Methods to measure the kinetics of protease inhibition by serpins

Anita J Horvath, Bernadine GC Lu, Robert N Pike, Stephen Paul Bottomley

Research output: Chapter in Book/Report/Conference proceedingChapter (Book)Researchpeer-review

Abstract

The serpin molecule has evolved an unusual mechanism of inhibition, involving an exposed reactive center loop (RCL) and conformational change to covalently trap a target protease. Successful inhibition of the protease is dependent on the rate of serpin-protease association and the efficiency with which the RCL inserts into beta-sheet A, translocating the covalently bound protease and thereby completing the inhibition process. This chapter describes the kinetic methods used for determining the rate of protease inhibition (k(a)) and the stoichiometry of inhibition. These kinetic variables provide a means to examine different serpin-protease pairings, assess the effects of mutations within a serpin on protease inhibition, and determine the physiologically cognate protease of a serpin.
Original languageEnglish
Title of host publicationMethods in Enzymology, Volume 501: Serpin Structure and Evolution
EditorsJames C Whisstock, Phillip I Bird
Place of PublicationUSA
PublisherAcademic Press
Pages223 - 235
Number of pages13
ISBN (Print)9780123859501
DOIs
Publication statusPublished - 2011

Cite this

Horvath, A. J., Lu, B. GC., Pike, R. N., & Bottomley, S. P. (2011). Methods to measure the kinetics of protease inhibition by serpins. In J. C. Whisstock, & P. I. Bird (Eds.), Methods in Enzymology, Volume 501: Serpin Structure and Evolution (pp. 223 - 235). USA: Academic Press. https://doi.org/10.1016/B978-0-12-385950-1.00011-0
Horvath, Anita J ; Lu, Bernadine GC ; Pike, Robert N ; Bottomley, Stephen Paul. / Methods to measure the kinetics of protease inhibition by serpins. Methods in Enzymology, Volume 501: Serpin Structure and Evolution. editor / James C Whisstock ; Phillip I Bird. USA : Academic Press, 2011. pp. 223 - 235
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Horvath, AJ, Lu, BGC, Pike, RN & Bottomley, SP 2011, Methods to measure the kinetics of protease inhibition by serpins. in JC Whisstock & PI Bird (eds), Methods in Enzymology, Volume 501: Serpin Structure and Evolution. Academic Press, USA, pp. 223 - 235. https://doi.org/10.1016/B978-0-12-385950-1.00011-0

Methods to measure the kinetics of protease inhibition by serpins. / Horvath, Anita J; Lu, Bernadine GC; Pike, Robert N; Bottomley, Stephen Paul.

Methods in Enzymology, Volume 501: Serpin Structure and Evolution. ed. / James C Whisstock; Phillip I Bird. USA : Academic Press, 2011. p. 223 - 235.

Research output: Chapter in Book/Report/Conference proceedingChapter (Book)Researchpeer-review

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Horvath AJ, Lu BGC, Pike RN, Bottomley SP. Methods to measure the kinetics of protease inhibition by serpins. In Whisstock JC, Bird PI, editors, Methods in Enzymology, Volume 501: Serpin Structure and Evolution. USA: Academic Press. 2011. p. 223 - 235 https://doi.org/10.1016/B978-0-12-385950-1.00011-0