A Comparison Between the Initial Rate Expressions Obtained Under Strict Conditions and the Rapid Equilibrium Assumption Using, as Example, a Four Substrate Enzyme Reaction

Abstract

The software WinStes, developed by our group, is used to derive the strict steadystate initial rate equation of the reaction mechanism of CTP:sn-glycerol-3-phosphate cytidylyltransferase [EC 2.7.7.39] from Bacillus subtilis. This enzyme catalyzes a reaction with two substrates and operates by a random ordered binding mechanism with two molecules of each substrate. The accuracy of the steady-state rate equation derived is checked by comparing the rate values it provides with those obtained from the simulated progress curves. To analyze the kinetics of this enzyme using the strict steady-state initial rate equation, several curves for different substrate concentrations and different rate constants are generated. A comparison of these curves with the curves obtained from the rapid equilibrium initial rate equation, with different substrate concentration values, serves to analyze how the strict steady-state rate equation values are closer to those of rapid equilibrium rate equations when rapid equilibrium conditions are fulfilled.