Ultrarapid mixing experiments reveal that Im7 folds via an on-pathway intermediate

Nat Struct Biol. 2001 Jan;8(1):68-72. doi: 10.1038/83074.

Abstract

Many proteins populate partially organized structures during folding. Since these intermediates often accumulate within the dead time (2-5 ms) of conventional stopped-flow and quench-flow devices, it has been difficult to determine their role in the formation of the native state. Here we use a microcapillary mixing apparatus, with a time resolution of approximately 150 micros, to directly follow the formation of an intermediate in the folding of a four-helix protein, Im7. Quantitative kinetic modeling of folding and unfolding data acquired over a wide range of urea concentrations demonstrate that this intermediate ensemble lies on a direct path from the unfolded to the native state.

Publication types

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

MeSH terms

  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / metabolism*
  • Colicins*
  • Kinetics
  • Models, Chemical
  • Protein Denaturation / drug effects
  • Protein Folding*
  • Protein Renaturation
  • Protein Structure, Secondary
  • Spectrometry, Fluorescence
  • Thermodynamics
  • Urea / pharmacology

Substances

  • Bacterial Proteins
  • Colicins
  • colicin immunity proteins
  • Urea