Enzyme Kinetics Determined by Single-Injection Isothermal Titration Calorimetry

Colette F Quinn; Lee D Hansen

doi:10.1007/978-1-4939-9179-2_17

Enzyme Kinetics Determined by Single-Injection Isothermal Titration Calorimetry

Methods Mol Biol. 2019:1964:241-249. doi: 10.1007/978-1-4939-9179-2_17.

Authors

Colette F Quinn¹, Lee D Hansen²

Affiliations

¹ Applications Lab, TA Instruments, Lindon, UT, USA. cquinn@tainstruments.com.
² Department of Chemistry and Biochemistry, Brigham Young University, Provo, UT, USA.

PMID: 30929247
DOI: 10.1007/978-1-4939-9179-2_17

Abstract

This chapter describes how to collect Michaelis-Menten kinetic data on an enzyme-catalyzed reaction with the isothermal titration calorimetry (ITC) and the single-injection method. ITC measures the heat rate which is directly proportional to the reaction rate. The enthalpy change (Δ_rH), K_m, k_cat, and v_max are determined in a single assay that does not require labeling or immobilization.

Keywords: Calorimetry; Enthalpy; Enzyme; ITC; Kinetics; Michaelis-Menten; Single-injection method.

MeSH terms

Calorimetry / methods*
Catalysis*
Enzymes / chemistry*
Hot Temperature
Kinetics
Thermodynamics

Substances

Enzymes