title: Ultrafast signals in protein folding and the polypeptide contracted state.
abstract: To test the significance of ultrafast protein folding signals (<<1 msec), we studied cytochrome c (Cyt c) and two Cyt c fragments with major C-terminal segments deleted. The fragments remain unfolded under all conditions and so could be used to define the unfolded baselines for protein fluorescence and circular dichroism (CD) as a function of denaturant concentration. When diluted from high to low denaturant in kinetic folding experiments, the fragments readjust to their new baseline values in a "burst phase" within the mixing dead time. The fragment burst phase reflects a contraction of the polypeptide from a more extended unfolded condition at high denaturant to a more contracted unfolded condition in the poorer, low denaturant solvent. Holo Cyt c exhibits fluorescence and CD burst phase signals that are essentially identical to the fragment signals over the whole range of final denaturant concentrations, evidently reflecting the same solvent-dependent, relatively nonspecific contraction and not the formation of a specific folding intermediate. The significance of fast folding signals in Cyt c and other proteins is discussed in relation to the hypothesis of an initial rate-limiting search-nucleation-collapse step in protein folding [Sosnick, T. R., Mayne, L. & Englander, S. W. (1996) Proteins Struct. Funct. Genet. 24, 413-426].
authors.affiliation: Johnson Research Foundation, Department of Biochemistry and Biophysics, University of Pennsylvania, Philadelphia, PA 19104-6059, USA.
authors.forename: T R
authors.initials: TR
authors.lastname: Sosnick
authors.forename: M D
authors.initials: MD
authors.lastname: Shtilerman
authors.forename: L
authors.initials: L
authors.lastname: Mayne
authors.forename: S W
authors.initials: SW
authors.lastname: Englander
mesh_terms: D000818:Animals; D003574:Cytochrome c Group / Q000737:chemistry*; D006736:Horses; D010446:Peptide Fragments / Q000737:chemistry; D017510:Protein Folding
chemical_list: D003574:Cytochrome c Group; D010446:Peptide Fragments