Ultrafast folding of the villin subdomain

BIOT 18

Troy Cellmer, cellmert@niddk.nih.gov1, William A. Eaton, eaton@helix.nih.gov2, and James Hofrichter, jim@sunder.niddk.nih.gov2. (1) Laboratory of Chemical Physics, NIDDK,/NIH, 9000 Rockville Pike, Bethesda, MD 20892-0520, (2) Laboratory of Chemical Physics, NIDDK, National Institutes of Health, Building 5, Room 104, 9000 Rockville Pike, Bethesda, MD 20892-0520
The ultra-fast-folding subdomain of the chicken-villin headpiece (villin) is the smallest, naturally-occurring protein that folds autonomously, making it feasible to directly compare experimental folding kinetics with theoretical and computational studies. We have studied the folding kinetics of villin as a function of temperature and guanidinium chloride (GdmCl) concentration using nanosecond laser temperature-jump measurements. At low GdmCl concentrations, the observed relaxation is bi-exponential, consisting of a fast (~100ns) relaxation and a slow (~5micro) phase that corresponds to the unfolding/refolding transition. The relative amplitude of the fast phase decreases monotonically from > 50% in water to ~ 10% in 1M GdmCl. At high GdmCl concentrations there is only a single slow phase. The GdmCl dependence of the folding (slow) relaxation rate is very weak, indicating the presence of a very small barrier to folding. Various quantitative estimations of the barrier height will be presented, and the possibility of “downhill” or barrier-less folding discussed.