The structure of the major transition state for folding of an FF domain from experiment and simulation

J Mol Biol. 2005 Jul 8;350(2):363-78. doi: 10.1016/j.jmb.2005.04.067.

Abstract

We have analysed the transition state of folding of the four-helix FF domain from HYPA/FBP11 by high-resolution experiment and simulation as part of a continuing effort to understand the principles of folding and the refinement of predictive methods. The major transition state for folding was subjected to a Phi-value analysis utilising 50 mutants. The transition state contained a nucleus for folding centred around the end of helix 1 (H1) and the beginning of helix 2 (H2). Secondary structure in this region was fully formed (PhiF=0.9-1) and tertiary interactions were well developed. Interactions in the distal part of the native structure were weak (PhiF=0-0.2). The hydrophobic core and other parts of the protein displayed intermediate Phi-values, suggesting that interactions coalesce as the end of H1 and beginning of H2 are in the process of being formed. The distribution of Phi-values resembled that of barnase, which folds via an intermediate, rather than that of CI2 which folds by a concerted nucleation-condensation mechanism. The overall picture of the transition state structure identified in molecular dynamics simulations is in qualitative agreement, with the turn connecting H1 and H2 being formed, a loosened core, and H4 partially unfolded and detached from the core. There are some differences in the details and interpretation of specific Phi-values.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Carrier Proteins / chemistry
  • Carrier Proteins / genetics
  • Carrier Proteins / metabolism
  • Computer Simulation*
  • Glycine / genetics
  • Glycine / metabolism
  • Humans
  • Kinetics
  • Models, Molecular
  • Mutation / genetics
  • Protein Denaturation
  • Protein Engineering
  • Protein Folding*
  • Protein Renaturation
  • Protein Structure, Secondary
  • Protein Structure, Tertiary*
  • Thermodynamics

Substances

  • Carrier Proteins
  • PRPF40A protein, human
  • Glycine