Ubiquitin folds through a highly polarized transition state

Protein Eng Des Sel. 2005 May;18(5):229-37. doi: 10.1093/protein/gzi025. Epub 2005 Apr 27.

Abstract

The small alpha/beta protein ubiquitin has been used as a model system for experimental and computational studies on protein folding for many years. Here, we present a comprehensive phi-value analysis and characterize the structure and energetics of the transition state ensemble (TSE). Twenty-seven non-disruptive mutations are made throughout the structure and a range of phi-values from zero to one are observed. The values cluster such that medium and high values and found only in the N-terminal region of the protein, whilst the C-terminal region has consistently low phi-values. In the TSE, the main alpha-helix appears to be fully formed (two phi-values which specifically probe helical structure are one) and the helix is stabilized by packing against the first beta-turn, which is partially structured. In striking comparison, the phi-values in the C-terminal region are all very low, suggesting that this region of the protein is largely unstructured in the TSE. Data are consistent with a nucleation-condensation mechanism in which there is a highly polarized folding nucleus comprising the first beta-hairpin and the alpha-helix. Data presented from the protein engineering study and phi-value analysis are compared with results from other experimental studies and also computational studies.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Kinetics
  • Models, Molecular*
  • Mutagenesis
  • Point Mutation
  • Protein Engineering*
  • Protein Folding*
  • Protein Structure, Secondary
  • Thermodynamics
  • Ubiquitin / chemistry*
  • Ubiquitin / genetics
  • Ubiquitin / metabolism

Substances

  • Ubiquitin