Enlightening the molecular basis of trypanothione specificity in trypanosomatids: Mutagenesis of Leishmania infantum glyoxalase II

Barata, Lídia; Sousa Silva, Marta; Schuldt, Linda; Ferreira, António E.N.; Gomes, Ricardo A.; Tomás, Ana M.; Weiss, Manfred S.; Ponces Freire, Ana; Cordeiro, Carlos

Enlightening the molecular basis of trypanothione specificity in trypanosomatids: Mutagenesis of Leishmania infantum glyoxalase II

2011

Leishmania infantum glyoxalase II shows absolute specificity towards its trypanothione thioester substrate. In the previous work, we performed a comparative analysis of glyoxalase II structures determined by X-ray crystallography which revealed that Tyr291 and Cys294, absent in the human homologue, are essential for substrate binding. To validate this trypanothione specificity hypothesis we produced a mutant L. infantum GLO2 enzyme by replacing Tyr291 and Cys294 by arginine and lysine, respectively. This new enzyme is capable to use the glutathione thioester substrate, with kinetic parameters similar to the ones from the human enzyme. Substrate specificity is likely to be mediated by spermidine moiety binding, providing a primer for understanding the molecular basis of trypanothione specificity.

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