Two similar glutathione S-transferases (GSTs), which do not bind to glutathione– or S-hexylglutathione–agarose affinity resins, have been purified from the yeast Yarrowia lipolytica. An approx. 400-fold purification was obtained by a combination of DEAE-Sephadex, phenyl-Sepharose, hydroxyapatite and Mono-Q anion-exchange chromatography. The native molecular mass of both proteins was estimated as approx. 110 kDa by both Superose-12 gel-filtration chromatography and non-denaturing electrophoresis. SDS/PAGE indicated a subunit mass of 50 kDa. Reverse-phase HPLC of purified proteins gave a single, well-resolved, peak, suggesting that the proteins are homodimers. Identical behaviour on HPLC, native electrophoresis and SDS/PAGE, N-terminal sequencing, sensitivity to a panel of inhibitors and identical specific activities with 1-chloro-2,4-dinitrobenzene as substrate suggest that the two isoenzymes are very similar. The enzymes do not immunoblot with antisera to any of the main GST classes, and N-terminal sequencing suggests no clear relationship with previously characterized enzymes, such as that of the fungus, Phanerochaete chrysosporium [Dowd, Buckley and Sheehan (1997) Biochem. J. 324, 243–248]. It is possible that the two isoenzymes arise as a result of post-translational modification of a single GST isoenzyme.
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Research Article|
August 01 1998
Glutathione S-transferases of the yeast Yarrowia lipolytica have unusually large molecular mass
Vivienne FOLEY;
Vivienne FOLEY
1Department of Biochemistry, University College Cork, Lee Maltings, Prospect Row, Cork, Republic of Ireland
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David SHEEHAN
David SHEEHAN
1
1Department of Biochemistry, University College Cork, Lee Maltings, Prospect Row, Cork, Republic of Ireland
1To whom correspondence should be addressed (e-mail D.SHEEHAN@BUREAU.UCC.IE).
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Publisher: Portland Press Ltd
Received:
January 26 1998
Revision Received:
April 17 1998
Accepted:
May 14 1998
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London © 1998
1998
Biochem J (1998) 333 (3): 839–845.
Article history
Received:
January 26 1998
Revision Received:
April 17 1998
Accepted:
May 14 1998
Citation
Vivienne FOLEY, David SHEEHAN; Glutathione S-transferases of the yeast Yarrowia lipolytica have unusually large molecular mass. Biochem J 1 August 1998; 333 (3): 839–845. doi: https://doi.org/10.1042/bj3330839
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