Abstract
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.
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Demarse, N.A., Killian, M.C., Hansen, L.D., Quinn, C.F. (2013). Determining Enzyme Kinetics via Isothermal Titration Calorimetry. In: Samuelson, J. (eds) Enzyme Engineering. Methods in Molecular Biology, vol 978. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-293-3_2
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DOI: https://doi.org/10.1007/978-1-62703-293-3_2
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-292-6
Online ISBN: 978-1-62703-293-3
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