A complete thermodynamic analysis of enzyme turnover links the free energy landscape to enzyme catalysis

Hannah B.L. Jones, Stephen A. Wells, Erica J. Prentice, Anthony Kwok, Liyin L. Liang, Vickery L. Arcus, Christopher R. Pudney

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Our understanding of how enzymes work is colored by static structure depictions where the enzyme scaffold is presented as either immobile, or in equilibrium between well-defined static conformations. Proteins however exhibit a large degree of motion over a broad range of timescales and magnitudes and this is defined thermodynamically by the enzyme free energy landscape (FEL). The role and importance of enzyme motion is extremely contentious. Much of the challenge is in the experimental detection of so called 'conformational sampling' involved in enzyme turnover. Herein we apply combined pressure and temperature kinetics studies to elucidate the full suite of thermodynamic parameters defining an enzyme FEL as it relates to enzyme turnover. We find that the key thermodynamic parameters governing vibrational modes related to enzyme turnover are the isobaric expansivity term and the change in heat capacity for enzyme catalysis. Variation in the enzyme FEL affects these terms. Our analysis is supported by a range of biophysical and computational approaches that specifically capture information on protein vibrational modes and the FEL (all atom flexibility calculations, red edge excitation shift spectroscopy and viscosity studies) that provide independent evidence for our findings. Our data suggest that restricting the enzyme FEL may be a powerful strategy when attempting to rationally engineer enzymes, particularly to alter thermal activity. Moreover, we demonstrate how rational predictions can be made with a rapid computational approach. This article is protected by copyright. All rights reserved.

Original languageEnglish
Pages (from-to)2829-2842
Number of pages14
JournalFEBS Journal
Volume284
Issue number17
Early online date26 Jun 2017
DOIs
Publication statusPublished - 1 Sep 2017

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Catalysis
Thermodynamics
Enzymes
Hot Temperature
Viscosity
Spectrum Analysis
Proteins
Pressure
Temperature

Keywords

  • enzyme
  • expansivity
  • free energy landscape
  • heat capacity
  • pressure

Cite this

A complete thermodynamic analysis of enzyme turnover links the free energy landscape to enzyme catalysis. / Jones, Hannah B.L.; Wells, Stephen A.; Prentice, Erica J.; Kwok, Anthony; Liang, Liyin L.; Arcus, Vickery L.; Pudney, Christopher R.

In: FEBS Journal, Vol. 284, No. 17, 01.09.2017, p. 2829-2842.

Research output: Contribution to journalArticle

Jones, Hannah B.L. ; Wells, Stephen A. ; Prentice, Erica J. ; Kwok, Anthony ; Liang, Liyin L. ; Arcus, Vickery L. ; Pudney, Christopher R. / A complete thermodynamic analysis of enzyme turnover links the free energy landscape to enzyme catalysis. In: FEBS Journal. 2017 ; Vol. 284, No. 17. pp. 2829-2842.
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