Common enzymological experiments allow free energy profile determination.

ABSTRACT: The determination of a complete set of rate constants [free energy profiles (FEPs)] for a complex kinetic mechanism is challenging. Enzymologists have devised a variety of informative steady-state kinetic experiments (e.g., Michaelis-Menten kinetics, viscosity dependence of kinetic parameters, kinetic isotope effects, etc.) that each provide distinct information regarding a particular kinetic system. A simple method for combining steady-state experiments in a single analysis is presented here, which allows microscopic rate constants and intrinsic kinetic isotope effects to be determined. It is first shown that Michaelis-Menten kinetic parameters (kcat and Km values), kinetic isotope efffets, solvent viscosity effects, and intermediate partitioning measurements are sufficient to define the

SUBMITTER: Toney MD 

PROVIDER: S-EPMC4286415 | biostudies-literature | 2013 Aug

REPOSITORIES: biostudies-literature