Abstract
The rate of inactivation of F1-ATPase, isolated from beef heart mitochondria, by the active acidic form of the natural inhibitor protein depends on the ATP concentration. An increase in concentration of ATP to ∼ 20 μM leads to a decrease in that of the inhibitor protein inducing 50% inhibition of the F1-ATPase during 5 s preincubation (C 50); further increase in ATP concentration to 1 mM causes little, if any, change in C 50. However, the C 50 values show a rise at ATP concentrations higher than 1 mM. This ATP dependence of the inhibitor action may be in agreement with a version of the alternating-site binding-change mechanism, which assumes that the two-site catalytic cycle intermediates possessing (i) the products (ADP + Pi) bound in the low-affinity state at one of the active sites and (ii) an ATP molecule at the other active site are the targets for the acidic form of the inhibitor protein.

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