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


Kocabiyik S. , Erduran I., Russel R., Danson M., Hough D.

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, cilt.224, ss.224-228, 1996 (SCI İndekslerine Giren Dergi) identifier identifier identifier

  • Cilt numarası: 224 Konu: 1
  • Basım Tarihi: 1996
  • Doi Numarası: 10.1006/bbrc.1996.1011
  • Dergi Adı: BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
  • Sayfa Sayıları: ss.224-228

Özet

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.