Disulfide bonds and protein folding

Biochemistry. 2000 Apr 18;39(15):4207-16. doi: 10.1021/bi992922o.

Abstract

The applications of disulfide-bond chemistry to studies of protein folding, structure, and stability are reviewed and illustrated with bovine pancreatic ribonuclease A (RNase A). After surveying the general properties and advantages of disulfide-bond studies, we illustrate the mechanism of reductive unfolding with RNase A, and discuss its application to probing structural fluctuations in folded proteins. The oxidative folding of RNase A is then described, focusing on the role of structure formation in the regeneration of the native disulfide bonds. The development of structure and conformational order in the disulfide intermediates during oxidative folding is characterized. Partially folded disulfide species are not observed, indicating that disulfide-coupled folding is highly cooperative. Contrary to the predictions of "rugged funnel" models of protein folding, misfolded disulfide species are also not observed despite the potentially stabilizing effect of many nonnative disulfide bonds. The mechanism of regenerating the native disulfide bonds suggests an analogous scenario for conformational folding. Finally, engineered covalent cross-links may be used to assay for the association of protein segments in the folding transition state, as illustrated with RNase A.

Publication types

  • Research Support, U.S. Gov't, P.H.S.
  • Review

MeSH terms

  • Animals
  • Cattle
  • Disulfides / chemistry*
  • Disulfides / metabolism*
  • Enzyme Stability
  • Models, Molecular
  • Oxidation-Reduction
  • Protein Conformation
  • Protein Denaturation
  • Protein Engineering
  • Protein Folding*
  • Protein Renaturation
  • Ribonuclease, Pancreatic / chemistry*
  • Ribonuclease, Pancreatic / metabolism*

Substances

  • Disulfides
  • Ribonuclease, Pancreatic