title: The stoichiometry of proton translocation through H+-ATPase of rat-liver mitochondria.
abstract: 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.
mesh_terms: D000251:Adenosine Triphosphatases / Q000378:metabolism*; D000818:Animals; D001692:Biological Transport; D004789:Enzyme Activation / Q000187:drug effects; D006863:Hydrogen-Ion Concentration; D007476:Ionophores / Q000494:pharmacology; D008930:Mitochondria, Liver / Q000201:enzymology*; D008956:Models, Chemical; D009570:Nitriles / Q000494:pharmacology; D006180:Proton-Translocating ATPases; D011522:Protons; D051381:Rats; D014634:Valinomycin / Q000494:pharmacology
chemical_list: D007476:Ionophores; D009570:Nitriles; D011522:Protons; D014634:Valinomycin; C001784:SF 6847; D000251:Adenosine Triphosphatases; D006180:Proton-Translocating ATPases