A novel method for study of protein folding kinetics by monitoring diffusion coefficient in time domain.

Abstract:

:Molecular diffusion process after the photo-induced electron injection to ferric cytochrome c (Fe(III) cyt c) in guanidine hydrochloride (GdnHCl) 3.5 M buffer solution is studied by the time-resolved transient grating technique. Circular dichroism studies have revealed that Fe(III) cyt c is unfolded under this condition but the reduced form, Fe(II) cyt c, is folded. Hence, this pulsed laser-induced reduction should initiate the folding process of cyt c. The observed transient grating signal shows prominent features, which have never been observed before. Based on several characteristic points, we concluded that the apparent diffusion coefficient (D) of Fe(II) cyt c after the reduction is time dependent, which must be associated with the protein folding dynamics. This time-dependent apparent D should reflect either the continuous time development of the hydrodynamic radius or population change of the unfolded and folded states during the folding dynamics. This is the first observation of the time-dependent apparent D during any chemical reaction, and this time-dependent measurement of D should be a unique and powerful way to study the protein folding kinetics from a viewpoint of the protein's shape or the protein-water intermolecular interaction.

journal_name

Biophys J

journal_title

Biophysical journal

authors

Nada T,Terazima M

doi

10.1016/S0006-3495(03)74615-3

subject

Has Abstract

pub_date

2003-09-01 00:00:00

pages

1876-81

issue

3

eissn

0006-3495

issn

1542-0086

pii

S0006-3495(03)74615-3

journal_volume

85

pub_type

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