorporated into liposomes and demonstrates ATP-dependent proton pumping activity. Hexahistine tags placed at the N-terminus, in contrast, were all inactive. These experiments demonstrate the feasibility of tagging the c subunit for further studies of the F0 and suggest an important role for the N-terminus of the c subunit in either assembly or function of the protein
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Biochemistry
MH  - COMPLEX
MH  - Dicyclohexylcarbodiimide
MH  - Enzyme Inhibitors
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - F1
MH  - function
MH  - Histidine
MH  - inhibitor
MH  - liposome
MH  - Liposomes
MH  - protein
MH  - Protein Subunits
MH  - proton
MH  - Proton Pump
MH  - Proton-Translocating ATPases
MH  - Protons
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 21262977LA - engRN - 0 (Enzyme Inhibitors)RN - 0 (Liposomes)RN - 0 (Plasmids)RN - 0 (Protein Subunits)RN - 0 (Proton Pumps)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - 56-65-5 (Adenosine Triphosphate)RN - 71-00-1 (Histidine)RN - EC 3.6.1.- (mitochondrial ATPase subunit c)RN - EC 3.6.3.14 (Proton-Translocating ATPases)PT - Journal ArticleID - F32-GM20422/GM/NIGMSID - R01-GM54108/GM/NIGMSDA - 20010523IS - 0003-9861SB - IMCY - United States
UR  - PM:11370839
SO  - Arch Biochem Biophys 2001 Mar 15 ;387(2):180-187

1848
UI  - 679
AU  - Tozawa K
AU  - Yagi H
AU  - Hisamatsu K
AU  - Ozawa K
AU  - Yoshida M
AU  - Akutsu H
AD  - Department of Chemistry and Biotechnology, Faculty of Engineering, Yokohama National University, Hodogaya-ku, Yokohama 240-8501, Japan akutsu@proteinosaka-uacjp
TI  - Functions and ATP-Binding Responses of the Twelve Histidine Residues in the TF(1)-ATPase beta Subunit
AB  - The C2 proton signals of all (twelve) histidine residues of the TF(1) beta subunit in the (1)H-NMR spectrum have been identified and assigned by means of pH change experiments and site-directed substitution of histidines by glutamines. pH and ligand titration experiments were carried out for these signals. Furthermore, the ATPase activity of the reconstituted alpha(3)beta(3)gamma complex was examined for the twelve mutant beta subunits. Two of three conserved histidines, namely, His- 119 and 324, were found to be important for expression of the ATPase activity. The former fixes the N-terminal domain to the central domain. His-324 is involved in the formation of the interface essential for the alpha(3)beta(3)gamma complex assembly. The other conserved residue, His- 363, showed a very low pK(a), suggesting that it is involved in the tertiary structure formation. On the binding of a nucleotide, only the signals of His-173, 179, 200, and 324 shifted. These histidines are located in the hinge region, and its proximity, of the beta subunit. This observation provided further support for the conformational change of the beta monomer from the open to the closed form on the binding of a nucleotide proposed by us [Yagi et al. (1999) Biophys. J. 77, 2175- 2183]. This conformational change should be one of the essential driving forces in the rotation of the alpha(3)beta(3)gamma complex
RP  - NOT IN FILE
NT  - UI - 21458708LA - engPT - Journal ArticleDA - 20010927IS - 0021-924XSB - IMCY - JapanJC - HIF
UR  - PM:11574072
SO  - J Biochem (Tokyo ) 2001 Oct ;130(4):527-533

1849
UI  - 130
AU  - Tsunoda SP
AU  - Aggeler R
AU  - Yoshida M
AU  - Capaldi RA
AD  - Institute of Molecular Biology, University of Oregon, Eugene, OR 97403- 1229, USA
TI  - Rotation of the c subunit oligomer in fully functional F1Fo ATP synthase
AB  - The F(1)F(o)-type ATP synthase is the smallest motor enzyme known. Previous studies had established that the central gamma and varepsilon subunits of the F(1) part rotate relative to a stator of alpha(3)beta(3) and delta subunits during catalysis. We now show that the ring of c subunits in the F(o) part moves along with the gamma and varepsilon subunits. This was demonstrated by linking the three rotor subunits with disulfide bridges between cysteine residues introduced genetically at the interfaces between the gamma, varepsilon, and c subunits. Essentially complete cross-linking of the gamma, varepsilon, and c subunits was achieved by using CuCl(2) to induce oxidation. This fixing of the three subunits together had no significant effect on ATP hydrolysis, proton translocation, or ATP synthesis, and each of these functions retained inhibitor sensitivity. These results unequivocally place the c subunit oligomer in the rotor part of this molecular machine
RP  - NOT IN FILE
NT  - UI - 21107643LA - engRN - 0 (Macromolecular Systems)RN - 0 (Protein Subunits)RN - 0 (Recombinant Proteins)RN - 28380-24-7 (Nigericin)RN - 538-75-0 (Dicyclohexylcarbodiimide)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - HL24526/HL/NHLBIDA - 20010222IS - 0027-8424SB - IMCY - United StatesJC - PV3
UR  - PM:11158567
SO  - Proc Natl Acad Sci U S A 2001 Jan 30 ;98(3):898-902

1850
UI  - 128
AU  - Tsunoda SP
AU  - Rodgers AJ
AU  - Aggeler R
AU  - Wilce MC
AU  - Yoshida M
AU  - Capaldi RA
AD  - Institute of Molecular Biology, University of Oregon, Eugene, OR 97403- 1229, USA
TI  - Large conformational changes of the epsilon subunit in the bacterial F1F0 ATP synthase provide a ratchet action to regulate this rotary motor enzyme
AB  - The F(1)F(0) ATP synthase is the smallest motor enzyme known. Previous studies had established that the central stalk, made of the gamma and epsilon subunits in the F(1) part and c subunit ring in the F(0) part, rotates relative to a stator composed of alpha(3)beta(3)deltaab(2) during ATP hydrolysis and synthesis. How this rotation is regulated has been less clear. Here, we show that the epsilon subunit plays a key role by acting as a switch of this motor. Two different arrangements of the epsilon subunit have been visualized recently. The first has been observed in beef heart mitochondrial F(1)-ATPase where the C-terminal portion is arranged as a two-alpha-helix hairpin structure that extends away from the alpha(3)beta(3) region, and toward the position of the c subunit ring in the intact F(1)F(0). The second arrangement was observed in a structure determination of a complex of the gamma and epsilon subunits of the Escherichia coli F(1)-ATPase. In this, the two C-terminal helices are apart and extend along the gamma to interact with the alpha and beta subunits in the intact complex. We have been able to trap these two arrangements by cross-linking after introducing appropriate Cys residues in E. coli F(1)F(0), confirming that both conformations of the epsilon subunit exist in the enzyme complex. With the C-terminal domain of varepsilon toward the F(0), ATP hydrolysis is activated, but the enzyme is fully coupled in both ATP hydrolysis and synthesis. With the C-terminal domain toward the F(1) part, ATP hydrolysis is inhibited and yet the enzyme is fully functional in ATP synthesis; i.e., it works in one direction only. These results help explain the inhibitory action of the varepsilon subunit in the F(1)F(0) complex and argue for a ratchet function of this subunit
RP  - NOT IN FILE
NT  - UI - 21287255LA - engRN - 0 (Protein Subunits)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleID - HL24526/HL/NHLBIDA - 20010606IS - 0027-8424SB - IMCY - United StatesJC - PV3
UR  - PM:11381110
SO  - Proc Natl Acad Sci U S A 2001 Jun 5 ;98(12):6560-6564

1851
UI  - 400
AU  - Weber J
AU  - Senior AE
AD  - Department of Biochemistry and Biophysics, University of Rochester Medical Center, Rochester, New York 14642
TI  - Bi-site catalysis in f1-atpase: does it exist?
AB  - The mechanism of action of F(1)F(0)-ATP synthase is controversial. Some favor a tri-site mechanism, where substrate must fill all three catalytic sites for activity, others a bi-site mechanism, where one of the three sites is always unoccupied. New approaches were applied to examine this question. First, ITP was used as hydrolysis substrate; lower binding affinities of ITP versus ATP enable more accurate assessment of sites occupancy. Second, distributions of all eight possible enzyme species (with zero, one, two or three sites filled) as fraction of total enzyme population at each ITP concentration were calculated, and compared with measured ITPase activity. Confirming data were obtained with ATP as substrate. Third, we performed a theoretical analysis of possible bi-site mechanisms. The results argue convincingly that bi-site hydrolysis activity is negligible, and may not even exist. Effectively, tri-site hydrolysis is the only mechanism. We argue that only tri-site hydrolysis drives subunit rotation. Theoretical analyses of possible bi-site mechanisms reveal serious flaws, not previously recognized. One is that, in bi-site catalysis, the predicted direction of subunit rotation is the same for both ATP synthesis and hydrolysis; a second is that infrequently occurring enzyme species are required
RP  - NOT IN FILE
NT  - UI - 21443722LA - engPT - Journal ArticleDA - 20010917IS - 0021-9258SB - IMCY - United StatesJC - HIV
UR  - PM:11451960
SO  - J Biol Chem 2001 Sep 21 ;276(38):35422-35428

1852
UI  - 21385
AU  - Wilkens S
TI  - Structure of the vacuolar adenosine triphosphatases
AB  - Vacuolar adenosine triphosphatases (V-ATPases) represent an important class of proton pumps found in endomembrane systems of eucaryotic cells, where they are involved in pH regulation. Progress has been made in the structure determination of this large, membrane-bound multisubunit enzyme complex. Electron microscopy of the V-ATPase has revealed a ball-and-stalk-like structure similar to F1F0-type ATP synthase, to which the V-ATPase is evolutionary related. Aside from the overall structural similarity of the V-ATPase and F-ATP synthase, a number of distinct structural differences exist between the two related enzymes, giving clues to their different function and regulation in the organism
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - ADENOSINE-TRIPHOSPHATASE
MH  - Animal
MH  - atp
MH  - ATP synthase
MH  - Biochemistry
MH  - Brain
MH  - Cattle
MH  - cell
MH  - Cells
MH  - Chemistry
MH  - COMPLEX
MH  - electron
MH  - electron microscopy
MH  - ELECTRON-MICROSCOPY
MH  - enzyme
MH  - Enzymes
MH  - function
MH  - Hydrogen-Ion Concentration
MH  - Hydrolysis
MH  - metabolism
MH  - Microscopy
MH  - Microscopy,Electron
MH  - Models,Biological
MH  - pH
MH  - physiology
MH  - Protein Conformation
MH  - Protein Structure,Tertiary
MH  - proton
MH  - Proton Pump
MH  - Proton Pumps
MH  - Proton-Translocating ATPases
MH  - Protons
MH  - regulation
MH  - structure
MH  - Support,U.S.Gov't,P.H.S.
MH  - SYNTHASE
MH  - SYSTEM
MH  - SYSTEMS
MH  - ultrastructure
MH  - universities
RP  - NOT IN FILE
NT  - University of California, Riverside, Department of Biochemistry, 92521, USA stephanwilkens@ucreduFAU - Wilkens, S
SO  - Cell Biochem Biophys 2001  ;34(2):191-208

1853
UI  - 682
AU  - Yasuda R
AU  - Noji H
AU  - Yoshida M
AU  - Kinosita K
AU  - Itoh H
AD  - CREST 'Genetic Programming' Team 13, Teikyo University Biotechnology Center 3F, Nogawa 907, Miyamae-Ku, Kawasaki 216-0001, Japan
TI  - Resolution of distinct rotational substeps by submillisecond kinetic analysis of F1-ATPase
AB  - The enzyme F1-ATPase has been shown to be a rotary motor in which the central gamma-subunit rotates inside the cylinder made of alpha3beta3 subunits. At low ATP concentrations, the motor rotates in discrete 120 degrees steps, consistent with sequential ATP hydrolysis on the three beta-subunits. The mechanism of stepping is unknown. Here we show by high-speed imaging that the 120 degrees step consists of roughly 90 degrees and 30 degrees substeps, each taking only a fraction of a millisecond. ATP binding drives the 90 degrees substep, and the 30 degrees substep is probably driven by release of a hydrolysis product. The two substeps are separated by two reactions of about 1 ms, which together occupy most of the ATP hydrolysis cycle. This scheme probably applies to rotation at full speed ( approximately 130 revolutions per second at saturating ATP) down to occasional stepping at nanomolar ATP concentrations, and supports the binding-change model for ATP synthesis by reverse rotation of F1-ATPase
RP  - NOT IN FILE
NT  - UI - 21206300LA - engRN - 0 (Molecular Motors)RN - 0 (Protein Subunits)RN - 56-65-5 (Adenosine Triphosphate)RN - EC 3.6.1.34 (H(+)-Transporting ATP Synthase)PT - Journal ArticleDA - 20010419IS - 0028-0836SB - IMCY - EnglandJC - NSC
UR  - PM:11309608
SO  - Nature 2001 Apr 19 ;410(6831):898-904

1854
UI  - 680
AU  - Yoshida M
AU  - Muneyuki E
AU  - Hisabori T
AD  - [1] Chemical Resources Laboratory, Tokyo Institute of Technology, Nagatsuta 4259, Yokohama 226-8503, Japan [2] myoshida@restitechacjp
TI  - Atp synthase - a marvellous rotary engine of the cell
AB  - ATP synthase can be thought of as a complex of two motors - the ATP- driven F1 motor and the proton-driven Fo motor - that rotate in opposite directions. The mechanisms by which rotation and catalysis are coupled in the working enzyme are now being unravelled on a molecular scale
RP  - NOT IN FILE
NT  - UI - 21424808LA - engPT - Journal ArticleDA - 20010904IS - 1471-0072SB - IMCY - EnglandJC - D0L
UR  - PM:11533724
SO  - Nat Rev Mol Cell Biol 2001 Sep ;2(9):669-677

1855
UI  - 21437
AU  - Zhang X
AU  - Moye-Rowley WS
TI  - Saccharomyces cerevisiae multidrug resistance gene expression inversely correlates with the status of the F(0) component of the mitochondrial ATPase
AB  - Loss of the mitochondrial genome (rho(0) cell) or elimination of the mitochondrial inner membrane protein Oxa1p causes a dramatic increase in expression of the ATP binding cassette transporter-encoding gene PDR5 in the yeast Saccharomyces cerevisiae. This increase in gene expression occurs via activation of the function of the Cys(6)-Zn(II)(2) cluster transcription factor Pdr3p, which in turn autoregulates expression of its structural gene. Surprisingly, the acquisition of PDR5-dependent multidrug resistance occurs at a very high frequency, consistent with the appearance of rho(-) cells in a fermentatively growing culture (approximately 2%). The degree of activation of Pdr3p target genes was found to vary considerably and to be influenced by the presence of the homologous protein, Pdr1p. Mutagenesis and overexpression studies provided evidence that the control of Pdr3p expression was the major control point of this transcription factor by mitochondrial retrograde signaling. Because both rho(0) and oxa1 mutant cells have multiple defects including loss of normal respiratory chain function and oxidative phosphorylation, a series of mutant strains with more selective defects in mitochondrial function was employed to identify the molecular signal that triggers PDR5 transcriptional activation. Only mutations that influenced the functional status of the F(0) subunit of the mitochondrial ATPase were found to lead to activation of PDR5 expression
MH  - A
MH  - ACTIVATION
MH  - Adenosinetriphosphatase
MH  - atp
MH  - ATP-Binding Cassette Transporters
MH  - ATPase
MH  - Base Sequence
MH  - BINDING
MH  - biology
MH  - cell
MH  - Cells
MH  - Cycloheximide
MH  - DNA Primers
MH  - DNA-Binding Proteins
MH  - Drug Resistance,Multiple
MH  - function
MH  - Gene Deletion
MH  - Gene Expression
MH  - Genes,Fungal
MH  - genetics
MH  - membrane
MH  - Membrane Proteins
MH  - metabolism
MH  - mutagenesis
MH  - mutant
MH  - Mutation
MH  - Oxidative Phosphorylation
MH  - pharmacology
MH  - Phosphorylation
MH  - Promoter Regions (Genetics)
MH  - protein
MH  - Proton-Translocating ATPases
MH  - RESISTANCE
MH  - Saccharomyces cerevisiae
MH  - SUBUNIT
MH  - Support,U.S.Gov't,P.H.S.
MH  - Transcription Factors
MH  - universities
MH  - YEAST
RP  - NOT IN FILE
NT  - Molecular Biology PhD Program, University of Iowa, Iowa City, Iowa 52242, USAFAU - Zhang, X
SO  - J Biol Chem 2001 Dec 21 ;276(51):47844-47852

1856
UI  - 21337
AU  - Ackerman SH
TI  - Atp11p and Atp12p are chaperones for F(1)-ATPase biogenesis in mitochondria
AB  - The bioenergetic needs of aerobic cells are met principally through the action of the F(1)F(0) ATP synthase, which catalyzes ATP synthesis during oxidative phosphorylation. The catalytic unit of the enzyme (F(1)) is a multimeric protein of the subunit composition alpha(3)beta(3)(gamma)(delta) epsilon. Our work, which employs the yeast Saccharomyces cerevisiae as a model system for studies of mitochondrial function, has provided evidence that assembly of the mitochondrial alpha and beta subunits into the F(1) oligomer requires two molecular chaperone proteins called Atp11p and Atp12p. Comprehensive knowledge of Atp11p and Atp12p activities in mitochondria bears relevance to human physiology and disease as these chaperone actions are now known to exist in mitochondria of human cells
MH  - A
MH  - alpha
MH  - Amino Acid Sequence
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - BETA
MH  - BETA-SUBUNIT
MH  - Binding Sites
MH  - biosynthesis
MH  - catalytic
MH  - cell
MH  - Cells
MH  - Chemistry
MH  - Comparative Study
MH  - DNA,Complementary
MH  - enzyme
MH  - enzymology
MH  - function
MH  - fungal proteins
MH  - Human
MH  - Hydrophobicity
MH  - isolation & purification
MH  - metabolism
MH  - Mitochondria
MH  - model
MH  - Molecular Chaperones
MH  - Molecular Sequence Data
MH  - Oxidative Phosphorylation
MH  - Phosphorylation
MH  - physiology
MH  - protein
MH  - Protein Structure,Tertiary
MH  - Proteins
MH  - Proton-Translocating ATPases
MH  - Saccharomyces cerevisiae
MH  - Sequence Alignment
MH  - Sequence Homology,Amino Acid
MH  - SUBUNIT
MH  - SYNTHASE
MH  - synthesis
MH  - SYSTEM
MH  - universities
MH  - YEAST
MH  - Yeasts
RP  - NOT IN FILE
NT  - Department of Surgery, Wayne State University School of Medicine, Detroit, MI 48201, USA sackerm@medwayneeduFAU - Ackerman, Sharon H
SO  - Biochim Biophys Acta 2002 Sep 10 ;1555(1-3):101-105

1857
UI  - 21353
AU  - Aggeler R
AU  - Coons J
AU  - Taylor SW
AU  - Ghosh SS
AU  - Garcia JJ
AU  - Capaldi RA
AU  - Marusich MF
TI  - A Functionally Active Human F1F0 ATPase Can be Purified by Immunocapture from Heart Tissue and Fibroblast Cell Lines. SUBUNIT STRUCTURE AND ACTIVITY STUDIES
AB  - Human mitochondrial F(1)F(0) ATP synthase was isolated with a one-step immunological approach, using a monoclonal antibody against F(1) in a 96-well microplate activity assay system, to establish a method for fast high throughput screening of inhibitors, toxins, and drugs with very small amounts of enzyme. For preparative purification, mitochondria from human heart tissue as well as cultured fibroblasts were solubilized with dodecyl-beta-d-maltoside, and the F(1)F(0) was isolated with anti-F(1) monoclonal antibody coupled to protein G-agarose beads. The immunoprecipitated F(1)F(0) contained a full complement of subunits that were identified with specific antibodies against five of the subunits (alpha, beta, OSCP, d, and IF(1)) and by MALDI-TOF and/or LC/MS/MS for all subunits except subunit c, which could not be resolved by these methods because of the limits of detection. Microscale immunocapture of F(1)F(0) from detergent-solubilized mitochondria or whole cell fibroblast extracts was performed using anti-F(1) monoclonal antibody immobilized on 96-well microplates. The captured complex V displayed ATP hydrolysis activity that was fully oligomycin and inhibitor protein IF(1)-sensitive. Moreover, IF(1) could be co-isolated with F(1)F(0) when the immunocapture procedure was carried out at pH 6.5 but was absent when the ATP synthase was isolated at pH 8.0. Immunocaptured F(1)F(0) lacking IF(1) could be inhibited by more than 90% by addition of recombinant inhibitor protein, and conversely, F(1)F(0) containing IF(1) could be activated more than 10-fold by brief exposure to pH 8.0, inducing the release of inhibitor protein. With this microplate system an ATP hydrolysis assay of complex V could be carried out with as little as 10 ng of heart mitochondria/well and as few as 3 x 10(4) cells/well from fibroblast cultures. The system is therefore suitable to screen patient-derived samples for alterations in amount or functionality of both the F(1)F(0) ATPase and IF(1)
MH  - A
MH  - ACTIVE
MH  - alpha
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BETA
MH  - biology
MH  - cell
MH  - Cell Line
MH  - COMPLEX
MH  - enzyme
MH  - Human
MH  - Hydrolysis
MH  - inhibitor
MH  - INHIBITOR PROTEIN
MH  - inhibitors
MH  - method
MH  - Methods
MH  - Mitochondria
MH  - MONOCLONAL-ANTIBODIES
MH  - oligomycin
MH  - pH
MH  - protein
MH  - purification
MH  - structure
MH  - SUBUNIT
MH  - SYNTHASE
MH  - SYSTEM
MH  - universities
RP  - NOT IN FILE
NT  - Institute of Molecular Biology, University of Oregon, Eugene, Oregon 97403, MitoKor, San Diego, California 92121, and the Instituto Nacional de Cardiologia, Departamento de Bioquimica, Juan Badiano 1 Col Seccion XVI, 14080, Mexico D F, MexicoFAU - Aggeler, Robert
SO  - J Biol Chem 2002 Sep 13 ;277(37):33906-33912

1858
UI  - 21428
AU  - Akagi M
AU  - Inui K
AU  - Tsukamoto H
AU  - Sakai N
AU  - Muramatsu T
AU  - Yamada M
AU  - Matsuzaki K
AU  - Goto Y
AU  - Nonaka I
AU  - Okada S
TI  - A point mutation of mitochondrial ATPase 6 gene in Leigh syndrome
AB  - A T-to-G transition at nucleotide 9176 (T9176G) in the mitochondrial adenosine triphosphate 6 gene (MTATP6) was detected in two siblings with Leigh syndrome. Heteroplasmy was observed in the mother's leukocytes. The T9176G mutation changes a highly conserved leucine residue to an arginine in subunit 6 of ATPase and is maternally inherited like mutations in the other mitochondrial genes. Another mutation in the same codon (T9176C) has been previously reported in Leigh syndrome. This gives strong support to the relevance of MTATP6 dysfunction in Leigh syndrome and the importance of leucine at that position
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - Arginine
MH  - ATPase
MH  - Case Report
MH  - Child
MH  - Child,Preschool
MH  - enzymology
MH  - Family Health
MH  - Fatal Outcome
MH  - Female
MH  - genetics
MH  - Human
MH  - Leigh Disease
MH  - Leucine
MH  - Male
MH  - metabolism
MH  - Mitochondria
MH  - Mitochondrial Proton-Translocating ATPases
MH  - Mutation
MH  - nucleotide
MH  - Pedigree
MH  - point mutation
MH  - RESIDUE
MH  - SUBUNIT
MH  - universities
RP  - NOT IN FILE
NT  - Department of Developmental Medicine (Pediatrics), Osaka University, Graduate School of Medical Science, 2-2 Yamadaoka, Suita, Osaka, JapanFAU - Akagi, Motohiro
SO  - Neuromuscul Disord 2002 Jan ;12(1):53-55

1859
UI  - 21358
AU  - Allchin D
TI  - To err and win a nobel prize: Paul Boyer, ATP synthase and the emergence of bioenergetics
AB  - Paul Boyer shared a Nobel Prize in 1997 for his work on the mechanism of ATP synthase. His earlier work, though (which contributed indirectly to his triumph), included major errors, both experimental and theoretical. Two benchmark cases offer insight into how scientists err and how they deal with error. Boyer's work also parallels and illustrates the emergence of bioenergetics in the second half of the twentieth century, rivaling achievements in evolution and molecular biology
MH  - A
MH  - atp
MH  - ATP synthase
MH  - ATP Synthetase Complexes
MH  - bioenergetics
MH  - biology
MH  - Chemistry
MH  - Energy Metabolism
MH  - Evolution
MH  - history
MH  - History of Medicine,20th Cent.
MH  - mechanism
MH  - Nobel Prize
MH  - Research Design
MH  - SYNTHASE
MH  - united states
MH  - universities
RP  - NOT IN FILE
NT  - Program in the History of Science and Technology, University of Minnesota, MN 55455, USA allchin@pclinkcomFAU - Allchin, Douglas
SO  - J Hist Biol 2002  ;35(1):149-172

1860
UI  - 21430
AU  - Arata Y
AU  - Baleja JD
AU  - Forgac M
TI  - Cysteine-directed cross-linking to subunit B suggests that subunit E forms part of the peripheral stalk of the vacuolar H+-ATPase
AB  - We have employed a combination of site-directed mutagenesis and covalent cross-linking to identify subunits in close proximity to subunit B in the vacuolar H(+)-ATPase (V-ATPase) complex. Unique cysteine residues were introduced into a Cys-less form of subunit B, and the V-ATPase complex in isolated vacuolar membranes from each mutant strain was reacted with the bifunctional, photoactivable maleimide reagent 4-(N-maleimido)benzophenone. Photoactivation resulted in cross-linking of the unique sulfhydryl groups on subunit B with other subunits in the complex. Four of the eight mutants constructed containing a unique cysteine residue at Ala(15), Lys(45), Glu(494), or Thr(501) resulted in the formation of cross-linked products, which were recognized by Western blot analysis using antibodies against both subunits B and E. These products had a molecular mass of 84 kDa, consistent with a cross-linked product of subunits B and E. Molecular modeling of subunit B places Ala(15) and Lys(45) near the top of the V(1) structure (i.e. farthest from the membrane), whereas Glu(494) and Thr(501) are predicted to reside near the bottom of V(1), with all four residues predicted to be oriented toward the external surface of the complex. A model incorporating these and previous data is presented in which subunit E exists in an extended conformation on the outer surface of the A(3)B(3) hexamer that forms the core of the V(1) domain. This location for subunit E suggests that this subunit forms part of the peripheral stalk of the V-ATPase that links the V(1) and V(0) domains
MH  - A
MH  - analysis
MH  - Animal
MH  - Biochemistry
MH  - Chemistry
MH  - COMPLEX
MH  - conformation
MH  - CROSS-LINKING
MH  - Cross-Linking Reagents
MH  - Cysteine
MH  - data
MH  - enzymology
MH  - H(+)ATPase
MH  - H+-ATPase
MH  - Immunoglobulin G
MH  - membrane
MH  - Membranes
MH  - metabolism
MH  - Mice
MH  - model
MH  - Models,Molecular
MH  - Molecular Conformation
MH  - mutagenesis
MH  - Mutagenesis,Site-Directed
MH  - mutant
MH  - physiology
MH  - Protein Subunits
MH  - Rabbits
MH  - Recombinant Proteins
MH  - RESIDUE
MH  - Saccharomyces cerevisiae
MH  - site-directed
MH  - stalk
MH  - structure
MH  - SUBUNIT
MH  - Support,Non-U.S.Gov't
MH  - Support,U.S.Gov't,P.H.S.
MH  - SURFACE
MH  - universities
MH  - Vacuolar Proton-Translocating ATPases
MH  - vacuoles
RP  - NOT IN FILE
NT  - Departments of Physiology and Biochemistry, Tufts University School of Medicine, Boston, Massachusetts 02111, USAFAU - Arata, Yoichiro
SO  - J Biol Chem 2002 Feb 1 ;277(5):3357-3363

1861
UI  - 21334
AU  - Arata Y
AU  - Baleja JD
AU  - Forgac M
TI  - Localization of Subunits D, E, and G in the Yeast V-ATPase Complex Using Cysteine-Mediated Cross-Linking to Subunit B
AB  - Using a combination of cysteine mutagenesis and covalent cross-linking, we have identified subunits in close proximity to specific sites within subunit B of the vacuolar (H(+))-ATPase (V-ATPase) of yeast. Unique cysteine residues were introduced into subunit B by site-directed mutagenesis, and the resultant V-ATPase complexes were reacted with the bifunctional, photoactivatable maleimide reagent 4-(N-maleimido)benzophenone (MBP) followed by irradiation. Cross-linked products were identified by Western blot using subunit-specific antibodies. Introduction of cysteine residues at positions Glu(106) and Asp(199) led to cross-linking of subunits B and E, at positions Asp(341) and Ala(424) to cross-linking of subunits B and D, and at positions Ala(15) and Lys(45) to cross-linking of subunits B and G. Using a molecular model of subunit B constructed on the basis of sequence homology between the V- and F-ATPases, the X-ray coordinates of the F(1)-ATPase, and energy minimization, Glu(106), Asp(199), Ala(15), and Lys(45) are all predicted to be located on the outer surface of the complex, with Ala(15) and Lys(45) located near the top of the complex furthest from the membrane. By contrast, Asp(341) and Ala(424) are predicted to face the interior of the A(3)B(3) hexamer. These results suggest that subunits E and G form part of a peripheral stalk connecting the V(1) and V(0) domains whereas subunit D forms part of a central stalk. Subunit D is thus the most likely homologue to the gamma subunit of F(1), which undergoes rotation during ATP hydrolysis and serves an essential function in rotary catalysis
MH  - A
MH  - atp
MH  - Biochemistry
MH  - Catalysis
MH  - COMPLEX
MH  - CROSS-LINKING
MH  - Cysteine
MH  - energy
MH  - F-ATPASE
MH  - function
MH  - Hydrolysis
MH  - membrane
MH  - model
MH  - mutagenesis
MH  - physiology
MH  - RESIDUE
MH  - rotation
MH  - Site
MH  - site-directed
MH  - stalk
MH  - SUBUNIT
MH  - SURFACE
MH  - universities
MH  - YEAST
RP  - NOT IN FILE
NT  - Departments of Physiology and Biochemistry, Tufts University School of Medicine, 136 Harrison Avenue, Boston, Massachusetts 02111FAU - Arata, Yoichiro
SO  - Biochemistry 2002 Sep 17 ;41(37):11301-11307

1862
UI  - 21382
AU  - Arechaga I
AU  - Butler PJ
AU  - Walker JE
TI  - Self-assembly of ATP synthase subunit c rings
AB  - Subunit c of the H(+) transporting ATP synthase is an essential part of its membrane domain that participates in transmembrane proton conduction. The annular architecture of the subunit c from different species has been previously reported. However, little is known about the type of interactions that affect the formation of c-rings in the ATPase complex. Here we report that subunit c over-expressed in Escherichia coli and purified in non-ionic detergent solutions self-assembles into annular structures in the absence of other subunits of the complex. The results suggest that the ability of subunit c to form rings is determined by its primary structure
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Bacterial Proton-Translocating ATPases
MH  - Chemistry
MH  - COMPLEX
MH  - Detergents
MH  - Dimethylamines
MH  - enzymology
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - Human
MH  - Image Processing,Computer-Assisted
MH  - INTERACTION
MH  - isolation & purification
MH  - membrane
MH  - Methods
MH  - Microscopy,Electron
MH  - Molecular Weight
MH  - Nutrition
MH  - physiology
MH  - primary
MH  - Protein Binding
MH  - Protein Subunits
MH  - proton
MH  - Recombinant Proteins
MH  - Solutions
MH  - structure
MH  - SUBUNIT
MH  - SYNTHASE
MH  - Ultracentrifugation
MH  - ultrastructure
RP  - NOT IN FILE
NT  - The Medical Research Council Dunn Human Nutrition Unit, Hills Road, CB2 2YK, Cambridge, UKFAU - Arechaga, Ignacio
SO  - FEBS Lett 2002 Mar 27 ;515(1-3):189-193

1863
UI  - 21376
AU  - Ariga T
AU  - Masaike T
AU  - Noji H
AU  - Yoshida M
TI  - Stepping rotation of F(1)-ATPase with one, two, or three altered catalytic sites that bind ATP only slowly
AB  - F(1)-ATPase is an ATP hydrolysis-driven motor in which the gamma subunit rotates in the stator cylinder alpha(3)beta(3). To know the coordination of three catalytic beta subunits during catalysis, hybrid F(1)-ATPases, each containing one, two, or three "slow" mutant beta subunits that bind ATP very slowly, were prepared, and the rotations were observed with a single molecule level. Each hybrid made one, two, or three steps per 360 degrees revolution, respectively, at 5 microm ATP where the wild-type enzyme rotated continuously without step under the same observing conditions. The observed dwell times of the steps are explained by the slow binding rate of ATP. Except for the steps, properties of rotation, such as the torque forces exerted during rotary movement, were not significantly changed from those of the wild-type enzyme. Thus, it appears that the presence of the slow beta subunit(s) does not seriously affect other normal beta subunit(s) in the same F(1)-ATPase molecule and that the order of sequential catalytic events is faithfully maintained even when ATP binding to one or two of the catalytic sites is retarded
MH  - A
MH  - Adenosine Triphosphate
MH  - atp
MH  - Base Sequence
MH  - BETA
MH  - BETA-SUBUNIT
MH  - BINDING
MH  - Catalysis
MH  - catalytic
MH  - catalytic domain
MH  - Chemistry
MH  - DNA Primers
MH  - enzyme
MH  - genetics
MH  - isolation & purification
MH  - Kinetics
MH  - metabolism
MH  - Movement
MH  - Mutagenesis,Site-Directed
MH  - mutant
MH  - Protein Binding
MH  - Proton-Translocating ATPases
MH  - rotation
MH  - Site
MH  - SUBUNIT
MH  - Support,Non-U.S.Gov't
MH  - Time
RP  - NOT IN FILE
NT  - Chemical Resources Laboratory, Tokyo Institute of Technology, 4259 Nagatsuta, Yokohama, 226-8503, JapanFAU - Ariga, Takayuki
SO  - J Biol Chem 2002 Jul 12 ;277(28):24870-24874

1864
UI  - 21315
AU  - Azarashvili TS
AU  - Tyynela J
AU  - Odinokova IV
AU  - Grigorjev PA
AU  - Baumann M
AU  - Evtodienko YV
AU  - Saris NE
TI  - Phosphorylation of a peptide related to subunit c of the F0F1-ATPase/ATP synthase and relationship to permeability transition pore opening in mitochondria
AB  - A phosphorylated polypeptide (ScIRP) from the inner membrane of rat liver mitochondria with an apparent molecular mass of 3.5 kDa was found to be immunoreactive with specific antibodies against subunit c of F0F1-ATPase/ATP synthase (Azarashvily, T. S., Tyynela, J., Baumann, M., Evtodienko, Yu. V., and Saris, N.-E. L. (2000). Biochem. Biophys. Res. Commun. 270, 741-744. In the present paper we show that the dephosphorylation of ScIRP was promoted by the Ca2+-induced mitochondrial permeability transition (MPT) and prevented by cyclosporin A. Preincubation of ScIRP isolated in its dephosphorylated form with the mitochondrial suspension decreased the membrane potential (delta psiM) and the Ca2+-uptake capacity by promoting MPT. Incorporation of ScIRP into black-lipid membranes increased the membrane conductivity by inducing channel formation that was also suppressed by antibodies to subunit c. These data indicate that the phosphorylation level of ScIRP is influenced by the MPT pore state, presumably by stimulation of calcineurin phosphatase by the Ca2+ used to induce MPT. The possibility of ScIRP being part of the MPT pore assembly is discussed in view of its capability to induced channel activity
MH  - A
MH  - Biophysics
MH  - data
MH  - delta
MH  - Liver
MH  - M
MH  - membrane
MH  - Membrane Potential
MH  - Membranes
MH  - Mitochondria
MH  - peptide
MH  - Permeability
MH  - phosphatase
MH  - Phosphorylation
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - Institute of Theoretical and Experimental Biophysics, Russian Academy of Sciences, Pushchino, Moscow RegionFAU - Azarashvili, Tamara S
SO  - J Bioenerg Biomembr 2002 Aug ;34(4):279-284

1865
UI  - 21394
AU  - Bandyopadhyay S
AU  - Ren H
AU  - Wang CS
AU  - Allison WS
TI  - The (alpha F(357)C)(3)(beta R(372)C)(3)gamma subcomplex of the F(1)-ATPase from the thermophilic Bacillus PS3 has altered ATPase activity after cross-linking alpha and beta subunits at noncatalytic site interfaces
AB  - In crystal structures of bovine MF(1), the side chains of alpha F(357) and beta R(372) are near the adenines of nucleotides bound to noncatalytic sites. To determine if during catalysis these side chains must pass through the different arrangements in which they are present in crystal structures, the catalytic properties of the (alpha F(357)C)(3)(beta R(372)C)(3)gamma subcomplex of the TF(1)-ATPase were characterized before and after cross-linking the introduced cysteines with CuCl(2). The unmodified mutant enzyme hydrolyzes MgATP at 50% the rate exhibited by wild type. Detailed comparison of the catalytic properties of the double mutant enzyme before and after cross-linking with those of the wild-type subcomplex revealed the following. Before cross-linking, the (alpha F(357)C)(3)(beta R(372)C)(3)gamma subcomplex has less tendency than wild type to release inhibitory MgADP entrapped in a catalytic site during turnover when MgATP binds to noncatalytic sites. Following cross-linking, ATPase activity is reduced 5-fold, and inhibitory MgADP entrapped in a catalytic site during turnover does not release under conditions wherein binding of ATP to noncatalytic sites of the wild-type enzyme promotes release of MgADP from the affected catalytic site. When assayed in the presence of lauryldimethylamine oxide, which prevents turnover-dependent entrapment of inhibitory MgADP in a catalytic site, ATPase activity of the cross-linked form is 47% that of the unmodified mutant enzyme. These results suggest that, during catalysis, the side chains of alpha F(357) and beta R(372) do not pass through the extremely different relative positions in which they exist at the three noncatalytic site interfaces in crystal structures
MH  - A
MH  - Adenosine Diphosphate
MH  - alpha
MH  - Amino Acid Substitution
MH  - atp
MH  - ATPase
MH  - Bacillus
MH  - BETA
MH  - BETA-SUBUNIT
MH  - BINDING
MH  - Biochemistry
MH  - Catalysis
MH  - catalytic
MH  - Chemistry
MH  - CROSS-LINKING
MH  - Cross-Linking Reagents
MH  - Cysteine
MH  - enzyme
MH  - Enzyme Activation
MH  - enzymology
MH  - genetics
MH  - interfaces
MH  - metabolism
MH  - mutant
MH  - Mutation
MH  - NONCATALYTIC SITES
MH  - nucleotide
MH  - Nucleotides
MH  - Protein Subunits
MH  - Proton-Translocating ATPases
MH  - PS3
MH  - Site
MH  - structure
MH  - SUBUNIT
MH  - Support,U.S.Gov't,P.H.S.
MH  - THERMOPHILIC
MH  - THERMOPHILIC BACILLUS PS3
MH  - turnover
MH  - universities
RP  - NOT IN FILE
NT  - Department of Chemistry and Biochemistry, University of California at San Diego, La Jolla, California 92093-0601, USAFAU - Bandyopadhyay, Sanjay
SO  - Biochemistry 2002 Mar 5 ;41(9):3226-3234

1866
UI  - 21399
AU  - Baracca A
AU  - Barogi S
AU  - Paolini S
AU  - Lenaz G
AU  - Solaini G
TI  - Fluorescence resonance energy transfer between coumarin-derived mitochondrial F(1)-ATPase gamma subunit and pyrenylmaleimide-labelled fragments of IF(1) and c subunit
AB  - We introduced a reporting group into a critical position of the mitochondrial F(1)-ATPase in order to gain structural information about enzyme-ligand complexes. Incubation of 7-diethylamino-3-(4'-maleimidylphenyl)-4-methylcoumarin (CPM) with bovine heart mitochondrial F(1)-ATPase pretreated with 1 nM sodium arsenite modified the only cysteine residue in the gamma subunit (gamma-Cys(78)), resulting in an enzyme-CPM fluorescent complex (CPM-F(1)) with an ATPase activity similar to that of the native enzyme. Transferred fluorescence of F(1)-bound CPM occurred when different peptide fragments of naturally binding polypeptides carrying a pyrenylmaleimide (PM) moiety were bound to the enzyme. Fluorescence resonance energy transfer (RET) from PM bound to cysteine residues associated with Glu(40), Lys(47) and Lys(58) of fragments of the inhibitor protein (IF(1)) with CPM-F(1) occurred with an efficiency of approx. 20, 21 and 3% respectively. The distance at which the efficiency of energy transfer was 50%, R(0), for the CPM and PM donor/acceptor pair was 4.1 nm, indicating that the three IF(1) fragments must be within 6.7 nm of gamma-Cys(78). RET from the PM-bound hydrophilic fragment of c subunit (residues 37-42) of the F(1)F(0)-ATPase complex and CPM-bound gamma-Cys(78) occurred with an efficiency of approx. 30%, indicating a distance of 4.7 nm between the two fluorophores. Based on previous observations and on the present RET measurements, the hydrophilic loop of c subunit was located at the base of the F(1) foot, and the N-terminal region of IF(1) was located on the surface of F(1) in the lower part of the alpha(3)beta(3) hexamer ring
MH  - A
MH  - Amino Acid Sequence
MH  - Animal
MH  - ATPase
MH  - BASE
MH  - BINDING
MH  - Cattle
MH  - Chemistry
MH  - COMPLEX
MH  - Coumarins
MH  - Cysteine
MH  - Electrophoresis,Polyacrylamide Gel
MH  - energy
MH  - Energy Transfer
MH  - enzyme
MH  - enzymology
MH  - fluorescence
MH  - Fluorescent Dyes
MH  - inhibitor
MH  - INHIBITOR PROTEIN
MH  - metabolism
MH  - Mitochondria
MH  - Models,Molecular
MH  - Molecular Sequence Data
MH  - peptide
MH  - Peptide Fragments
MH  - protein
MH  - Protein Conformation
MH  - Proton-Translocating ATPases
MH  - RESIDUE
MH  - Sodium
MH  - Spectrometry,Fluorescence
MH  - SUBUNIT
MH  - Sulfhydryl Compounds
MH  - Support,Non-U.S.Gov't
MH  - SURFACE
MH  - TRANSFER
RP  - NOT IN FILE
NT  - Dipartimento di Biochimica G Moruzzi, Universita degli Studi di Bologna, via Irnerio 48, 40126 Bologna, ItalyFAU - Baracca, Alessandra
SO  - Biochem J 2002 Feb 15 ;362(Pt 1):165-171

1867
UI  - 21321
AU  - Belogrudov G
TI  - Factor B is essential for ATP synthesis by mitochondria
AB  - Factor B is a water-soluble protein, which is required for the coupled activity of the mitochondrial ATP synthase complex. Specific removal of factor B from well-coupled bovine heart submitochondrial particles (SMP) results in uncoupling and the loss of ATP-driven membrane potential formation and reverse electron transfer from succinate to NAD. Addition of recombinant human factor B (molecular mass 20,341Da) to factor B-depleted SMP (AE-SMP) restores these properties [G.I. Belogrudov, and Y. Hatefi, (2002) J. Biol. Chem. 277, 6097-6103]. This paper shows that extraction and purification of ATP synthase complex (complex V) from bovine heart mitochondria results in extensive loss of factor B. Addition of recombinant human factor B to AE-SMP completely restores the lost oxidative phosphorylation and ATP-32P(i) exchange activities of the particles and increases the ATP-32P(i) exchange activity of complex V by 2.5-fold. These results further indicate that factor B is an essential component of the mammalian ATP synthase complex
MH  - A
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - Biochemistry
MH  - COMPLEX
MH  - electron
MH  - electron transfer
MH  - Electron-transfer
MH  - Human
MH  - membrane
MH  - Membrane Potential
MH  - Mitochondria
MH  - Nad
MH  - Oxidative Phosphorylation
MH  - Phosphorylation
MH  - protein
MH  - purification
MH  - Submitochondrial Particles
MH  - succinate
MH  - SYNTHASE
MH  - synthesis
MH  - TRANSFER
RP  - NOT IN FILE
NT  - Division of Biochemistry, Department of Molecular and Experimental Medicine, The Scripps Research Institute, 92037, La Jolla, CA, USAFAU - Belogrudov, Grigory
SO  - Arch Biochem Biophys 2002 Oct 15 ;406(2):271

1868
UI  - 21423
AU  - Belogrudov GI
AU  - Hatefi Y
TI  - Factor B and the mitochondrial ATP synthase complex
AB  - Factor B is a subunit of the mammalian ATP synthase complex, whose existence has been controversial. This paper describes the molecular and functional properties of a recombinant human factor B, which when added to bovine submitochondrial particles depleted of their factor B restores the energy coupling activity of the ATP synthase complexes. The mature human factor B has 175 amino acids and a molecular mass of 20,341 Da. The preparation is water-soluble, monomeric, and is inactivated by monothiol- and especially dithiol-modifying reagents, probably reacting at its cysteine residues Cys-92 and Cys-94. A likely factor B gene composed of 5 exons has been identified on chromosome 14q21.3, and the functional role of factor B in the mammalian ATP synthase complex has been discussed
MH  - A
MH  - ACID
MH  - Amino Acid Sequence
MH  - Amino Acids
MH  - Animal
MH  - atp
MH  - ATP synthase
MH  - Base Sequence
MH  - Binding Sites
MH  - Biochemistry
MH  - Cattle
MH  - Chemistry
MH  - Chromosome Mapping
MH  - Cloning,Molecular
MH  - COMPLEX
MH  - coupling
MH  - Cysteine
MH  - drug effects
MH  - Electron Transport
MH  - energy
MH  - enzymology
MH  - Exons
MH  - genetics
MH  - Human
MH  - isolation & purification
MH  - Kinetics
MH  - metabolism
MH  - Mitochondria,Heart
MH  - Mitochondrial Proton-Translocating ATPases
MH  - Molecular Sequence Data
MH  - Molecular Weight
MH  - Oligomycins
MH  - pharmacology
MH  - Protein Subunits
MH  - Recombinant Proteins
MH  - RESIDUE
MH  - Submitochondrial Particles
MH  - SUBUNIT
MH  - Sulfhydryl Compounds
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - Division of Biochemistry, Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, California 92037, USAFAU - Belogrudov, Grigory I
SO  - J Biol Chem 2002 Feb 22 ;277(8):6097-6103

1869
UI  - 21361
AU  - Berger K
AU  - Sivars U
AU  - Winzell MS
AU  - Johansson P
AU  - Hellman U
AU  - Rippe C
AU  - Erlanson-Albertsson C
TI  - Mitochondrial ATP synthase--a possible target protein in the regulation of energy metabolism in vitro and in vivo
AB  - The increasing prevalence of obesity in the Western world has stimulated an intense search for mechanisms regulating food intake and energy balance. A number of appetite-regulating peptides have been identified, their receptors cloned and the intracellular events characterized. One possible energy-dissipating mechanism is the mitochondrial uncoupling of ATP-synthesis from respiratory chain oxidation through uncoupling proteins, whereby energy derived from food could be dissipated as heat, instead of stored as ATP. The exact role of the uncoupling proteins in energy balance is, however, uncertain. We show here that mitochondrial F1F0-ATP synthase itself is a target protein for an anorectic peptide, enterostatin, demonstrated both after affinity purification of rat brain membranes and through a direct physical interaction between enterostatin and purified F1-ATP synthase. In insulinoma cells (INS-1) enterostatin was found to target F1F0-ATP synthase, causing an inhibition of ATP production, an increased thermogenesis and increased oxygen consumption. The experiments suggest a role of mitochondrial F1F0-ATP synthase in the suppressed insulin secretion induced by enterostatin. It could be speculated that this targeting mechanism is involved in the decreased energy efficiency following enterostatin treatment in rat
MH  - A
MH  - affinity
MH  - atp
MH  - ATP production
MH  - ATP synthesis
MH  - biology
MH  - Brain
MH  - cell
MH  - Cells
MH  - energy
MH  - Energy Metabolism
MH  - F1F0-ATP SYNTHASE
MH  - Heat
MH  - In Vitro
MH  - INTERACTION
MH  - mechanism
MH  - MECHANISMS
MH  - membrane
MH  - Membranes
MH  - metabolism
MH  - Obesity
MH  - Oxygen
MH  - Oxygen Consumption
MH  - peptide
MH  - peptides
MH  - protein
MH  - Proteins
MH  - purification
MH  - regulation
MH  - secretion
MH  - SYNTHASE
MH  - universities
RP  - NOT IN FILE
NT  - Department of Cell and Molecular Biology, Biomedical Center, University of Lund, Sweden KarinBerger@medkemluseFAU - Berger, Karin
SO  - Nutr Neurosci 2002 Jun ;5(3):201-210

1870
UI  - 21398
AU  - Bockmann RA
AU  - Grubmuller H
TI  - Nanoseconds molecular dynamics simulation of primary mechanical energy transfer steps in F1-ATP synthase
AB  - The mitochondrial membrane protein FoF1-ATP synthase synthesizes adenosine triphosphate (ATP), the universal currency of energy in the cell. This process involves mechanochemical energy transfer from a rotating asymmetric gamma-'stalk' to the three active sites of the F1 unit, which drives the bound ATP out of the binding pocket. Here, the primary structural changes associated with this energy transfer in F1-ATP synthase were studied with multi-nanosecond molecular dynamics simulations. By forced rotation of the gamma-stalk that mimics the effect of proton motive Fo-rotation during ATP synthesis, a time-resolved atomic model for the structural changes in the F1 part in terms of propagating conformational motions is obtained. For these, different time scales are found, which allows the separation of nanosecond from microsecond conformational motions. In the simulations, rotation of the gamma-stalk lowers the ATP affinity of the betaTP binding pocket and triggers fast, spontaneous closure of the empty betaE subunit. The simulations explain several mutation studies and the reduced hydrolysis rate of gamma-depleted F1-ATPase
MH  - A
MH  - ACTIVE
MH  - ACTIVE SITE
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - affinity
MH  - Animal
MH  - atp
MH  - ATP synthesis
MH  - BINDING
MH  - Binding Sites
MH  - Biophysics
MH  - Catalysis
MH  - Cattle
MH  - cell
MH  - Chemistry
MH  - Computer Simulation
MH  - energy
MH  - Energy Transfer
MH  - enzymology
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - Hydrolysis
MH  - Kinetics
MH  - membrane
MH  - metabolism
MH  - microsecond
MH  - Mitochondria,Heart
MH  - model
MH  - Models,Molecular
MH  - MOLECULAR-DYNAMICS
MH  - Mutation
MH  - primary
MH  - protein
MH  - Protein Conformation
MH  - Protein Subunits
MH  - proton
MH  - Proton-Translocating ATPases
MH  - rotation
MH  - SIMULATION
MH  - SIMULATIONS
MH  - Site
MH  - SUBUNIT
MH  - SYNTHASE
MH  - synthesis
MH  - Time
MH  - TRANSFER
RP  - NOT IN FILE
NT  - Theoretical Molecular Biophysics Group, Max-Planck-Institute for Biophysical Chemistry, Am Fassberg 11, 37077 Gottingen, GermanyFAU - Bockmann, Rainer A
SO  - Nat Struct Biol 2002 Mar ;9(3):198-202

1871
UI  - 21043
AU  - Bockmann RA
AU  - Grubmuller H
TI  - Nanoseconds molecular dynamics simulation of primary mechanical energy transfer steps in F1-ATP synthase
AB  - The mitochondrial membrane protein FoF1-ATP synthase synthesizes adenosine triphosphate (ATP), the universal currency of energy in the cell. This process involves mechanochemical energy transfer from a rotating asymmetric [gamma]-'stalk' to the three active sites of the F1 unit, which drives the bound ATP out of the binding pocket. Here, the primary structural changes associated with this energy transfer in F1-ATP synthase were studied with multi-nanosecond molecular dynamics simulations. By forced rotation of the [gamma]-stalk that mimics the effect of proton motive Fo-rotation during ATP synthesis, a time-resolved atomic model for the structural changes in the F1 part in terms of propagating conformational motions is obtained. For these, different time scales are found, which allows the separation of nanosecond from microsecond conformational motions. In the simulations, rotation of the [gamma]-stalk lowers the ATP affinity of the [beta]TP binding pocket and triggers fast, spontaneous closure of the empty [beta]E subunit. The simulations explain several mutation studies and the reduced hydrolysis rate of [gamma]-depleted F1-ATPase
MH  - A
MH  - ACTIVE
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthesis
MH  - BINDING
MH  - Biophysics
MH  - Chemistry
MH  - Energy Transfer
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - Hydrolysis
MH  - membrane
MH  - microsecond
MH  - model
MH  - MOLECULAR-DYNAMICS
MH  - protein
MH  - proton
MH  - rotation
MH  - SIMULATION
MH  - SIMULATIONS
MH  - Site
MH  - SUBUNIT
MH  - SYNTHASE
MH  - synthesis
MH  - Time
MH  - TRANSFER
RP  - NOT IN FILE
NT  - Theoretical Molecular Biophysics Group, Max-Planck-Institute for Biophysical Chemistry, Am Fassberg 11, 37077 Gottingen, Germany
SO  - Nat Struct Biol 2002 Feb 11 ;.():

1872
UI  - 21310
AU  - Bon C
AU  - Dianoux AJ
AU  - Ferrand M
AU  - Lehmann MS
AD  - Institut Laue Langevin, BP156, 38042 Grenoble Cedex 9, France
TI  - A model for water motion in crystals of lysozyme based on an incoherent quasielastic neutron-scattering study
AB  - This paper reports an incoherent quasielastic neutron scattering study of the single particle, diffusive motions of water molecules surrounding a globular protein, the hen egg-white lysozyme. For the first time such an analysis has been done on protein crystals. It can thus be directly related and compared with a recent structural study of the same sample. The measurement temperature ranged from 100 to 300 K, but focus was on the room temperature analysis. The very good agreement between the structural and dynamical studies suggested a model for the dynamics of water in triclinic crystals of lysozyme in the time range ~330 ps and at 300 K. Herein, the dynamics of all water molecules is affected by the presence of the protein, and the water molecules can be divided into two populations. The first mainly corresponds to the first hydration shell, in which water molecules reorient themselves fivefold to 10-fold slower than in bulk solvent, and diffuse by jumps from hydration site to hydration site. The long-range diffusion coefficient is five to sixfold less than for bulk solvent. The second group corresponds to water molecules further away from the surface of the protein, in a second incomplete hydration layer, confined between hydrated macromolecules. Within the time scale probed they undergo a translational diffusion with a self-diffusion coefficient reduced ~50- fold compared with bulk solvent. As protein crystals have a highly crowded arrangement close to the packing of macromolecules in cells, our conclusion can be discussed with respect to solvent behavior in intracellular media: as the mobility is highest next to the surface, it suggests that under some crowding conditions, a two-dimensional motion for the transport of metabolites can be dominant
MH  - A
MH  - analysis
MH  - cell
MH  - Cells
MH  - Diffusion
MH  - model
MH  - protein
MH  - Site
MH  - SOLVENT
MH  - SURFACE
MH  - Temperature
MH  - Time
MH  - transport
MH  - united states
MH  - Water
RP  - NOT IN FILE
NT  - UI - 22190590LA - engPT - Journal ArticleDA - 20020830IS - 0006-3495SB - IMCY - United States
UR  - PM:12202382
SO  - Biophys J 2002 Sep ;83(3):1578-1588

1873
UI  - 21341
AU  - Borthwick KJ
AU  - Karet FE
TI  - Inherited disorders of the H+-ATPase
AB  - PURPOSE OF REVIEW The alpha-intercalated cell in the distal nephron shares a number of molecular features with the osteoclast, including site-limited proton pumps that are present at high density. These are multisubunit H -ATPases, which are essential for acid-base homeostasis and for the maintenance of normal bone turnover. In recent years it has become evident that some rare inherited human disorders are due to pump dysfunction in kidney or in bone; these are reviewed here.RECENT FINDINGS The present review provides an overview of acid secretion in both kidney and bone, and describes the recently identified diseases that are associated with mutations in tissue-specific subunits of these pumps.SUMMARY Elucidation of the molecular bases of a number of inherited renal acidopathies and bone disorders raises the possibility that additional tissue-specific subunits of these important pumps will be identified, gives hope for a better understanding of normal function at the molecular level, and may have implications for future therapeutic development
MH  - A
MH  - ACID
MH  - ATPase
MH  - BASE
MH  - cell
MH  - development
MH  - function
MH  - genetics
MH  - H+-ATPase
MH  - Homeostasis
MH  - Human
MH  - Kidney
MH  - proton
MH  - Proton Pump
MH  - review
MH  - secretion
MH  - SUBUNIT
MH  - turnover
MH  - universities
RP  - NOT IN FILE
NT  - Departments of Medical Genetics and Nephrology, University of Cambridge, Cambridge, UKFAU - Borthwick, Katherine J
SO  - Curr Opin Nephrol Hypertens 2002 Sep ;11(5):563-568

1874
UI  - 21378
AU  - Bosetti F
AU  - Brizzi F
AU  - Barogi S
AU  - Mancuso M
AU  - Siciliano G
AU  - Tendi EA
AU  - Murri L
AU  - Rapoport SI
AU  - Solaini G
TI  - Cytochrome c oxidase and mitochondrial F1F0-ATPase (ATP synthase) activities in platelets and brain from patients with Alzheimer's disease
AB  - Evidence suggests that mitochondrial dysfunction is prominent in Alzheimer's disease (AD). A failure of one or more of the mitochondrial electron transport chain enzymes or of F(1)F(0)-ATPase (ATP synthase) could compromise brain energy stores, generate damaging reactive oxygen species (ROS), and lead to neuronal death. In the present study, cytochrome c oxidase (COX) and F(1)F(0)-ATPase activities of isolated mitochondria from platelets and postmortem motor cortex and hippocampus from AD patients and age-matched control subjects were assayed. Compared with controls, COX activity was decreased significantly in platelets (-30%, P < 0.01, n = 20) and hippocampus (-35 to -40%, P < 0.05, n = 6), but not in motor cortex from the AD patients. In contrast, in AD platelets and brain tissues, F(1)F(0)-ATP hydrolysis activity was not significantly changed. Moreover, the ATP synthesis rate was similar in mitochondria of platelets from AD patients and controls. These results demonstrate that COX but not F(1)F(0)-ATPase is a mitochondrial target in AD, in both a brain association area and in platelets. A reduced COX activity may make the tissue vulnerable to excitotoxicity or reduced oxygen availability
MH  - A
MH  - Adenosine Triphosphate
MH  - Aged
MH  - Aged,80 and over
MH  - Alzheimer Disease
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - biosynthesis
MH  - Blood Platelets
MH  - Brain
MH  - Comparative Study
MH  - cytochrome
MH  - Cytochrome c
MH  - Cytochrome-c Oxidase
MH  - electron
MH  - Electron Transport
MH  - energy
MH  - enzyme
MH  - Enzymes
MH  - enzymology
MH  - Female
MH  - Human
MH  - Hydrolysis
MH  - Male
MH  - metabolism
MH  - Middle Age
MH  - Mitochondria
MH  - Mitochondrial Proton-Translocating ATPases
MH  - Oxygen
MH  - P
MH  - pathology
MH  - Support,Non-U.S.Gov't
MH  - SYNTHASE
MH  - synthesis
MH  - transport
RP  - NOT IN FILE
NT  - Scuola Superiore di Studi Universitari e di Perfezionamento S Anna, Via G Carducci 40, 56127 Pisa, ItalyFAU - Bosetti, Francesca
SO  - Neurobiol Aging 2002 May ;23(3):371-376

1875
UI  - 21431
AU  - Bowman BJ
AU  - Bowman EJ
TI  - Mutations in subunit C of the vacuolar ATPase confer resistance to bafilomycin and identify a conserved antibiotic binding site
AB  - Bafilomycin A1, a potent inhibitor of vacuolar H(+)-ATPases (V-ATPase), inhibited growth of Neurospora crassa in medium adjusted to alkaline pH. Ninety-eight mutant strains were selected for growth on medium (pH 7.2) containing 0.3 or 1.0 microm bafilomycin. Three criteria suggested that 11 mutant strains were altered in the V-ATPase: 1) these strains accumulated high amounts of arginine when grown at pH 5.8 in the presence of bafilomycin, 2) the mutation mapped to the locus of vma-3, which encodes the proteolipid subunit c of the V-ATPase, and 3) V-ATPase activity in purified vacuolar membranes was resistant to bafilomycin. Sequencing of the genomic DNA encoding vma-3 identified the following mutations: T32I (two strains), F136L (two strains), Y143H (two strains), and Y143N (five strains). Characterization of V-ATPase activity in the four kinds of mutant strains showed that the enzyme was resistant to bafilomycin in vitro, with half-maximal inhibition obtained at 80-400 nm compared with 6.3 nm for the wild-type enzyme. Surprisingly, the mutant enzymes showed only weak resistance to concanamycin. Interestingly, the positions of two mutations corresponded to positions of oligomycin-resistant mutations in the c subunit of F(1)F(0)-ATP synthases (F-ATPases), suggesting that bafilomycin and oligomycin utilize a similar binding site and mechanism of inhibition in the related F- and V-ATPases
MH  - A
MH  - A1
MH  - Amino Acid Sequence
MH  - antagonists & inhibitors
MH  - ANTIBIOTIC
MH  - antibiotics,macrolide
MH  - Arginine
MH  - ATPase
MH  - Base Sequence
MH  - BINDING
MH  - BINDING SITE
MH  - Chemistry
MH  - Dicyclohexylcarbodiimide
MH  - Dna
MH  - DNA Primers
MH  - drug effects
MH  - enzyme
MH  - Enzyme Inhibitors
MH  - Enzymes
MH  - F-ATPASE
MH  - fungal proteins
MH  - genetics
MH  - growth & development
MH  - H(+)ATPase
MH  - In Vitro
MH  - inhibitor
MH  - mechanism
MH  - membrane
MH  - Membranes
MH  - Molecular Sequence Data
MH  - mutant
MH  - Mutation
MH  - Neurospora
MH  - Neurospora crassa
MH  - oligomycin
MH  - pH
MH  - pharmacology
MH  - proteolipid
MH  - RESISTANCE
MH  - Sequence Homology,Amino Acid
MH  - Site
MH  - SUBUNIT
MH  - Support,U.S.Gov't,P.H.S.
MH  - SYNTHASE
MH  - universities
MH  - Vacuolar Proton-Translocating ATPases
RP  - NOT IN FILE
NT  - Department of Molecular, University of California, Santa Cruz, California 95064, USA bowman@biologyucsceduFAU - Bowman, Barry J
SO  - J Biol Chem 2002 Feb 8 ;277(6):3965-3972

1876
UI  - 21395
AU  - Boyer PD
TI  - Catalytic site occupancy during ATP synthase catalysis
AB  - An early proposal was that for rapid ATP synthesis by the rotational ATP synthase, a specific second site must bind ADP and P(i), and for rapid ATP hydrolysis a different second site must bind ATP. Such bi-site activation was considered to occur whether or not an ADP or ATP was at a third site. In contrast, a more recent proposal is that rapid ATP hydrolysis requires that all three sites have bound ADP or ATP present. However, discovery that one second site binds ADP better than ATP, together with other data and considerations support the earlier proposal. The retention or rebinding of ADP can explain why three sites fill during hydrolysis as ATP concentration is increased although bi-site activation still prevails
MH  - A
MH  - ACTIVATION
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - ADP
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - biology
MH  - Catalysis
MH  - catalytic
MH  - catalytic domain
MH  - data
MH  - Enzyme Activation
MH  - Hydrolysis
MH  - metabolism
MH  - Models,Biological
MH  - Proton-Translocating ATPases
MH  - Site
MH  - SYNTHASE
MH  - synthesis
MH  - universities
RP  - NOT IN FILE
NT  - Molecular Biology Institute, Boyer Hall, University of California, Los Angeles, CA 90095-1570, USA pdboyer@uclaeduFAU - Boyer, Paul D
SO  - FEBS Lett 2002 Feb 13 ;512(1-3):29-32

1877
UI  - 21343
AU  - Boyer PD
TI  - A research journey with ATP synthase
MH  - A
MH  - atp
MH  - ATP synthase
MH  - biology
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - Molecular Biology Institute, UCLA, Los Angeles, California 90095-1570FAU - Boyer, Paul D
SO  - J Biol Chem 2002 Oct 18 ;277(42):39045-39061

1878
UI  - 21319
AU  - Brunner S
AU  - Everard-Gigot V
AU  - Stuart RA
TI  - Su e of the yeast F1Fo-ATP synthase forms homo-dimers
AB  - The yeast F(1)F(o)-ATP synthase forms a dimeric complex in the mitochondrial inner membrane. Dimerization of two F(1)F(o)-monomeric complexes involves the physical association of two membrane-embedded F(o)-sectors and in a manner, which is dependent on the F(o)-subunit, Su e. Sequence analysis of Su e protein family members indicated the presence of a conserved coiled-coil motif. As this motif is often the basis for protein homo-dimerization events, it was hypothesized that Su e forms homo-dimers in the inner membrane, and that formation Su e dimers between two neighboring F(o)-complexes would facilitate dimerization of the F(1)F(o)-ATP synthase complex (Arnold et al., (1998) EMBO J., 17, 7170-78). Using a histidine-tagged derivative of yeast Su e, Su e-His12, combined with cross-linking and affinity purification approaches, we have directly demonstrated the ability of the yeast Su e protein to form homo-dimers. Functionality of the Su e-His12 derivative was confirmed by its ability to assemble into the ATP synthase complex and to support its dimerization in the su e null mutant yeast cells. The close association of two neighboring Su e proteins was also demonstrated using cross-linking with Cu(++), which binds and cross-links a unique Cys residue in neighboring Su e proteins. Finally, we propose a model for the molecular basis of the homo-dimerization of the Su e proteins
MH  - A
MH  - affinity
MH  - analysis
MH  - atp
MH  - ATP synthase
MH  - cell
MH  - Cells
MH  - COMPLEX
MH  - CROSS-LINKING
MH  - Dimerization
MH  - membrane
MH  - model
MH  - mutant
MH  - protein
MH  - Proteins
MH  - purification
MH  - RESIDUE
MH  - SYNTHASE
MH  - universities
MH  - YEAST
RP  - NOT IN FILE
NT  - Department of Biological Sciences, Marquette University, Milwaukee, WI 53201-1881
SO  - J Biol Chem 2002 Oct 10 ;107(1).):

1879
UI  - 21342
AU  - Cabezon E
AU  - Butler PJ
AU  - Runswick MJ
AU  - Carbajo RJ
AU  - Walker JE
TI  - Homologous and heterologous inhibitory effects of ATPase inhibitor proteins on F-ATPases
AB  - In Saccharomyces cerevisiae, at least three proteins: IF1, STF1 and STF2 appear to be involved in the regulation of ATP synthase. Both IF1 and STF1 inhibit F1, whereas the proposed function for STF2 is to facilitate the binding of IF1 and STF1 to F1. The oligomerization properties of yeast IF1 and STF1 have been investigated by sedimentation equilibrium analytical ultracentrifugation and by covalent cross-linking. Both techniques confirm that IF1 and STF1 oligomerize in opposite directions in relation to pH, suggesting that both proteins might regulate yeast F1F0-ATPase under different conditions. Their effects on bovine F-ATPases are also described. Whereas bovine IF1 inhibits yeast F1-ATPase even better than yeast IF1 or STF1, the capability of yeast IF1 to inhibit the bovine enzyme is very low and it decreases with time. Such an effect is also observed in the study of the homologous inhibition of yeast F1-ATPase. Yeast inhibitors are not as effective as their bovine counterpart and the complex seems to dissociate gradually
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BINDING
MH  - COMPLEX
MH  - CROSS-LINKING
MH  - enzyme
MH  - equilibrium
MH  - F-ATPASE
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - function
MH  - Human
MH  - inhibitor
MH  - INHIBITOR PROTEIN
MH  - inhibitors
MH  - Nutrition
MH  - pH
MH  - protein
MH  - Proteins
MH  - regulation
MH  - Saccharomyces cerevisiae
MH  - SYNTHASE
MH  - Time
MH  - YEAST
RP  - NOT IN FILE
NT  - MRC-Dunn Human Nutrition Unit, MRC, Cambridge CB2 2XY
SO  - J Biol Chem 2002 Aug 17 ;.():

1880
UI  - 21389
AU  - Capaldi RA
AU  - Aggeler R
TI  - Mechanism of the F(1)F(0)-type ATP synthase, a biological rotary motor
AB  - The F(1)F(0)-type ATP synthase is a key enzyme in cellular energy interconversion. During ATP synthesis, this large protein complex uses a proton gradient and the associated membrane potential to synthesize ATP. It can also reverse and hydrolyze ATP to generate a proton gradient. The structure of this enzyme in different functional forms is now being rapidly elucidated. The emerging consensus is that the enzyme is constructed as two rotary motors, one in the F(1) part that links catalytic site events with movements of an internal rotor, and the other in the F(0) part, linking proton translocation to movements of this F(0) rotor. Although both motors can work separately, they must be connected together to interconvert energy. Evidence for the function of the rotary motor, from structural, genetic and biophysical studies, is reviewed here, and some uncertainties and remaining mysteries of the enzyme mechanism are also discussed
MH  - A
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - Bacteria
MH  - biology
MH  - catalytic
MH  - Chemistry
MH  - COMPLEX
MH  - energy
MH  - enzyme
MH  - enzymology
MH  - function
MH  - mechanism
MH  - membrane
MH  - Membrane Potential
MH  - Membrane Potentials
MH  - metabolism
MH  - Models,Molecular
MH  - Molecular Motors
MH  - Movement
MH  - physiology
MH  - protein
MH  - Protein Conformation
MH  - Protein Folding
MH  - proton
MH  - PROTON GRADIENT
MH  - Proton-Translocating ATPases
MH  - rotation
MH  - Site
MH  - structure
MH  - Support,Non-U.S.Gov't
MH  - Support,U.S.Gov't,P.H.S.
MH  - SYNTHASE
MH  - synthesis
MH  - translocation
MH  - universities
RP  - NOT IN FILE
NT  - Institute of Molecular Biology, University of Oregon, Eugene, OR 97403-1229, USA rcapaldi@oregonuoregoneduFAU - Capaldi, Roderick A
SO  - Trends Biochem Sci 2002 Mar ;27(3):154-160

1881
UI  - 21391
AU  - Cipriano DJ
AU  - Bi Y
AU  - Dunn SD
TI  - Genetic fusions of globular proteins to the epsilon subunit of the Escherichia coli ATP synthase: Implications for in vivo rotational catalysis and epsilon subunit function
AB  - The rotational mechanism of ATP synthase was investigated by fusing three proteins from Escherichia coli, the 12-kDa soluble cytochrome b(562), the 20-kDa flavodoxin, and the 28-kDa flavodoxin reductase, to the C terminus of the epsilon subunit of the enzyme. According to the concept of rotational catalysis, because epsilon is part of the rotor a large domain added at this site should sterically clash with the second stalk, blocking rotation and fully inhibiting the enzyme. E. coli cells expressing the cytochrome b(562) fusion in place of wild-type epsilon grew using acetate as the energy source, indicating their capacity for oxidative phosphorylation. Cells expressing the larger flavodoxin or flavodoxin reductase fusions failed to grow on acetate. Immunoblot analysis showed that the fusion proteins were stable in the cells and that they had no effect on enzyme assembly. These results provide initial evidence supporting rotational catalysis in vivo. In membrane vesicles, the cytochrome b(562) fusion caused an increase in the apparent ATPase activity but a minor decrease in proton pumping. Vesicles bearing ATP synthase containing the larger fusion proteins showed reduced but significant levels of ATPase activity that was sensitive to inhibition by dicyclohexylcarbodiimide (DCCD) but no proton pumping. Thus, all fusions to epsilon generated an uncoupled component of ATPase activity. These results imply that a function of the C terminus of epsilon in F(1)F(0) is to increase the efficiency of the enzyme by specifically preventing the uncoupled hydrolysis of ATP. Given the sensitivity to DCCD, this uncoupled ATP hydrolysis may arise from rotational steps of gammaepsilon in the inappropriate direction after ATP is bound at the catalytic site. It is proposed that the C-terminal domain of epsilon functions to ensure that rotation occurs only in the direction of ATP synthesis when ADP is bound and only in the direction of hydrolysis when ATP is bound
MH  - A
MH  - acetate
MH  - Adenosine Triphosphate
MH  - Adenosinetriphosphatase
MH  - ADP
MH  - analysis
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - ATP Synthetase Complexes
MH  - ATPase
MH  - Binding Sites
MH  - Biochemistry
MH  - Blotting,Western
MH  - Catalysis
MH  - catalytic
MH  - catalytic domain
MH  - cell
MH  - Cell Division
MH  - Cells
MH  - Chemistry
MH  - COLI ATP SYNTHASE
MH  - cytochrome
MH  - Cytochrome b
MH  - Dicyclohexylcarbodiimide
MH  - Dna
MH  - Dose-Response Relationship,Drug
MH  - energy
MH  - enzyme
MH  - enzymology
MH  - EPSILON-SUBUNIT
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - function
MH  - Hydrolysis
MH  - Immunoblotting
MH  - mechanism
MH  - membrane
MH  - membrane vesicles
MH  - metabolism
MH  - Models,Biological
MH  - Mutagenesis,Site-Directed
MH  - Oxidative Phosphorylation
MH  - Oxygen
MH  - pharmacology
MH  - Phosphorylation
MH  - Plasmids
MH  - protein
MH  - Protein Binding
MH  - Protein Structure,Tertiary
MH  - Proteins
MH  - proton
MH  - Protons
MH  - Recombinant Fusion Proteins
MH  - rotation
MH  - Site
MH  - stalk
MH  - SUBUNIT
MH  - Support,Non-U.S.Gov't
MH  - SYNTHASE
MH  - synthesis
MH  - Temperature
MH  - Time Factors
MH  - universities
MH  - vesicles
RP  - NOT IN FILE
NT  - Department of Biochemistry, University of Western Ontario, London, Ontario N6A 5C1, CanadaFAU - Cipriano, Daniel J
SO  - J Biol Chem 2002 May 10 ;277(19):16782-16790

1882
UI  - 21356
AU  - Cook RW
AU  - Jolly RD
AU  - Palmer DN
AU  - Tammen I
AU  - Broom MF
AU  - McKinnon R
TI  - Neuronal ceroid lipofuscinosis in Merino sheep
AB  - OBJECTIVE: To characterise neuronal ceroid lipofuscinosis (NCL) in Merino sheep. DESIGN: A prospective clinical, pathological, biochemical and genetic study. PROCEDURE: NCL cases were studied from a medium-wool Merino flock, the stud of origin of its replacement rams, and an experimental flock established at the University of Sydney. RESULTS: Behavioural changes and visual impairment were first detected at 7 to 12 months of age and progressed, with associated motor disturbances and at later stages seizures, to premature death by 27 months of age. At necropsy there was severe cerebrocortical atrophy associated with neuronal loss, astrocytosis and the presence in neurons of eosinophilic intracytoplasmic storage bodies with the characteristics of a lipopigment. In the retina there was progressive loss of photoreceptor cells. Storage bodies isolated from fresh brain, liver and pancreas formed electron-dense aggregates and coarse multilamellar and fine fingerprint profiles ultrastructurally, and consisted mainly of the hydrophobic protein, subunit c of mitochondrial ATP synthase. A homozygosity mapping approach localised the gene causing the disease in Merino sheep to the chromosomal region (OAR7q13-15) associated with NCL in South Hampshire sheep. CONCLUSION: NCL in Merino sheep is a subunit c-storing disease, clinically and pathologically similar to NCL in South Hampshire sheep. We propose that the disease in both breeds represents mutation at the same gene locus in chromosomal region OAR7q13-15
MH  - A
MH  - Animal
MH  - atp
MH  - ATP synthase
MH  - Blotting,Western
MH  - Brain
MH  - cell
MH  - Cells
MH  - complications
MH  - Electrophoresis,Gel,Two-Dimensional
MH  - epidemiology
MH  - etiology
MH  - Female
MH  - genetics
MH  - Genotype
MH  - Homozygote
MH  - Liver
MH  - Male
MH  - Neuronal Ceroid-Lipofuscinosis
MH  - New South Wales
MH  - pathology
MH  - Polymerase Chain Reaction
MH  - Prospective Studies
MH  - protein
MH  - Seizures
MH  - Sheep
MH  - Sheep Diseases
MH  - SUBUNIT
MH  - Support,Non-U.S.Gov't
MH  - Support,U.S.Gov't,P.H.S.
MH  - SYNTHASE
MH  - ultrastructure
MH  - universities
MH  - veterinary
RP  - NOT IN FILE
NT  - NSW Agriculture, Regional Veterinary Laboratory, WollongbarFAU - Cook, R W
SO  - Aust Vet J 2002 May ;80(5):292-297

1883
UI  - 21050
AU  - Curtis KK
AU  - Francis SA
AU  - Oluwatosin Y
AU  - Kane PM
TI  - Mutational analysis of the subunit C (Vma5p) of the yeast vacuolar H+-ATPase
AB  - Subunit C is a V1 sector subunit found in all V-ATPases that may be part of the peripheral stalk connecting the peripheral V1 sector with the membrane-bound Vo sector of the enzyme (Wilkens et al. (1999) J. Biol. Chem. 274, 31804-31810). To elucidate subunit C function we performed random and site-directed mutagenesis of the yeast VMA5 gene. Site-directed mutations in the most highly conserved region of Vma5p, residues 305-325, decreased catalytic activity of the V-ATPase by up to 48% without affecting assembly. A truncation mutant (K360stop), identified by random mutagenesis, suggested a small region near the C-terminus of the protein (amino acids 382-388) might be important for subunit stability. Site-directed mutagenesis revealed that three aromatic amino acids in this region (Y382, F385, and Y388), in addition to four other conserved aromatic amino acids (F260, Y262, F296, F300), are essential for stable assembly of V1 with V0, although alanine substitutions at these positions support some activity in vivo. Surprisingly, three mutations (F260A, Y262A, and F385A) greatly decrease the stability of the V-ATPase in vitro but increase its kcat for ATP hydrolysis and proton transport by at least three-fold. The peripheral stalk of V-ATPases must balance the stability essential for productive catalysis with the dynamic instability involved in regulation; these three mutations may perturb that balance
MH  - A
MH  - ACID
MH  - Amino Acids
MH  - analysis
MH  - atp
MH  - Biochemistry
MH  - Catalysis
MH  - function
MH  - H+-ATPase
MH  - Hydrolysis
MH  - In Vitro
MH  - mutagenesis
MH  - mutant
MH  - protein
MH  - proton
MH  - regulation
MH  - RESIDUE
MH  - site-directed
MH  - stalk
MH  - SUBUNIT
MH  - transport
RP  - NOT IN FILE
NT  - Biochemistry and Molecular Biology, SUNY Upstate Medical University, Syracuse, NY 13210
SO  - J Biol Chem 2002 Jan 3 ;.():

1884
UI  - 21300
AU  - Daizadeh I
AU  - Medvedev DM
AU  - Stuchebrukhov AA
AD  - Department of Molecular and Cellular Biology, Harvard University, Cambridge, MA, USA
TI  - Electron transfer in ferredoxin: are tunneling pathways evolutionarily conserved?
AB  - A theoretical study of electron transfer (ET) pathways in a recently crystallized Clostridium acidurici ferredoxin is reported. The electronic structure of the protein complex is treated at the semiempirical extended Huckel level, and the tunneling pathways are calculated with the rigorous quantum mechanical method of tunneling currents. The model predicts two pathways between the two [4Fe-4S] cubanes: a strong one running directly from Cys(14) to Cys(43) and a weaker one from Cys(14) via Ile(23) to Cys(18), whereas other amino acids do not play a significant role in the electron tunneling. The cysteine ligands conduct almost all of the current when Ile(23) is mutated to valine in silico, so that there is no appreciable change in the ET rate. The calculated value of the transfer matrix element is consistent with the experimentally determined rate of transfer. Results of the sequence analysis performed on this ferredoxin reveal that Ile(23) is a highly variable amino acid compared with the cubane-ligating cysteine amino acids, even though Ile(23) lies directly between the donor and acceptor complexes. We further argue that the homologous proteins with a [3Fe-4S] cofactor, which does not have one of the four cysteine ligands, use the same tunneling pathways as those in this ferredoxin, on the basis of the high homology as well as the absolute conservation of Cys(14) and Cys(43) which serve as the main tunneling conduit. Our results explain why mutation of amino acids around and between the donor and acceptor cubane clusters, including that of Ile(23), does not appreciably affect the rate of transfer and add support to the proposal that there exist evolutionarily conserved electron tunneling pathways in biological ET reactions
MH  - A
MH  - acceptor
MH  - ACID
MH  - Amino Acids
MH  - analysis
MH  - biology
MH  - COMPLEX
MH  - Cysteine
MH  - electron
MH  - electron transfer
MH  - Electron-transfer
MH  - Ligands
MH  - method
MH  - model
MH  - protein
MH  - Proteins
MH  - structure
MH  - TRANSFER
MH  - united states
MH  - universities
RP  - NOT IN FILE
NT  - UI - 21918418DA - 20020328IS - 0737-4038LA - engID - GM 54052-02/GM/NIGMSPT - Journal ArticleCY - United StatesRN - 0 (Amino Acids)RN - 0 (Ferredoxins)SB - IM
UR  - PM:11919281
SO  - Mol Biol Evol 2002 Apr ;19(4):406-415

1885
UI  - 21365
AU  - Del Rizzo PA
AU  - Bi Y
AU  - Dunn SD
AU  - Shilton BH
TI  - The "second stalk" of Escherichia coli ATP synthase: structure of the isolated dimerization domain
AB  - The b subunit of E. coli F(0)F(1)-ATPase links the peripheral F(1) subunits to the membrane-integral F(0) portion and functions as a "stator", preventing rotation of F(1). The b subunit is present as a dimer in ATP synthase, and residues 62-122 are required to mediate dimerization. To understand how the b subunit dimer is formed, we have studied the structure of the isolated dimerization domain, b(62-122). Analytical ultracentrifugation and solution small-angle X-ray scattering (SAXS) indicate that the b(62-122) dimer is extremely elongated, with a frictional ratio of 1.60, a maximal dimension of 95 A, and a radius of gyration of 27 A, values that are consistent with an alpha-helical coiled-coil structure. The crystal structure of b(62-122) has been solved and refined to 1.55 A. The protein crystallized as an isolated, monomeric alpha helix with a length of 90 A. Combining the crystal structure of monomeric b(62-122) with SAXS data from the dimer in solution, we have constructed a model for the b(62-122) dimer in which the two helices form a coiled coil with a right-handed superhelical twist. Analysis of b sequences from E. coli and other prokaryotes indicates conservation of an undecad repeat, which is characteristic of a right-handed coiled coil and consistent with our structural model. Mutation of residue Arg-83, which interrupts the undecad pattern, to alanine markedly stabilized the dimer, as expected for the proposed two-stranded, right-handed coiled-coil structure
MH  - A
MH  - alpha
MH  - Amino Acid Sequence
MH  - analysis
MH  - Arginine
MH  - atp
MH  - ATP synthase
MH  - Biochemistry
MH  - Chemistry
MH  - COLI ATP SYNTHASE
MH  - Crystallography,X-Ray
MH  - data
MH  - Dimerization
MH  - enzymology
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - function
MH  - genetics
MH  - model
MH  - Models,Molecular
MH  - Molecular Sequence Data
MH  - Mutation
MH  - protein
MH  - Protein Conformation
MH  - Protein Structure,Tertiary
MH  - Protein Subunits
MH  - Proton-Translocating ATPases
MH  - RESIDUE
MH  - rotation
MH  - structure
MH  - SUBUNIT
MH  - Support,Non-U.S.Gov't
MH  - Support,U.S.Gov't,Non-P.H.S.
MH  - Support,U.S.Gov't,P.H.S.
MH  - SYNTHASE
MH  - universities
RP  - NOT IN FILE
NT  - Department of Biochemistry, University of Western Ontario, London, ON N6A 5C1, CanadaFAU - Del Rizzo, Paul A
SO  - Biochemistry 2002 May 28 ;41(21):6875-6884

1886
UI  - 21276
AU  - Dmitriev OY
AU  - Abildgaard F
AU  - Markley JL
AU  - Fillingame RH
AD  - Department of Biomolecular Chemistry, University of Wisconsin Medical School, and Department of Biochemistry, University of Wisconsin, Madison, Wisconsin 53706
TI  - Structure of Ala24/Asp61 --> Asp24/Asn61 Substituted Subunit c of Escherichia coli ATP Synthase: Implications for the Mechanism of Proton Transport and Rotary Movement in the F(o) Complex
AB  - The structure of the A24D/D61N substituted subunit c of Escherichia coli ATP synthase, in which the essential carboxylate has been switched from residue 61 of the second transmembrane helix (TMH) to residue 24 of the first TMH, has been determined by heteronuclear multidimensional NMR in a monophasic chloroform/methanol/water (4:4:1) solvent mixture. As in the case of the wild-type protein, A24D/D61N substituted subunit c forms a hairpin of two extended alpha-helices (residues 5-39 and 46- 78), with residues 40-45 forming a connecting loop at the center of the protein. The structure was determined at pH 5, where Asp24 is fully protonated. The relative orientation of the two extended helices in the A24D/D61N structure is different from that in the protonated form of the wild-type protein, also determined at pH 5. The C-terminal helix is rotated by 150 degrees relative to the wild-type structure, and the N- terminal helix is rotated such that the essential Asp24 carboxyl group packs on the same side of the molecule as Asp61 in the wild-type protein. The changes in helix-helix orientation lead to a structure that is quite similar to that of the deprotonated form of wild-type subunit c, determined at pH 8. When a decameric ring of c subunits was modeled from the new structure, the Asp24 carboxyl group was found to pack in a cavity at the interface between two subunits that is similar to the cavity in which Asp61 of the wild-type protein is predicted to pack. The interacting faces of the packed subunits in this model are also similar to those in the wild-type model. The results provide further evidence that subunit c is likely to fold in at least two conformational states differing most notably in the orientation of the C-terminal helix. Based upon the structure, a mechanistic model is discussed that indicates how the wild-type and A24D/D61N subunits could utilize similar helical movements during H(+) transport-coupled rotation of the decameric c ring
MH  - A
MH  - atp
MH  - ATP synthase
MH  - Biochemistry
MH  - Chemistry
MH  - COLI ATP SYNTHASE
MH  - COMPLEX
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - mechanism
MH  - model
MH  - Movement
MH  - NMR
MH  - pH
MH  - protein
MH  - proton
MH  - RESIDUE
MH  - rotation
MH  - SOLVENT
MH  - structure
MH  - SUBUNIT
MH  - SYNTHASE
MH  - transport
RP  - NOT IN FILE
NT  - UI - 21966003LA - engPT - Journal ArticleDA - 20020423IS - 0006-2960SB - IMCY - United States
UR  - PM:11969414
SO  - Biochemistry 2002 Apr 30 ;41(17):5537-5547

1887
UI  - 21374
AU  - Dmitriev OY
AU  - Abildgaard F
AU  - Markley JL
AU  - Fillingame RH
TI  - Structure of Ala24/Asp61 --> Asp24/Asn61 substituted subunit c of Escherichia coli ATP synthase: implications for the mechanism of proton transport and rotary movement in the F0 complex
AB  - The structure of the A24D/D61N substituted subunit c of Escherichia coli ATP synthase, in which the essential carboxylate has been switched from residue 61 of the second transmembrane helix (TMH) to residue 24 of the first TMH, has been determined by heteronuclear multidimensional NMR in a monophasic chloroform/methanol/water (4:4:1) solvent mixture. As in the case of the wild-type protein, A24D/D61N substituted subunit c forms a hairpin of two extended alpha-helices (residues 5-39 and 46-78), with residues 40-45 forming a connecting loop at the center of the protein. The structure was determined at pH 5, where Asp24 is fully protonated. The relative orientation of the two extended helices in the A24D/D61N structure is different from that in the protonated form of the wild-type protein, also determined at pH 5. The C-terminal helix is rotated by 150 degrees relative to the wild-type structure, and the N-terminal helix is rotated such that the essential Asp24 carboxyl group packs on the same side of the molecule as Asp61 in the wild-type protein. The changes in helix-helix orientation lead to a structure that is quite similar to that of the deprotonated form of wild-type subunit c, determined at pH 8. When a decameric ring of c subunits was modeled from the new structure, the Asp24 carboxyl group was found to pack in a cavity at the interface between two subunits that is similar to the cavity in which Asp61 of the wild-type protein is predicted to pack. The interacting faces of the packed subunits in this model are also similar to those in the wild-type model. The results provide further evidence that subunit c is likely to fold in at least two conformational states differing most notably in the orientation of the C-terminal helix. Based upon the structure, a mechanistic model is discussed that indicates how the wild-type and A24D/D61N subunits could utilize similar helical movements during H(+) transport-coupled rotation of the decameric c ring
MH  - A
MH  - Alanine
MH  - Amino Acid Sequence
MH  - Amino Acid Substitution
MH  - Asparagine
MH  - Aspartic Acid
MH  - atp
MH  - ATP synthase
MH  - Bacterial Proton-Translocating ATPases
MH  - Chemistry
MH  - COLI ATP SYNTHASE
MH  - COMPLEX
MH  - Computer Simulation
MH  - Crystallography,X-Ray
MH  - Escherichia coli
MH  - Escherichia coli Proteins
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - genetics
MH  - mechanism
MH  - Mitochondrial Proton-Translocating ATPases
MH  - model
MH  - Models,Molecular
MH  - Molecular Sequence Data
MH  - Movement
MH  - Mutagenesis,Site-Directed
MH  - NMR
MH  - Nuclear Magnetic Resonance,Biomolecular
MH  - pH
MH  - protein
MH  - proton
MH  - RESIDUE
MH  - rotation
MH  - SOLVENT
MH  - structure
MH  - SUBUNIT
MH  - Support,Non-U.S.Gov't
MH  - Support,U.S.Gov't,Non-P.H.S.
MH  - Support,U.S.Gov't,P.H.S.
MH  - SYNTHASE
MH  - Thermodynamics
MH  - transport
MH  - universities
RP  - NOT IN FILE
NT  - Department of Biomolecular Chemistry, University of Wisconsin Medical School, Madison, Wisconsin 53706, USAFAU - Dmitriev, Oleg Y
SO  - Biochemistry 2002 Apr 30 ;41(17):5537-5547

1888
UI  - 21406
AU  - Dong K
AU  - Ren H
AU  - Allison WS
TI  - The fluorescence spectrum of the introduced tryptophans in the alpha 3(beta F155W)3gamma subcomplex of the F1-ATPase from the thermophilic Bacillus PS3 cannot be used to distinguish between the number of nucleoside di- and triphosphates bound to catalytic sites
AB  - It has been reported that shifts in the fluorescence emission spectrum of the introduced tryptophans in the betaF155W mutant of Escherichia coli F(1) (bovine heart mitochondria F(1) residue number) can quantitatively distinguish between the number of catalytic sites occupied with ADP and ATP during steady-state ATP hydrolysis (Weber, J., Bowman, C., and Senior, A. E. (1996) J. Biol. Chem. 271, 18711--18718). In contrast, addition of MgADP, Mg-5'-adenylyl beta,gamma-imidophosphate (MgAMP-PNP), and MgATP in 1:1 ratios to the alpha(3)(betaF155W)(3)gamma subcomplex of thermophilic Bacillus PS3 F(1) (TF(1)) induced nearly identical blue shifts in the fluorescence emission maximum that was accompanied by quenching. Addition of 2 mm MgADP induced a slightly greater blue shift and a slight increase in intensity over those observed with 1:1 MgADP. However, addition of 2 mm MgAMP-PNP or MgATP induced a much greater blue shift and substantially enhanced fluorescence intensity over those observed in the presence of stoichiometric MgADP or MgAMP-PNP. It is clear from these results that the fluorescence spectrum of the introduced tryptophans in the betaF155W mutant of TF(1) does not respond in regular increments at any wavelength as catalytic sites are filled with nucleotides. The fluorescence spectrum observed after entrapping MgADP-fluoroaluminate complexes in two catalytic sites of the betaF155W subcomplex indicates that the fluorescence emission spectrum of the enzyme is maximally perturbed when nucleotides are bound to two catalytic sites. This finding is consistent with accumulating evidence suggesting that only two beta subunits in the alpha(3)beta(3)gamma subcomplex of TF(1) can simultaneously exist in the completely closed conformation
MH  - A
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - Adenylyl Imidodiphosphate
MH  - ADP
MH  - alpha
MH  - atp
MH  - Bacillus
MH  - BETA
MH  - BETA-SUBUNIT
MH  - Biochemistry
MH  - catalytic
MH  - catalytic domain
MH  - Chemistry
MH  - COMPLEX
MH  - conformation
MH  - enzyme
MH  - enzymology
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - fluorescence
MH  - Hydrolysis
MH  - metabolism
MH  - Mitochondria
MH  - mutant
MH  - Mutation
MH  - nucleotide
MH  - Nucleotides
MH  - Protein Subunits
MH  - Proton-Translocating ATPases
MH  - PS3
MH  - RESIDUE
MH  - Site
MH  - spectra
MH  - Spectrometry,Fluorescence
MH  - SUBUNIT
MH  - Support,U.S.Gov't,P.H.S.
MH  - THERMOPHILIC
MH  - THERMOPHILIC BACILLUS PS3
MH  - tryptophan
MH  - universities
RP  - NOT IN FILE
NT  - Department of Chemistry and Biochemistry, University of California at San Diego, La Jolla, California 92093-0601, USAFAU - Dong, Ken
SO  - J Biol Chem 2002 Mar 15 ;277(11):9540-9547

1889
UI  - 21368
AU  - Dopson M
AU  - Lindstrom EB
AU  - Hallberg KB
TI  - ATP generation during reduced inorganic sulfur compound oxidation by Acidithiobacillus caldus is exclusively due to electron transport phosphorylation
AB  - The synthesis of adenosine 5-triphosphate (ATP) (increase in phosphorylation potential) during the oxidation of reduced inorganic sulfur compounds was studied in the moderately thermophilic acidophileAcidithiobacillus caldus (strain KU) (formerly Thiohacillus caldus). The phosphorylation potential increased during the oxidation of all reduced inorganic sulfur compounds tested compared with resting cells. The generation of ATP in whole cells was inhibited by the F0F1 ATPase inhibitor oligomycin, electron transport chain inhibitors, valinomycin and potassium ions. There was no increase in the phosphorylation potential, nor synthesis of ATP. in the absence of electron transport. An apparent lack of substrate-level phosphorylation was indicated by the lack of adenosine 5-phosphosulfate reductase in tetrathionate-grown At. caldus. Studies were also performed on the synthesis of ATP by membrane vesicles of At. caldus when presented with an artificial proton gradient. Complete inhibition of ATP synthesis in these vesicles occurred when they were loaded with N,N-dicyclohexylcarbodiimide (DCCD), but not when they were loaded with oligomycin, vanadate or electron transport chain inhibitors. The data presented here suggest that during the oxidation of reduced inorganic sulfur compounds by At. caldus, all ATP is synthesized by oxidative phosphorylation via a membrane-bound F0F1 ATPase driven by a proton gradient
MH  - A
MH  - Adenosine
MH  - atp
MH  - ATP synthesis
MH  - ATPase
MH  - cell
MH  - Cells
MH  - data
MH  - electron
MH  - Electron Transport
MH  - F0F1
MH  - inhibitor
MH  - inhibitors
MH  - ion
MH  - Ions
MH  - membrane
MH  - membrane vesicles
MH  - oligomycin
MH  - Oxidative Phosphorylation
MH  - Phosphorylation
MH  - Potassium
MH  - proton
MH  - PROTON GRADIENT
MH  - synthesis
MH  - THERMOPHILIC
MH  - transport
MH  - universities
MH  - Valinomycin
MH  - vesicles
RP  - NOT IN FILE
NT  - Department of Microbiology, Umea University, SwedenFAU - Dopson, Mark
SO  - Extremophiles 2002 Apr ;6(2):123-129

1890
UI  - 21354
AU  - Drobny M
AU  - Schnolzer M
AU  - Fiedler S
AU  - Luttge U
AU  - Fischer-Schliebs E
AU  - Christian AL
AU  - Ratajczak R
TI  - Phenotypic subunit composition of the tobacco (Nicotiana tabacum L.) vacuolar-type H(+)-translocating ATPase
AB  - The model plant tobacco (Nicotiana tabacum L.) was chosen for a survey of the subunit composition of the V-ATPase at the protein level. V-ATPase was purified from tobacco leaf cell tonoplasts by solubilization with the nonionic detergent Triton X-100 and immunoprecipitation. In the purified fraction 12 proteins were present. By matrix-assisted laser-desorption ionization mass spectrometry (MALDI-MS) and amino acid sequencing 11 of these polypeptides could be identified as subunits A, B, C, D, F, G, c, d and three different isoforms of subunit E. The polypeptide which could not be identified by MALDI analysis might represent subunit H. The data presented here, for the first time, enable an unequivocal identification of V-ATPase subunits after gel electrophoresis and open the possibility to assign changes in polypeptide composition to variations in respective V-ATPase subunits occurring as a response to environmental conditions or during plant development
MH  - A
MH  - ACID
MH  - Amino Acid Sequence
MH  - analysis
MH  - ATPase
MH  - cell
MH  - Chemistry
MH  - data
MH  - development
MH  - Electrophoresis
MH  - enzymology
MH  - genetics
MH  - isolation & purification
MH  - model
MH  - Models,Molecular
MH  - Molecular Sequence Data
MH  - Molecular Weight
MH  - Peptide Fragments
MH  - Phenotype
MH  - plant
MH  - protein
MH  - Protein Subunits
MH  - Proteins
MH  - Spectrometry,Mass,Matrix-Assisted Laser Desorption-Ionization
MH  - SUBUNIT
MH  - Support,Non-U.S.Gov't
MH  - Time
MH  - Tobacco
MH  - Trypsin
MH  - universities
MH  - Vacuolar Proton-Translocating ATPases
RP  - NOT IN FILE
NT  - Darmstadt University of Technology, Institute of Botany, Schnittspahnstrasse 3-5, D-64287, Darmstadt, GermanyFAU - Drobny, Martina
SO  - Biochim Biophys Acta 2002 Aug ;%19;1564(1):243-255

1891
UI  - 21258
AU  - Dutzler R
AU  - Campbell EB
AU  - Cadene M
AU  - Chait BT
AU  - MacKinnon R
AD  - Howard Hughes Medical Institute, Laboratory of Molecular Neurobiology and Biophysics, Rockefeller University, 1230 York Avenue, New York, New York 10021, USA
TI  - X-ray structure of a ClC chloride channel at 3.0 A reveals the molecular basis of anion selectivity
AB  - The ClC chloride channels catalyse the selective flow of Cl- ions across cell membranes, thereby regulating electrical excitation in skeletal muscle and the flow of salt and water across epithelial barriers. Genetic defects in ClC Cl- channels underlie several familial muscle and kidney diseases. Here we present the X-ray structures of two prokaryotic ClC Cl- channels from Salmonella enterica serovar typhimurium and Escherichia coli at 3.0 and 3.5 A, respectively. Both structures reveal two identical pores, each pore being formed by a separate subunit contained within a homodimeric membrane protein. Individual subunits are composed of two roughly repeated halves that span the membrane with opposite orientations. This antiparallel architecture defines a selectivity filter in which a Cl- ion is stabilized by electrostatic interactions with alpha-helix dipoles and by chemical coordination with nitrogen atoms and hydroxyl groups. These findings provide a structural basis for further understanding the function of ClC Cl- channels, and establish the physical and chemical basis of their anion selectivity
MH  - A
MH  - Anions
MH  - Bacterial Proteins
MH  - Biophysics
MH  - Cell Membrane
MH  - England
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - function
MH  - ion
MH  - Ions
MH  - Kidney
MH  - membrane
MH  - Membranes
MH  - protein
MH  - Proteins
MH  - structure
MH  - SUBUNIT
MH  - Water
RP  - NOT IN FILE
NT  - UI - 21655566LA - engRN - 0 (Anions)RN - 0 (Bacterial Proteins)RN - 0 (Chloride Channels)RN - 0 (Escherichia coli Proteins)RN - 0 (Recombinant Proteins)PT - Journal ArticleDA - 20020117IS - 0028-0836SB - IMCY - England
UR  - PM:11796999
SO  - Nature 2002 Jan 17 ;415(6869):287-294

1892
UI  - 21154
AU  - Feniouk BA
AU  - Cherepanov DA
AU  - Voskoboynikova NE
AU  - Mulkidjanian AY
AU  - Junge W
AD  - Division of Biophysics, Faculty of Biology/Chemistry, University of Osnabruck, D-49069 Osnabruck, Germany
TI  - Chromatophore Vesicles of Rhodobacter capsulatus Contain on Average One F(O)F(1)-ATP Synthase Each
AB  - ATP synthase is a unique rotary machine that uses the transmembrane electrochemical potential difference of proton (Delta(H(+))) to synthesize ATP from ADP and inorganic phosphate. Charge translocation by the enzyme can be most conveniently followed in chromatophores of phototrophic bacteria (vesicles derived from invaginations of the cytoplasmic membrane). Excitation of chromatophores by a short flash of light generates a step of the proton-motive force, and the charge transfer, which is coupled to ATP synthesis, can be spectrophotometrically monitored by electrochromic absorption transients of intrinsic carotenoids in the coupling membrane. We assessed the average number of functional enzyme molecules per chromatophore vesicle. Kinetic analysis of the electrochromic transients plus/minus specific ATP synthase inhibitors (efrapeptin and venturicidin) showed that the extent of the enzyme-related proton transfer dropped as a function of the inhibitor concentration, whereas the time constant of the proton transfer changed only marginally. Statistical analysis of the kinetic data revealed that the average number of proton-conducting F(O)F(1)-molecules per chromatophore was approximately one. Thereby chromatophores of Rhodobacter capsulatus provide a system where the coupling of proton transfer to ATP synthesis can be studied in a single enzyme/single vesicle mode
MH  - A
MH  - absorption
MH  - ADP
MH  - analysis
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - Bacteria
MH  - Biophysics
MH  - capsulatus
MH  - carotenoid
MH  - Carotenoids
MH  - chromatophore
MH  - chromatophores
MH  - CONSTANT
MH  - coupling
MH  - flash
MH  - function
MH  - inhibitor
MH  - INORGANIC-PHOSPHATE
MH  - Light
MH  - membrane
MH  - proton
MH  - Proton transfer
MH  - Proton-Motive Force
MH  - Proton-transfer
MH  - Rhodobacter
MH  - rhodobacter capsulatus
MH  - RHODOBACTER-CAPSULATUS
MH  - SYNTHASE
MH  - synthesis
MH  - SYSTEM
MH  - Time
MH  - TRANSFER
MH  - translocation
MH  - vesicles
RP  - NOT IN FILE
NT  - UI - 21855976LA - engPT - Journal ArticleDA - 20020227IS - 0006-3495SB - IMCY - United States
UR  - PM:11867431
SO  - Biophys J 2002 Mar ;82(3):1115-1122

1893
UI  - 21355
AU  - Ferrandiz MJ
AU  - de la Campa AG
TI  - The membrane-associated F(0)F(1) ATPase is essential for the viability of Streptococcus pneumoniae
AB  - Genetic studies aimed at eliminating expression of the atp operon (F(0)F(1) H(+)-ATPase) of Streptococcus pneumoniae by genetic disruption of atpC, the first gene of the operon, with a chloramphenicol-resistance cassette were performed. Resistant transformants were obtained only when the recipient strain had a duplication of atpC, recombination occurring in such a way that transcription of the operon from its own promoter was allowed. These results imply that the atp operon is essential for the viability of the cells
MH  - A
MH  - atp
MH  - ATPase
MH  - cell
MH  - Cell Membrane
MH  - Cells
MH  - Chloramphenicol O-Acetyltransferase
MH  - enzymology
MH  - Gene Deletion
MH  - Genes,Essential
MH  - genetics
MH  - growth & development
MH  - metabolism
MH  - Mitochondrial Proton-Translocating ATPases
MH  - Proton-Translocating ATPases
MH  - Recombinant Fusion Proteins
MH  - Recombination,Genetic
MH  - Streptococcus
MH  - Streptococcus pneumoniae
MH  - Support,Non-U.S.Gov't
MH  - Transformation,Bacterial
RP  - NOT IN FILE
NT  - Unidad de Genetica Bacteriana (Consejo Superior de Investigaciones Cientificas), Centro Nacional de Microbiologia, Instituto de Salud Carlos III, 28220 Majadahonda, Madrid, SpainFAU - Ferrandiz, Maria Jose
SO  - FEMS Microbiol Lett 2002 Jun 18 ;212(1):133-138

1894
UI  - 21338
AU  - Fillingame RH
AU  - Angevine CM
AU  - Dmitriev OY
TI  - Coupling proton movements to c-ring rotation in F(1)F(o) ATP synthase: aqueous access channels and helix rotations at the a-c interface
AB  - F(1)F(o) ATP synthases generate ATP by a rotary catalytic mechanism in which H(+) transport is coupled to rotation of a ring of c subunits within the transmembrane sector of the enzyme. Protons bind to and then are released from the aspartyl-61 residue of subunit c at the center of the membrane. Proton access channels to and from aspartyl-61 are thought to form in subunit a of the F(o) sector. Here, we summarize new information on the structural organization of subunit a and the mapping of aqueous accessible residues in the fourth and fifth transmembrane helices (TMHs). Cysteine substituted residues, lying on opposite faces of aTMH-4, preferentially react with either N-ethyl-maleimide (NEM) or Ag(+). We propose that aTMH-4 rotates to alternately expose each helical face to aspartyl-61 of subunit c during the proton transport cycle. The concerted helical rotation of aTMH-4 and cTMH-2 are proposed to be coupled to the stepwise mechanical movement of the c-rotor
MH  - A
MH  - Aquaporins
MH  - atp
MH  - ATP synthase
MH  - catalytic
MH  - Chemistry
MH  - coupling
MH  - Cysteine
MH  - enzyme
MH  - Escherichia coli
MH  - Ethylmaleimide
MH  - Hydrogen-Ion Concentration
MH  - Magnetic Resonance Spectroscopy
MH  - mechanism
MH  - membrane
MH  - Molecular Motors
MH  - Molecular Structure
MH  - Movement
MH  - proton
MH  - Proton-Translocating ATPases
MH  - Protons
MH  - RESIDUE
MH  - rotation
MH  - SUBUNIT
MH  - SYNTHASE
MH  - transport
MH  - universities
RP  - NOT IN FILE
NT  - Department of Biomolecular Chemistry, University of Wisconsin Medical School, Madison 53706-1532, USA rhfillin@facstaffwisceduFAU - Fillingame, Robert H
SO  - Biochim Biophys Acta 2002 Sep 10 ;1555(1-3):29-36

1895
UI  - 21320
AU  - Gaballo A
AU  - Zanotti F
AU  - Papa S
TI  - Structures and Interactions of Proteins Involved in the Coupling Function of the Protonmotive F(o)F(1)-ATP Synthase
AB  - The mitochondrial F(1)F(o) ATP synthase complex has a key role in cellular energy metabolism. The general architecture of the enzyme is conserved among species and consists of a globular catalytic moiety F(1), protruding out of the inner side of the membrane, a membrane integral proton translocating moiety F(o), and a stalk connecting F(1) to F(o). The X-ray crystallographic analysis of the structure of the bovine mitochondrial F(1) ATPase has provided a structural basis for the binding-change rotary mechanism of the catalytic process in F(1), in which the gamma subunit rotates in the central cavity of the F(1) alpha3/beta3 hexamer. Rotation of gamma and eta subunits in the E. coli enzyme and of, gamma and delta subunits in the mitochondrial enzyme, is driven, during ATP synthesis, by proton motive rotation of an oligomer of c subunits (10-12 copies) within the F(o) base piece. Average analysis of electron microscopy images and cross-linking results have revealed that, in addition to a central stalk, contributed by gamma and delta/eta subunits, there is a second lateral one connecting the peripheries of F(o) and F(1). To gain deeper insight into the mechanism of coupling between proton translocation and catalytic activity (ATP synthesis and hydrolysis), studies have been undertaken on the role of F(1) and F(o) subunits which contribute to the structural and functional connection between the catalytic sector F(1) and the proton translocating moiety F(o). These studies, which employed limited proteolysis, chemical cross-linking and functional analysis of the native and reconstituted F(1)F(o) complex, as well as isolated F(1), have shown that the N-terminus of alpha subunits, located at the top of the F(1) hexamer is essential for energy coupling in the F(1)F(o) complex. The alpha N-terminus domain appears to be connected to F(o) by OSCP (F(o) subunit conferring sensitivity of the complex to oligomycin). In turn, OSCP contacts F(o)I-PVP(b) and d subunits, with which it constitutes a structure surrounding the central gamma and delta rotary shaft. Cross-linking of F(o)I-PVP(b) and gamma subunits causes a dramatic enhancement of downhill proton translocation decoupled from ATP synthesis but is without effect on ATP driven uphill proton transport. This would indicate the existence of different rate-limiting steps in the two directions of proton translocation through F(o). In mitochondria, futile ATP hydrolysis by the F(1)F(o) complex is inhibited by the ATPase inhibitor protein (IF(1)), which reversibly binds at one side of the F(1)F(o) connection. The trans-membrane DeltapH component of the respiratory Deltap displaces IF(1) from the complex; in particular the matrix pH is the critical factor for IF(1)association and its related inhibitory activity. The 42L-58K segment of the IF(1) has been shown to be the most active segment of the protein; it interacts with the surface of one alpha/beta pairs of F(1), thus inhibiting, with the same pH dependence as the natural IF(1), the conformational interconversions of the catalytic sites involved in ATP hydrolysis. IF(1) has a relevant physiopathological role for the conservation of the cellular ATP pool in ischemic tissues. Under these conditions IF(1), which appears to be over expressed, prevents dissipation of the glycolytic ATP
MH  - A
MH  - ACTIVE
MH  - alpha
MH  - ALPHA-SUBUNIT
MH  - analysis
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - ATPase
MH  - BASE
MH  - bioenergetics
MH  - catalytic
MH  - COMPLEX
MH  - coupling
MH  - CROSS-LINKING
MH  - delta
MH  - DELTA-SUBUNIT
MH  - DEPENDENCE
MH  - electron
MH  - electron microscopy
MH  - ELECTRON-MICROSCOPY
MH  - energy
MH  - Energy Metabolism
MH  - enzyme
MH  - function
MH  - Hydrolysis
MH  - inhibitor
MH  - INHIBITOR PROTEIN
MH  - INTERACTION
MH  - mechanism
MH  - membrane
MH  - metabolism
MH  - Microscopy
MH  - Mitochondria
MH  - oligomycin
MH  - pH
MH  - protein
MH  - Proteins
MH  - proton
MH  - rotation
MH  - Site
MH  - stalk
MH  - structure
MH  - SUBUNIT
MH  - SURFACE
MH  - SYNTHASE
MH  - synthesis
MH  - translocation
MH  - transport
MH  - universities
RP  - NOT IN FILE
NT  - Institute of Biomembranes and Bioenergetics, Consiglio Nazionale delle Ricerche, University of Bari, Bari, Italy agaballo@biochemunibaitFAU - Gaballo, A
SO  - Curr Protein Pept Sci 2002 Aug ;3(4):451-460

1896
UI  - 21364
AU  - Georgievskii Y
AU  - Medvedev ES
AU  - Stuchebrukhov AA
TI  - Proton transport via the membrane surface
AB  - Some proton pumps, such as cytochrome c oxidase (C(c)O), translocate protons across biological membranes at a rate that considerably exceeds the rate of proton transport to the entrance of the proton-conducting channel via bulk diffusion. This effect is usually ascribed to a proton-collecting antenna surrounding the channel entrance. In this paper, we consider a realistic phenomenological model of such an antenna. In our model, a homogeneous membrane surface, which can mediate proton diffusion toward the channel entrance, is populated with protolytic groups that are in dynamic equilibrium with the solution. Equations that describe coupled surface-bulk proton diffusion are derived and analyzed. A general expression for the rate constant of proton transport via such a coupled surface-bulk diffusion mechanism is obtained. A rigorous criterion is formulated of when proton diffusion along the surface enhances the transport. The enhancement factor is found to depend on the ratio of the surface and bulk diffusional constants, pK(a) values of surface protolytic groups, and their concentration. A capture radius for a proton on the surface and an effective size of the antenna are found. The theory also predicts the effective distance that a proton can migrate on the membrane surface between a source (such as CcO) and a sink (such as ATP synthase) without fully equilibrating with the bulk. In pure aqueous solutions, protons can travel over long distances (microns). In buffered solutions, the travel distance is much shorter (nanometers); still the enhancement effect of the surface diffusion on the proton flow to a target on the surface can be tens to hundreds at physiological buffer concentrations. These results are discussed in a general context of chemiosmotic theory
MH  - A
MH  - atp
MH  - ATP synthase
MH  - Biophysics
MH  - buffer
MH  - Buffers
MH  - chemiosmotic theory
MH  - Chemistry
MH  - CONSTANT
MH  - cytochrome
MH  - Cytochrome c
MH  - Cytochrome-c Oxidase
MH  - Diffusion
MH  - equilibrium
MH  - Ion Channels
MH  - Ion Transport
MH  - Kinetics
MH  - Mathematics
MH  - mechanism
MH  - membrane
MH  - Membranes
MH  - metabolism
MH  - model
MH  - Models,Biological
MH  - proton
MH  - Proton Pump
MH  - Proton Pumps
MH  - Protons
MH  - rate constant
MH  - Solutions
MH  - SURFACE
MH  - SYNTHASE
MH  - theory
MH  - transport
MH  - universities
MH  - Water
RP  - NOT IN FILE
NT  - Department of Chemistry, University of California, Davis, California 95616, USAFAU - Georgievskii, Yuri
SO  - Biophys J 2002 Jun ;82(6):2833-2846

1897
UI  - 21301
AU  - Georgievskii Y
AU  - Medvedev ES
AU  - Stuchebrukhov AA
AD  - Department of Chemistry, University of California, Davis, California 95616 USA
TI  - Proton Transport via the Membrane Surface
AB  - Some proton pumps, such as cytochrome c oxidase (C(c)O), translocate protons across biological membranes at a rate that considerably exceeds the rate of proton transport to the entrance of the proton-conducting channel via bulk diffusion. This effect is usually ascribed to a proton-collecting antenna surrounding the channel entrance. In this paper, we consider a realistic phenomenological model of such an antenna. In our model, a homogeneous membrane surface, which can mediate proton diffusion toward the channel entrance, is populated with protolytic groups that are in dynamic equilibrium with the solution. Equations that describe coupled surface-bulk proton diffusion are derived and analyzed. A general expression for the rate constant of proton transport via such a coupled surface-bulk diffusion mechanism is obtained. A rigorous criterion is formulated of when proton diffusion along the surface enhances the transport. The enhancement factor is found to depend on the ratio of the surface and bulk diffusional constants, pK(a) values of surface protolytic groups, and their concentration. A capture radius for a proton on the surface and an effective size of the antenna are found. The theory also predicts the effective distance that a proton can migrate on the membrane surface between a source (such as CcO) and a sink (such as ATP synthase) without fully equilibrating with the bulk. In pure aqueous solutions, protons can travel over long distances (microns). In buffered solutions, the travel distance is much shorter (nanometers); still the enhancement effect of the surface diffusion on the proton flow to a target on the surface can be tens to hundreds at physiological buffer concentrations. These results are discussed in a general context of chemiosmotic theory
MH  - A
MH  - atp
MH  - ATP synthase
MH  - buffer
MH  - chemiosmotic theory
MH  - Chemistry
MH  - CONSTANT
MH  - cytochrome
MH  - Cytochrome c
MH  - Diffusion
MH  - equilibrium
MH  - mechanism
MH  - membrane
MH  - Membranes
MH  - model
MH  - proton
MH  - Proton Pump
MH  - Protons
MH  - rate constant
MH  - Solutions
MH  - SURFACE
MH  - SYNTHASE
MH  - theory
MH  - transport
MH  - united states
MH  - universities
RP  - NOT IN FILE
NT  - UI - 22017736DA - 20020522IS - 0006-3495LA - engPT - Journal ArticleCY - United StatesSB - IM
UR  - PM:12023208
SO  - Biophys J 2002 Jun ;82(6):2833-2846

1898
UI  - 21362
AU  - Gibson LC
AU  - Cadwallader G
AU  - Finbow ME
TI  - Evidence that there are two copies of subunit c" in V0 complexes in the vacuolar H+-ATPase
AB  - The proton-translocating core of eukaryotic vacuolar H(+)-ATPase (V-ATPase), V(0) consists of a hexameric arrangement of transmembrane alpha-helices formed from the related polypeptides, subunit c and subunit c". The former is comprised of four transmembrane alpha-helices, whilst the latter has an extra transmembrane domain at its N-terminus. In addition, the fungal form of V(0) contains a minor subunit c-related polypeptide, subunit c'. All three are required for activity of the proton pump in Saccharomyces cerevisiae. We have introduced cysteine residues in the N-terminal extension of subunit c" in a cysteine-free form. All mutant forms are active in the V-ATPase from S. cerevisiae. Oxidation of vacuolar membranes containing the cysteine-replaced forms gave a cross-linked product of 42000Da. Analysis of this species showed it to be a dimeric form of subunit c", and further studies confirmed there are two copies of subunit c" in the V-ATPases in which it is present. Co-expression of double cysteine-replaced forms of both subunit c and c" gave rise to only homotypic cross-linked forms. Also, subunit c oligomeric complexes are present in vacuolar membranes in the absence of subunit c", consistent with previous observations showing hexameric arrangements of subunit c in gap-junction-like membranes. In vitro studies showed subunit c" can bind to subunit c and itself. The extent of binding can be increased by removal of the N-terminal domain of subunit c". This domain may therefore function to limit the copy number of subunit c" in V(0). A deletion study shows that the domain is essential for the activity of subunit c". The results can be combined into a model of V(0) which contains two subunit c" protomers with the extra transmembrane domain located toward the central pore. Thus the predicted stoichiometry of V(0) in which subunit c" is present is subunit c(3):subunit c(')(1):subunit c(")(2). On the basis of the mutational and binding studies, it seems likely that two copies of subunit c" are next to each other
MH  - A
MH  - ACTIVE
MH  - analysis
MH  - BINDING
MH  - COMPLEX
MH  - Cysteine
MH  - function
MH  - H(+)ATPase
MH  - H+-ATPase
MH  - In Vitro
MH  - membrane
MH  - Membranes
MH  - model
MH  - mutant
MH  - proton
MH  - Proton Pump
MH  - RESIDUE
MH  - Saccharomyces cerevisiae
MH  - stoichiometry
MH  - SUBUNIT
MH  - universities
RP  - NOT IN FILE
NT  - School of Biological and Biomedical Sciences, Glasgow Caledonian University, Cowcaddens Road, Glasgow, G4 0BA, UK and CRC Beatson Laboratories, Beatson Institute for Cancer Research, Garscube Estate, Switchback Road, Bearsden, Glasgow, G61 1BD, UKFAU - Gibson, Lucien C D
SO  - Biochem J 2002 Sep 15 ;366(Pt 3):911-919

1899
UI  - 21335
AU  - Giraud MF
AU  - Paumard P
AU  - Soubannier V
AU  - Vaillier J
AU  - Arselin G
AU  - Salin B
AU  - Schaeffer J
AU  - Brethes D
AU  - di Rago JP
AU  - Velours J
TI  - Is there a relationship between the supramolecular organization of the mitochondrial ATP synthase and the formation of cristae?
AB  - Blue native polyacrylamide gel electrophoresis (BN-PAGE) analyses of detergent mitochondrial extracts have provided evidence that the yeast ATP synthase could form dimers. Cross-linking experiments performed on a modified version of the i-subunit of this enzyme indicate the existence of such ATP synthase dimers in the yeast inner mitochondrial membrane. We also show that the first transmembrane segment of the eukaryotic b-subunit (bTM1), like the two supernumerary subunits e and g, is required for dimerization/oligomerization of ATP synthases. Unlike mitochondria of wild-type cells that display a well-developed cristae network, mitochondria of yeast cells devoid of subunits e, g, or bTM1 present morphological alterations with an abnormal proliferation of the inner mitochondrial membrane. From these observations, we postulate that an anomalous organization of the inner mitochondrial membrane occurs due to the absence of ATP synthase dimers/oligomers. We provide a model in which the mitochondrial ATP synthase is a key element in cristae morphogenesis
MH  - A
MH  - atp
MH  - ATP synthase
MH  - cell
MH  - Cells
MH  - Chemistry
MH  - CROSS-LINKING
MH  - Dimerization
MH  - Electrophoresis
MH  - enzyme
MH  - enzymology
MH  - Intracellular Membranes
MH  - membrane
MH  - Microscopy,Electron
MH  - Microscopy,Fluorescence
MH  - Mitochondria
MH  - Mitochondrial Proton-Translocating ATPases
MH  - model
MH  - Morphogenesis
MH  - Saccharomyces cerevisiae
MH  - SUBUNIT
MH  - Support,Non-U.S.Gov't
MH  - SYNTHASE
MH  - ultrastructure
MH  - YEAST
RP  - NOT IN FILE
NT  - Institut de Biochimie et Genetique Cellulaires du CNRS, Universite Victor Segalen, Bordeaux, France marie-francegiraud@ibgcu-bordeaux2frFAU - Giraud, Marie-France
SO  - Biochim Biophys Acta 2002 Sep 10 ;1555(1-3):174-180

1900
UI  - 21381
AU  - Groth G
AU  - Hisabori T
AU  - Lill H
AU  - Bald D
TI  - Substitution of a single amino acid switches the tentoxin-resistant thermophilic F1-ATPase into a tentoxin-sensitive enzyme
AB  - In contrast to the homologous bacterial and mitochondrial enzymes the chloroplast F(1)-ATPase (CF(1)) is strongly affected by the phytopathogenic inhibitor tentoxin. Based on structural information obtained from crystals of a CF(1)-tentoxin co-complex (Groth, G. (2002) Proc. Natl. Acad. Sci. U. S. A. 99, 3464-3468) we have replaced residues betaSer(66) and alphaArg(132) in the alpha(3)beta(3)gamma subcomplex of the thermophilic F(1)-ATPase from Bacillus PS3 by the corresponding residues of the chloroplast ATPase to confer tentoxin sensitivity to the thermophilic enzyme. The mutation alphaArg(132) --> Pro, proposed to relieve steric constraints on tentoxin binding, did not have any significant effect. However, mutation betaSer(66) --> Ala, predicted to provide a crucial hydrogen bond with the inhibitor, resulted in tentoxin inhibition of ATP hydrolysis comparable with the situation found with the chloroplast enzyme
MH  - A
MH  - ACID
MH  - Adenosine Triphosphate
MH  - Amino Acid Substitution
MH  - atp
MH  - ATPase
MH  - Bacillus
MH  - Base Sequence
MH  - BINDING
MH  - Biochemistry
MH  - Chemistry
MH  - chloroplast
MH  - DNA Primers
MH  - drug effects
MH  - enzyme
MH  - Enzymes
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - Hydrogen
MH  - Hydrolysis
MH  - inhibitor
MH  - metabolism
MH  - Models,Molecular
MH  - Mutation
MH  - Peptides,Cyclic
MH  - pharmacology
MH  - plant
MH  - Protein Conformation
MH  - Proton-Translocating ATPases
MH  - PS3
MH  - RESIDUE
MH  - Support,Non-U.S.Gov't
MH  - switch
MH  - tentoxin
MH  - THERMOPHILIC
RP  - NOT IN FILE
NT  - Department of Plant Biochemistry, Heinrich-Heine Universitat, D-40225 Dusseldorf, GermanyFAU - Groth, Georg
SO  - J Biol Chem 2002 Jun 7 ;277(23):20117-20119

1901
UI  - 21384
AU  - Groth G
TI  - Structure of spinach chloroplast F1-ATPase complexed with the phytopathogenic inhibitor tentoxin
AB  - Tentoxin, a natural cyclic tetrapeptide produced by phytopathogenic fungi from the Alternaria species affects the catalytic function of the chloroplast F(1)-ATPase in certain sensitive species of plants. In this study, we show that the uncompetitive inhibitor tentoxin binds to the alphabeta-interface of the chloroplast F(1)-ATPase in a cleft localized at betaAsp-83. Most of the binding site is located on the noncatalytic alpha-subunit. The crystal structure of the tentoxin-inhibited CF(1)-complex suggests that the inhibitor is hydrogen bonded to Asp-83 in the catalytic beta-subunit but forms hydrophobic contacts with residues Ile-63, Leu-65, Val-75, Tyr-237, Leu-238, and Met-274 in the adjacent alpha-subunit. Except for minor changes around the tentoxin-binding site, the structure of the chloroplast alpha(3)beta(3)-core complex is the same as that determined with the native chloroplast ATPase. Tentoxin seems to act by inhibiting inter-subunit contacts at the alphabeta-interface and by blocking the interconversion of binding sites in the catalytic mechanism
MH  - A
MH  - ALPHA-SUBUNIT
MH  - Animal
MH  - antagonists & inhibitors
MH  - ATPase
MH  - BETA-SUBUNIT
MH  - BINDING
MH  - BINDING SITE
MH  - Binding Sites
MH  - catalytic
MH  - Cattle
MH  - Chemistry
MH  - chloroplast
MH  - Chloroplasts
MH  - COMPLEX
MH  - Crystallography,X-Ray
MH  - cyclic
MH  - enzymology
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - function
MH  - Fungi
MH  - Hydrogen
MH  - Hydrogen Bonding
MH  - Hydrophobicity
MH  - inhibitor
MH  - mechanism
MH  - metabolism
MH  - Mitochondria
MH  - Models,Molecular
MH  - Peptides,Cyclic
MH  - pharmacology
MH  - plant
MH  - Plants
MH  - Protein Structure,Quaternary
MH  - Protein Structure,Secondary
MH  - Protein Subunits
MH  - Proton-Translocating ATPases
MH  - RESIDUE
MH  - Site
MH  - Spinach
MH  - structure
MH  - Structure-Activity Relationship
MH  - Support,Non-U.S.Gov't
MH  - tentoxin
RP  - NOT IN FILE
NT  - Heinrich-Heine-Universitat, Biochemie der Pflanzen, Universitatsstrasse 1, D-40225 Duesseldorf, Germany georggroth@uni-duesseldorfdeFAU - Groth, Georg
SO  - Proc Natl Acad Sci U S A 2002 Mar ;%19;99(6):3464-3468

1902
UI  - 21360
AU  - Gumbiowski K
AU  - Panke O
AU  - Junge W
AU  - Engelbrecht S
TI  - Rotation of the c subunit oligomer in EF(0)EF(1) mutant cD61N
AB  - ATP synthases (F(0)F(1)-ATPases) mechanically couple ion flow through the membrane-intrinsic portion, F(0), to ATP synthesis within the peripheral portion, F(1). The coupling most probably occurs through the rotation of a central rotor (subunits c(10)epsilon gamma) relative to the stator (subunits ab(2)delta(alpha beta)(3)). The translocation of protons is conceived to involve the rotation of the ring of c subunits (the c oligomer) containing the essential acidic residue cD61 against subunits ab(2). In line with this notion, the mutants cD61N and cD61G have been previously reported to lack proton translocation. However, it has been surprising that the membrane-bound mutated holoenzyme hydrolyzed ATP but without translocating protons. Using detergent-solubilized and immobilized EF(0)F(1) and by application of the microvideographic assay for rotation, we found that the c oligomer, which carried a fluorescent actin filament, rotates in the presence of ATP in the mutant cD61N just as in the wild type enzyme. This observation excluded slippage among subunit gamma, the central rotary shaft, and the c oligomer and suggested free rotation without proton pumping between the oligomer and subunit a in the membrane-bound enzyme
MH  - A
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - Base Sequence
MH  - Chemistry
MH  - coupling
MH  - DNA Primers
MH  - enzyme
MH  - Escherichia coli
MH  - genetics
MH  - ion
MH  - Kinetics
MH  - metabolism
MH  - Molecular Sequence Data
MH  - Mutagenesis,Site-Directed
MH  - mutant
MH  - Plasmids
MH  - Polymerase Chain Reaction
MH  - Protein Subunits
MH  - Protein Transport
MH  - proton
MH  - Proton-Translocating ATPases
MH  - Protons
MH  - Recombinant Proteins
MH  - RESIDUE
MH  - Restriction Mapping
MH  - rotation
MH  - SUBUNIT
MH  - Support,Non-U.S.Gov't
MH  - SYNTHASE
MH  - synthesis
MH  - translocation
RP  - NOT IN FILE
NT  - Universitat Osnabruck, FB Biologie, Abt Biophysik, Barbarastrasse 11, 49076 Osnabruck, GermanyFAU - Gumbiowski, Karin
SO  - J Biol Chem 2002 Aug 30 ;277(35):31287-31290

1903
UI  - 21326
AU  - Haines T
AU  - Dencher N
TI  - Cardiolipin: a proton trap for oxidative phosphorylation
AB  - The role of specific lipid structures in biological membranes has been elusive. There are hundreds of them in nature. Why has nature made them? How do they aid in the functioning of membrane proteins? Genetics with its 'knock out' organisms declares that functions persist in the absence of any particular lipid. Nonetheless some lipids, such as cardiolipin (CL), are associated with particular functions in the cell. It may merely expand the variety of culture conditions (pH, temperature, etc.) under which the wild-type organism survives. This article explores a unique role of CL as a proton trap within membranes that conduct oxidative phosphorylation and therefore the synthesis of ATP. CL's pK(2) (above 8.0) provides a role for it as a headgroup proton trap for oxidative phosphorylation. It suggests why CL is found in membranes that pump protons. The high pK(2) also indicates that the headgroup has but one negative charge in the neutral pH range. Data on the binding of CL to all of the oxidative phosphorylation proteins suggest that the CL may aggregate the oxidative phosphorylation proteins into a patch while it restricts pumped protons within its headgroup domain - supplying protons to the ATP synthase with minimal changes in the bulk phase pH
MH  - A
MH  - atp
MH  - ATP synthase
MH  - BINDING
MH  - cell
MH  - Chemistry
MH  - data
MH  - function
MH  - genetics
MH  - Lipids
MH  - membrane
MH  - Membrane Proteins
MH  - Membranes
MH  - Oxidative Phosphorylation
MH  - pH
MH  - Phosphorylation
MH  - protein
MH  - Proteins
MH  - proton
MH  - Protons
MH  - structure
MH  - SYNTHASE
MH  - synthesis
MH  - Temperature
MH  - universities
RP  - NOT IN FILE
NT  - Department of Chemistry, City College of the City University of New York, 10031, New York, NY, USAFAU - Haines, Thomas
SO  - FEBS Lett 2002 Sep 25 ;528(1-3):35

1904
UI  - 21313
AU  - Hazard A
AU  - Montemagno C
TI  - Improved purification for thermophilic F(1)F(0) ATP synthase using n-dodecyl beta-D-maltoside
AB  - Here we report a fast, simple purification for thermophilic F(1)F(0) ATP synthase (TF(1)F(0)) that utilizes a cocktail of stabilizing reagents and the detergent n-dodecyl beta-D-maltoside to yield enzyme with an ATPase activity of 41&mgr;mol/min/mg, 2.5-fold higher than that previously reported. ATPase activity was 80% inhibited by the F(0)-reactive reagent dicyclohexylcarbodiimide, indicating that F(1)-F(0) interactions were largely intact. To measure ATP-driven proton pumping activity, purified TF(1)F(0) was incorporated into liposomes, and the ATP-induced change in internal pH was measured using the fluorescent probe pyranine. In the presence of valinomycin, a maximum ATP-driven DeltapH of 0.8 units was obtained. To measure ATP synthesis activity, TF(1)F(0) was incorporated into liposomes with the light-dependent proton pump bacteriorhodopsin. Proteoliposomes were illuminated to generate an electrochemical gradient, after which ADP and inorganic phosphate were added to initiate ATP synthesis. A steady state ATP synthesis activity of 490nmol/min/mg was achieved after an initial approximately 30-min lag phase
MH  - A
MH  - ADP
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - ATPase
MH  - Bacteriorhodopsin
MH  - Dicyclohexylcarbodiimide
MH  - enzyme
MH  - INORGANIC-PHOSPHATE
MH  - INTERACTION
MH  - liposome
MH  - Liposomes
MH  - pH
MH  - phosphate
MH  - proteoliposome
MH  - proton
MH  - Proton Pump
MH  - purification
MH  - SYNTHASE
MH  - synthesis
MH  - THERMOPHILIC
MH  - universities
MH  - Valinomycin
RP  - NOT IN FILE
NT  - Department of Biological and Environmental Engineering, Cornell University, 14853, Ithaca, NY, USA
SO  - Arch Biochem Biophys 2002 Nov 1 ;407(1):117

1905
UI  - 21327
AU  - Herbert SK
TI  - A new regulatory role for the chloroplast ATP synthase
MH  - A
MH  - atp
MH  - ATP synthase
MH  - chloroplast
MH  - SYNTHASE
MH  - universities
RP  - NOT IN FILE
NT  - Department of Botany, University of Wyoming, Aven Nelson 215, Laramie, WY 82071-3165FAU - Herbert, Stephen K
SO  - Proc Natl Acad Sci U S A 2002 Oct 1 ;99(20):12518-12519

1906
UI  - 21325
AU  - Hisabori T
AU  - Konno H
AU  - Ichimura H
AU  - Strotmann H
AU  - Bald D
TI  - Molecular devices of chloroplast F(1)-ATP synthase for the regulation
AB  - In chloroplasts, synthesis of ATP is energetically coupled with the utilization of a proton gradient formed by photosynthetic electron transport. The involved enzyme, the chloroplast ATP synthase, can potentially hydrolyze ATP when the magnitude of the transmembrane electrochemical potential difference of protons (Delta(micro)H(+)) is small, e.g. at low light intensity or in the dark. To prevent this wasteful consumption of ATP, the activity of chloroplast ATP synthase is regulated as the occasion may demand. As regulation systems Delta(micro)H(+) activation, thiol modulation, tight binding of ADP and the role of the intrinsic inhibitory subunit epsilon is well documented. In this article, we discuss recent progress in understanding of the regulation system of the chloroplast ATP synthase at the molecular level
MH  - A
MH  - ACTIVATION
MH  - Adenosine Triphosphate
MH  - ADP
MH  - Amino Acid Sequence
MH  - atp
MH  - ATP synthase
MH  - BINDING
MH  - Chemistry
MH  - chloroplast
MH  - Chloroplast Proton-Translocating ATPases
MH  - Chloroplasts
MH  - Comparative Study
MH  - Disulfides
MH  - electron
MH  - Electron Transport
MH  - enzyme
MH  - Enzyme Activation
MH  - Light
MH  - Molecular Sequence Data
MH  - Oxidation-Reduction
MH  - Photosynthesis
MH  - Protein Conformation
MH  - proton
MH  - PROTON GRADIENT
MH  - Protons
MH  - regulation
MH  - rotation
MH  - Sequence Alignment
MH  - SUBUNIT
MH  - SUBUNIT-EPSILON
MH  - Support,Non-U.S.Gov't
MH  - SYNTHASE
MH  - synthesis
MH  - SYSTEM
MH  - SYSTEMS
MH  - transport
RP  - NOT IN FILE
NT  - Chemical Resources Laboratory, Tokyo Institute of Technology, Yokohama, Kanagawa, Japan thisabor@restitechacjpFAU - Hisabori, Toru
SO  - Biochim Biophys Acta 2002 Sep 10 ;1555(1-3):140-146

1907
UI  - 21344
AU  - Hong S
AU  - Pedersen PL
TI  - ATP synthase of yeast: structural insight into the different inhibitory potencies of two regulatory peptides and identification of a new potential regulator
AB  - Mitochondrial ATP synthases, the major producers of ATP in higher eukaryotic cells, are known to be regulated by a peptide designated IF(1). In contrast, in yeast three such peptides have been identified, IF(1) and STF(1), which inhibit the reverse ATPase reaction, and STF(2), a modulator of the action of these inhibitors. Despite significant homology to IF(1), STF(1) exhibits less than half ( approximately 40%) its inhibitory potency. The two-fold purpose of this bioinformatic study was to gain structural insight into the different inhibitory potencies of IF(1) and STF(1) and to determine to what extent yeast are unique in employing multiple peptides to regulate the ATP synthase. Sequence and secondary structural analyses and comparison with the known structure of bovine IF(1) predicted a dimeric structure for yeast STF(1) in which the C-terminal regions form a coiled-coil. Moreover, sequence comparisons showed that within this C-terminal region a conserved acidic residue (Asp 59) in yeast IF(1) is replaced by Asn in STF(1). In the known structure of bovine IF(1), predicted to be very similar to that of yeast IF(1), the residue Glu 68 corresponding to Asp 59 participates in the formation of a four-residue conserved acidic cluster in the middle of the coiled-coil in the C-terminal region. It is deduced here that this acidic cluster is likely to be important in the regulation of IF(1)'s inhibitory capacity and that replacement of conserved Asp 59 by Asn in STF(1) may reduce its potency. Although other homologs to the inhibitors IF(1) and STF(1) were not found in searches of available eukaryotic genomes, including human, a new homolog, named STF(3), with 65% identity to the modulator STF(2), was discovered within the yeast genome and identified to be expressed by searching the yeast EST database. Thus, yeast appears unique in regulating the ATP synthase by involving multiple peptides (IF(1), STF(1), STF(2), and perhaps STF(3))
MH  - A
MH  - Amino Acid Sequence
MH  - Animal
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Cattle
MH  - cell
MH  - Cell Membrane
MH  - Cells
MH  - Chemistry
MH  - Dimerization
MH  - enzymology
MH  - Expressed Sequence Tags
MH  - fungal proteins
MH  - Human
MH  - inhibitor
MH  - inhibitors
MH  - metabolism
MH  - Mitochondria
MH  - Mitochondrial Proton-Translocating ATPases
MH  - Models,Biological
MH  - Models,Molecular
MH  - Molecular Sequence Data
MH  - Mutagenesis,Site-Directed
MH  - peptide
MH  - peptides
MH  - physiology
MH  - Protein Structure,Secondary
MH  - Protein Structure,Tertiary
MH  - regulation
MH  - RESIDUE
MH  - secondary
MH  - Sequence Homology,Amino Acid
MH  - structure
MH  - Support,U.S.Gov't,P.H.S.
MH  - SYNTHASE
MH  - universities
MH  - YEAST
RP  - NOT IN FILE
NT  - Department of Biological Chemistry, Johns Hopkins University School of Medicine, 725 N Wolfe Street, Baltimore, MD 21205-2185, USAFAU - Hong, Sangjin
SO  - Arch Biochem Biophys 2002 Sep 1 ;405(1):38-43

1908
UI  - 21357
AU  - Hunziker RW
AU  - Escher BI
AU  - Schwarzenbach RP
TI  - Acute toxicity of triorganotin compounds: different specific effects on the energy metabolism and role of pH
AB  - Triorganotin compounds exhibit several modes of toxic action on the energy metabolism in energy-transducing membranes. The inhibition of the adenosine triphosphate (ATP) synthase and the hydroxide/chloride-antiport have been extensively investigated, but debate still exists on whether further mechanisms are relevant. In this work, two possible further effects have been investigated: inhibition of the bc1 complex and the hydroxide uniport, and in addition, the overall inhibition of the ATP synthesis was investigated in chromatophores of the photosynthetic purple bacterium Rhodobacter sphaeroides at pH = 7.5 and pH = 6.1. Experimental conditions were chosen in order to exclude the hydroxide/anion antiport as a possible effect. Inhibition of the cytochromes bc1 complex was detected, but at such high concentrations that it is not relevant for acute toxicity. Tributyltin was found to induce a decrease of the membrane potential, which can be attributed to a hydroxide uniport, whereas for triphenyltin no such activity was observed. For both compounds, inhibition of the ATP synthesis was higher at pH = 6.1 than at pH = 7.5. Also the hydroxide uniport activity of tributyltin was higher at lower pH. The contribution of the hydroxide uniport of tributyltin to the overall inhibition of the ATP synthesis cannot be quantified; however, hydroxide uniport occurred in the same concentration range as inhibition of the ATP synthesis. For triphenyltin, inhibition of the ATP synthesis can be attributed to the inhibition of the ATP synthase. It was concluded that chromatophores of R. sphaeroides are a useful system to discriminate various effects of toxicants on the energy metabolism of a cell
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - Bacteria
MH  - bc1
MH  - cell
MH  - chromatophore
MH  - chromatophores
MH  - COMPLEX
MH  - cytochrome
MH  - Cytochromes
MH  - energy
MH  - Energy Metabolism
MH  - mechanism
MH  - MECHANISMS
MH  - membrane
MH  - Membrane Potential
MH  - Membranes
MH  - metabolism
MH  - pH
MH  - Rhodobacter
MH  - Rhodobacter sphaeroides
MH  - sphaeroides
MH  - SYNTHASE
MH  - synthesis
MH  - SYSTEM
RP  - NOT IN FILE
NT  - Swiss Federal Institute for Environmental Science and Technology, DuebendorfFAU - Hunziker, Rene W
SO  - Environ Toxicol Chem 2002 Jun ;21(6):1191-1197

1909
UI  - 21323
AU  - Ichikawa N
AU  - Nakabayashi K
AU  - Hashimoto T
TI  - A yeast mitochondrial ATPase inhibitor interacts with three proteins that are easy to dissociate from the mitochondrial inner membrane
AB  - A mitochondrial ATPase inhibitor is a 7.4 kDa protein that regulates the catalytic activity of ATP synthase (F(1)F(o)-ATPase). In the present study, we examined the binding sites of the inhibitor on the mitochondrial membrane using chemical cross-linkers, disuccinimidyl suberate (DSS) and N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ). Most of the inhibitors were recovered from the inner membrane fraction of mitochondria, indicating that the inhibitor binds to the membrane. Seven different cross-linked products that reacted with the antibody against the inhibitor were detected. The apparent molecular masses of the products were 61, 58, 47, 41, 28, 27, and 26 kDa. The 61 and 58 kDa products were attributed to the inhibitor+alpha and inhibitor+beta adducts on immunoblotting. The proteins cross-linked to the inhibitor in the 28, 27, and 26 kDa products were distinguished from subunit 4 (23 kDa), oligomycin sensitivity conferring protein (21 kDa), and subunit d (20 kDa) of F(1)F(o)-ATPase by analysis of the cross-linked products of mutant mitochondria in which the three proteins were replaced by hemagglutinin-tagged versions. The 28, 27, and 26 kDa products could be gradually dissociated from the mitochondrial membrane by increasing the salt concentration. These results shows that the endogenous inhibitor binds not only to the catalytic part of the enzyme, but also to the 19-21 kDa proteins that loosely associate with the mitochondrial inner membrane
MH  - A
MH  - analysis
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - BINDING
MH  - BINDING SITE
MH  - Binding Sites
MH  - catalytic
MH  - enzyme
MH  - Human
MH  - inhibitor
MH  - inhibitors
MH  - membrane
MH  - Mitochondria
MH  - mutant
MH  - Nutrition
MH  - oligomycin
MH  - protein
MH  - Proteins
MH  - Site
MH  - SUBUNIT
MH  - SYNTHASE
MH  - universities
MH  - YEAST
RP  - NOT IN FILE
NT  - Department of Food and Nutrition, Faculty of Human Life Science, Osaka City University, Sugimoto, Sumiyoshi, Osaka 558-8585, Japan ichikawa@lifeosaka-cuacjpFAU - Ichikawa, Naoki
SO  - J Biochem (Tokyo ) 2002 Oct ;132(4):649-654

1910
UI  - 21317
AU  - Itoi S
AU  - Kinoshita S
AU  - Kikuchi K
AU  - Watabe S
TI  - Changes of carp FoF1-ATPase in association with temperature acclimation
AB  - Previously we have shown, using two-dimensional electrophoresis, that mitochondrial ATP synthase (FoF1-ATPase) ss-subunit is the 55-kDa protein increased in cold-acclimated carp Cyprinus carpio (Kikuchi K., Itoi S, and Watabe S, Fisheries Sci 65: 629-636, 1999). To clarify the coordinate expression in various subunits of carp FoF1-ATPase with temperature acclimation, we examined the differences in mRNA levels of mitochondrial proteins encoded by both nuclear and mitochondrial genes in fast muscle of carp acclimated to 10 and 30( degrees )C. The mRNA levels of nuclear genes per unit weight of total RNA were nearly two fold higher in the 10( degrees )C- than 30( degrees )C-acclimated carp. However, the transcripts of mitochondrial genes for the 10( degrees )C-acclimated carp in terms of the same comparing unit were 6- 7 times as much as those for the 30( degrees )C-acclimated carp. The FoF1-ATPase activities measured at 10, 25 and 30( degrees )C were nearly two fold higher for the cold-acclimated fish than their warm-acclimated counterparts. Such quantitative and qualitative changes in carp FoF1-ATPase may contribute to extra ATP production required to compensate for energy balance at suboptimal temperatures
MH  - atp
MH  - ATP production
MH  - ATP synthase
MH  - biology
MH  - Electrophoresis
MH  - energy
MH  - protein
MH  - Proteins
MH  - SUBUNIT
MH  - SYNTHASE
MH  - Temperature
MH  - Time
MH  - universities
RP  - NOT IN FILE
NT  - Laboratory of Aquatic Moleculer Biology and Biotechnology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, Japan
SO  - Am J Physiol Regul Integr Comp Physiol 2002 Sep 12 ;.():

1911
UI  - 21396
AU  - Jolly RD
AU  - Brown S
AU  - Das AM
AU  - Walkley SU
TI  - Mitochondrial dysfunction in the neuronal ceroid-lipofuscinoses (Batten disease)
AB  - There are at least eight genetic entities known as the ceroid-lipofuscinoses in humans which share clinical and pathological features that have caused them to be grouped together under the eponym of Batten disease. They present pathologically as lysosomal storage diseases but are also characterised by severe neurodegeneration. Although the biochemical defects appear primarily centred on lysosomes and defects in proteolysis, the link between this and pathogenesis of neuronal death is poorly understood.The pathogenesis of neurodegeneration has been studied particularly in two animal models these being the English setter dog and the New Zealand Southhampshire sheep (OCL6). In these, and some of the human entities, there is evidence of mitochondrial dysfunction. This includes the accumulation of subunit c of ATP synthase as a component of storage material in at least six of eight genetic forms of the disease; structural abnormalities of mitochondria and selective loss of neurons in areas of the brain that are particularly metabolically active. Direct evidence of dysfunction comes from mitochondrial function tests in fibroblasts and, in animal models, isolated liver mitochondria. Supporting evidence of mitochondrial dysfunction was shown by disturbances in proportions of energy-rich phosphates in fibroblasts in some of these diseases. If these various defects were reflected in neurons, then it would support the hypothesis that neuron death was associated with energy-linked excitotoxicity
MH  - A
MH  - ACTIVE
MH  - Animal
MH  - atp
MH  - ATP synthase
MH  - Brain
MH  - Dogs
MH  - Energy Metabolism
MH  - function
MH  - Human
MH  - Liver
MH  - metabolism
MH  - Mitochondria
MH  - model
MH  - Nerve Degeneration
MH  - Neuronal Ceroid-Lipofuscinosis
MH  - pathology
MH  - phosphate
MH  - Phosphates
MH  - physiology
MH  - Sheep
MH  - SUBUNIT
MH  - Support,Non-U.S.Gov't
MH  - Support,U.S.Gov't,P.H.S.
MH  - SYNTHASE
MH  - universities
MH  - veterinary
RP  - NOT IN FILE
NT  - Institute of Veterinary, Animal and Biomedical Science, Massey University, Palmerston North, New Zealand rdjolly@masseyacnzFAU - Jolly, R D
SO  - Neurochem Int 2002 May ;40(6):565-571

1912
UI  - 21349
AU  - Kaim G
AU  - Prummer M
AU  - Sick B
AU  - Zumofen G
AU  - Renn A
AU  - Wild UP
AU  - Dimroth P
TI  - Coupled rotation within single F0F1 enzyme complexes during ATP synthesis or hydrolysis
AB  - F0F1 ATP synthases are the smallest rotary motors in nature and work as ATP factories in bacteria, plants and animals. Here we report on the first observation of intersubunit rotation in fully coupled single F0F1 molecules during ATP synthesis or hydrolysis. We investigate the Na+-translocating ATP synthase of Propionigenium modestum specifically labeled by a single fluorophore at one c subunit using polarization-resolved confocal microscopy. Rotation during ATP synthesis was observed with the immobilized enzyme reconstituted into proteoliposomes after applying a diffusion potential, but not with a Na+ concentration gradient alone. During ATP hydrolysis, stepwise rotation of the labeled c subunit was found in the presence of 2 mM NaCl, but not without the addition of Na+ ions. Moreover, upon the incubation with the F0-specific inhibitor dicyclohexylcarbodiimide the rotation was severely inhibited
MH  - A
MH  - Adenosine Triphosphate
MH  - Animal
MH  - antagonists & inhibitors
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - Bacteria
MH  - Chemistry
MH  - COMPLEX
MH  - Dicyclohexylcarbodiimide
MH  - Diffusion
MH  - diffusion potential
MH  - drug effects
MH  - enzyme
MH  - Enzyme Inhibitors
MH  - Enzymes,Immobilized
MH  - enzymology
MH  - F0F1
MH  - Fluorescent Dyes
MH  - Fusobacterium
MH  - genetics
MH  - Hydrolysis
MH  - inhibitor
MH  - ion
MH  - Ions
MH  - Liposomes
MH  - Macromolecular Systems
MH  - metabolism
MH  - Microscopy
MH  - Microscopy,Confocal
MH  - Models,Molecular
MH  - Molecular Motors
MH  - Mutagenesis,Site-Directed
MH  - pharmacology
MH  - plant
MH  - Plants
MH  - proteoliposome
MH  - Proton-Translocating ATPases
MH  - rotation
MH  - Sodium
MH  - SUBUNIT
MH  - SYNTHASE
MH  - synthesis
RP  - NOT IN FILE
NT  - Institute of Microbiology, ETH Zurich, Schmelzbergstrasse 7, CH-8092, Zurich, SwitzerlandFAU - Kaim, Georg
SO  - FEBS Lett 2002 Aug 14 ;525(1-3):156-163

1913
UI  - 21352
AU  - Knauf U
AU  - Hachtel W
TI  - The genes encoding subunits of ATP synthase are conserved in the reduced plastid genome of the heterotrophic alga Prototheca wickerhamii
AB  - The heterotrophic unicellular alga Prototheca wickerhamii is closely related to the photoautotrophic Chlorella vulgaris but has a 54,100-bp plastid DNA (ptDNA) that is much smaller than the chloroplast DNA of C. vulgaris (150,613 bp). The nucleotide sequence of 28,093 bp of the Prototheca ptDNA has been determined. No genes for photosynthetic functions have been found, except for sequences encoding six subunits of the ATP synthase ( atpA, atpB, atpE, atpF, atpH, and atpI). Transcripts of these atp genes have also been detected. Whether the leucoplasts of Prototheca contain a functional ATP synthase has still to be elucidated. Identified genes further include tufA, minD, cysT, and genes coding for three rRNAs, 22 tRNAs, and 12 ribosomal proteins. The results support the idea that, in the reduced plastid genome of Prototheca, genes coding for components of the plastid translational apparatus have been preferentially retained, and might be needed for the expression of the atp genes and some unassigned ORFs
MH  - A
MH  - atp
MH  - ATP synthase
MH  - Chemistry
MH  - chloroplast
MH  - Chloroplast Proton-Translocating ATPases
MH  - Dna
MH  - enzymology
MH  - Evolution,Molecular
MH  - function
MH  - genetics
MH  - nucleotide
MH  - Plastids
MH  - protein
MH  - Protein Subunits
MH  - Proteins
MH  - Prototheca
MH  - Restriction Mapping
MH  - RNA,Ribosomal
MH  - RNA,Transfer
MH  - SUBUNIT
MH  - SYNTHASE
MH  - Transcription,Genetic
RP  - NOT IN FILE
NT  - Botanisches Institut der Universitat Bonn, Karlrobert-Kreiten-Strasse 13, 53115 Bonn, GermanyFAU - Knauf, U
SO  - Mol Genet Genomics 2002 Jun ;267(4):492-497

1914
UI  - 21346
AU  - Kuhlbrandt W
AU  - Zeelen J
AU  - Dietrich J
TI  - Structure, mechanism, and regulation of the Neurospora plasma membrane H+-ATPase
AB  - Proton pumps in the plasma membrane of plants and yeasts maintain the intracellular pH and membrane potential. To gain insight into the molecular mechanisms of proton pumping, we built an atomic homology model of the proton pump based on the 2.6 angstrom x-ray structure of the related Ca2+ pump from rabbit sarcoplasmic reticulum. The model, when fitted to an 8 angstrom map of the Neurospora proton pump determined by electron microscopy, reveals the likely path of the proton through the membrane and shows that the nucleotide-binding domain rotates by approximately 70 degrees to deliver adenosine triphosphate (ATP) to the phosphorylation site. A synthetic peptide corresponding to the carboxyl-terminal regulatory domain stimulates ATPase activity, suggesting a mechanism for proton transport regulation
MH  - A
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - Amino Acid Sequence
MH  - atp
MH  - ATPase
MH  - Cell Membrane
MH  - Chemistry
MH  - CRYOELECTRON MICROSCOPY
MH  - electron
MH  - electron microscopy
MH  - ELECTRON-MICROSCOPY
MH  - Enzyme Activation
MH  - enzymology
MH  - H+-ATPase
MH  - mechanism
MH  - MECHANISMS
MH  - membrane
MH  - Membrane Potential
MH  - metabolism
MH  - Microscopy
MH  - model
MH  - Models,Molecular
MH  - Molecular Sequence Data
MH  - Neurospora
MH  - nucleotide binding
MH  - peptide
MH  - Peptide Fragments
MH  - pH
MH  - Phosphorylation
MH  - plant
MH  - Plants
MH  - Protein Conformation
MH  - Protein Structure,Tertiary
MH  - proton
MH  - Proton Pump
MH  - Proton-Translocating ATPases
MH  - regulation
MH  - Sarcoplasmic Reticulum
MH  - Site
MH  - structure
MH  - transport
MH  - YEAST
MH  - Yeasts
RP  - NOT IN FILE
NT  - Max-Planck-Institut fur Biophysik, Heinrich-Hoffmann-Str 7, 60528 Frankfurt am Main, GermanyFAU - Kuhlbrandt, Werner
SO  - Science 2002 Sep 6 ;297(5587):1692-1696

1915
UI  - 21367
AU  - Long JC
AU  - DeLeon-Rangel J
AU  - Vik SB
TI  - Characterization of the first cytoplasmic loop of subunit a of the Escherichia coli ATP synthase by surface labeling, cross-linking, and mutagenesis
AB  - The first cytoplasmic loop of subunit a of the Escherichia coli ATP synthase has been analyzed by cysteine substitution mutagenesis. 13 of the 26 residues tested were found to be accessible to the reaction with 3-(N-maleimidylpropionyl)-biocytin. The other 13 residues predominantly found in the central region of the polypeptide chain between the two transmembrane spans were more resistant to labeling by 3-(N-maleimidylpropionyl)-biocytin while in membrane vesicle preparations. This region of subunit a contains a conserved residue Glu-80, which when mutated to lysine resulted in a significant loss of ATP-driven proton translocation. Other substitutions including glutamine, alanine, and leucine were much less detrimental to function. Cross-linking studies with a photoactive cross-linking reagent were carried out. One mutant, K74C, was found to generate distinct cross-links to subunit b, and the cross-linking had little effect on proton translocation. The results indicate that the first transmembrane span (residues 40-64) of subunit a is probably near one or both of the b subunits and that a less accessible region of the first cytoplasmic loop (residues 75-90) is probably near the cytoplasmic surface, perhaps in contact with b subunits
MH  - A
MH  - Adenosine Triphosphate
MH  - Alanine
MH  - Amino Acid Sequence
MH  - atp
MH  - ATP synthase
MH  - Bacterial Proton-Translocating ATPases
MH  - Chemistry
MH  - COLI ATP SYNTHASE
MH  - CROSS-LINKING
MH  - Cross-Linking Reagents
MH  - Cysteine
MH  - Cytoplasm
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - function
MH  - Glutamine
MH  - Immunoblotting
MH  - Leucine
MH  - membrane
MH  - metabolism
MH  - Molecular Sequence Data
MH  - mutagenesis
MH  - Mutagenesis,Site-Directed
MH  - mutant
MH  - Mutation
MH  - pharmacology
MH  - Plasmids
MH  - Protein Conformation
MH  - Protein Structure,Secondary
MH  - Protein Structure,Tertiary
MH  - proton
MH  - RESIDUE
MH  - Spectrometry,Fluorescence
MH  - SUBUNIT
MH  - Support,Non-U.S.Gov't
MH  - Support,U.S.Gov't,P.H.S.
MH  - SURFACE
MH  - SYNTHASE
MH  - Time Factors
MH  - translocation
MH  - universities
RP  - NOT IN FILE
NT  - Department of Biological Sciences, Southern Methodist University, Dallas, TX 75275, USAFAU - Long, Julie C
SO  - J Biol Chem 2002 Jul 26 ;277(30):27288-27293

1916
UI  - 21348
AU  - Lu M
AU  - Vergara S
AU  - Zhang L
AU  - Holliday LS
AU  - Aris J
AU  - Gluck SL
TI  - The Amino-terminal Domain of the E Subunit of Vacuolar H+-ATPase (V-ATPase) Interacts with the H Subunit and Is Required for V-ATPase Function
AB  - Vacuolar H(+)-ATPases (V-ATPases) are highly conserved proton pumps that couple hydrolysis of cytosolic ATP to proton transport out of the cytosol. Although it is generally believed that V-ATPases transport protons by a rotary catalytic mechanism analogous to that used by F(1)F(0)-ATPases, the structure and subunit composition of the central or peripheral stalk of the multisubunit complex are not well understood. We searched for proteins that bind to the E subunit of V-ATPase using the yeast two-hybrid assay and identified the H subunit as an interacting partner. Physical association between the E and H subunits of V-ATPase was confirmed in vitro by precipitation assays. Deletion mapping analysis revealed that a 78-amino acid fragment at the amino terminus of the E subunit was sufficient for binding to the H subunit. Expression of the amino-terminal fragments of the E subunits from human and yeast as dominant-negative mutants resulted in dramatic decreases in bafilomycin A(1)-sensitive ATP hydrolysis and proton transport activities of V-ATPase. Our data demonstrate the physiological significance of the interaction between the E and H subunits of V-ATPase and extend previous studies on the arrangement of subunits on the peripheral stalk of V-ATPase
MH  - A
MH  - ACID
MH  - analysis
MH  - atp
MH  - BINDING
MH  - biology
MH  - catalytic
MH  - cell
MH  - COMPLEX
MH  - data
MH  - function
MH  - H(+)ATPase
MH  - H+-ATPase
MH  - Human
MH  - Hydrolysis
MH  - In Vitro
MH  - INTERACTION
MH  - mechanism
MH  - mutant
MH  - protein
MH  - Proteins
MH  - proton
MH  - Proton Pump
MH  - Protons
MH  - stalk
MH  - structure
MH  - SUBUNIT
MH  - transport
MH  - universities
MH  - YEAST
RP  - NOT IN FILE
NT  - Departments of Medicine and Anatomy and Cell Biology, University of Florida College of Medicine, and the Department of Orthodontics, University of Florida College of Dentistry, Gainesville, Florida 32610FAU - Lu, Ming
SO  - J Biol Chem 2002 Oct 11 ;277(41):38409-38415

1917
UI  - 21350
AU  - Malyan A
AU  - Allison W
TI  - Properties of noncatalytic sites of thioredoxin-activated chloroplast coupling factor 1
AB  - Nucleotide binding properties of two vacant noncatalytic sites of thioredoxin-activated chloroplast coupling factor 1 (CF(1)) were studied. Kinetics of nucleotide binding to noncatalytic sites is described by the first-order equation that allows for two nucleotide binding sites that differ in kinetic features. Dependence of the nucleotide binding rate on nucleotide concentration suggests that tight nucleotide binding is preceded by rapid reversible binding of nucleotides. ADP binding is cooperative. The preincubation of CF(1) with Mg(2+) produces only slight effect on the rate of ADP binding and decreases the ATP binding rate. The ATP and ADP dissociation from noncatalytic sites is described by the first-order equation for similar sites with dissociation rate constants k(-2)(ADP)=1.5 x 10(-1) min(-1) and k(-2)(ATP) congruent with 10(-3) min(-1), respectively. As follows from the study, the noncatalytic sites of CF(1) are not homogeneous. One of them retains the major part of endogenous ADP after CF(1) precipitation with ammonium sulfate. Its other two sites can bind both ADP and ATP but have different kinetic parameters and different affinity for nucleotides
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - ADP
MH  - ADP BINDING
MH  - affinity
MH  - atp
MH  - BINDING
MH  - BINDING SITE
MH  - Binding Sites
MH  - bioenergetics
MH  - Catalysis
MH  - Chemistry
MH  - chloroplast
MH  - Chloroplast Proton-Translocating ATPases
MH  - CONSTANT
MH  - coupling
MH  - COUPLING FACTOR
MH  - DEPENDENCE
MH  - Dithiothreitol
MH  - Kinetics
MH  - Magnesium
MH  - metabolism
MH  - NONCATALYTIC SITES
MH  - nucleotide
MH  - nucleotide binding
MH  - nucleotide binding sites
MH  - NUCLEOTIDE-BINDING-SITES
MH  - Nucleotides
MH  - pharmacology
MH  - Photosynthesis
MH  - rate constant
MH  - Site
MH  - Support,Non-U.S.Gov't
MH  - Support,U.S.Gov't,P.H.S.
MH  - Thioredoxin
RP  - NOT IN FILE
NT  - Laboratory of Bioenergetics of Photosynthesis, Institute of Basic Biological Problems, Russian Academy of Sciences, Pushchino, Region 142290, Moscow, Russia malyan@isspserpukhovsuFAU - Malyan, Alexander
SO  - Biochim Biophys Acta 2002 Jul 1 ;1554(3):153-158

1918
UI  - 21366
AU  - Martin-Galiano AJ
AU  - Gorgojo B
AU  - Kunin CM
AU  - de la Campa AG
TI  - Mefloquine and new related compounds target the F(0) complex of the F(0)F(1) H(+)-ATPase of Streptococcus pneumoniae
AB  - The activities of mefloquine (MFL) and related compounds against previously characterized Streptococcus pneumoniae strains carrying defined amino acid substitutions in the c subunit of the F(0)F(1) H(+)-ATPase were studied. In addition, a series of MFL-resistant (Mfl(r)) strains were isolated and characterized. A good correlation was observed between inhibition of growth and inhibition of the membrane-associated F(0)F(1) H(+)-ATPase activity. MFL was about 10-fold more active than optochin and about 200-fold more active than quinine in inhibiting both the growth and the ATPase activities of laboratory pneumococcal strain R6. Mutant strains were inhibited by the different compounds to different degrees, depending on their specific mutations in the c subunit. The resistant strains studied had point mutations that changed amino acid residues in either the c subunit or the a subunit of the F(0) complex. Changes in the c subunit were located in one of the two transmembrane alpha helices: residues M13, G14, G20, M23, and N24 of helix 1 and residues M44, G47, V48, A49, and V57 of helix 2. Changes in the a subunit were also found in either of the transmembrane alpha helices, helix 5 or 6: residue L186 of helix 5 and residues W206, F209, and S214 of helix 6. These results suggest that the transmembrane helices of the c and a subunits interact and that the mutated residues are important for the structure of the F(0) complex and proton translocation
MH  - A
MH  - ACID
MH  - ACTIVE
MH  - alpha
MH  - Amino Acid Sequence
MH  - Amino Alcohols
MH  - Antimalarials
MH  - ATPase
MH  - Cell Membrane
MH  - COMPLEX
MH  - DNA,Bacterial
MH  - drug effects
MH  - enzymology
MH  - genetics
MH  - H(+)ATPase
MH  - Mefloquine
MH  - Microbial Sensitivity Tests
MH  - Molecular Sequence Data
MH  - mutant
MH  - Mutation
MH  - pharmacology
MH  - point mutation
MH  - proton
MH  - Proton-Translocating ATPases
MH  - RESIDUE
MH  - Reverse Transcriptase Polymerase Chain Reaction
MH  - Streptococcus
MH  - Streptococcus pneumoniae
MH  - structure
MH  - SUBUNIT
MH  - Support,Non-U.S.Gov't
MH  - translocation
RP  - NOT IN FILE
NT  - Unidad de Genetica Bacteriana (Consejo Superior de Investigaciones Cientificas), Centro Nacional de Microbiologia, Instituto de Salud Carlos III, 28220 Majadahonda, Madrid, SpainFAU - Martin-Galiano, Antonio Javier
SO  - Antimicrob Agents Chemother 2002 Jun ;46(6):1680-1687

1919
UI  - 21390
AU  - Masaike T
AU  - Muneyuki E
AU  - Noji H
AU  - Kinosita K
AU  - Yoshida M
TI  - F1-ATPase changes its conformations upon phosphate release
AB  - Motor proteins, myosin, and kinesin have gamma-phosphate sensors in the switch II loop that play key roles in conformational changes that support motility. Here we report that a rotary motor, F1-ATPase, also changes its conformations upon phosphate release. The tryptophan mutation was introduced into Arg-333 in the beta subunit of F1-ATPase from thermophilic Bacillus PS3 as a probe of conformational changes. This residue interacts with the switch II loop (residues 308-315) of the beta subunit in a nucleotide-bound conformation. The addition of ATP to the mutant F1 subcomplex alpha3beta(R333W)3gamma caused transient increase and subsequent decay of the Trp fluorescence. The increase was caused by conformational changes on ATP binding. The rate of decay agreed well with that of phosphate release monitored by phosphate-binding protein assays. This is the first evidence that the beta subunit changes its conformation upon phosphate release, which may share a common mechanism of exerting motility with other motor proteins
MH  - A
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - Arginine
MH  - atp
MH  - Bacillus
MH  - BETA
MH  - BETA-SUBUNIT
MH  - BINDING
MH  - Chemistry
MH  - conformation
MH  - conformational change
MH  - conformational changes
MH  - Dose-Response Relationship,Drug
MH  - enzymology
MH  - Escherichia coli
MH  - F1
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - fluorescence
MH  - Hydrolysis
MH  - mechanism
MH  - metabolism
MH  - Models,Chemical
MH  - Models,Molecular
MH  - mutant
MH  - Mutation
MH  - phosphate
MH  - Phosphates
MH  - protein
MH  - Protein Binding
MH  - Protein Conformation
MH  - Proteins
MH  - Proton-Translocating ATPases
MH  - PS3
MH  - RESIDUE
MH  - Spectrometry,Fluorescence
MH  - SUBUNIT
MH  - Support,Non-U.S.Gov't
MH  - switch
MH  - THERMOPHILIC
MH  - THERMOPHILIC BACILLUS PS3
MH  - Time Factors
MH  - tryptophan
RP  - NOT IN FILE
NT  - Chemical Resources Laboratory, Tokyo Institute of Technology, 4259 Nagatsuta, Yokohama, 226-8503, JapanFAU - Masaike, Tomoko
SO  - J Biol Chem 2002 Jun 14 ;277(24):21643-21649

1920
UI  - 21380
AU  - Matthey U
AU  - Braun D
AU  - Dimroth P
TI  - NMR investigations of subunit c of the ATP synthase from Propionigenium modestum in chloroform/methanol/water (4 : 4 : 1)
AB  - The subunit c from the ATP synthase of Propionigenium modestum was studied by NMR in chloroform/methanol/water (4 : 4 : 1). In this solvent, subunit c consists of two helical segments, comprised of residues L5 to I26 and G29 to N82, respectively. On comparing the secondary structure of subunit c from P. modestum in the organic solvent mixture with that in dodecylsulfate micelles several deviations became apparent: in the organic solvent, the interruption of the alpha helical structure within the conserved GXGXGXGX motif was shortened from five to two residues, the prominent interruption of the alpha helical structure in the cystoplasmic loop region was not apparent, and neither was there a break in the alpha helix after the sodium ion-binding Glu65 residue. The folding of subunit c of P. modestum in the organic solvent also deviated from that of Escherichia coli in the same environment, the most important difference being that subunit c of P. modestum did not adopt a stable hairpin structure like subunit c of E. coli
MH  - A
MH  - alpha
MH  - Amino Acid Sequence
MH  - atp
MH  - ATP synthase
MH  - Chemistry
MH  - Chloroform
MH  - enzymology
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - Fusobacterium
MH  - Methanol
MH  - Micelles
MH  - Molecular Sequence Data
MH  - NMR
MH  - Nuclear Magnetic Resonance,Biomolecular
MH  - P
MH  - Protein Structure,Secondary
MH  - Proton-Translocating ATPases
MH  - RESIDUE
MH  - secondary
MH  - Sequence Homology,Amino Acid
MH  - Sodium
MH  - SOLVENT
MH  - structure
MH  - SUBUNIT
MH  - SYNTHASE
MH  - Water
RP  - NOT IN FILE
NT  - Institut fur Mikrobiologie, Eidgenossische Technische Hochschule, Zurich, Switzerland matthey@microbiolethzchFAU - Matthey, Ulrich
SO  - Eur J Biochem 2002 Apr ;269(7):1942-1946

1921
UI  - 1
AU  - Meier T
AU  - Matthey U
AU  - Henzen F
AU  - Dimroth P
AU  - Muller DJ
TI  - The central plug in the reconstituted undecameric c cylinder of a bacterial ATP synthase consists of phospholipids
AB  - The isolated rotor cylinder of the ATP synthase from Ilyobacter tartaricus was reconstituted into two-dimensional crystalline arrays. Atomic force microscopy imaging indicated a central cavity on one side of the rotor and a central plug protruding from the other side. Upon incubation with phospholipase C, the plug disappeared, but the appearance of the surrounding c subunit oligomer was not affected. This indicates that the plug consists of phospholipids. As the detergent- purified c cylinder is completely devoid of phospholipids, these are incorporated into the central hole from one side of the cylinder during the reconstitution procedure
RP  - NOT IN FILE
NT  - UI - 21460503LA - engPT - Journal ArticleDA - 20010928IS - 0014-5793SB - IMCY - NetherlandsJC - EUH
UR  - PM:11576527
SO  - FEBS Lett 2002 Sep 21 ;505(3):353-356

1922
UI  - 21322
AU  - Minoletti C
AU  - Santolini J
AU  - Haraux F
AU  - Pothier J
AU  - Andre F
TI  - Rebuilt 3D structure of the chloroplast f1 ATPase-tentoxin complex
AB  - The F(1) part of the chloroplast H(+) adenosine triphosphate (ATP)-synthase (CF(1)) strongly interacts with tentoxin, a natural fungous cyclic tetrapeptide known to inhibit the chloroplast enzyme and not the mammalian mitochondrial enzyme. Whereas the synthesis or the hydrolysis of ATP requires the stepwise rotation of the protein rotor gamma within the (alphabeta)(3) crown, only one molecule of tentoxin is needed to fully inhibit the complex. With the help of an original homology modeling technique, based on robust distance geometry protocols, we built a tridimensional model of the alpha(3)beta(3)gamma CF(1) subcomplex (3200 residues), in which we introduced three different nucleotide occupancies to check their possible influence on the tentoxin binding site. Simultaneous comparison of three available high-resolution X-ray structures of F(1), performed with a local structural alignment search tool, led to characterizing common structural blocks and the distorsions experienced by the complex during the catalytic turnover. The common structural blocks were used as a starting point of the spinach CF(1) structure rebuilding. Finally, tentoxin was docked into its putative binding site of the reconstructed structure. The docking method was initially validated in the mitochondrial enzyme by its ability to relocate nucleotides into their original position in the crystal. Tentoxin binding was found possible to the two alpha/beta interfaces associated with the empty and adenosine diphosphate (ADP)-loaded catalytic sites, but not to the one associated with the ATP-loaded site. These results suggest a mechanism of CF(1) inhibition by one molecule of tentoxin, by the impossibility of the alpha/beta interface bearing tentoxin to pass through the ATP-loaded state. Proteins 2002;49:302-320
MH  - A
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - atp
MH  - BINDING
MH  - BINDING SITE
MH  - catalytic
MH  - chloroplast
MH  - COMPLEX
MH  - cyclic
MH  - enzyme
MH  - F1
MH  - Hydrolysis
MH  - interfaces
MH  - mechanism
MH  - method
MH  - model
MH  - nucleotide
MH  - Nucleotides
MH  - protein
MH  - Proteins
MH  - RESIDUE
MH  - rotation
MH  - Site
MH  - Spinach
MH  - structure
MH  - synthesis
MH  - tentoxin
MH  - turnover
RP  - NOT IN FILE
NT  - CNRS URA 2096, Proteines Membranaires Transductrices d'Energie, Section de Bioenergetique, Departement de Biologie Cellulaire et Moleculaire, CEA-SACLAY, FranceFAU - Minoletti, Claire
SO  - Proteins 2002 Nov 15 ;49(3):302-320

1923
UI  - 21405
AU  - Mitome N
AU  - Ono S
AU  - Suzuki T
AU  - Shimabukuro K
AU  - Muneyuki E
AU  - Yoshida M
TI  - The presence of phosphate at a catalytic site suppresses the formation of the MgADP-inhibited form of F(1)-ATPase
AB  - F1-ATPase is inactivated by entrapment of MgADP in catalytic sites and reactivated by MgATP or P(i). Here, using a mutant alpha(3)beta(3)gamma complex of thermophilic F(1)-ATPase (alpha W463F/beta Y341W) and monitoring nucleotide binding by fluorescence quenching of an introduced tryptophan, we found that P(i) interfered with the binding of MgATP to F(1)-ATPase, but binding of MgADP was interfered with to a lesser extent. Hydrolysis of MgATP by F(1)-ATPase during the experiments did not obscure the interpretation because another mutant, which was able to bind nucleotide but not hydrolyse ATP (alpha W463F/beta E190Q/beta Y341W), also gave the same results. The half-maximal concentrations of P(i) that suppressed the MgADP-inhibited form and interfered with MgATP binding were both approximately 20 mm. It is likely that the presence of P(i) at a catalytic site shifts the equilibrium from the MgADP-inhibited form to the enzyme-MgADP-P(i) complex, an active intermediate in the catalytic cycle
MH  - A
MH  - ACTIVE
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - alpha
MH  - antagonists & inhibitors
MH  - atp
MH  - BINDING
MH  - catalytic
MH  - catalytic domain
MH  - Chemistry
MH  - COMPLEX
MH  - equilibrium
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - fluorescence
MH  - Hydrolysis
MH  - intermediate
MH  - metabolism
MH  - mutant
MH  - nucleotide
MH  - nucleotide binding
MH  - pharmacology
MH  - phosphate
MH  - Phosphates
MH  - Proton-Translocating ATPases
MH  - Site
MH  - THERMOPHILIC
MH  - tryptophan
RP  - NOT IN FILE
NT  - Chemical Resources Laboratory, Tokyo Institute of Technology, Yokohama, JapanFAU - Mitome, Noriyo
SO  - Eur J Biochem 2002 Jan ;269(1):53-60

1924
UI  - 21369
AU  - Morstadt L
AU  - Graber P
AU  - De Pascalis L
AU  - Kleinig H
AU  - Speth V
AU  - Beyer P
TI  - Chemiosmotic ATP synthesis in photosynthetically inactive chromoplasts from Narcissus pseudonarcissus L. linked to a redox pathway potentially also involved in carotene desaturation
AB  - Mature chromoplasts from daffodil (Narcissus pseudonarcissus) flowers, although devoid of thylakoid structures, contain immunologically detectable alpha-subunits of ATP-synthase (H(+)-transporting ATP phosphohydrolase; EC 3.6.3.14). To show the presence of the entire functional protein complex, chromoplast membrane proteins were solubilized and reconstituted in phosphatidylcholine liposomes. The membranes were energized by an acid-base transition in the presence of a K(+)/valinomycin diffusion potential, and the initial rate of ATP synthesis was measured with a luciferin/luciferase assay. In addition, by demonstrating NADPH-dependent ATP synthesis, we show that an NAD(P)H-dependent respiratory redox pathway in chromoplasts, previously identified as an important constituent of the carotene desaturation system, proceeds concomitant with membrane energization
MH  - A
MH  - Adenosine Triphosphate
MH  - ALPHA-SUBUNIT
MH  - Ammonium Chloride
MH  - Angiosperms
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - biosynthesis
MH  - Carotenoids
MH  - COMPLEX
MH  - Diffusion
MH  - diffusion potential
MH  - Dinitrophenols
MH  - Dinucleoside Phosphates
MH  - drug effects
MH  - liposome
MH  - Liposomes
MH  - membrane
MH  - Membrane Proteins
MH  - Membranes
MH  - metabolism
MH  - Microscopy,Electron
MH  - Nadp
MH  - Osmotic Pressure
MH  - Oxidation-Reduction
MH  - Oxygen Consumption
MH  - pharmacology
MH  - Photosynthesis
MH  - photosynthetic reaction center,plant
MH  - physiology
MH  - Plastids
MH  - protein
MH  - Proteins
MH  - Proton-Translocating ATPases
MH  - redox
MH  - structure
MH  - Support,Non-U.S.Gov't
MH  - synthesis
MH  - SYSTEM
MH  - thylakoid
MH  - ultrastructure
RP  - NOT IN FILE
NT  - Zentrum fur Angewandte Biowissenschaften, Universitat Freiburg, Schanzlestrasse 1, 79104 Freiburg, GermanyFAU - Morstadt, Lucia
SO  - Planta 2002 May ;215(1):134-140

1925
UI  - 21377
AU  - Muller M
AU  - Panke O
AU  - Junge W
AU  - Engelbrecht S
TI  - F1-ATPase, the C-terminal end of subunit gamma is not required for ATP hydrolysis-driven rotation
AB  - ATP hydrolysis by the isolated F(1)-ATPase drives the rotation of the central shaft, subunit gamma, which is located within a hexagon formed by subunits (alphabeta)(3). The C-terminal end of gamma forms an alpha-helix which properly fits into the "hydrophobic bearing" provided by loops of subunits alpha and beta. This "bearing" is expected to be essential for the rotary function. We checked the importance of this contact region by successive C-terminal deletions of 3, 6, 9, 12, 15, and 18 amino acid residues (Escherichia coli F(1)-ATPase). The ATP hydrolysis activity of a load-free ensemble of F(1) with 12 residues deleted decreased to 24% of the control. EF(1) with deletions of 15 or 18 residues was inactive, probably because it failed to assemble. The average torque generated by a single molecule of EF(1) when loaded by a fluorescent actin filament was, however, unaffected by deletions of up to 12 residues, as was their rotational behavior (all samples rotated during 60 +/- 19% of the observation time). Activation energy analysis with the ensemble revealed a moderate decrease from 54 kJ/mol for EF(1) (full-length gamma) to 34 kJ/mol for EF(1)(gamma-12). These observations imply that the intactness of the C terminus of subunit gamma provides structural stability and/or routing during assembly of the enzyme, but that it is not required for the rotary action under load, proper
MH  - A
MH  - ACID
MH  - ACTIVATION
MH  - Adenosine Triphosphate
MH  - alpha
MH  - Amino Acid Sequence
MH  - analysis
MH  - atp
MH  - Base Sequence
MH  - BETA
MH  - Chemistry
MH  - energy
MH  - enzyme
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - function
MH  - genetics
MH  - Hydrolysis
MH  - Kinetics
MH  - metabolism
MH  - Molecular Sequence Data
MH  - physiology
MH  - Proton-Translocating ATPases
MH  - RESIDUE
MH  - rotation
MH  - SUBUNIT
MH  - Support,Non-U.S.Gov't
MH  - Time
RP  - NOT IN FILE
NT  - Universitat Osnabruck, FB Biologie, Abt Biophysik, Barbarastrasse 11, 49076 Osnabruck, GermanyFAU - Muller, Martin
SO  - J Biol Chem 2002 Jun 28 ;277(26):23308-23313

1926
UI  - 21306
AU  - Nachliel E
AU  - Gutman M
AU  - Tittor J
AU  - Oesterhelt D
AD  - Laser Laboratory for Fast Reactions in Biology, Department of Biochemistry, Tel Aviv University, Tel Aviv 69978, Israel
TI  - Proton transfer dynamics on the surface of the late m state of bacteriorhodopsin
AB  - The cytoplasmic surface of the BR (initial) state of bacteriorhodopsin is characterized by a cluster of three carboxylates that function as a proton-collecting antenna. Systematic replacement of most of the surface carboxylates indicated that the cluster is made of D104, E161, and E234 (Checover, S., Y. Marantz, E. Nachliel, M. Gutman, M. Pfeiffer, J. Tittor, D. Oesterhelt, and N. Dencher. 2001. Biochemistry. 40:4281-4292), yet the BR state is a resting configuration; thus, its proton-collecting antenna can only indicate the presence of its role in the photo-intermediates where the protein is re-protonated by protons coming from the cytoplasmic matrix. In the present study we used the D96N and the triple (D96G/F171C/F219L) mutant for monitoring the proton-collecting properties of the protein in its late M state. The protein was maintained in a steady M state by continuous illumination and subjected to reversible pulse protonation caused by repeated excitation of pyranine present in the reaction mixture. The re-protonation dynamics of the pyranine anion was subjected to kinetic analysis, and the rate constants of the reaction of free protons with the surface groups and the proton exchange reactions between them were calculated. The reconstruction of the experimental signal indicated that the late M state of bacteriorhodopsin exhibits an efficient mechanism of proton delivery to the unoccupied-most basic-residue on its cytoplasmic surface (D38), which exceeds that of the BR configuration of the protein. The kinetic analysis was carried out in conjunction with the published structure of the M state (Sass, H., G. Buldt, R. Gessenich, D. Hehn, D. Neff, R. Schlesinger, J. Berendzen, and P. Ormos. 2000. Nature. 406:649-653), the model that resolves most of the cytoplasmic surface. The combination of the kinetic analysis and the structural information led to identification of two proton-conducting tracks on the protein's surface that are funneling protons to D38. One track is made of the carboxylate moieties of residues D36 and E237, while the other is made of D102 and E232. In the late M state the carboxylates of both tracks are closer to D38 than in the BR (initial) state, accounting for a more efficient proton equilibration between the bulk and the protein's proton entrance channel. The triple mutant resembles in the kinetic properties of its proton conducting surface more the BR-M state than the initial state confirming structural similarities with the BR-M state and differences to the BR initial state
MH  - A
MH  - analysis
MH  - Bacteriorhodopsin
MH  - Biochemistry
MH  - biology
MH  - CONSTANT
MH  - equilibration
MH  - function
MH  - M
MH  - mechanism
MH  - model
MH  - mutant
MH  - P
MH  - protein
MH  - proton
MH  - Proton transfer
MH  - Proton-transfer
MH  - protonation
MH  - Protons
MH  - rate constant
MH  - RESIDUE
MH  - structure
MH  - SURFACE
MH  - TRANSFER
MH  - united states
MH  - universities
RP  - NOT IN FILE
NT  - UI - 22074785DA - 20020624IS - 0006-3495LA - engPT - Journal ArticleCY - United StatesSB - IM
UR  - PM:12080130
SO  - Biophys J 2002 Jul ;83(1):416-426

1927
UI  - 21373
AU  - Nath S
TI  - The molecular mechanism of ATP synthesis by F1F0-ATP synthase: a scrutiny of the major possibilities
AB  - A critical goal of metabolism in living cells is the synthesis of adenosine triphosphate (ATP). ATP is synthesized by the enzyme F1F0-ATP synthase. This enzyme, the smallest-known molecular machine, couples proton translocation through its membrane-embedded, hydrophobic domain, F0, to the synthesis of ATP from adenosine diphosphate (ADP) and inorganic phosphate (Pi) in its soluble, hydrophilic headpiece, F1. Animals, plants and microorganisms all capture and utilize energy by this important chemical reaction. How does it occur? The binding change mechanism and the torsional mechanism of energy transduction and ATP synthesis are two mechanisms that have been proposed in the literature. According to the binding change mechanism (which considers reversible catalysis and site-site cooperativity), energy is required primarily for release of synthesized ATP, but not for its synthesis. On the other hand, according to the torsional mechanism (which considers an irreversible mode of catalysis and absence of cooperativity), all the elementary steps require energy, and the ion-protein interaction energy obtained from the ion gradients is used to synthesize ATP, for Pi binding, and for straining the beta-epsilon bond in order to enable ADP to bind. The energy to release preformed ATP from the tight catalytic site (betaDP) is provided by the formation of the beta-epsilon ester linkage. First, the central features of these mechanisms are clearly delineated. Then, a critical scrutiny of these mechanisms is undertaken. The predictions of the torsional mechanism are listed. In particular, how the torsional mechanism deals with the specific difficulties associated with other mechanisms, and how it seeks to explain a wealth of structural, spectroscopic, and biochemical data is discussed in detail. Recent experimental data in support of the mechanism are presented. Finally, in view of the molecular machine nature of energy transduction, the indispensability of applying engineering tools at the molecular level is highlighted. This paves the way for the development of a new field: Molecular Physiological Engineering
MH  - A
MH  - Adenosine
MH  - Adenosine Diphosphate
MH  - Adenosine Triphosphate
MH  - ADP
MH  - Animal
MH  - atp
MH  - ATP synthesis
MH  - BINDING
MH  - biosynthesis
MH  - Catalysis
MH  - catalytic
MH  - cell
MH  - Cells
MH  - CHANGE MECHANISM
MH  - data
MH  - development
MH  - energy
MH  - enzyme
MH  - F0
MH  - F1
MH  - F1F0-ATP SYNTHASE
MH  - FIELD
MH  - INORGANIC-PHOSPHATE
MH  - INTERACTION
MH  - ion
MH  - mechanism
MH  - MECHANISMS
MH  - metabolism
MH  - phosphate
MH  - plant
MH  - Plants
MH  - proton
MH  - Proton-Translocating ATPases
MH  - Site
MH  - Support,Non-U.S.Gov't
MH  - SYNTHASE
MH  - synthesis
MH  - translocation
RP  - NOT IN FILE
NT  - Department of Biochemical Engineering and Biotechnology, Indian Institute of Technology, Hauz Khas, New Delhi sunath@dbebiitdernetinFAU - Nath, Sunil
SO  - Adv Biochem Eng Biotechnol 2002  ;74:65-98.():65-98

1928
UI  - 20981
AU  - Naumann R
AU  - Baumgart T
AU  - Graber P
AU  - Jonczyk A
AU  - Offenhausser A
AU  - Knoll W
AD  - Max Planck Institute of Polymer Research, Ackermannweg 10, D-55128, Mainz, Germany
TI  - Proton transport through a peptide-tethered bilayer lipid membrane by the H(+)-ATP synthase from chloroplasts measured by impedance spectroscopy
AB  - A lipid membrane was tethered to a gold film by a peptide spacer molecule terminated by a sulfhydryl group. Membranes were formed by fusion of liposomes prepared from egg phosphatidylcholine on self assembled monolayers of the thiolipopeptide Myr-Lys(Myr)-Ser-Ser-Pro- Ala-Ser-Ser-Ala-Ala-Ser-Ala-Cys-amide mixed with mercaptoethanol as a diluent molecule or lateral spacer. These mixed films, although not representing a perfect lipid bilayer, have been shown to retain the activity of incorporated H(+)-ATP synthases from chloroplasts in contrast to films prepared from the pure thiolipopeptide. The activity of the protein was demonstrated by impedance spectroscopy. The resistance decreased due to proton transport across the lipid film, which occurs as a consequence of adenosine triphosphate (ATP) hydrolysis. Several effects previously determined from kinetic measurements of the enzyme reconstituted in liposomes such as saturation with respect to the substrate (ATP), inhibition by venturicidin, activation by a positive potential pulse and increase of the proton current as a function of increasingly negative potentials have been confirmed also for this tethered membrane system. Changes in the impedance spectra due to the addition of ATP were fully reversible
MH  - A
MH  - ACTIVATION
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - chloroplast
MH  - Chloroplasts
MH  - England
MH  - function
MH  - GOLD
MH  - Hydrolysis
MH  - Liposomes
MH  - membrane
MH  - Membranes
MH  - protein
MH  - proton
MH  - spectra
MH  - spectroscopy
MH  - SYNTHASE
MH  - SYSTEM
MH  - transport
RP  - NOT IN FILE
NT  - UI - 21607606LA - engPT - Journal ArticleDA - 20011214IS - 0956-5663SB - IMCY - England
UR  - PM:11742732
SO  - Biosens Bioelectron 2002 Jan ;17(1-2):25-34

1929
UI  - 21425
AU  - Naumann R
AU  - Baumgart T
AU  - Graber P
AU  - Jonczyk A
AU  - Offenhausser A
AU  - Knoll W
TI  - Proton transport through a peptide-tethered bilayer lipid membrane by the H(+)-ATP synthase from chloroplasts measured by impedance spectroscopy
AB  - A lipid membrane was tethered to a gold film by a peptide spacer molecule terminated by a sulfhydryl group. Membranes were formed by fusion of liposomes prepared from egg phosphatidylcholine on self assembled monolayers of the thiolipopeptide Myr-Lys(Myr)-Ser-Ser-Pro-Ala-Ser-Ser-Ala-Ala-Ser-Ala-Cys-amide mixed with mercaptoethanol as a diluent molecule or lateral spacer. These mixed films, although not representing a perfect lipid bilayer, have been shown to retain the activity of incorporated H(+)-ATP synthases from chloroplasts in contrast to films prepared from the pure thiolipopeptide. The activity of the protein was demonstrated by impedance spectroscopy. The resistance decreased due to proton transport across the lipid film, which occurs as a consequence of adenosine triphosphate (ATP) hydrolysis. Several effects previously determined from kinetic measurements of the enzyme reconstituted in liposomes such as saturation with respect to the substrate (ATP), inhibition by venturicidin, activation by a positive potential pulse and increase of the proton current as a function of increasingly negative potentials have been confirmed also for this tethered membrane system. Changes in the impedance spectra due to the addition of ATP were fully reversible
MH  - A
MH  - ACTIVATION
MH  - Adenosine
MH  - Adenosine Triphosphate
MH  - atp
MH  - chloroplast
MH  - Chloroplasts
MH  - Electric Impedance
MH  - enzyme
MH  - enzymology
MH  - function
MH  - GOLD
MH  - Hydrolysis
MH  - Lipid Bilayers
MH  - liposome
MH  - Liposomes
MH  - membrane
MH  - Membranes
MH  - metabolism
MH  - Microscopy,Fluorescence
MH  - peptide
MH  - protein
MH  - proton
MH  - Proton-Translocating ATPases
MH  - RESISTANCE
MH  - spectra
MH  - spectroscopy
MH  - spectrum analysis
MH  - Surface Plasmon Resonance
MH  - SYNTHASE
MH  - SYSTEM
MH  - transport
MH  - venturicidin
RP  - NOT IN FILE
NT  - Max Planck Institute of Polymer Research, Ackermannweg 10, D-55128 Mainz, Germany naumannr@mpip-mainzmpgdeFAU - Naumann, R
SO  - Biosens Bioelectron 2002 Jan ;17(1-2):25-34

1930
UI  - 21324
AU  - Nishio K
AU  - Iwamoto-Kihara A
AU  - Yamamoto A
AU  - Wada Y
AU  - Futai M
TI  - Subunit rotation of ATP synthase embedded in membranes: a or beta subunit rotation relative to the c subunit ring
AB  - ATP synthase F(o)F(1) (alpha(3)beta(3)gammadelta varepsilon ab(2)c(10-14)) couples an electrochemical proton gradient and a chemical reaction through the rotation of its subunit assembly. In this study, we engineered F(o)F(1) to examine the rotation of the catalytic F(1) beta or membrane sector F(o) a subunit when the F(o) c subunit ring was immobilized; a biotin-tag was introduced onto the beta or a subunit, and a His-tag onto the c subunit ring. Membrane fragments were obtained from Escherichia coli cells carrying the recombinant plasmid for the engineered F(o)F(1) and were immobilized on a glass surface. An actin filament connected to the beta or a subunit rotated counterclockwise on the addition of ATP, and generated essentially the same torque as one connected to the c ring of F(o)F(1) immobilized through a His-tag linked to the alpha or beta subunit. These results established that the gamma varepsilon c(10-14) and alpha(3)beta(3)deltaab(2) complexes are mechanical units of the membrane-embedded enzyme involved in rotational catalysis
MH  - A
MH  - alpha
MH  - atp
MH  - ATP synthase
MH  - BETA
MH  - BETA-SUBUNIT
MH  - Catalysis
MH  - catalytic
MH  - cell
MH  - Cells
MH  - COMPLEX
MH  - enzyme
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - membrane
MH  - Membranes
MH  - proton
MH  - PROTON GRADIENT
MH  - rotation
MH  - SUBUNIT
MH  - SURFACE
MH  - SYNTHASE
MH  - universities
RP  - NOT IN FILE
NT  - Division of Biological Sciences, Institute of Scientific and Industrial Research, Osaka University, Core Research for Evolutional Science and Technology (CREST) of the Japan Science and Technology Corporation, Osaka 567-0047, JapanFAU - Nishio, Kazuaki
SO  - Proc Natl Acad Sci U S A 2002 Oct 15 ;99(21):13448-13452

1931
UI  - 21316
AU  - Pacheco-Moises F
AU  - Minauro-Sanmiguel F
AU  - Bravo C
AU  - Garcia JJ
TI  - Sulfite inhibits the F1F0-ATP synthase and activates the F1F0-ATPase of Paracoccus denitrificans
AB  - The F1F0 complex of Paracoccus denitrificans (PdF1F0) is the fastest ATP synthase but the slowest ATPase. Sulfite exerts maximal activation of the PdF1F0-ATPase (Pacheco-Moises, F., Garcia, J. J., Rodriguez-Zavala, J. S., and Moreno-Sanchez, R. (2000). Eur J. Biochem. 267, 993-1000) but its effect on the PdF1F0-ATP synthase activity remains unknown. Therefore, we studied the effect of sulfite on ATP synthesis and 32Pi <--> ATP exchange reactions of inside-out membrane vesicles of P. denitrificans. Sulfite inhibited both reactions under conditions of maximal delta pH and normal sensitivity to dicyclohexylcarbodiimide. Sulfite increased by 10- and 5-fold the K0.5 for Mg2+-ADP and Pi during ATP synthesis, respectively, and by 4-fold the IC50 of Mg2+-ADP for inhibition of the PdF1F0-ATPase activity. Thus, sulfite exerts opposite effects on the forward and reverse functioning of the PdF1F0 complex. These effects are not due to membrane or PdF1F0 uncoupling. Kinetic and structural modifications that could account for these results are discussed
MH  - ACTIVATION
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - ATPase
MH  - COMPLEX
MH  - delta
MH  - DELTA-PH
MH  - Dicyclohexylcarbodiimide
MH  - F1F0-ATP SYNTHASE
MH  - inside-out
MH  - membrane
MH  - membrane vesicles
MH  - P
MH  - pH
MH  - SYNTHASE
MH  - synthesis
MH  - vesicles
RP  - NOT IN FILE
NT  - Departamento de Bioquimica, Instituto Nacional de Cardiologia, Ignacio Chavez, Mexico, DF, MexicoFAU - Pacheco-Moises, Fermin
SO  - J Bioenerg Biomembr 2002 Aug ;34(4):269-278

1932
UI  - 21400
AU  - Paumard P
AU  - Vaillier J
AU  - Coulary B
AU  - Schaeffer J
AU  - Soubannier V
AU  - Mueller DM
AU  - Brethes D
AU  - di Rago JP
AU  - Velours J
TI  - The ATP synthase is involved in generating mitochondrial cristae morphology
AB  - The inner membrane of the mitochondrion folds inwards, forming the cristae. This folding allows a greater amount of membrane to be packed into the mitochondrion. The data in this study demonstrate that subunits e and g of the mitochondrial ATP synthase are involved in generating mitochondrial cristae morphology. These two subunits are non-essential components of ATP synthase and are required for the dimerization and oligomerization of ATP synthase. Mitochondria of yeast cells deficient in either subunits e or g were found to have numerous digitations and onion-like structures that correspond to an uncontrolled biogenesis and/or folding of the inner mitochondrial membrane. The present data show that there is a link between dimerization of the mitochondrial ATP synthase and cristae morphology. A model is proposed of the assembly of ATP synthase dimers, taking into account the oligomerization of the yeast enzyme and earlier data on the ultrastructure of mitochondrial cristae, which suggests that the association of ATP synthase dimers is involved in the control of the biogenesis of the inner mitochondrial membrane
MH  - A
MH  - atp
MH  - ATP synthase
MH  - cell
MH  - Cells
MH  - Chemistry
MH  - data
MH  - Dimerization
MH  - enzyme
MH  - enzymology
MH  - Intracellular Membranes
MH  - membrane
MH  - Microscopy,Electron
MH  - Mitochondria
MH  - Mitochondrial Proton-Translocating ATPases
MH  - model
MH  - Models,Molecular
MH  - physiology
MH  - Saccharomyces cerevisiae
MH  - structure
MH  - SUBUNIT
MH  - Support,Non-U.S.Gov't
MH  - Support,U.S.Gov't,P.H.S.
MH  - SYNTHASE
MH  - ultrastructure
MH  - YEAST
RP  - NOT IN FILE
NT  - Institut de Biochimie et Genetique Cellulaires du CNRS, Universite Victor Segalen, Bordeaux 2, 1 rue Camille Saint Saens, 33077 Bordeaux cedex, FranceFAU - Paumard, Patrick
SO  - EMBO J 2002 Feb 1 ;21(3):221-230

1933
UI  - 21345
AU  - Paumard P
AU  - Arselin G
AU  - Vaillier J
AU  - Chaignepain S
AU  - Bathany K
AU  - Schmitter JM
AU  - Brethes D
AU  - Velours J
TI  - Two ATP synthases can be linked through subunits i in the inner mitochondrial membrane of Saccharomyces cerevisiae
AB  - Cross-linking experiments showed that the supernumerary subunit i is close to the interface between two ATP synthases. These data were used to demonstrate the presence of ATP synthase dimers in the inner mitochondrial membrane of Saccharomyces cerevisiae. A cysteine residue was introduced into the inter-membrane space located C-terminal part of subunit i. Cross-linking experiments revealed a dimerization of subunit i. This cross-linking occurred only with the dimeric form of the enzyme after incubating intact mitochondria with a bis-maleimide reagent, thus indicating an inter-ATP synthase cross-linking, whereas the monomeric form of the enzyme exhibited only an intra-ATP synthase cross-linking with subunit 6, another component of the membranous domain of the ATP synthase
MH  - A
MH  - Amino Acid Sequence
MH  - atp
MH  - ATP synthase
MH  - Chemistry
MH  - CROSS-LINKING
MH  - Cross-Linking Reagents
MH  - Cysteine
MH  - data
MH  - Dimerization
MH  - enzyme
MH  - enzymology
MH  - Intracellular Membranes
MH  - Maleimides
MH  - membrane
MH  - Mitochondria
MH  - Mitochondrial Proton-Translocating ATPases
MH  - Molecular Sequence Data
MH  - Proton-Translocating ATPases
MH  - RESIDUE
MH  - Saccharomyces cerevisiae
MH  - Saccharomyces cerevisiae Proteins
MH  - SUBUNIT
MH  - Support,Non-U.S.Gov't
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - Institut de Biochimie et Genetique Cellulaires du CNRS, Universite Victor Segalen, Bordeaux 2, 1 rue Camille Saint-Saens 33077 Bordeaux Cedex, FranceFAU - Paumard, Patrick
SO  - Biochemistry 2002 Aug ;%20;41(33):10390-10396

1934
UI  - 21314
AU  - Radresa O
AU  - Ogata K
AU  - Wodak S
AU  - Ruysschaert JM
AU  - Goormaghtigh E
TI  - Modeling the three-dimensional structure of H+-ATPase of Neurospora crassa
AB  - Homology modeling in combination with transmembrane topology predictions are used to build the atomic model of Neurospora crassa plasma membrane H+-ATPase, using as template the 2.6 A crystal structure of rabbit sarcoplasmic reticulum Ca2+-ATPase [Toyoshima, C., Nakasako, M., Nomura, H. & Ogawa, H. (2000) Nature 405, 647-655]. Comparison of the two calcium-binding sites in the crystal structure of Ca2+-ATPase with the equivalent region in the H+-ATPase model shows that the latter is devoid of most of the negatively charged groups required to bind the cations, suggesting a different role for this region. Using the built model, a pathway for proton transport is then proposed from computed locations of internal polar cavities, large enough to contain at least one water molecule. As a control, the same approach is applied to the high-resolution crystal structure of halorhodopsin and the proton pump bacteriorhodopsin. This revealed a striking correspondence between the positions of internal polar cavities, those of crystallographic water molecules and, in the case of bacteriorhodopsin, the residues mediating proton translocation. In our H+-ATPase model, most of these cavities are in contact with residues previously shown to affect coupling of proton translocation to ATP hydrolysis. A string of six polar cavities identified in the cytoplasmic domain, the most accurate part of the model, suggests a proton entry path starting close to the phosphorylation site. Strikingly, members of the haloacid dehalogenase superfamily, which are close structural homologs of this domain but do not share the same function, display only one polar cavity in the vicinity of the conserved catalytic Asp residue
MH  - A
MH  - atp
MH  - Bacteriorhodopsin
MH  - catalytic
MH  - Cations
MH  - conformation
MH  - coupling
MH  - function
MH  - H+-ATPase
MH  - Hydrolysis
MH  - M
MH  - membrane
MH  - Membranes
MH  - model
MH  - Neurospora
MH  - Neurospora crassa
MH  - Phosphorylation
MH  - proton
MH  - Proton Pump
MH  - RESIDUE
MH  - Sarcoplasmic Reticulum
MH  - Site
MH  - structure
MH  - translocation
MH  - transport
MH  - Water
RP  - NOT IN FILE
NT  - Service de Structure et Fonction des Membranes Biologiques, Universite Libre de Bruxelles, Bruxelles, Belgium; Unite de Conformation des Macromolecules Biologiques, Universite Libre de Bruxelles, Bruxelles, BelgiumFAU - Radresa, Olivier
SO  - Eur J Biochem 2002 Nov ;269(21):5246-5258

1935
UI  - 21375
AU  - Revington M
AU  - Dunn SD
AU  - Shaw GS
TI  - Folding and stability of the b subunit of the F(1)F(0) ATP synthase
AB  - The F(1)F(0) ATP synthase is a reversible molecular motor that employs a rotary catalytic cycle to couple a chemiosmotic membrane potential to the formation/hydrolysis of ATP. The multisubunit enzyme contains two copies of the b subunit that form a homodimer as part of a narrow, peripheral stalk structure that connects the membrane (F(0)) and soluble (F(1)) sectors. The three-dimensional structure of the b subunit is unknown making the nature of any interactions or conformational changes within the F(1)F(0) complex difficult to interpret. We have used circular dichroism and analytical ultracentrifugation analyses of a series of N- and C-terminal truncated b proteins to investigate its stability and structure. Thermal denaturation of the b constructs exhibited distinct two-state, cooperative unfolding with T(m) values between 30 and 40 degrees C. CD spectra for the region comprising residues 53-122 (b(53-122)) showed theta;(222)/theta;(208) = 0.99, which reduced to 0.92 in the presence of the hydrophobic solvent trifluoroethanol. Thermodynamic parameters for b(53-122) (DeltaG, DeltaH and DeltaC(p)) were similar to those reported for several nonideal, coiled-coil proteins. Together these results are most consistent with a noncanonical and unstable parallel coiled-coil at the interface of the b dimer
MH  - A
MH  - Amino Acid Sequence
MH  - atp
MH  - ATP synthase
MH  - Biochemistry
MH  - catalytic
MH  - Chemistry
MH  - Circular Dichroism
MH  - COMPLEX
MH  - conformational change
MH  - conformational changes
MH  - enzyme
MH  - INTERACTION
MH  - membrane
MH  - Membrane Potential
MH  - metabolism
MH  - Molecular Sequence Data
MH  - protein
MH  - Protein Conformation
MH  - Protein Folding
MH  - Protein Structure,Secondary
MH  - Proteins
MH  - Proton-Translocating ATPases
MH  - RESIDUE
MH  - Sequence Alignment
MH  - Sodium Chloride
MH  - SOLVENT
MH  - spectra
MH  - stalk
MH  - structure
MH  - SUBUNIT
MH  - Support,Non-U.S.Gov't
MH  - SYNTHASE
MH  - Temperature
MH  - Thermodynamics
MH  - Trifluoroethanol
MH  - Ultracentrifugation
MH  - universities
RP  - NOT IN FILE
NT  - Department of Biochemistry, The University of Western Ontario, London, Ontario, Canada, N6A 5C1FAU - Revington, Matthew
SO  - Protein Sci 2002 May ;11(5):1227-1238

1936
UI  - 21233
AU  - Rocchia W
AU  - Sridharan S
AU  - Nicholls A
AU  - Alexov E
AU  - Chiabrera A
AU  - Honig B
TI  - Rapid Grid-Based Construction of the Molecular Surface and the Use of Induced Surface Charge to Calculate Reaction Field Energies: Applications to the Molecular Systems and Geometric Objects. 
MH  - SURFACE
MH  - FIELD
MH  - SYSTEMS
MH  - SYSTEM
RP  - NOT IN FILE
SO  - J Comp Chem 2002  ;23():128-137

1937
UI  - 21339
AU  - Rubinstein J
AU  - Walker J
TI  - ATP Synthase from Saccharomyces cerevisiae: Location of the OSCP Subunit in the Peripheral Stalk Region
AB  - A biotinylation signal has been fused to the C terminus of the oligomycin sensitivity conferral protein (OSCP) of the ATP synthase complex from Saccharomyces cerevisiae. The signal is biotinylated in vivo and the biotinylated complex binds avidin in vitro. By electron microscopy of negatively stained particles of the ATP synthase-avidin complex, the bound avidin has been localised close to the F(1) domain. The images were subjected to multi-reference alignment and classification. Because of the presence of a flexible linker between the OSCP and the biotinylation signal, the class-averages differ in the position of the avidin relative to the F(1) domain. These positions lie on an arc, and its centre indicates the position of the C terminus of the OSCP on the surface of the F(1) domain. Since the N-terminal region of the OSCP is known to interact with the N-terminal regions of alpha-subunits, which are on top of the F(1) domain distal from the F(o) membrane domain, the OSCP extends almost 10nm along the surface of F(1) down towards F(o) where it interacts with the C terminus of the b subunit, which extends up from F(o). The labelling technique has also allowed a reliable 2D projection map to be developed for the intact ATP synthase from S.cerevisiae. The map reveals a marked asymmetry in the F(o) part of the complex that can be attributed to subunits in the F(o) domain
MH  - A
MH  - ALPHA-SUBUNIT
MH  - atp
MH  - ATP synthase
MH  - COMPLEX
MH  - electron
MH  - electron microscopy
MH  - ELECTRON-MICROSCOPY
MH  - Human
MH  - In Vitro
MH  - membrane
MH  - Microscopy
MH  - Nutrition
MH  - oligomycin
MH  - protein
MH  - Saccharomyces cerevisiae
MH  - stalk
MH  - SUBUNIT
MH  - SURFACE
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - MRC Dunn Human Nutrition Unit, Hills Road, CB2 2XY, Cambridge, UKFAU - Rubinstein, John
SO  - J Mol Biol 2002 Aug 23 ;321(4):613

1938
UI  - 21372
AU  - Santolini J
AU  - Minoletti C
AU  - Gomis JM
AU  - Sigalat C
AU  - Andre F
AU  - Haraux F
TI  - An insight into the mechanism of inhibition and reactivation of the F(1)-ATPases by tentoxin
AB  - The mechanism of inhibition and reactivation of chloroplast ATP-synthase by the fungal cyclotetrapeptide tentoxin was investigated by photolabeling experiments, binding studies, and kinetic analysis using synthetic analogues of tentoxin. The alpha-subunit of chloroplast F(1)-ATPase (CF(1)) was specifically labeled by a photoactivatable tentoxin derivative, providing the first direct evidence of tentoxin binding to the alpha-subunit, and 3D homology modeling was used to locate tentoxin in its putative binding site at the alpha/beta interface. The non-photosynthetic F(1)-ATPase from thermophilic bacterium (TF(1)) proved to be also tentoxin-sensitive, and enzyme turnover dramatically increased the rate of tentoxin binding to its inhibitory site, contrary to what was previously observed with epsilon-depleted CF(1) [Santolini, J., Haraux, F., Sigalat, C., Moal, G., and Andre, F. (1999) J. Biol. Chem. 274, 849-858]. We propose that tentoxin preferentially binds to an ADP-loaded alpha beta pair, and mechanically blocks the catalytic cycle, perhaps by the impossibility of converting this alpha beta pair into an ATP-loaded alpha beta pair. Using (14)C-tentoxin and selected synthetic analogues, we found that toxin binding to the tight inhibitory site of CF(1) exerts some cooperative effect on the loose reactivatory site, but that no reciprocal effect exists. When the two tentoxin-binding sites are filled in reactivated F(1)-ATPase, they do not exchange their role during catalytic turnover, indicating an impairment between nucleotide occupancy and the shape of tentoxin-binding pocket. This analysis provides a mechanical interpretation of the inhibition of F(1)-ATPase by tentoxin and a clue for understanding the reactivation process
MH  - A
MH  - Affinity Labels
MH  - alpha
MH  - ALPHA-SUBUNIT
MH  - analysis
MH  - antagonists & inhibitors
MH  - ATP synthase
MH  - Bacillus
MH  - Bacteria
MH  - BETA
MH  - BINDING
MH  - BINDING SITE
MH  - Binding Sites
MH  - catalytic
MH  - Chemistry
MH  - chloroplast
MH  - Chloroplasts
MH  - enzyme
MH  - Enzyme Inhibitors
MH  - Enzyme Reactivators
MH  - enzymology
MH  - Kinetics
MH  - mechanism
MH  - metabolism
MH  - Models,Biological
MH  - Models,Molecular
MH  - nucleotide
MH  - Peptides,Cyclic
MH  - pharmacology
MH  - Protein Conformation
MH  - Protein Structure,Secondary
MH  - Proton-Translocating ATPases
MH  - Site
MH  - Support,Non-U.S.Gov't
MH  - tentoxin
MH  - THERMOPHILIC
MH  - turnover
RP  - NOT IN FILE
NT  - Service de Bioenergetique, DBJC, CEA Saclay, 91191 Gif-sur-Yvette Cedex, FranceFAU - Santolini, Jerome
SO  - Biochemistry 2002 May 14 ;41(19):6008-6018

1939
UI  - 21336
AU  - Schagger H
TI  - Respiratory chain supercomplexes of mitochondria and bacteria
AB  - Respiratory chain complexes are fragments of larger structural and functional units, the respiratory chain supercomplexes or "respirasomes", which exist in bacterial and mitochondrial membranes. Supercomplexes of mitochondria and bacteria contain complexes III, IV, and complex I, with the notable exception of Saccharomyces cerevisiae, which does not possess complex I. These supercomplexes often are stable to sonication but sensitive to most detergents except digitonin. In S. cerevisiae, a major component linking complexes III and IV together is cardiolipin.In Paracoccus denitrificans, complex I itself is rather detergent-sensitive and thus could not be obtained in detergent-solubilized form so far. However, it can be isolated as part of a supercomplex. Stabilization of complex I by binding to complex III was also found in human mitochondria. Further functional roles of the organization in a supercomplex are catalytic enhancement by reducing diffusion distances of substrates or, depending on the organism, channelling of the substrates quinone and cytochrome c. This makes redox reactions less dependent of midpoint potentials of substrates, and permits electron flow at low degree of substrate reduction.A dimeric state of ATP synthase seems to be specific for mitochondria. Exclusively, monomeric ATP synthase was found in Acetobacterium woodii, in P. denitrificans, and in spinach chloroplasts
MH  - A
MH  - Animal
MH  - atp
MH  - ATP synthase
MH  - ATP Synthetase Complexes
MH  - Bacteria
MH  - Bacterial Proton-Translocating ATPases
MH  - BINDING
MH  - Cardiolipins
MH  - catalytic
MH  - Cattle
MH  - Chemistry
MH  - chloroplast
MH  - Chloroplast Proton-Translocating ATPases
MH  - Chloroplasts
MH  - Comparative Study
MH  - COMPLEX
MH  - cytochrome
MH  - Cytochrome c
MH  - Detergents
MH  - Diffusion
MH  - Dimerization
MH  - electron
MH  - enzymology
MH  - Human
MH  - isolation & purification
MH  - membrane
MH  - Membranes
MH  - midpoint potential
MH  - Mitochondria
MH  - Mitochondria,Heart
MH  - Mitochondrial Proton-Translocating ATPases
MH  - Models,Molecular
MH  - NADH,NADPH Oxidoreductases
MH  - Oxidation-Reduction
MH  - Oxidative Phosphorylation
MH  - P
MH  - Paracoccus denitrificans
MH  - quinone
MH  - redox
MH  - Saccharomyces cerevisiae
MH  - Spinach
MH  - SPINACH-CHLOROPLASTS
MH  - Support,Non-U.S.Gov't
MH  - SYNTHASE
MH  - Ubiquinol-Cytochrome-c Reductase
RP  - NOT IN FILE
NT  - Biochemie I, Zentrum der Biologischen Chemie, Universitatsklinikum Frankfurt, Theodor-Stern-Kai 7, Haus 25B, D-60590 Frankfort on the Main, Germany schagger@zbcklinikuni-frankfurtdeFAU - Schagger, Hermann
SO  - Biochim Biophys Acta 2002 Sep 10 ;1555(1-3):154-159

1940
UI  - 21359
AU  - Scheidt KA
AU  - Bannister TD
AU  - Tasaka A
AU  - Wendt MD
AU  - Savall BM
AU  - Fegley GJ
AU  - Roush WR
TI  - Total synthesis of (-)-bafilomycin A(1)
AB  - A highly stereoselective total synthesis of (-)-bafilomycin A(1), the naturally occurring enantiomer of this potent vacuolar ATPase inhibitor, is described. The synthesis features the highly stereoselective aldol reaction of methyl ketone 8b and aldehyde 60c and a Suzuki cross-coupling reaction of the highly functionalized advanced intermediates 12 and 39. Vinyl iodide 12 was synthesized by a 14-step sequence starting from the readily available beta-alkoxy aldehyde 14, while the vinylboronic acid component 39 was synthesized by a nine-step sequence from beta-hydroxy-alpha-methyl butyrate 44 via a sequence involving the alpha-methoxypropargylation of chiral aldehyde 49 with the alpha-methoxypropargylstannane reagent 54. Syntheses of fragments 12 and 39 also feature diastereoselective double asymmetric crotylboration reactions to set several of the critical stereocenters. The Suzuki cross-coupling of 12 and 39 provided seco ester 40, which following conversion to the seco acid underwent smooth macrolactonization to give 41. The success of the macrocyclization required that C(7)-OH be unprotected. The Mukaiyama aldol reaction between aldehyde 60c and the TMS enol ether generated from 8b provided aldol 65 with high diastereoselectivity. Finally, all silicon protecting groups were removed by treatment of the penultimate intermediate 65 with TAS-F (tris(dimethylamino)sulfonium difluorotrimethylsilicate), thereby completing the total synthesis of (-)-bafilomycin A(1)
MH  - A
MH  - ACID
MH  - antagonists & inhibitors
MH  - Antibiotics,Antifungal
MH  - antibiotics,macrolide
MH  - ATPase
MH  - chemical synthesis
MH  - Chemistry
MH  - Enzyme Inhibitors
MH  - inhibitor
MH  - intermediate
MH  - Stereoisomerism
MH  - Support,U.S.Gov't,P.H.S.
MH  - synthesis
MH  - universities
MH  - Vacuolar Proton-Translocating ATPases
RP  - NOT IN FILE
NT  - Department of Chemistry, University of Michigan, Ann Arbor, Michigan 48109, USAFAU - Scheidt, Karl A
SO  - J Am Chem Soc 2002 Jun ;%19;124(24):6981-6990

1941
UI  - 21371
AU  - Senior AE
AU  - Nadanaciva S
AU  - Weber J
TI  - The molecular mechanism of ATP synthesis by F1F0-ATP synthase
AB  - ATP synthesis by oxidative phosphorylation and photophosphorylation, catalyzed by F1F0-ATP synthase, is the fundamental means of cell energy production. Earlier mutagenesis studies had gone some way to describing the mechanism. More recently, several X-ray structures at atomic resolution have pictured the catalytic sites, and real-time video recordings of subunit rotation have left no doubt of the nature of energy coupling between the transmembrane proton gradient and the catalytic sites in this extraordinary molecular motor. Nonetheless, the molecular events that are required to accomplish the chemical synthesis of ATP remain undefined. In this review we summarize current state of knowledge and present a hypothesis for the molecular mechanism of ATP synthesis
MH  - A
MH  - Adenosine Triphosphate
MH  - Animal
MH  - atp
MH  - ATP synthesis
MH  - Binding Sites
MH  - Biochemistry
MH  - Biophysics
MH  - biosynthesis
MH  - Catalysis
MH  - catalytic
MH  - cell
MH  - chemical synthesis
MH  - Chemistry
MH  - coupling
MH  - Crystallography,X-Ray
MH  - DNA Mutational Analysis
MH  - energy
MH  - Escherichia coli
MH  - F1F0-ATP SYNTHASE
MH  - Human
MH  - Magnetic Resonance Spectroscopy
MH  - mechanism
MH  - metabolism
MH  - Models,Molecular
MH  - Molecular Motors
MH  - mutagenesis
MH  - Oxidative Phosphorylation
MH  - Phosphorylation
MH  - Photophosphorylation
MH  - Plants
MH  - proton
MH  - PROTON GRADIENT
MH  - Proton-Translocating ATPases
MH  - resolution
MH  - review
MH  - rotation
MH  - Site
MH  - structure
MH  - SUBUNIT
MH  - Support,U.S.Gov't,P.H.S.
MH  - SYNTHASE
MH  - synthesis
MH  - universities
RP  - NOT IN FILE
NT  - Department of Biochemistry and Biophysics, University of Rochester Medical Center, Box 712, 601 Elmwood Avenue, Rochester, NY 14642, USA alan_senior@urmcrochestereduFAU - Senior, Alan E
SO  - Biochim Biophys Acta 2002 Feb 15 ;1553(3):188-211

1942
UI  - 21312
AU  - Smondyrev AM
AU  - Voth GA
AD  - Department of Chemistry and Henry Eyring Center for Theoretical Chemistry, University of Utah, Salt Lake City, Utah 84112-0850, USA
TI  - Molecular dynamics simulation of proton transport near the surface of a phospholipid membrane
AB  - The structural and dynamical properties of a hydrated proton near the surface of DMPC membrane were studied using a molecular dynamics simulation. The proton transport between water molecules was modeled using the second generation multistate empirical valence bond model. The proton diffusion was found to be inhibited at the membrane surface. The potential of mean force for the proton adsorption to the membrane surface and its release back into the bulk water was also determined, yielding a small barrier in each direction. An efficient algorithm for Ewald summation calculations for the multistate empirical valence bond model is also introduced
MH  - A
MH  - Chemistry
MH  - Diffusion
MH  - Lipids
MH  - membrane
MH  - model
MH  - MOLECULAR-DYNAMICS
MH  - Phospholipids
MH  - proton
MH  - Protons
MH  - SIMULATION
MH  - SURFACE
MH  - transport
MH  - united states
MH  - universities
MH  - Water
RP  - NOT IN FILE
NT  - UI - 21856006LA - engRN - 0 (Lipids)RN - 0 (Phospholipids)RN - 0 (Protons)RN - 13699-48-4 (Dimyristoylphosphatidylcholine)RN - 7732-18-5 (Water)PT - Journal ArticleID - GM53148/GM/NIGMSDA - 20020227IS - 0006-3495SB - IMCY - United States
UR  - PM:11867461
SO  - Biophys J 2002 Mar ;82(3):1460-1468

1943
UI  - 21045
AU  - Soubannier V
AU  - Vaillier J
AU  - Paumard P
AU  - Coulary B
AU  - Schaeffer J
AU  - Velours J
TI  - In the absence of the first membrane-spanning segment of subunit 4 (b), the yeast ATP synthase is functional but does not dimerize or oligomerize
AB  - The N-terminal portion of the mitochondrial b-subunit is anchored in the inner mitochondrial membrane by two hydrophobic segments. We investigated the role of the first membrane-spanning segment, which is absent in prokaryotic and chloroplastic enzymes. In the absence of the first membrane-spanning segment of the yeast subunit (subunit 4), a strong decrease in the amount of subunit g was found. The mutant ATP synthase did not dimerize or oligomerize and mutant cells displayed anomalous mitochondrial morphologies with onion-like structures. This phenotype is similar to that of the null mutant in the ATP20 gene that encodes subunit g, a component involved in the dimerization / oligomerization of ATP synthase. Our data indicate that the first membrane-spanning segment of the mitochondrial b-subunit is not essential for the function of the enzyme since its removal did not directly alter the oxidative phosphorylation. It is proposed that the unique membrane-spanning segment of subunit g and the first membrane-spanning segment of subunit 4 interact, as shown by cross-linking experiments. We hypothesize that in eukaryotic cells the b-subunit has evolved to accommodate the interaction with the g-subunit, an associated ATP synthase component only present in the mitochondrial enzyme
MH  - A
MH  - atp
MH  - ATP synthase
MH  - Cells
MH  - CROSS-LINKING
MH  - Enzymes
MH  - function
MH  - membrane
MH  - mutant
MH  - Oxidative Phosphorylation
MH  - Phosphorylation
MH  - structure
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - Institut de Biochimie et Gntique Cellulaires, CNRS, Bordeaux 33077
SO  - J Biol Chem 2002 Jan 17 ;.():

1944
UI  - 21318
AU  - Steinbrecher T
AU  - Hucke O
AU  - Steigmiller S
AU  - Borsch M
AU  - Labahn A
TI  - Binding affinities and protein ligand complex geometries of nucleotides at the F(1) part of the mitochondrial ATP synthase obtained by ligand docking calculations
AB  - F(0)F(1) ATP synthases utilize a transmembrane electrochemical potential difference to synthesize ATP from ADP and phosphate. In this work, the binding modes of ADP, ATP and ATP analogues to the catalytic sites of the F(1) part of the mitochondrial ATP synthase were investigated with ligand docking calculations. Binding geometries of ATP and ADP at the three catalytic sites agree with X-ray crystal data; their binding free energies suggest an assignment to the 'tight', 'open' and 'loose' states. The rates of multi-site hydrolysis for two fluorescent ATP derivatives were measured using a fluorescence assay. Reduced hydrolysis rates compared to ATP can be explained by the ligand docking calculations
MH  - A
MH  - ADP
MH  - affinity
MH  - atp
MH  - ATP synthase
MH  - BINDING
MH  - catalytic
MH  - COMPLEX
MH  - data
MH  - energy
MH  - fluorescence
MH  - Hydrolysis
MH  - nucleotide
MH  - Nucleotides
MH  - phosphate
MH  - protein
MH  - Site
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - Institut fur Physikalische Chemie der Albert-Ludwigs-Universitat Freiburg, Albertstrasse 23a, D-79104, Freiburg, GermanyFAU - Steinbrecher, Thomas
SO  - FEBS Lett 2002 Oct 23 ;530(1-3):99

1945
UI  - 21392
AU  - Sun-Wada GH
AU  - Imai-Senga Y
AU  - Yamamoto A
AU  - Murata Y
AU  - Hirata T
AU  - Wada Y
AU  - Futai M
TI  - A proton pump ATPase with testis-specific E1-subunit isoform required for acrosome acidification
AB  - The vacuolar-type H(+)-ATPases (V-ATPases) are a family of multimeric proton pumps involved in a wide variety of physiological processes. We have identified two novel mouse genes, Atp6e1 and Atp6e2, encoding testis-specific (E1) and ubiquitous (E2) V-ATPase subunit E isoforms, respectively. The E1 transcript appears about 3 weeks after birth, corresponding to the start of meiosis, and is expressed specifically in round spermatids in seminiferous tubules. Immunohistochemistry with isoform-specific antibodies revealed that the V-ATPase with E1 and a2 isoforms is located specifically in developing acrosomes of spermatids and acrosomes in mature sperm. In contrast, the E2 isoform was expressed in all tissues examined and present in the perinuclear compartments of spermatocytes. The E1 isoform exhibits 70% identity with the E2, and both isoforms functionally complemented a null mutation of the yeast counterpart VMA4, indicating that they are bona fide V-ATPase subunits. The chimeric enzymes showed slightly lower K(m)(ATP) than yeast V-ATPase. Consistent with the temperature-sensitive growth of Deltavma4-expressing E1 isoform, vacuolar membrane vesicles exhibited temperature-sensitive coupling between ATP hydrolysis and proton transport. These results suggest that E1 isoform is essential for energy coupling involved in acidification of acrosome
MH  - A
MH  - Acrosome
MH  - Amino Acid Sequence
MH  - Animal
MH  - atp
MH  - ATPase
MH  - Blotting,Northern
MH  - Chemistry
MH  - coupling
MH  - energy
MH  - enzyme
MH  - Enzymes
MH  - enzymology
MH  - Fertility
MH  - genetics
MH  - H(+)ATPase
MH  - Hydrogen-Ion Concentration
MH  - Hydrolysis
MH  - In Situ Hybridization
MH  - Kinetics
MH  - Male
MH  - membrane
MH  - membrane vesicles
MH  - metabolism
MH  - Mice
MH  - Molecular Sequence Data
MH  - Mutation
MH  - Protein Conformation
MH  - proton
MH  - Proton Pump
MH  - Proton Pumps
MH  - Seminiferous Tubules
MH  - Sequence Alignment
MH  - SUBUNIT
MH  - Support,Non-U.S.Gov't
MH  - Testis
MH  - transport
MH  - universities
MH  - Vacuolar Proton-Translocating ATPases
MH  - vesicles
MH  - YEAST
MH  - Yeasts
RP  - NOT IN FILE
NT  - Division of Biological Sciences, Institute of Scientific and Industrial Research, Osaka University, Core Research for Evolutional Science and Technology (CREST) of the Japan Science and Technology Corp, Osaka 567-0047, JapanFAU - Sun-Wada, Ge-Hong
SO  - J Biol Chem 2002 May 17 ;277(20):18098-18105

1946
UI  - 21401
AU  - Suzuki T
AU  - Ueno H
AU  - Mitome N
AU  - Suzuki J
AU  - Yoshida M
TI  - F(0) of ATP synthase is a rotary proton channel. Obligatory coupling of proton translocation with rotation of c-subunit ring
AB  - Coupling of proton flow and rotation in the F(0) motor of ATP synthase was investigated using the thermophilic Bacillus PS3 enzyme expressed functionally in Escherichia coli cells. Cysteine residues introduced into the N-terminal regions of subunits b and c of ATP synthase (bL2C/cS2C) were readily oxidized by treating the expressing cells with CuCl(2) to form predominantly a b-c cross-link with b-b and c-c cross-links being minor products. The oxidized ATP synthases, either in the inverted membrane vesicles or in the reconstituted proteoliposomes, showed drastically decreased proton pumping and ATPase activities compared with the reduced ones. Also, the oxidized F(0), either in the F(1)-stripped inverted vesicles or in the reconstituted F(0)-proteoliposomes, hardly mediated passive proton translocation through F(0). Careful analysis using single mutants (bL2C or cS2C) as controls indicated that the b-c cross-link was responsible for these defects. Thus, rotation of the c-oligomer ring relative to subunit b is obligatory for proton translocation; if there is no rotation of the c-ring there is no proton flow through F(0)
MH  - A
MH  - analysis
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Bacillus
MH  - Base Sequence
MH  - cell
MH  - Cells
MH  - Chemistry
MH  - coupling
MH  - Cysteine
MH  - DNA Primers
MH  - enzyme
MH  - enzymology
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - Ion Transport
MH  - isolation & purification
MH  - membrane
MH  - membrane vesicles
MH  - metabolism
MH  - mutant
MH  - proteoliposome
MH  - proton
MH  - Proton-Translocating ATPases
MH  - Protons
MH  - PS3
MH  - Recombinant Proteins
MH  - RESIDUE
MH  - rotation
MH  - SUBUNIT
MH  - SYNTHASE
MH  - THERMOPHILIC
MH  - THERMOPHILIC BACILLUS PS3
MH  - translocation
MH  - vesicles
RP  - NOT IN FILE
NT  - Chemical Resources Laboratory, Tokyo Institute of Technology, Nagatsuta 4259, Yokohama 226-8503, JapanFAU - Suzuki, Toshiharu
SO  - J Biol Chem 2002 Apr 12 ;277(15):13281-13285

1947
UI  - 21044
AU  - Suzuki T
AU  - Ueno H
AU  - Mitome N
AU  - Suzuki J
AU  - Yoshida M
TI  - Fo of ATP synthase is a rotary proton channel: Obligatory coupling of proton translocation with rotation of c-subunit ring
AB  - Coupling of proton flow and rotation in F(o) motor of ATP synthase was investigated using the thermophilic Bacillus PS3 enzyme expressed functionally in Escherichia coli cells. Cysteine residues introduced into the N-terminal regions of subunits b and c of ATP synthase (bL2C/cS2C) were readily oxidized by treating the expressing cells with CuCl(2) to form predominantly a b-c cross-link with b-b and c-c cross-links being minor products. The oxidized ATP synthases, either in the inverted membrane vesicles or in the reconstituted proteoliposomes, showed drastically decreased proton pumping and ATPase activities compared with the reduced ones. Also, the oxidized F(o), either in the F(1)-stripped inverted vesicles or in the reconstituted F(o)-proteoliposomes, hardly mediated passive proton translocation through F(o). Careful analysis using single mutants (bL2C or cS2C) as controls indicated that b-c cross-link was responsible for these defects. Thus, rotation of the c-oligomer ring relative to subunit b is obligatory for proton translocation; no rotation of the c-ring, no proton flow through F(o)
MH  - A
MH  - analysis
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Bacillus
MH  - Cells
MH  - coupling
MH  - Cysteine
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - membrane
MH  - membrane vesicles
MH  - mutant
MH  - proteoliposome
MH  - proton
MH  - PS3
MH  - RESIDUE
MH  - rotation
MH  - SUBUNIT
MH  - SYNTHASE
MH  - THERMOPHILIC BACILLUS PS3
MH  - translocation
MH  - vesicles
RP  - NOT IN FILE
NT  - Chemical resources laboratory, Tokyo Institute of Technology, Yokohama, Kanagawa 226-8503
SO  - J Biol Chem 2002 Jan 28 ;.():

1948
UI  - 21379
AU  - Turina P
AU  - Melandri BA
TI  - A point mutation in the ATP synthase of Rhodobacter capsulatus results in differential contributions of Delta(pH) and Delta(phi) in driving the ATP synthesis reaction
AB  - The interface between the c-subunit oligomer and the a subunit in the F0 sector of the ATP synthase is believed to form the core of the rotating motor powered by the protonic flow. Besides the essential cAsp61 and aArg210 residues (Escherichia coli numbering), a few other residues at this interface, although nonessential, show a high degree of conservation, among these aGlu219. The homologous residue aGlu210 in the ATP synthase of the photosynthetic bacterium Rhodobacter capsulatus has been substituted by a lysine. Inner membranes prepared from the mutant strain showed approximately half of the ATP synthesis activity when driven both by light and by acid-base transitions. As estimated with the ACMA assay, proton pumping rates in the inner membranes were also reduced to a similar extent in the mutant. The most striking impairment of ATP synthesis in the mutant, a decrease as low as 12 times as compared to the wild-type, was observed in the absence of a transmembrane electrical membrane potential (Delta(phi)) at low transmembrane pH difference (Delta(pH)). Therefore, the mutation seems to affect both the mechanism responsible for coupling F1 with proton translocation by F0, and the mechanism determining the relative contribution of Delta(pH) and Delta(phi) in driving ATP synthesis
MH  - A
MH  - Adenosine Triphosphate
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - Bacteria
MH  - Base Sequence
MH  - Biochemistry
MH  - biology
MH  - Biophysics
MH  - biosynthesis
MH  - Blotting,Western
MH  - capsulatus
MH  - coupling
MH  - DNA Primers
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F0
MH  - F1
MH  - genetics
MH  - Light
MH  - mechanism
MH  - membrane
MH  - Membrane Potential
MH  - Membranes
MH  - metabolism
MH  - Mutagenesis,Site-Directed
MH  - mutant
MH  - Mutation
MH  - pH
MH  - point mutation
MH  - proton
MH  - Proton-Translocating ATPases
MH  - RESIDUE
MH  - Rhodobacter
MH  - rhodobacter capsulatus
MH  - RHODOBACTER-CAPSULATUS
MH  - SUBUNIT
MH  - Support,Non-U.S.Gov't
MH  - SYNTHASE
MH  - synthesis
MH  - Time
MH  - translocation
MH  - universities
RP  - NOT IN FILE
NT  - Department of Biology, Laboratory of Biochemistry and Biophysics, University of Bologna, ItalyFAU - Turina, Paola
SO  - Eur J Biochem 2002 Apr ;269(7):1984-1992

1949
UI  - 21370
AU  - Valiyaveetil F
AU  - Hermolin J
AU  - Fillingame RH
TI  - pH dependent inactivation of solubilized F1F0 ATP synthase by dicyclohexylcarbodiimide: pK(a) of detergent unmasked aspartyl-61 in Escherichia coli subunit c
AB  - The pH dependence of the reaction of dicyclohexylcarbodiimide with the essential aspartyl-61 residue in subunit c of Escherichia coli ATP synthase was compared in membranes and in a detergent dispersed preparation of the enzyme. The rate of reaction was estimated by measuring the inactivation of ATPase activity. The reaction with the detergent dispersed form of the enzyme proved to be pH sensitive with the essential aspartyl group titrating with a pK(a)=8. However, when measured with E. coli membranes, the reaction proved to be pH insensitive. The results suggest that the reacting aspartyl-61 residues are shielded from the bulk aqueous solvent when in the membrane, but then become aqueous-accessible following detergent solubilization
MH  - A
MH  - antagonists & inhibitors
MH  - Aspartic Acid
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Cell Membrane
MH  - Chemistry
MH  - COLI ATP SYNTHASE
MH  - DEPENDENCE
MH  - Detergents
MH  - Dicyclohexylcarbodiimide
MH  - drug effects
MH  - enzyme
MH  - Enzyme Inhibitors
MH  - enzymology
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - Hydrogen-Ion Concentration
MH  - isolation & purification
MH  - Kinetics
MH  - membrane
MH  - Membranes
MH  - pH
MH  - pharmacology
MH  - Polyethylene Glycols
MH  - Proton-Translocating ATPases
MH  - RESIDUE
MH  - SOLVENT
MH  - SUBUNIT
MH  - Support,U.S.Gov't,P.H.S.
MH  - SYNTHASE
MH  - universities
RP  - NOT IN FILE
NT  - Department of Biomolecular Chemistry, University of Wisconsin Medical School, 1300 University Avenue, Madison, WI 53706, USAFAU - Valiyaveetil, Francis
SO  - Biochim Biophys Acta 2002 Feb 15 ;1553(3):296-301

1950
UI  - 21275
AU  - Valiyaveetil F
AU  - Hermolin J
AU  - Fillingame RH
AD  - Department of Biomolecular Chemistry, University of Wisconsin Medical School, 1300 University Avenue, 53706, Madison, WI, USA
TI  - pH dependent inactivation of solubilized F(1)F(0) ATP synthase by dicyclohexylcarbodiimide: pK(a) of detergent unmasked aspartyl-61 in Escherichia coli subunit c
AB  - The pH dependence of the reaction of dicyclohexylcarbodiimide with the essential aspartyl-61 residue in subunit c of Escherichia coli ATP synthase was compared in membranes and in a detergent dispersed preparation of the enzyme. The rate of reaction was estimated by measuring the inactivation of ATPase activity. The reaction with the detergent dispersed form of the enzyme proved to be pH sensitive with the essential aspartyl group titrating with a pK(a)=8. However, when measured with E. coli membranes, the reaction proved to be pH insensitive. The results suggest that the reacting aspartyl-61 residues are shielded from the bulk aqueous solvent when in the membrane, but then become aqueous-accessible following detergent solubilization
MH  - A
MH  - atp
MH  - ATP synthase
MH  - ATPase
MH  - Chemistry
MH  - COLI ATP SYNTHASE
MH  - DEPENDENCE
MH  - Dicyclohexylcarbodiimide
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - membrane
MH  - Membranes
MH  - pH
MH  - RESIDUE
MH  - SOLVENT
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 21992410LA - engPT - Journal ArticleDA - 20020508IS - 0006-3002SB - IMCY - Netherlands
UR  - PM:11997138
SO  - Biochim Biophys Acta 2002 Feb 15 ;1553(3):296-301

1951
UI  - 21397
AU  - Van Der HN
AU  - Docampo R
TI  - Proton and sodium pumps regulate the plasma membrane potential of different stages of Trypanosoma cruzi
AB  - We studied the plasma membrane potential (DeltaPsi) in different stages of Trypanosoma cruzi using the potentiometric fluorescent dye bisoxonol. Our results clearly demonstrate that a proton pump plays a significant role in the regulation of DeltaPsi in all stages of T. cruzi as evidenced by depolarization of the DeltaPsi by H(+)-ATPase inhibitors dicyclohexylcarbo-diimide, N-ethylmaleimide, diethylstilbestrol, and bafilomycin A(1). The H(+)-ATPase appeared to be activated by acidic conditions in trypomastigotes as evidenced by hyperpolarization of the DeltaPsi upon addition of acid. In contrast to epimastigotes and amastigotes, the DeltaPsi of trypomastigotes was markedly sensitive to extracellular Na(+) and K(+) concentrations and evidence was provided for an outward directed Cl(minus sign) channel in this stage as well. Additionally, for the first time, the existence of an ouabain-sensitive functional sodium pump was demonstrated in T. cruzi based on the depolarization of the DeltaPsi in trypomastigotes by ouabain in the presence of Na(+). In the epimastigotes, ouabain had no effect on the DeltaPsi in a sodium rich buffer. However, ouabain induced an additional significant depolarization in these stages when their DeltaPsi was already partially depolarized by the H(+)-ATPase inhibitor dicyclohexylcarbo-diimide, supporting the presence of an ouabain-sensitive sodium pump whose activity is masked by the H(+)-ATPase. In the amastigotes no evidence for a functional sodium pump could be found. In support of an inwardly directed K(+) channel, the DeltaPsi was hyperpolarized by K(+) free buffer in trypomastigotes and epimastigotes and by Ba(2+) in epimastigotes and amastigotes. The presence of K(+) channels in amastigotes and a sodium pump in trypomastigotes, in addition to the H(+)-ATPase, could provide important new targets for trypanocidal drug development
MH  - A
MH  - ACID
MH  - Animal
MH  - antagonists & inhibitors
MH  - Barium
MH  - buffer
MH  - Cell Membrane
MH  - development
MH  - Dicyclohexylcarbodiimide
MH  - drug effects
MH  - DYE
MH  - growth & development
MH  - H(+)ATPase
MH  - Hydrogen-Ion Concentration
MH  - inhibitor
MH  - inhibitors
MH  - membrane
MH  - Membrane Potential
MH  - Membrane Potentials
MH  - metabolism
MH  - Na(+)-K(+)-Exchanging ATPase
MH  - Ouabain
MH  - parasitology
MH  - pharmacology
MH  - physiology
MH  - Potassium
MH  - proton
MH  - Proton Pump
MH  - Proton Pumps
MH  - Proton-Translocating ATPases
MH  - regulation
MH  - Sodium
MH  - Support,Non-U.S.Gov't
MH  - Support,U.S.Gov't,P.H.S.
MH  - Time
MH  - Trypanosoma cruzi
MH  - universities
MH  - veterinary
RP  - NOT IN FILE
NT  - Laboratory of Molecular Parasitology, Department of Pathobiology, College of Veterinary Medicine, University of Illinois at Urbana-Champaign, 2001 South Lincoln Avenue, Urbana, IL 61802, USAFAU - Van Der Heyden, Nicole
SO  - Mol Biochem Parasitol 2002 Mar ;120(1):127-139

1952
UI  - 21433
AU  - von Ballmoos C
AU  - Appoldt Y
AU  - Brunner J
AU  - Granier T
AU  - Vasella A
AU  - Dimroth P
TI  - Membrane topography of the coupling ion binding site in Na+-translocating F1F0 ATP synthase
AB  - A carbodiimide with a photoactivatable diazirine substituent was synthesized and incubated with the Na(+)-translocating F(1)F(0) ATP synthase from both Propionigenium modestum and Ilyobacter tartaricus. This caused severe inhibition of ATP hydrolysis activity in the absence of Na(+) ions but not in its presence, indicating the specific reaction with the Na(+) binding c-Glu(65) residue. Photocross-linking was investigated with the substituted ATP synthase from both bacteria in reconstituted 1-palmitoyl-2-oleyl-sn-glycero-3-phosphocholine (POPC)-containing proteoliposomes. A subunit c/POPC conjugate was found in the illuminated samples but no a-c cross-links were observed, not even after ATP-induced rotation of the c-ring. Our substituted diazirine moiety on c-Glu(65) was therefore in close contact with phospholipid but does not contact subunit a. Na(+)in/(22)Na(+)out exchange activity of the ATP synthase was not affected by modifying the c-Glu(65) sites with the carbodiimide, but upon photoinduced cross-linking, this activity was abolished. Cross-linking the rotor to lipids apparently arrested rotational mobility required for moving Na(+) ions back and forth across the membrane. The site of cross-linking was analyzed by digestions of the substituted POPC using phospholipases C and A(2) and by mass spectroscopy. The substitutions were found exclusively at the fatty acid side chains, which indicates that c-Glu(65) is located within the core of the membrane
MH  - A
MH  - ACID
MH  - Adenosinetriphosphatase
MH  - atp
MH  - ATP synthase
MH  - Bacteria
MH  - BINDING
MH  - BINDING SITE
MH  - Binding Sites
MH  - Carbodiimides
MH  - Cation Transport Proteins
MH  - Cell Membrane
MH  - chemical synthesis
MH  - Chemistry
MH  - coupling
MH  - CROSS-LINKING
MH  - enzymology
MH  - Fusobacterium
MH  - Hydrolysis
MH  - ion
MH  - Ions
MH  - isolation & purification
MH  - Lipids
MH  - Liposomes
MH  - membrane
MH  - metabolism
MH  - Molecular Conformation
MH  - Phosphatidylcholines
MH  - Protein Subunits
MH  - proteoliposome
MH  - Proton-Translocating ATPases
MH  - RESIDUE
MH  - rotation
MH  - Site
MH  - Sodium
MH  - Spectrometry,Mass,Matrix-Assisted Laser Desorption-Ionization
MH  - spectroscopy
MH  - SUBUNIT
MH  - SYNTHASE
MH  - ultrastructure
RP  - NOT IN FILE
NT  - Institut fur Mikrobiologie der Eidgenossischen Technischen Hochschule, ETH Zentrum, CH-8092 Zurich, SwitzerlandFAU - von Ballmoos, Christoph
SO  - J Biol Chem 2002 Feb 1 ;277(5):3504-3510

1953
UI  - 21027
AU  - von Ballmoos C
AU  - Appoldt Y
AU  - Brunner J
AU  - Granier T
AU  - Vasella A
AU  - Dimroth P
AD  - Institut fur Mikrobiologie der Eidgenossischen Technischen Hochschule, ETH Zentrum, CH-8092 Zurich, Switzerland
TI  - Membrane topography of the coupling ion binding site in Na+- translocating F1F0 ATP synthase
AB  - A carbodiimide with a photoactivatable diazirine substituent was synthesized and incubated with the Na(+)-translocating F(1)F(0) ATP synthase from both Propionigenium modestum and Ilyobacter tartaricus. This caused severe inhibition of ATP hydrolysis activity in the absence of Na(+) ions but not in its presence, indicating the specific reaction with the Na(+) binding c-Glu(65) residue. Photocross-linking was investigated with the substituted ATP synthase from both bacteria in reconstituted 1-palmitoyl-2-oleyl-sn-glycero-3-phosphocholine (POPC)- containing proteoliposomes. A subunit c/POPC conjugate was found in the illuminated samples but no a-c cross-links were observed, not even after ATP-induced rotation of the c-ring. Our substituted diazirine moiety on c-Glu(65) was therefore in close contact with phospholipid but does not contact subunit a. Na(+)in/(22)Na(+)out exchange activity of the ATP synthase was not affected by modifying the c-Glu(65) sites with the carbodiimide, but upon photoinduced cross-linking, this activity was abolished. Cross-linking the rotor to lipids apparently arrested rotational mobility required for moving Na(+) ions back and forth across the membrane. The site of cross-linking was analyzed by digestions of the substituted POPC using phospholipases C and A(2) and by mass spectroscopy. The substitutions were found exclusively at the fatty acid side chains, which indicates that c-Glu(65) is located within the core of the membrane
MH  - A
MH  - ACID
MH  - atp
MH  - ATP synthase
MH  - Bacteria
MH  - BINDING
MH  - coupling
MH  - CROSS-LINKING
MH  - Hydrolysis
MH  - ion
MH  - Ions
MH  - Lipids
MH  - membrane
MH  - proteoliposome
MH  - RESIDUE
MH  - rotation
MH  - Site
MH  - spectroscopy
MH  - SUBUNIT
MH  - SYNTHASE
RP  - NOT IN FILE
NT  - UI - 21671363LA - engPT - Journal ArticleDA - 20020128IS - 0021-9258SB - IMCY - United States
UR  - PM:11719523
SO  - J Biol Chem 2002 Feb 1 ;277(5):3504-3510

1954
UI  - 21351
AU  - Vonck J
AU  - von Nidda TK
AU  - Meier T
AU  - Matthey U
AU  - Mills DJ
AU  - Kuhlbrandt W
AU  - Dimroth P
TI  - Molecular architecture of the undecameric rotor of a bacterial Na+-ATP synthase
AB  - The sodium ion-translocating F(1)F(0) ATP synthase from the bacterium Ilyobacter tartaricus contains a remarkably stable rotor ring composed of 11 c subunits. The rotor ring was isolated, crystallised in two dimensions and analysed by electron cryo-microscopy. Here, we present an alpha-carbon model of the c-subunit ring. Each monomeric c subunit of 89 amino acid residues folds into a helical hairpin consisting of two membrane-spanning helices and a cytoplasmic loop. The 11 N-terminal helices are closely spaced within an inner ring surrounding a cavity of approximately 17A (1.7 nm). The tight helix packing leaves no space for side-chains and is accounted for by a highly conserved motif of four glycine residues in the inner, N-terminal helix. Each inner helix is connected by a clearly visible loop to an outer C-terminal helix. The outer helix has a kink near the position of the ion-binding site residue Glu65 in the centre of the membrane and another kink near the C terminus. Two helices from the outer ring and one from the inner ring form the ion-binding site in the middle of the membrane and a potential access channel from the binding site to the cytoplasmic surface. Three possible inter-subunit ion-bridges are likely to account for the remarkable temperature stability of I.tartaricus c-rings compared to those of other organisms
MH  - A
MH  - ACID
MH  - Amino Acid Sequence
MH  - atp
MH  - ATP synthase
MH  - Bacteria
MH  - BINDING
MH  - BINDING SITE
MH  - Binding Sites
MH  - Biophysics
MH  - Chemistry
MH  - Conserved Sequence
MH  - CRYOELECTRON MICROSCOPY
MH  - Cytoplasm
MH  - electron
MH  - ELECTRON CRYOMICROSCOPY
MH  - enzymology
MH  - Ions
MH  - membrane
MH  - metabolism
MH  - model
MH  - Models,Molecular
MH  - Molecular Sequence Data
MH  - Phospholipids
MH  - Protein Structure,Quaternary
MH  - Protein Structure,Secondary
MH  - Protein Subunits
MH  - Proton-Translocating ATPases
MH  - RESIDUE
MH  - Site
MH  - Sodium
MH  - SUBUNIT
MH  - Support,Non-U.S.Gov't
MH  - SURFACE
MH  - SYNTHASE
MH  - Temperature
MH  - ultrastructure
RP  - NOT IN FILE
NT  - Max-Planck-Institute of Biophysics, Heinrich-Hoffmann-Str 7, Frankfurt, GermanyFAU - Vonck, Janet
SO  - J Mol Biol 2002 Aug 9 ;321(2):307-316

1955
UI  - 21403
AU  - Walz D
AU  - Caplan SR
TI  - Bacterial flagellar motor and H(+)/ATP synthase: two proton-driven rotary molecular devices with different functions
AB  - Both the bacterial flagellar motor and the H(+)/ATP synthase are membrane-bound macromolecular complexes in which the movement of protons through channels across the membrane is coupled to the rotation of a part of the complex around an axis perpendicular to the membrane. Despite this similarity, the two devices are designed for quite different functions. The flagellar motor is responsible for a practically smooth rotation of the flagellar filament in order to propel the cell. Smooth rotation is not essential for the H(+)/ATP synthase, which accumulates torque by twisting a rod-shaped structure. Possible mechanisms for generating torque in the two devices are presented, based on the models which have been proposed. The performances of the various mechanisms are discussed
MH  - A
MH  - Bacteria
MH  - cell
MH  - COMPLEX
MH  - enzymology
MH  - function
MH  - mechanism
MH  - MECHANISMS
MH  - membrane
MH  - metabolism
MH  - model
MH  - Models,Biological
MH  - Movement
MH  - proton
MH  - Proton-Translocating ATPases
MH  - Protons
MH  - rotation
MH  - structure
MH  - SYNTHASE
MH  - universities
RP  - NOT IN FILE
NT  - Biozentrum, University of Basel, Klinger bergstrasse 70, CH-4056 Basel, Switzerland DieterWalz@unibaschFAU - Walz, Dieter
SO  - Bioelectrochemistry 2002 Jan ;55(1-2):89-92

1956
UI  - 21393
AU  - Weber J
AU  - Wilke-Mounts S
AU  - Senior AE
TI  - Quantitative determination of binding affinity of delta-subunit in Escherichia coli F1-ATPase: effects of mutation, Mg2+, and pH on Kd
AB  - To study the stator function in ATP synthase, a fluorimetric assay has been devised for quantitative determination of binding affinity of delta-subunit to Escherichia coli F(1)-ATPase. The signal used is that of the natural tryptophan at residue delta28, which is enhanced by 50% upon binding of delta-subunit to alpha(3)beta(3)gammaepsilon complex. K(d) for delta binding is 1.4 nm, which is energetically equivalent (50.2 kJ/mol) to that required to resist the rotor strain. Only one site for delta binding was detected. The deltaW28L mutation increased K(d) to 4.6 nm, equivalent to a loss of 2.9 kJ/mol binding energy. While this was insufficient to cause detectable functional impairment, it did facilitate preparation of delta-depleted F(1). The alphaG29D mutation reduced K(d) to 26 nm, equivalent to a loss of 7.2 kJ/mol binding energy. This mutation did cause serious functional impairment, referable to interruption of binding of delta to F(1). Results with the two mutants illuminate how finely balanced is the stator resistance function. delta' fragment, consisting of residues delta1-134, bound with the same K(d) as intact delta, showing that, at least in absence of F(o) subunits, the C-terminal domain of delta contributes zero binding energy. Mg(2+) ions had a strong effect on increasing delta binding affinity, supporting the possibility of bridging metal ion involvement in stator function. High pH environment greatly reduced delta binding affinity, suggesting the involvement of protonatable side-chains in the binding site
MH  - A
MH  - affinity
MH  - atp
MH  - ATP synthase
MH  - Base Sequence
MH  - BINDING
MH  - BINDING SITE
MH  - Biochemistry
MH  - Biophysics
MH  - COLI F1 ATPASE
MH  - COMPLEX
MH  - delta
MH  - DELTA-SUBUNIT
MH  - DNA Primers
MH  - energy
MH  - enzymology
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - F1 ATPASE
MH  - F1-ATPASE
MH  - function
MH  - genetics
MH  - ion
MH  - Ions
MH  - isolation & purification
MH  - Magnesium
MH  - metabolism
MH  - Mutagenesis,Site-Directed
MH  - mutant
MH  - Mutation
MH  - pH
MH  - Proton-Translocating ATPases
MH  - RESIDUE
MH  - RESISTANCE
MH  - Site
MH  - Spectrometry,Fluorescence
MH  - SUBUNIT
MH  - SYNTHASE
MH  - tryptophan
MH  - universities
RP  - NOT IN FILE
NT  - Department of Biochemistry and Biophysics, University of Rochester Medical Center, Rochester, New York 14642, USAFAU - Weber, Joachim
SO  - J Biol Chem 2002 May 24 ;277(21):18390-18396

1957
UI  - 21422
AU  - Weber J
AU  - Bijol V
AU  - Wilke-Mounts S
AU  - Senior AE
TI  - Cysteine-reactive fluorescence probes of catalytic sites of ATP synthase
AB  - We searched for new fluorescent probes of catalytic-site nucleotide binding in F(1)F(0)-ATP synthase by introducing Cys mutations at positions in or close to catalytic sites and then reacting Cys-mutant F(1) with thiol-reactive fluorescent probes. Four suitable mutant/probe combinations were identified. beta F410C labeled by 7-fluorobenz-2-oxa-1,3-diazole-4-sulfonamide (ABD-F) gave very large signal changes in response to nucleotide, allowing facile measurement of fluorescence and nucleotide-binding parameters, not only in F(1) but also in F(1)F(0). The results are consistent with the presence of three asymmetric catalytic sites of widely different affinities, with similar properties in both enzymes, and revealed a unique probe environment at the high-affinity site 1. beta Y331C F(1) labeled by ABD-F gave a large signal which monitored catalytic site polarity changes that occur along the ATP hydrolysis pathway. Two other mutant/probe combinations with significant nucleotide-responsive signals were beta Y331C labeled by 5-((((2-iodoacetyl)amino)ethyl)amino)naphthaline-1-sulfonic acid and alpha F291C labeled by 2-4'-(iodoacetamido)anilino)naphthalene-6-sulfonic acid. The signal of the latter responds differentially to nucleoside diphosphate versus triphosphate bound in catalytic sites
MH  - A
MH  - ACID
MH  - Adenosine Triphosphate
MH  - affinity
MH  - alpha
MH  - atp
MH  - ATP synthase
MH  - ATP Synthetase Complexes
MH  - BETA
MH  - BINDING
MH  - Binding Sites
MH  - Biochemistry
MH  - Biophysics
MH  - catalytic
MH  - catalytic domain
MH  - CATALYTIC SITE NUCLEOTIDE
MH  - Chemistry
MH  - Cysteine
MH  - Dose-Response Relationship,Drug
MH  - enzyme
MH  - Enzymes
MH  - Escherichia coli
MH  - fluorescence
MH  - Fluorescent Dyes
MH  - genetics
MH  - Hydrolysis
MH  - Kinetics
MH  - Magnesium
MH  - metabolism
MH  - Methods
MH  - Mutation
MH  - nucleotide
MH  - nucleotide binding
MH  - Nucleotides
MH  - Oxygen
MH  - Phosphorylation
MH  - Protein Binding
MH  - Site
MH  - Spectrometry,Fluorescence
MH  - Support,U.S.Gov't,P.H.S.
MH  - SYNTHASE
MH  - universities
RP  - NOT IN FILE
NT  - Department of Biochemistry and Biophysics, University of Rochester Medical Center, Rochester, New York 14642, USAFAU - Weber, Joachim
SO  - Arch Biochem Biophys 2002 Jan 1 ;397(1):1-10

1958
UI  - 21340
AU  - Wehrle F
AU  - Kaim G
AU  - Dimroth P
TI  - Molecular Mechanism of the ATP Synthase's F(o) Motor Probed by Mutational Analyses of Subunit a
AB  - The most prominent residue of subunit a of the F(1)F(o) ATP synthase is a universally conserved arginine (aR227 in Propionigenium modestum), which was reported to permit no substitution with retention of ATP synthesis or H(+)-coupled ATP hydrolysis activity. We show here that ATP synthases with R227K or R227H mutations in the P.modestum a subunit catalyse ATP-driven Na(+) transport above or below pH 8.0, respectively. Reconstituted F(o) with either mutation catalysed 22Na(+)(out)/Na(+)(in) exchange with similar pH profiles as found in ATP-driven Na(+) transport. ATP synthase with an aR227A substitution catalysed Na(+)-dependent ATP hydrolysis, which was completely inhibited by dicyclohexylcarbodiimide, but not coupled to Na(+) transport. This suggests that in the mutant the dissociation of Na(+) becomes more difficult and that the alkali ions remain therefore permanently bound to the c subunit sites. The reconstituted mutant enzyme was also able to synthesise ATP in the presence of a membrane potential, which stopped at elevated external Na(+) concentrations. These observations reinforce the importance of aR227 to facilitate the dissociation of Na(+) from approaching rotor sites. This task of aR227 was corroborated by other results with the aR227A mutant: (i) after reconstitution into liposomes, F(o) with the aR227A mutation did not catalyse 22Na(+)(out)/Na(+)(in) exchange at high internal sodium concentrations, and (ii) at a constant DeltapNa(+), 22Na(+) uptake was inhibited at elevated internal Na(+) concentrations. Hence, in mutant aR227A, sodium ions can only dissociate from their rotor sites into a reservoir of low sodium ion concentration, whereas in the wild-type the positively charged aR227 allows the dissociation of Na(+) even into compartments of high Na(+) concentration
MH  - A
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - CONSTANT
MH  - Dicyclohexylcarbodiimide
MH  - enzyme
MH  - Hydrolysis
MH  - ion
MH  - Ions
MH  - liposome
MH  - Liposomes
MH  - mechanism
MH  - membrane
MH  - Membrane Potential
MH  - mutant
MH  - pH
MH  - reconstitution
MH  - RESIDUE
MH  - Site
MH  - Sodium
MH  - SUBUNIT
MH  - SYNTHASE
MH  - synthesis
MH  - transport
RP  - NOT IN FILE
NT  - Institut fur Mikrobiologie, Eidgenossische Technische Hochschule, Schmelzbergstr 7, CH-8092, Zurich, SwitzerlandFAU - Wehrle, Franziska
SO  - J Mol Biol 2002 Sep 13 ;322(2):369

1959
UI  - 21363
AU  - Wehrle F
AU  - Appoldt Y
AU  - Kaim G
AU  - Dimroth P
TI  - Reconstitution of Fo of the sodium ion translocating ATP synthase of Propionigenium modestum from its heterologously expressed and purified subunits
AB  - The atpB and atpF genes of Propionigenium modestum were cloned as His-tag fusion constructs and expressed in Escherichia coli. Both recombinant subunits a and b were purified via Ni(2+) chelate affinity chromatography. A functionally active Fo complex was reassembled in vitro from subunits a, b and c, and incorporated into liposomes. The F(o) liposomes catalysed (22)Na(+) uptake in response to an inside negative potassium diffusion potential, and the uptake was prevented by modification of the c subunits with N,N'-dicyclohexylcarbodiimide (DCCD). In the absence of a membrane potential the Fo complexes catalysed (22)Na(+)(out)/Na(+)(in)-exchange. After F(1) addition the F(1)F(o) complex was formed and the holoenzyme catalysed ATP synthesis, ATP dependent Na(+) pumping, and ATP hydrolysis, which was inhibited by DCCD. Functional F(o) hybrids were reconstituted with recombinant subunits a and b from P. modestum and c(11) from Ilyobacter tartaricus. These Fo hybrids had Na(+) translocation activities that were not distinguishable from that of P. modestum F(o)
MH  - A
MH  - ACTIVE
MH  - affinity
MH  - atp
MH  - ATP synthase
MH  - ATP synthesis
MH  - Cloning,Molecular
MH  - COMPLEX
MH  - Diffusion
MH  - diffusion potential
MH  - enzymology
MH  - Escherichia coli
MH  - ESCHERICHIA-COLI
MH  - FO
MH  - genetics
MH  - Gram-Negative Anaerobic Bacteria
MH  - Hydrolysis
MH  - In Vitro
MH  - ion
MH  - Ion Transport
MH  - liposome
MH  - Liposomes
MH  - membrane
MH  - Membrane Potential
MH  - metabolism
MH  - Methods
MH  - P
MH  - Potassium
MH  - Proton-Translocating ATPases
MH  - Recombinant Fusion Proteins
MH  - reconstitution
MH  - Sodium
MH  - SUBUNIT
MH  - Support,Non-U.S.Gov't
MH  - SYNTHASE
MH  - synthesis
MH  - translocation
RP  - NOT IN FILE
NT  - Institut fur Mikrobiologie, Eidgenossische Technische Hochschule, Zurich, SwitzerlandFAU - Wehrle, Franziska
SO  - Eur J Biochem 2002 May ;269(10):2567-2573

1960
UI  - 21311
AU  - Zhang J
AU  - Unwin PR
AD  - Contribution from the Department of Chemistry, University of Warwick, Coventry CV4 7AL, UK
TI  - Proton diffusion at phospholipid assemblies
AB  - A new scanning electrochemical microscopy proton feedback method has been developed for investigating lateral proton diffusion at phospholipid assemblies: specifically Langmuir monolayers at the water/air interface. In this approach, a base is electrogenerated by the reduction of a weak acid (producing hydrogen) at a "submarine" ultramicroelectrode (UME) placed in the aqueous subphase of a Langmuir trough close to a monolayer. The electrogenerated base diffuses to and titrates monolayer-bound protons and is converted back to its initial form, so enhancing the current response at the UME. Local deprotonation of the monolayer creates a concentration gradient for lateral proton diffusion. A numerical model has been developed, taking into account the potential-dependent association/dissociation constant of the interfacial acid groups. A comparison is made of monolayers comprising either acidic DL-alpha-phosphatidyl-L-serine, dipalmitoyl (DPPS) or zwitterionic L-alpha-phosphatidylcholine, dipalmitoyl (DPPC) monolayers at a range of surface pressures. It is demonstrated that lateral proton fluxes at DPPS are significant, but the lateral proton diffusion coefficient is lower than in bulk solution. In contrast, lateral proton diffusion cannot be detected at DPPC, suggesting that the acid/base character of the phospholipid is important in determining the magnitude of interfacial proton fluxes
MH  - A
MH  - ACID
MH  - BASE
MH  - Chemistry
MH  - CONSTANT
MH  - Diffusion
MH  - Hydrogen
MH  - method
MH  - Microscopy
MH  - model
MH  - Phospholipids
MH  - proton
MH  - Protons
MH  - REDUCTION
MH  - SCANNING ELECTROCHEMICAL MICROSCOPY
MH  - SURFACE
MH  - united states
MH  - universities
RP  - NOT IN FILE
NT  - UI - 21871177LA - engRN - 0 (Phospholipids)RN - 0 (Protons)PT - Journal ArticleDA - 20020306IS - 0002-7863SB - IMCY - United States
UR  - PM:11878995
SO  - J Am Chem Soc 2002 Mar 13 ;124(10):2379-2383
