Hydration of the folding transition state ensemble of a protein

Ludovic Brun, Daniel Isom, Priya Velu, Bertrand García-Moreno, Catherine Ann Royer

Research output: Contribution to journalArticle

37 Citations (Scopus)

Abstract

A complete description of the mechanisms of protein folding requires knowledge of the structural and physical character of the folding transition state ensembles (TSEs). A key question concerning the role of hydration of the hydrophobic core in determining folding mechanisms remains. To address this, we probed the state of hydration of the TSE of staphylococcal nuclease (SNase) by examining the fluorescence-detected pressure-jump relaxation behavior of six SNase variants in which a residue in the hydrophobic core, Val-66, was replaced with polar or ionizable residues (Lys, Arg, His, Asp, Glu, and Asn). Because of a large positive activation volume for folding, the major effect of pressure on the wild-type protein is to decrease the folding rate. By the time wild-type SNase reaches the folding transition state, most water has already been expelled from its hydrophobic core. In contrast, the major effect of pressure on the variant proteins is an increase in the unfolding rate due to a large negative activation volume for unfolding. This results from a significant increase in the level of hydration of the TSE when an internal ionizable group is present. These data confirm that the role of water in the folding reaction can differ from protein to protein and that even a single substitution in a critical position can modulate significantly the properties of the TSE.

Original languageEnglish (US)
Pages (from-to)3473-3480
Number of pages8
JournalBiochemistry
Volume45
Issue number11
DOIs
StatePublished - Mar 21 2006
Externally publishedYes

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Micrococcal Nuclease
Hydration
Pressure
Proteins
Chemical activation
Protein folding
Water
Protein Folding
Substitution reactions
Fluorescence

ASJC Scopus subject areas

  • Biochemistry

Cite this

Brun, L., Isom, D., Velu, P., García-Moreno, B., & Royer, C. A. (2006). Hydration of the folding transition state ensemble of a protein. Biochemistry, 45(11), 3473-3480. https://doi.org/10.1021/bi052638z

Hydration of the folding transition state ensemble of a protein. / Brun, Ludovic; Isom, Daniel; Velu, Priya; García-Moreno, Bertrand; Royer, Catherine Ann.

In: Biochemistry, Vol. 45, No. 11, 21.03.2006, p. 3473-3480.

Research output: Contribution to journalArticle

Brun, L, Isom, D, Velu, P, García-Moreno, B & Royer, CA 2006, 'Hydration of the folding transition state ensemble of a protein', Biochemistry, vol. 45, no. 11, pp. 3473-3480. https://doi.org/10.1021/bi052638z
Brun L, Isom D, Velu P, García-Moreno B, Royer CA. Hydration of the folding transition state ensemble of a protein. Biochemistry. 2006 Mar 21;45(11):3473-3480. https://doi.org/10.1021/bi052638z
Brun, Ludovic ; Isom, Daniel ; Velu, Priya ; García-Moreno, Bertrand ; Royer, Catherine Ann. / Hydration of the folding transition state ensemble of a protein. In: Biochemistry. 2006 ; Vol. 45, No. 11. pp. 3473-3480.
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