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

被引：6

作者：

Barata, Lidia ^{[1
,2
]}

Silva, Marta Sousa ^{[1
,3
]}

Schuldt, Linda ^{[2
]}

Ferreira, Antonio E. N. ^{[1
]}

Gomes, Ricardo A. ^{[1
]}

Tomas, Ana M. ^{[3
,4
]}

Weiss, Manfred S. ^{[2
]}

Freire, Ana Ponces ^{[1
]}

Cordeiro, Carlos ^{[1
]}

机构：

[1] Univ Lisbon, Ctr Quim & Bioquim, Dept Quim & Bioquim, Fac Ciencias, P-1749016 Lisbon, Portugal

[2] DESY, European Mol Biol Lab, Hamburg Outstn, D-22603 Hamburg, Germany

[3] Univ Porto, IBMC, P-4150180 Oporto, Portugal

[4] Univ Porto, ICBAS, P-4099003 Oporto, Portugal

来源：

EXPERIMENTAL PARASITOLOGY | 2011年 / 129卷 / 04期

关键词：

Leishmania infantum; Glyoxalase II; Trypanothione; Enzyme specificity shift; Enzyme kinetics; SUBSTRATE-SPECIFICITY; SWISS-MODEL; GLUTATHIONE; DETOXIFICATION; PATHWAY; SYSTEM;

D O I：

10.1016/j.exppara.2011.08.008

中图分类号：

R38 [医学寄生虫学]; Q [生物科学];

学科分类号：

07 ; 0710 ; 09 ; 100103 ;

摘要：

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. (C) 2011 Elsevier Inc. All rights reserved.

引用

页码：402 / 408

页数：7

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