Quantitative modelfree analysis of urea binding to unfolded ubiquitin using a combination of small angle X-ray and neutron scattering

J Am Chem Soc. 2009 Jul 1;131(25):8769-71. doi: 10.1021/ja9013248.

Abstract

Characterization of the conformational properties of denatured proteins is essential to our understanding of the molecular basis of protein folding and stability. Here we combine small angle neutron and X-ray scattering to study the interaction of urea with the protein ubiquitin. Comparing coherent intensities scattered at zero angle, and exploiting the scattering densities of H(2)O, D(2)O, ubiquitin, and urea for X-rays and neutrons, we quantitatively determine the number of urea molecules preferentially bound during unfolding of ubiquitin. We find that a pH change from 6.5 to 2.5 triggers recruitment of approximately 20 urea molecules from bulk solution per ubiquitin molecule during the unfolding process.

MeSH terms

  • Hydrogen-Ion Concentration
  • Models, Molecular
  • Protein Binding
  • Protein Conformation
  • Protein Denaturation
  • Protein Folding
  • Scattering, Small Angle
  • Ubiquitin / chemistry
  • Ubiquitin / metabolism*
  • Urea / chemistry
  • Urea / metabolism*
  • X-Ray Diffraction

Substances

  • Ubiquitin
  • Urea