Studies on the mechanism of oxidative phosphorylation. Different effects of F0 inhibitors on unisite and multisite ATP hydrolysis by bovine submitochondrial particles.
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.
Matsuno-Yagi
A
A
Department of Molecular and Experimental Medicine, Scripps Research Institute, La Jolla, California 92037.
Hatefi
Y
Y
D000255:Adenosine Triphosphate / Q000378:metabolism*; D000818:Animals; D002417:Cattle; D004024:Dicyclohexylcarbodiimide / Q000494:pharmacology; D006160:Guanosine Triphosphate / Q000494:pharmacology; D006868:Hydrolysis; D007700:Kinetics; D008929:Mitochondria, Heart / Q000648:ultrastructure*; D009840:Oligomycins / Q000494:pharmacology; D010085:Oxidative Phosphorylation; D006180:Proton-Translocating ATPases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D013367:Submitochondrial Particles / Q000378:metabolism*; D014220:Trialkyltin Compounds / Q000494:pharmacology; D014698:Venturicidins / Q000494:pharmacology
D009840:Oligomycins; D014220:Trialkyltin Compounds; D014698:Venturicidins; C011559:tributyltin; D004024:Dicyclohexylcarbodiimide; D006160:Guanosine Triphosphate; D000255:Adenosine Triphosphate; D006180:Proton-Translocating ATPases