Utilization of integrated Michaelis-Menten equation to determine kinetic constants

Abstract

Students of biochemistry and related biosciences are urged to solve problems where kinetic parameters are calculated from initial rates obtained at different substrate concentrations. Troubles begin when they go to the laboratory to perform kinetic experiments and realize that usual laboratory instruments do not measure initial rates but only substrate or product concentrations as a function of reaction time. To overcome this problem we present a methodology which uses the integrated form of Michaelis-Menten equation. The method presented has a theoretical and pedagogic basis which is not as arbitrary as other approaches. Here we present and describe the methodology for analyzing time course data together with some examples of the essential computer procedures to implement these analyses. To simplify the understanding of this methodology the experimental examples are confined to linear inhibitions and experimental points utilized are the same from which the initial rates are determined. © 2007 by The International Union of Biochemistry and Molecular Biology.