A dynamic knockout reveals that conformational fluctuations influence the chemical step of enzyme catalysis

Gira Bhabha, Jeeyeon Lee, Damian C. Ekiert, Jongsik Gam, Ian A. Wilson, H. Jane Dyson, Stephen J. Benkovic, Peter E. Wright

Research output: Contribution to journalArticle

300 Citations (Scopus)

Abstract

Conformational dynamics play a key role in enzyme catalysis. Although protein motions have clear implications for ligand flux, a role for dynamics in the chemical step of enzyme catalysis has not been clearly established. We generated a mutant of Escherichia coli dihydrofolate reductase that abrogates millisecond-time-scale fluctuations in the enzyme active site without perturbing its structural and electrostatic preorganization. This dynamic knockout severely impairs hydride transfer. Thus, we have found a link between conformational fluctuations on the millisecond time scale and the chemical step of an enzymatic reaction, with broad implications for our understanding of enzyme mechanisms and for design of novel protein catalysts.

Original languageEnglish (US)
Pages (from-to)234-238
Number of pages5
JournalScience
Volume332
Issue number6026
DOIs
StatePublished - Apr 8 2011

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Catalysis
Enzymes
Tetrahydrofolate Dehydrogenase
Static Electricity
Catalytic Domain
Proteins
Escherichia coli
Ligands

All Science Journal Classification (ASJC) codes

  • General

Cite this

Bhabha, G., Lee, J., Ekiert, D. C., Gam, J., Wilson, I. A., Dyson, H. J., ... Wright, P. E. (2011). A dynamic knockout reveals that conformational fluctuations influence the chemical step of enzyme catalysis. Science, 332(6026), 234-238. https://doi.org/10.1126/science.1198542
Bhabha, Gira ; Lee, Jeeyeon ; Ekiert, Damian C. ; Gam, Jongsik ; Wilson, Ian A. ; Dyson, H. Jane ; Benkovic, Stephen J. ; Wright, Peter E. / A dynamic knockout reveals that conformational fluctuations influence the chemical step of enzyme catalysis. In: Science. 2011 ; Vol. 332, No. 6026. pp. 234-238.
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Bhabha, G, Lee, J, Ekiert, DC, Gam, J, Wilson, IA, Dyson, HJ, Benkovic, SJ & Wright, PE 2011, 'A dynamic knockout reveals that conformational fluctuations influence the chemical step of enzyme catalysis', Science, vol. 332, no. 6026, pp. 234-238. https://doi.org/10.1126/science.1198542

A dynamic knockout reveals that conformational fluctuations influence the chemical step of enzyme catalysis. / Bhabha, Gira; Lee, Jeeyeon; Ekiert, Damian C.; Gam, Jongsik; Wilson, Ian A.; Dyson, H. Jane; Benkovic, Stephen J.; Wright, Peter E.

In: Science, Vol. 332, No. 6026, 08.04.2011, p. 234-238.

Research output: Contribution to journalArticle

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