A glutathione S-transferase (GST) was purified to homogeneity from the white-rot fungus, Phanerochaete chrysosporium, by affinity chromatography on glutathione–agarose followed by Mono-Q ion-exchange FPLC. This protein immunoblotted with antisera to rat Theta class GST 5-5 and also showed N-terminal sequence similarity to the Theta class, including the presence of a conserved serine residue that has been specifically implicated in catalysis in this class [Wilce, Board, Feil and Parker (1995) EMBO J. 14, 2133–2143] and other residues conserved in plant sequences. Catalytic activity was found to be highly labile in the purified protein, although preliminary evidence for activity (approx. 120 m-units/mg) with 1,2-epoxy-3-(p-nitrophenoxy)propane was obtained in some preparations. The enzyme seems to be a dimer with a subunit molecular mass of 25 kDa by SDS/PAGE. The native molecular masses estimated by non-denaturing electrophoresis and by Superose-12 gel filtration were 58 and 45 kDa respectively. A second protein purified in this study also gave low level of activity with 1,2-epoxy-3-(p-nitrophenoxy)propane and had a subunit molecular mass of 28 kDa (native size 62–63 kDa), but did not immunoblot with any GST class and seemed to be N-terminally blocked.
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Research Article|
May 15 1997
Glutathione S-transferases from the white-rot fungus, Phanerochaete chrysosporium
Caitriona A. DOWD;
Caitriona A. DOWD
1Department of Biochemistry, University College Cork, Lee Maltings, Prospect Row, Cork, Ireland
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Catherine M. BUCKLEY;
Catherine M. BUCKLEY
1Department of Biochemistry, University College Cork, Lee Maltings, Prospect Row, Cork, Ireland
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David SHEEHAN
David SHEEHAN
*
1Department of Biochemistry, University College Cork, Lee Maltings, Prospect Row, Cork, Ireland
*To whom correspondence should be addressed.
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Publisher: Portland Press Ltd
Received:
September 13 1996
Revision Received:
January 06 1997
Accepted:
January 20 1997
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London © 1997
1997
Biochem J (1997) 324 (1): 243–248.
Article history
Received:
September 13 1996
Revision Received:
January 06 1997
Accepted:
January 20 1997
Citation
Caitriona A. DOWD, Catherine M. BUCKLEY, David SHEEHAN; Glutathione S-transferases from the white-rot fungus, Phanerochaete chrysosporium. Biochem J 15 May 1997; 324 (1): 243–248. doi: https://doi.org/10.1042/bj3240243
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