Coil-globule transition in the denatured state of a small protein

Eilon Sherman, Gilad Haran*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

264 Scopus citations

Abstract

Upon transfer from strongly denaturing to native conditions, proteins undergo a collapse that either precedes folding or occurs simultaneously with it. This collapse is similar to the well known coil-globule transition of polymers. Here we employ single-molecule fluorescence methods to fully characterize the equilibrium coil-globule transition in the denatured state of the IgG-binding domain of protein L. By using FRET measurements on freely diffusing individual molecules, we determine the radius of gyration of the protein, which shows a gradual expansion as the concentration of the denaturant, guanidinium hydrochloride, is increased all the way up to 7 M. This expansion is observed also in fluorescence correlation spectroscopy measurements of the hydrodynamic radius of the protein. We analyze the radius of gyration measurements using the theory of the coil-globule transition of Sanchez [Sanchez, I. C. (1979) Macromolecules 12, 980-988], which balances the excluded volume entropy of the chain with the average interresidue interaction energy. In particular, we calculate the solvation energy of the denatured protein, a property that is not readily accessible in other experiments. The dependence of this energy on denaturant concentration is nonlinear, contrasting with the common linear extrapolation method used to describe denaturation energy. Interestingly, a fit to the binding model of chemical denaturation suggests a single denaturant binding site per protein residue. The size of the denatured protein under native conditions can be extrapolated from the data as well, showing that the fully collapsed state of protein is only ≈10% larger than the folded state.

Original languageAmerican English
Pages (from-to)11539-11543
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume103
Issue number31
DOIs
StatePublished - 1 Aug 2006
Externally publishedYes

Keywords

  • Fluorescence correlation spectroscopy
  • Protein folding
  • Single-molecule fluorescence

Fingerprint

Dive into the research topics of 'Coil-globule transition in the denatured state of a small protein'. Together they form a unique fingerprint.

Cite this