Rapid Measurement of Inhibitor Binding Kinetics by Isothermal Titration Calorimetry

Overview

Journal Nat Commun

Specialty Biology

Date 2018 Mar 3

PMID 29497037

Citations 15

Authors

Justin M Di Trani

Stephane De Cesco

Rebecca OLeary

Jessica Plescia

Claudia Jorge do Nascimento

Nicolas Moitessier

Anthony K Mittermaier

Affiliations

Soon will be listed here.

Abstract

Although drug development typically focuses on binding thermodynamics, recent studies suggest that kinetic properties can strongly impact a drug candidate's efficacy. Robust techniques for measuring inhibitor association and dissociation rates are therefore essential. To address this need, we have developed a pair of complementary isothermal titration calorimetry (ITC) techniques for measuring the kinetics of enzyme inhibition. The advantages of ITC over standard techniques include speed, generality, and versatility; ITC also measures the rate of catalysis directly, making it ideal for quantifying rapid, inhibitor-dependent changes in enzyme activity. We used our methods to study the reversible covalent and non-covalent inhibitors of prolyl oligopeptidase (POP). We extracted kinetics spanning three orders of magnitude, including those too rapid for standard methods, and measured sub-nM binding affinities below the typical ITC limit. These results shed light on the inhibition of POP and demonstrate the general utility of ITC-based enzyme inhibition kinetic measurements.

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