Microsecond folding dynamics of the F13W G29A mutant of the B domain of staphylococcal protein A by laser-induced temperature jump

被引:60
作者
Dimitriadis, G
Drysdale, A
Myers, JK
Arora, P
Radford, SE
Oas, TG
Smith, DA [1 ]
机构
[1] Univ Leeds, Dept Phys & Astron, Leeds LS2 9JT, W Yorkshire, England
[2] Univ Leeds, Astbury Ctr Struct Mol Biol, Leeds LS2 9JT, W Yorkshire, England
[3] Univ Leeds, Sch Biochem & Microbiol, Leeds LS2 9JT, W Yorkshire, England
[4] Duke Univ, Dept Biochem, Durham, NC 27710 USA
[5] Vanderbilt Univ, Med Ctr, Ctr Struct Biol, Dept Biochem, Nashville, TN 37232 USA
基金
英国生物技术与生命科学研究理事会;
关键词
D O I
10.1073/pnas.0306433101
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
The small size (58 residues) and simple structure of the B domain of staphylococcal protein A (BdpA) have led to this domain being a paradigm for theoretical studies of folding. Experimental studies of the folding of BdpA have been limited by the rapidity of its folding kinetics. We report the folding kinetics of a fluorescent mutant of BdpA (G29A F13W), named F13W*, using nanosecond laser-induced temperature jump experiments. Automation of the apparatus has permitted large data sets to be acquired that provide excellent signal-to-noise ratio over a wide range of experimental conditions. By measuring the temperature and denaturant dependence of equilibrium and kinetic data for F13W*, we show that thermodynamic modeling of multidimensional equilibrium and kinetic surfaces is a robust method that allows reliable extrapolation of rate constants to regions of the folding landscape not directly accessible experimentally. The results reveal that F13W* is the fastest-folding protein of its size studied to date, with a maximum folding rate constant at 0 M guanidinium chloride and 45degreesC of 249,000 s(-1). Assuming the single-exponential kinetics represent barrier-limited folding, these data limit the value for the preexponential factor for folding of this protein to at least approximate to2 x 10(6) S-1.
引用
收藏
页码:3809 / 3814
页数:6
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