Monomer topology defines folding speed of heptamer

Protein Sci. 2004 May;13(5):1317-21. doi: 10.1110/ps.03559504. Epub 2004 Apr 9.

Abstract

Small monomeric proteins often fold in apparent two-state processes with folding speeds dictated by their native-state topology. Here we test, for the first time, the influence of monomer topology on the folding speed of an oligomeric protein: the heptameric cochaperonin protein 10 (cpn10), which in the native state has seven beta-barrel subunits noncovalently assembled through beta-strand pairing. Cpn10 is a particularly useful model because equilibrium-unfolding experiments have revealed that the denatured state in urea is that of a nonnative heptamer. Surprisingly, refolding of the nonnative cpn10 heptamer is a simple two-state kinetic process with a folding-rate constant in water (2.1 sec(-1); pH 7.0, 20 degrees C) that is in excellent agreement with the prediction based on the native-state topology of the cpn10 monomer. Thus, the monomers appear to fold as independent units, with a speed that correlates with topology, although the C and N termini are trapped in beta-strand pairing with neighboring subunits. In contrast, refolding of unfolded cpn10 monomers is dominated by a slow association step.

Publication types

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

MeSH terms

  • Chaperonin 10 / analysis
  • Chaperonin 10 / chemistry*
  • Fluorescence
  • Guanidine / chemistry
  • Humans
  • Kinetics
  • Protein Folding
  • Urea / chemistry

Substances

  • Chaperonin 10
  • Urea
  • Guanidine