title: Practical steady-state enzyme kinetics.
abstract: Enzymes are key components of most biological processes. Characterization of enzymes is therefore frequently required during the study of biological systems. Steady-state kinetics provides a simple and rapid means of assessing the substrate specificity of an enzyme. When combined with site-directed mutagenesis (see Site-Directed Mutagenesis), it can be used to probe the roles of particular amino acids in the enzyme in substrate recognition and catalysis. Effects of interaction partners and posttranslational modifications can also be assessed using steady-state kinetics. This overview explains the general principles of steady-state enzyme kinetics experiments in a practical, rather than theoretical, way. Any biochemistry textbook will have a section on the theory of Michaelis-Menten kinetics, including derivations of the relevant equations. No specific enzymatic assay is described here, although a method for monitoring product formation or substrate consumption over time (an assay) is required to perform the experiments described.
authors.affiliation: Department of Biophysics and Biophysical Chemistry, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Electronic address: jlorsch@jhmi.edu.
authors.forename: Jon R
authors.initials: JR
authors.lastname: Lorsch
mesh_terms: D000465:Algorithms; D055162:Biocatalysis; D057075:Enzyme Assays; D004791:Enzyme Inhibitors / Q000737:chemistry; D004798:Enzymes / Q000737:chemistry*; D007700:Kinetics; D008956:Models, Chemical; D013379:Substrate Specificity
chemical_list: D004791:Enzyme Inhibitors; D004798:Enzymes
keywords: Enzyme-catalyzed reaction; Kinetic parameters determination; Michaelis-Menten equation; Substrate concentration