Rapid measurement of inhibitor binding kinetics by isothermal titration calorimetry

Nature Communications
Justin M Di TraniAnthony K Mittermaier

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.

References

May 15, 1991·The Biochemical Journal·S Kalwant, A G Porter
Aug 10, 1999·Journal of Molecular Recognition : JMR·R R Gabdoulline, R C Wade
Sep 14, 2001·Analytical Biochemistry·M J Todd, J Gomez
Dec 3, 2002·Journal of Medicinal Chemistry·Per-Olof MarkgrenU Helena Danielson
Jun 26, 2004·The Biochemical Journal·Jarkko I VenäläinenPekka T Männistö
Apr 13, 2005·Biochemistry·James G Robertson
Aug 5, 2006·Nature Reviews. Drug Discovery·Robert A CopelandThomas D Meek
Apr 5, 2007·Nature Protocols·Adrian Velazquez-Campoy, Ernesto Freire
Apr 17, 2008·Biochemistry·Peter J Tummino, Robert A Copeland
Jul 8, 2008·Acta Biochimica Et Biophysica Sinica·Yi Liang
Jan 12, 2010·Journal of Medicinal Chemistry·Janice LawandiNicolas Moitessier
Jul 29, 2010·Current Opinion in Chemical Biology·Hao Lu, Peter J Tonge
Apr 2, 2011·Nature Reviews. Drug Discovery·Juswinder SinghAdrian Whitty
May 1, 2013·Proceedings of the National Academy of Sciences of the United States of America·Elisabeth V SchneiderLars Neumann
Dec 19, 2013·Proceedings of the National Academy of Sciences of the United States of America·Phillip A SchwartzBrion William Murray
Nov 30, 2014·Analytical Biochemistry·Petr KuzmičBrion W Murray
Jan 11, 2017·Breast Cancer : the Journal of the Japanese Breast Cancer Society·Satoshi TanakaMinoru Sakaguchi
Jun 8, 2017·Analytical Chemistry·Justin M Di TraniAnthony K Mittermaier
Jun 24, 2017·ACS Chemical Neuroscience·Peter J Tonge
Jun 27, 2017·European Journal of Medicinal Chemistry·Stephane De CescoNicolas Moitessier

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Citations

Sep 14, 2018·PloS One·Trung Hai NguyenDavid D L Minh
Dec 5, 2019·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Wilfried RozhonBrigitte Poppenberger
Oct 16, 2018·Frontiers in Pharmacology·Isabella A GuedesLaurent E Dardenne
Oct 4, 2020·Nature Communications·Hongzhang DengXiaoyuan Chen
May 12, 2021·Journal of Molecular Recognition : JMR·Robert J FalconerAnthony K Mittermaier
Oct 2, 2019·ACS Central Science·Kin Sing Stephen LeeBruce D Hammock
Jun 22, 2018·Analytical Chemistry·Justin M Di TraniAnthony K Mittermaier
Dec 18, 2018·ACS Infectious Diseases·Gerardo Andrés Libreros-ZúñigaMarcio Vinicius Bertacine Dias
Aug 21, 2019·Analytical Chemistry·Yun WangAnthony K Mittermaier

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Methods Mentioned

BETA
isothermal titration calorimetry
NMR

Software Mentioned

MATLAB scripts

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