A lysozyme folding intermediate revealed by solution X-ray scattering

J Mol Biol. 1996 Sep 6;261(5):658-71. doi: 10.1006/jmbi.1996.0491.

Abstract

Equilibrium unfolding of hen egg lysozyme as a function of urea concentration at pH 2.9 has been studied by solution X-ray scattering. Differences in the unfolding transition are observed as monitored by the radius of gyration Rg, and by far and near UV CD (circular dichroism) at 222 nm and 298 nm, respectively. This suggests the existence of a third unfolding species, in addition to the native and the unfolded states. A singular value decomposition (SVD) analysis was made of the scattering curves at different urea concentrations. This analysis shows clear evidence of a third basis component in the X-ray scattering curves, thus supporting the results of the Rg and CD measurements. The denaturant binding model was employed to estimate the thermodynamic parameters of denaturation for the intermediate and unfolded states. Use of these parameters to refine the SVD analysis allows us to reconstruct a scattering profile for the pure intermediate state. Simplified partially folded models, based on the crystal structure of hen lysozyme, support a working model for the intermediate, whose structure may be correlated with that of the kinetic intermediate found in the refolding pathway studied by Dobson and coworkers.

Publication types

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

MeSH terms

  • Circular Dichroism
  • Hydrogen-Ion Concentration
  • Mathematical Computing
  • Models, Molecular
  • Muramidase / chemistry*
  • Protein Conformation
  • Protein Denaturation
  • Protein Folding*
  • Scattering, Radiation
  • Solutions
  • Ultraviolet Rays
  • X-Rays

Substances

  • Solutions
  • Muramidase