Uses and misuses of progress curve analysis in enzyme kinetics

Abstract

AbstractProgress curve analysis is a convenient tool for the characterization of enzyme action: a single reaction mixture provides multiple experimental measured points for continuously varying amounts of substrates and products with exactly the same enzyme and modulator concentrations. The determination of kinetic parameters from the progress curves, however, requires complex mathematical evaluation of the time-course data. Some freely available programs (e.g. FITSIM, DYNAFIT) are widely applied to fit kinetic parameters to user-defined enzymatic mechanisms, but users often overlook the stringent requirements of the analytic procedures for appropriate design of the input experiments. Flaws in the experimental setup result in unreliable parameters with consequent misinterpretation of the biological phenomenon under study. The present commentary suggests some helpful mathematical tools to improve the analytic procedure in order to diagnose major errors in concept and design of kinetic experiments.