Contribution of inter-subunit interactions to the thermostability of Pyrococcus furiosus citrate synthase

Moore, V., Kanu, A., Byron, O. , Campbell, G., Danson, M.J., Hough, D.J. and Crennell, S.J. (2011) Contribution of inter-subunit interactions to the thermostability of Pyrococcus furiosus citrate synthase. Extremophiles, 15(3), pp. 327-336. (doi:10.1007/s00792-011-0363-6)

Moore, V., Kanu, A., Byron, O. , Campbell, G., Danson, M.J., Hough, D.J. and Crennell, S.J. (2011) Contribution of inter-subunit interactions to the thermostability of Pyrococcus furiosus citrate synthase. Extremophiles, 15(3), pp. 327-336. (doi:10.1007/s00792-011-0363-6)

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Publisher's URL: http://dx.doi.org/10.1007/s00792-011-0363-6

Abstract

Using citrate synthase from the hyperthermophile Pyrococcus furiosus (PfCS) as our test molecule, we show through guanidine hydrochloride-induced unfolding that the dimer separates into folded, but inactive, monomers before individual subunit unfolding takes place. Given that forces across the dimer interface are vital for thermostability, a robust computational method was derived that uses the University of Houston Brownian Dynamics (UHBD) program to calculate both the hydrophobic and electrostatic contribution to the dimerisation energy at 100°C. The results from computational and experimental determination of the lowered stability of interface mutants were correlated, being both of the same order of magnitude and placing the mutant proteins in the same order of stability. This computational method, optimised for hyperthermophilic molecules and tested in the laboratory, after further testing on other examples, could be of widespread use in the prediction of thermostabilising mutations in other oligomeric proteins for which dissociation is the first step in unfolding.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Campbell, Mr Gordon and Byron, Professor Olwyn
Authors: Moore, V., Kanu, A., Byron, O., Campbell, G., Danson, M.J., Hough, D.J., and Crennell, S.J.
College/School:College of Medical Veterinary and Life Sciences > School of Life Sciences
College of Medical Veterinary and Life Sciences > Institute of Infection Immunity and Inflammation
Journal Name:Extremophiles
ISSN:1431-0651

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