NMR Spectroscopic Characterization of Millisecond Protein Folding by Transverse Relaxation Dispersion Measurements
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- 作者：Markus Zeeb and Jochen Balbach
- 刊名：Journal of the American Chemical Society
- 出版年：2005
- 出版时间：September 28, 2005
- 年：2005
- 卷：127
- 期：38
- 页码：13207 - 13212
- 全文大小：213K
- 年卷期：v.127,no.38(September 28, 2005)
- ISSN：1520-5126
文摘
The cold shock protein CspB adopts its native and functional tertiary structure on the millisecondtime scale. We employed transverse relaxation NMR methods, which allow a quantitative measurement ofthe cooperativity of this fast folding reaction on a residue basis. Thereby, chemical exchange contributionsto the transverse relaxation rate (R2) were observed for every residue of CspB verifying the potential ofthis method to identify not only local dynamics but also to characterize global events. Toward this end, thehomogeneity of the transition state of folding was probed by comparing Chevron plots (i.e., dependence ofthe apparent folding rate on the denaturant concentration) determined by stopped-flow fluorescence withChevron plots of six residues acquired by R2 dispersion experiments. The coinciding results obtained forprobes at different locations in the three-dimensional structure of CspB indicate the ability and significanceof transverse relaxation NMR to determine Chevron plots on a residue-by-residue basis providing detailedinsights on the nature of the transition state of folding.