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


Reference:

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

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    Official 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 100A degrees 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.

    Details

    Item Type Articles
    CreatorsMoore, V., Kanu, A., Byron, O., Campbell, G., Danson, M. J., Hough, D. W. and Crennell, S. J.
    DOI10.1007/s00792-011-0363-6
    Uncontrolled Keywordsfluorescence, oligomeric stability, dimerisation energy, citrate synthase, uhbd, analytical ultracentrifugation, archaeon
    DepartmentsFaculty of Science > Biology & Biochemistry
    Research CentresCentre for Extremophile Research (CER)
    Publisher StatementCrennell_Extremophiles_2011_15_3_327.pdf: The original publication is available at www.springerlink.com; Crennell_Extremophiles_2011_15_3_327.doc: The original publication is available at www.springerlink.com
    RefereedYes
    StatusPublished
    ID Code23838

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