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

Dimitriadis, G., Drysdale, A., Myers, J.K., Arora, P., Radford, S.E., Oas, T.G. and Smith, D.A. (2004) Microsecond folding dynamics of the F13W G29A mutant of the B domain of staphylococcal protein A by laser-induced temperature jump. Proceedings of the National Academy of Sciences, 101 (11). pp. 3809-3814. ISSN 0027-8424

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Abstract

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 45°C 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 ≈2 x 10(6) s(-1).

Item Type: Article
Copyright, Publisher and Additional Information: Copyright © 2004 by the National Academy of Sciences
Academic Units: The University of Leeds > Faculty of Maths and Physical Sciences (Leeds) > School of Physics and Astronomy (Leeds)
The University of Leeds > University of Leeds Research Centres and Institutes > Astbury Centre for Structural Molecular Biology (Leeds)
Depositing User: Sherpa Assistant
Date Deposited: 01 Aug 2005
Last Modified: 08 Feb 2013 17:01
Published Version: http://www.pnas.org/cgi/content/full/101/11/3809
Status: Published
Refereed: Yes
Identification Number: 10.1073/pnas.0306433101
URI: http://eprints.whiterose.ac.uk/id/eprint/562

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