Early events, kinetic intermediates and the mechanism of protein folding in cytochrome C.
Kinetic studies of the early events in cytochrome c folding are reviewed with a focus on the evidence for folding intermediates on the submillisecond timescale. Evidence from time-resolved absorption, circular dichroism, magnetic circular dichroism, fluorescence energy and electron transfer, small-angle X-ray scattering and amide hydrogen exchange studies on the t < or = 1 ms timescale reveals a picture of cytochrome c folding that starts with the approximately 1-micros conformational diffusion dynamics of the unfolded chains. A fractional population of the unfolded chains collapses on the 1 - 100 micros timescale to a compact intermediate I(C) containing some native-like secondary structure. Although the existence and nature of I(C) as a discrete folding intermediate remains controversial, there is extensive high time-resolution kinetic evidence for the rapid formation of I(C) as a true intermediate, i.e., a metastable state separated from the unfolded state by a discrete free energy barrier. Final folding to the native state takes place on millisecond and longer timescales, depending on the presence of kinetic traps such as heme misligation and proline mis-isomerization. The high folding rates observed in equilibrium molten globule models suggest that I(C) may be a productive folding intermediate. Whether it is an obligatory step on the pathway to the high free energy barrier associated with millisecond timescale folding to the native state, however, remains to be determined.
Goldbeck
Robert A
RA
Department of Chemistry and Biochemistry, University of California, Santa Cruz, California 95046, USA.
Chen
Eefei
E
Department of Chemistry and Biochemistry, University of California, Santa Cruz, California 95046, USA.
Kliger
David S
DS
Department of Chemistry and Biochemistry, University of California, Santa Cruz, California 95046, USA.
D002942:Circular Dichroism; D045304:Cytochromes c / Q000737:chemistry / Q000378:metabolism*; D007700:Kinetics; D017510:Protein Folding; D017433:Protein Structure, Secondary; D058767:Protein Unfolding; D013816:Thermodynamics; D013997:Time Factors
D045304:Cytochromes c