Hostname: page-component-78c5997874-s2hrs Total loading time: 0 Render date: 2024-11-11T06:55:18.931Z Has data issue: false hasContentIssue false
  • English
  • Français

The effects of disulfide bonds on the denatured state of barnase

Published online by Cambridge University Press:  10 February 2001

JANE CLARKE ,
ANDREA M. HOUNSLOW ,
CHRIS J. BOND ,
ALAN R. FERSHT  and
VALERIE DAGGETT
JANE CLARKE
Affiliation:
Centre for Protein Engineering, University Chemical Laboratories, Lensfield Rd., Cambridge CB2 1EW, United Kingdom
ANDREA M. HOUNSLOW
Affiliation:
Centre for Protein Engineering, University Chemical Laboratories, Lensfield Rd., Cambridge CB2 1EW, United Kingdom Current address: Department of Molecular Biology and Biotechnology, University of Sheffield, Sheffield S10 2TN, United Kingdom.
CHRIS J. BOND
Affiliation:
Biomolecular Structure and Design Program, University of Washington, Seattle, Washington 98195-7610
ALAN R. FERSHT
Affiliation:
Centre for Protein Engineering, University Chemical Laboratories, Lensfield Rd., Cambridge CB2 1EW, United Kingdom
VALERIE DAGGETT
Affiliation:
Department of Medicinal Chemistry, University of Washington, Seattle, Washington 98195-7610 Biomolecular Structure and Design Program, University of Washington, Seattle, Washington 98195-7610
Get access

Abstract

The effects of engineered disulfide bonds on protein stability are poorly understood because they can influence the structure, dynamics, and energetics of both the native and denatured states. To explore the effects of two engineered disulfide bonds on the stability of barnase, we have conducted a combined molecular dynamics and NMR study of the denatured state of the two mutants. As expected, the disulfide bonds constrain the denatured state. However, specific extended β-sheet structure can also be detected in one of the mutant proteins. This mutant is also more stable than would be predicted. Our study suggests a possible cause of the very high stability conferred by this disulfide bond: the wild-type denatured ensemble is stabilized by a nonnative hydrophobic cluster, which is constrained from occurring in the mutant due to the formation of secondary structure.

Keywords

barnasedenatured statesdisulfide bondsmolecular dynamicsNMR
Type
Research Article
Information
Protein Science , Volume 9 , Issue 12 , December 2000 , pp. 2394 - 2404
Copyright
© 2000 The Protein Society

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)