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• W2085751015 abstract "Equations are derived for predicting the effects of substrate concentration on the inactivation rate constants of tight-binding competitive and uncompetitive inhibitors. These relationships are used to study the inhibition of mitochondrial oxidative phosphorylation by efrapeptin. The results show that the apparent rate constant for efrapeptin inactivation of ATP synthesis decreases with increase in phosphate concentration. The reciprocal of the observed rate constant varies linearly with changes in the level of phosphate as predicted for a competitive inhibitor. The concentrations of ADP during ATP synthesis and of ATP during ATP hydrolysis, on the other hand, have no effect on the rate of inactivation by efrapeptin. This is in contrast to previous observations that adenine nucleotide substrates influence the level of efrapeptin bound at equilibrium (R. L. Cross, and W. E. Kohlbrenner, 1978, J. Biol. Chem.253, 4865–4873). The results suggest that efrapeptin interacts primarily at the phosphate binding site and that adenine nucleotides may influence equilibrium binding of efrapeptin by affecting the rate of dissociation of the inhibitor. Studies of efrapeptin inhibition of ATP synthesis under pseudo-first-order conditions show that the onset of inhibition is first order with respect to efrapeptin. The maximum apparent rate constant for efrapeptin binding, obtained by extrapolation to zero phosphate concentration, is 1.5 × 105m−1 s−1. Also described is a computer program for calculating the concentrations of complexes formed in a mixture of interacting species. The program may be used for most multiple-equilibrium calculations and permits the estimation of the levels of protonated complexes at any pH. The program was used to select Mg2+ concentrations which ensure that a large and relatively constant fraction of added ADP is present as MgADP. In the range of phosphate and ADP concentrations commonly used in studies of oxidative phosphorylation a 3 mm excess of Mg2+ relative to ADP was found sufficient to maintain high levels of MgADP at pH 8.0." @default.
• W2085751015 created "2016-06-24" @default.
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• W2085751015 date "1979-12-01" @default.
• W2085751015 modified "2023-09-26" @default.
• W2085751015 title "The mode of inhibition of oxidative phosphorylation by efrapeptin (A23871): Measurement of substrate effects on rates of inactivation by a tight-binding inhibitor" @default.
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• W2085751015 doi "https://doi.org/10.1016/0003-9861(79)90536-8" @default.
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