Quantitative determination of site-specific conformational distributions in an unfolded protein by solid-state nuclear magnetic resonance.

ABSTRACT: Solid-state nuclear magnetic resonance (NMR) techniques are used to investigate the structure of the 35-residue villin headpiece subdomain (HP35) in folded, partially denatured, and fully denatured states. Experiments are carried out in frozen glycerol/water solutions, with chemical denaturation by guanidine hydrochloride (GdnHCl). Without GdnHCl, two-dimensional solid-state (13)C NMR spectra of samples prepared with uniform (13)C labeling of selected residues show relatively sharp cross-peaks at chemical shifts that are consistent with the known three-helix bundle structure of folded HP35. At high GdnHCl concentrations, most cross-peaks broaden and shift, qualitatively indicating disruption of the folded structure and development of static conformational disorder in the frozen denatured s

SUBMITTER: Hu KN 

PROVIDER: S-EPMC2754388 | biostudies-literature | 2009 Oct

REPOSITORIES: biostudies-literature