Practical steady-state enzyme kinetics

Jon R. Lorsch

Research output: Chapter in Book/Report/Conference proceedingChapter

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.

Original languageEnglish (US)
Title of host publicationLaboratory Methods in Enzymology
Subtitle of host publicationProtein Part A
PublisherAcademic Press Inc.
Pages3-15
Number of pages13
ISBN (Print)9780124200708
DOIs
StatePublished - Jan 1 2014

Publication series

NameMethods in Enzymology
Volume536
ISSN (Print)0076-6879
ISSN (Electronic)1557-7988

Keywords

  • Enzyme-catalyzed reaction
  • Kinetic parameters determination
  • Michaelis-Menten equation
  • Substrate concentration

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Fingerprint Dive into the research topics of 'Practical steady-state enzyme kinetics'. Together they form a unique fingerprint.

  • Cite this

    Lorsch, J. R. (2014). Practical steady-state enzyme kinetics. In Laboratory Methods in Enzymology: Protein Part A (pp. 3-15). (Methods in Enzymology; Vol. 536). Academic Press Inc.. https://doi.org/10.1016/B978-0-12-420070-8.00001-5