The glyoxalase system, comprizing glyoxalase I and glyoxalase II, is a ubiquitous pathway that detoxifies highly reactive aldehydes, such as methylglyoxal, using glutathione as a cofactor. Recent studies of Leishmania major glyoxalase I and Trypanosoma brucei glyoxalase II have revealed a unique dependence upon the trypanosomatid thiol trypanothione as a cofactor. This difference suggests that the trypanothione-dependent glyoxalase system may be an attractive target for rational drug design against the trypanosomatid parasites. Here we describe the cloning, expression and kinetic characterization of glyoxalase I from Trypanosoma cruzi. Like L. major glyoxalase I, recombinant T. cruzi glyoxalase I showed a preference for nickel as its metal cofactor. In contrast with the L. major enzyme, T. cruzi glyoxalase I was far less fast-idious in its choice of metal cofactor efficiently utilizing cobalt, manganese and zinc. T. cruzi glyoxalase I isomerized hemithio-acetal adducts of trypanothione more than 2400 times more efficiently than glutathione adducts, with the methylglyoxal adducts 2–3-fold better substrates than the equivalent phenylglyoxal adducts. However, glutathionylspermidine hemithioacetal adducts were most efficiently isomerized and the glutathionylspermidine-based inhibitor S-4-bromobenzylglutathionylspermidine was found to be a potent linear competitive inhibitor of the T. cruzi enzyme with a Ki of 5.4±0.6 μM. Prediction algorithms, combined with subcellular fractionation, suggest that T. cruzi glyoxalase I localizes not only to the cytosol but also the mitochondria of T. cruzi epimastigotes. The contrasting substrate specificities of human and trypanosomatid glyoxalase enzymes, confirmed in the present study, suggest that the glyoxalase system may be an attractive target for anti-trypanosomal chemotherapy.
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Research Article|
November 14 2006
Trypanothione-dependent glyoxalase I in Trypanosoma cruzi
Neil Greig;
Neil Greig
1Division of Biological Chemistry and Molecular Microbiology, Wellcome Trust Biocentre, College of Life Sciences, University of Dundee, Dundee, DD1 5EH, Scotland, U.K.
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Susan Wyllie;
Susan Wyllie
1Division of Biological Chemistry and Molecular Microbiology, Wellcome Trust Biocentre, College of Life Sciences, University of Dundee, Dundee, DD1 5EH, Scotland, U.K.
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Tim J. Vickers;
Tim J. Vickers
1
1Division of Biological Chemistry and Molecular Microbiology, Wellcome Trust Biocentre, College of Life Sciences, University of Dundee, Dundee, DD1 5EH, Scotland, U.K.
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Alan H. Fairlamb
Alan H. Fairlamb
2
1Division of Biological Chemistry and Molecular Microbiology, Wellcome Trust Biocentre, College of Life Sciences, University of Dundee, Dundee, DD1 5EH, Scotland, U.K.
2To whom correspondence should be addressed (email a.h.fairlamb@dundee.ac.uk).
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Publisher: Portland Press Ltd
Received:
June 13 2006
Revision Received:
September 04 2006
Accepted:
September 08 2006
Accepted Manuscript online:
September 08 2006
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London
2006
Biochem J (2006) 400 (2): 217–223.
Article history
Received:
June 13 2006
Revision Received:
September 04 2006
Accepted:
September 08 2006
Accepted Manuscript online:
September 08 2006
Citation
Neil Greig, Susan Wyllie, Tim J. Vickers, Alan H. Fairlamb; Trypanothione-dependent glyoxalase I in Trypanosoma cruzi. Biochem J 1 December 2006; 400 (2): 217–223. doi: https://doi.org/10.1042/BJ20060882
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