The Transient‐State Kinetics of Two‐Substrate Enzyme Systems Operating by an Ordered Ternary‐Complex Mechanism

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Abstract
Transient‐state kinetic equations, applicable to single‐turnover experiments with limiting amounts of substrate and catalytic‐suicide experiments with substrates in excess to enzyme, have been derived for two‐substrate enzyme reactions occurring by an ordered ternary‐complex mechanism. The relative magnitude and dependence on substrate concentrations of rates and amplitudes for the exponential transients corresponding to this mechanism are quantitatively defined and related to steady‐state kinetic parameters for the reaction mechanism. It is shown that ordered ternary‐complex reactions under standard assay conditions may exhibit a biphasic transient rate behaviour, and the precise conditions under which two transients can be detected are described and discussed.