Catalytic hydrolysis and synthesis of adenosine 5'-triphosphate by stereoisomers of covalently labeled F1-adenosinetriphosphatase and reconstituted submitochondrial particles. 1987

J H Wang, and J Cesana, and J C Wu
Bioenergetics Laboratory, State University of New York, Buffalo 14214-3094.

Bovine heart F1-adenosinetriphosphatase (F1) was labeled specifically and precisely with 7-chloro-4-nitro-2,1,3-[14C]benzoxadiazole ([14C]NBD-Cl). The stereospecifically labeled F1 (O-beta'-[14C]-NBD-F1) was partially reactivated by LiCl treatment, which could cause rearrangement of the beta subunits to form O-beta', beta''-[14C]NBD-F1. Both labeled enzymes were used to combine with F1-deficient submitochondrial particles (ASU) to form the reconstituted particles O-beta'-NBD-F1-ASU and O-beta', beta''-NBD-F1-ASU, respectively. A comparison of the observed steady-state rates of catalytic ATP hydrolysis and oxidative phosphorylation by these specifically labeled submitochondrial particles (SMP) with those of the unlabeled control samples suggests that oxidative phosphorylation involves more active sites of F1 than catalytic ATP hydrolysis. A comparison of the observed ATPase activity of uncoupled labeled SMP and the activity for ATP-driven reverse electron transport in coupled labeled SMP with the corresponding values of the unlabeled control samples shows that the observed fractional inhibition ATP hydrolysis is the same for both the coupled SMP and uncoupled SMP and is determined only by the state of stereospecific labeling of F1. The effect of preincubation under simulated oxidative phosphorylation conditions on the ATPase activity of the unperturbed, specifically NBD-labeled submitochondrial particles was also examined. The data show that respiration-generated proton flux does not cause the beta subunits in bovine heart proton-ATPase to continue switching places with each other during oxidative phosphorylation. Samples of NBD-F1 with specific labels on its nonhydrolytic beta'' subunits but none on its hydrolytic beta' subunit were prepared by a three-cycle process.(ABSTRACT TRUNCATED AT 250 WORDS)

UI MeSH Term Description Entries
D007700 Kinetics The rate dynamics in chemical or physical systems.
D008929 Mitochondria, Heart The mitochondria of the myocardium. Heart Mitochondria,Myocardial Mitochondria,Mitochondrion, Heart,Heart Mitochondrion,Mitochondria, Myocardial
D010085 Oxidative Phosphorylation Electron transfer through the cytochrome system liberating free energy which is transformed into high-energy phosphate bonds. Phosphorylation, Oxidative,Oxidative Phosphorylations,Phosphorylations, Oxidative
D002417 Cattle Domesticated bovine animals of the genus Bos, usually kept on a farm or ranch and used for the production of meat or dairy products or for heavy labor. Beef Cow,Bos grunniens,Bos indicus,Bos indicus Cattle,Bos taurus,Cow,Cow, Domestic,Dairy Cow,Holstein Cow,Indicine Cattle,Taurine Cattle,Taurus Cattle,Yak,Zebu,Beef Cows,Bos indicus Cattles,Cattle, Bos indicus,Cattle, Indicine,Cattle, Taurine,Cattle, Taurus,Cattles, Bos indicus,Cattles, Indicine,Cattles, Taurine,Cattles, Taurus,Cow, Beef,Cow, Dairy,Cow, Holstein,Cows,Dairy Cows,Domestic Cow,Domestic Cows,Indicine Cattles,Taurine Cattles,Taurus Cattles,Yaks,Zebus
D006180 Proton-Translocating ATPases Multisubunit enzymes that reversibly synthesize ADENOSINE TRIPHOSPHATE. They are coupled to the transport of protons across a membrane. ATP Dependent Proton Translocase,ATPase, F0,ATPase, F1,Adenosinetriphosphatase F1,F(1)F(0)-ATPase,F1 ATPase,H(+)-Transporting ATP Synthase,H(+)-Transporting ATPase,H(+)ATPase Complex,Proton-Translocating ATPase,Proton-Translocating ATPase Complex,Proton-Translocating ATPase Complexes,ATPase, F(1)F(0),ATPase, F0F1,ATPase, H(+),Adenosine Triphosphatase Complex,F(0)F(1)-ATP Synthase,F-0-ATPase,F-1-ATPase,F0F1 ATPase,F1-ATPase,F1F0 ATPase Complex,H(+)-ATPase,H(+)-Transporting ATP Synthase, Acyl-Phosphate-Linked,H+ ATPase,H+ Transporting ATP Synthase,H+-Translocating ATPase,Proton-Translocating ATPase, F0 Sector,Proton-Translocating ATPase, F1 Sector,ATPase Complex, Proton-Translocating,ATPase Complexes, Proton-Translocating,ATPase, H+,ATPase, H+-Translocating,ATPase, Proton-Translocating,Complex, Adenosine Triphosphatase,Complexes, Proton-Translocating ATPase,F 0 ATPase,F 1 ATPase,F0 ATPase,H+ Translocating ATPase,Proton Translocating ATPase,Proton Translocating ATPase Complex,Proton Translocating ATPase Complexes,Proton Translocating ATPase, F0 Sector,Proton Translocating ATPase, F1 Sector,Triphosphatase Complex, Adenosine
D006868 Hydrolysis The process of cleaving a chemical compound by the addition of a molecule of water.
D000255 Adenosine Triphosphate An adenine nucleotide containing three phosphate groups esterified to the sugar moiety. In addition to its crucial roles in metabolism adenosine triphosphate is a neurotransmitter. ATP,Adenosine Triphosphate, Calcium Salt,Adenosine Triphosphate, Chromium Salt,Adenosine Triphosphate, Magnesium Salt,Adenosine Triphosphate, Manganese Salt,Adenylpyrophosphate,CaATP,CrATP,Manganese Adenosine Triphosphate,MgATP,MnATP,ATP-MgCl2,Adenosine Triphosphate, Chromium Ammonium Salt,Adenosine Triphosphate, Magnesium Chloride,Atriphos,Chromium Adenosine Triphosphate,Cr(H2O)4 ATP,Magnesium Adenosine Triphosphate,Striadyne,ATP MgCl2
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D013237 Stereoisomerism The phenomenon whereby compounds whose molecules have the same number and kind of atoms and the same atomic arrangement, but differ in their spatial relationships. (From McGraw-Hill Dictionary of Scientific and Technical Terms, 5th ed) Molecular Stereochemistry,Stereoisomers,Stereochemistry, Molecular,Stereoisomer
D013367 Submitochondrial Particles The various filaments, granules, tubules or other inclusions within mitochondria. Particle, Submitochondrial,Particles, Submitochondrial,Submitochondrial Particle

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