title: Probing early events in ferrous cytochrome c folding with time-resolved natural and magnetic circular dichroism spectroscopies.
abstract: In a 1998 collaboration with Tony Fink, we coupled nanosecond circular dichroism methods (TRCD) with a CO-photolysis system for quickly triggering folding in cytochrome c (cyt c) in order to make the first time-resolved far-UV CD measurement of early secondary structure formation in a protein. The small signal observed in that initial study, approximately 10% of native helicity, became the seed for increasingly robust results from subsequent studies bringing additional natural and magnetic circular polarization dichroism and optical rotatory dispersion detection methods (e.g., TRORD, TRMCD, and TRMORD), coupled to fast photolysis and photoreduction triggers, to the study of early folding events. Nanosecond polarization methods are reviewed here in the context of the range of initiation methods and structure-sensitive probes currently available for fast folding studies. We also review the impact of experimental results from fast polarization studies on questions in folding dynamics such as the possibility of multiple folding pathways implied by energy landscape models, the sequence dependence of ultrafast helix formation, and the simultaneity of chain collapse and secondary structure formation implicit in molten globule models for kinetic folding intermediates.
authors.affiliation: Department of Chemistry & Biochemistry, University of California, Santa Cruz, California 95064, USA. chen@chemistry.ucsc.edu
authors.forename: Eefei
authors.initials: E
authors.lastname: Chen
authors.forename: Robert A
authors.initials: RA
authors.lastname: Goldbeck
authors.forename: David S
authors.initials: DS
authors.lastname: Kliger
mesh_terms: D000818:Animals; D002942:Circular Dichroism / Q000379:methods*; D045304:Cytochromes c / Q000737:chemistry*; D004867:Equipment Design; D006418:Heme / Q000737:chemistry*; D006801:Humans; D007501:Iron / Q000737:chemistry*; D011489:Protein Denaturation; D017510:Protein Folding; D017433:Protein Structure, Secondary; D013997:Time Factors; D014466:Ultraviolet Rays
chemical_list: D006418:Heme; D045304:Cytochromes c; D007501:Iron