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Affinity chromatography using trypanocidal arsenical drugs identifies a specific interaction between glycerol-3-phosphate dehydrogenase from Trypanosoma brucei and Cymelarsan.

H Denise·C Giroud·M P Barrett·T Baltz
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Abstract

A 36-kDa protein was isolated by affinity chromatography using Cymelarsan, an arsenical drug currently used in African trypanosomiasis treatment, as ligand. This protein was identified as glycerol-3-phosphate dehydrogenase. Trypanosomal glycerol-3-phosphate was bound covalently, whereas its counterpart from rabbit muscle bound by ionic interaction. Arsenical drugs inhibit the enzyme in a dose-dependent manner. Oxidation of cysteine residues protects against inactivation without significantly diminishing enzymic activity. Drug concentrations giving 50% inhibition of the dehydrogenase activity were determined for the enzyme from both Trypanosoma brucei and rabbit and indicate a higher sensitivity of the trypanosomal enzyme to arsenical drugs and thiol reagents. MS was used to identify residues of glycerol-3-phosphate dehydrogenase bound by Cymelarsan; they are not conserved in the mammalian enzyme.

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AnimalsArsenicalsBinding SitesChromatography, AffinityCloning, MolecularCross-Linking ReagentsCysteineEscherichia coliGlycerolphosphate DehydrogenaseMusclesPeptidesPolymerase Chain ReactionRabbitsRecombinant ProteinsSequence AnalysisTrypanocidal AgentsTrypanosoma brucei bruceiComparative StudyJournal ArticleResearch Support, Non-U.S. Gov't

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H Denise, C Giroud, M P Barrett, T Baltz. 1999. Affinity chromatography using trypanocidal arsenical drugs identifies a specific interaction between glycerol-3-phosphate dehydrogenase from Trypanosoma brucei and Cymelarsan.. https://doi.org/10.1046/j.1432-1327.1999.00048.x

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