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

V Moore, A Kanu, O Byron, G Campbell, Michael J Danson, David W Hough, Susan J Crennell

Research output: Contribution to journalArticle

4 Citations (Scopus)
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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.
Original languageEnglish
Pages (from-to)327-336
Number of pages10
JournalExtremophiles
Volume15
Issue number3
DOIs
Publication statusPublished - May 2011

Fingerprint

Pyrococcus furiosus
Citrate (si)-Synthase
Guanidine
Dimerization
Mutant Proteins
Static Electricity
Mutation
Proteins

Keywords

  • fluorescence
  • oligomeric stability
  • dimerisation energy
  • citrate synthase
  • UHBD
  • analytical ultracentrifugation
  • archaeon

Cite this

Contribution of inter-subunit interactions to the thermostability of Pyrococcus furiosus citrate synthase. / Moore, V; Kanu, A; Byron, O; Campbell, G; Danson, Michael J; Hough, David W; Crennell, Susan J.

In: Extremophiles, Vol. 15, No. 3, 05.2011, p. 327-336.

Research output: Contribution to journalArticle

Moore, V ; Kanu, A ; Byron, O ; Campbell, G ; Danson, Michael J ; Hough, David W ; Crennell, Susan J. / Contribution of inter-subunit interactions to the thermostability of Pyrococcus furiosus citrate synthase. In: Extremophiles. 2011 ; Vol. 15, No. 3. pp. 327-336.
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