TY - CHAP
T1 - Practical steady-state enzyme kinetics
AU - Lorsch, Jon R.
PY - 2014
Y1 - 2014
N2 - 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.
AB - 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.
KW - Enzyme-catalyzed reaction
KW - Kinetic parameters determination
KW - Michaelis-Menten equation
KW - Substrate concentration
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U2 - 10.1016/B978-0-12-420070-8.00001-5
DO - 10.1016/B978-0-12-420070-8.00001-5
M3 - Chapter
C2 - 24423262
AN - SCOPUS:84892418797
SN - 9780124200708
T3 - Methods in Enzymology
SP - 3
EP - 15
BT - Laboratory Methods in Enzymology
PB - Academic Press Inc.
ER -