Precautions when determining kinetically the order of inactivation of enzymes by functionally irreversible inhibitors.

The number of molecules of an irreversible inhibitor that are responsible for inactivation of a catalytic site is often deduced from the slope of a plot of the log of the apparent rate of inactivation (k') at different concentrations of inhibitor versus the log of the inhibitor concentrations. The purpose of this note is to urge caution in experimental design and interpretation if one attempts to utilize this kinetic technique to characterize the order of inactivation brought about by functionally irreversible inhibitors that initially bind reversibly to an enzyme in the process of inactivation. Representative literature cases which have utilized plots of log k' versus log [I] for this type of inactivation are discussed.