Determining Enzyme Kinetics via Isothermal Titration Calorimetry
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Isothermal titration calorimetry (ITC) has emerged as a powerful tool for determining the thermodynamic properties of chemical
or physical equilibria such as protein–protein, ligand–receptor, and protein–DNA binding interactions. The utility of ITC
for determining kinetic information, however, has not been fully recognized. Methods for collecting and analyzing data on
enzyme kinetics are discussed here. The step-by-step process of converting the raw heat output rate into the kinetic parameters
of the Michaelis–Menten equation is explicitly stated. The hydrolysis of sucrose by invertase is used to demonstrate the capability
of the instrument and method.
