The effect of cysteine-43 mutation on thermostability and kinetic properties of citrate synthase from Thermoplasma acidophilum

1996-07-05
Kocabıyık, Semra
Russel, RJM
Danson, MJ
Hough, DW
In this study, we have substituted serine-43 by cysteine in the recombinant citrate synthase from a moderately thermophilic Archaeon Thermoplasma acidophilum, for site-specific attachment of labels and have investigated the effects of this mutation on the biochemical properties and thermal stability of the enzyme. Both wild-type and the mutant enzymes were purified to homogenity using affinity chromatography on Matrex Gel Red A. The mutant Thermoplasma citrate synthase is very similar to wild-type citrate synthase in its substrate and co-factor specificities, pH profile and thermal stability. The mutation, however, has decreased the enzyme activity. The newly introduced reactive sulphydryl group could be easily modified by DTNB and labelled with 4-chloro-7-sulphobenzofuran, without loss of any activity. (C) 1996 Academic Press, Inc.
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS

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Citation Formats
S. Kocabıyık, R. Russel, M. Danson, and D. Hough, “The effect of cysteine-43 mutation on thermostability and kinetic properties of citrate synthase from Thermoplasma acidophilum,” BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, pp. 224–228, 1996, Accessed: 00, 2020. [Online]. Available: https://hdl.handle.net/11511/49070.