Purification and characterization of a trypanothione-glutathione thioltransferase from Trypanosoma cruzi.
Although trypanothione [T(S)2] is the major thiol component in trypanosomatidae, significant amounts of glutathione are present in Trypanosoma cruzi. This could be explained by the existence of enzymes using glutathione or both glutathione and T(S)2 as cofactors. To assess these hypotheses, a cytosolic fraction of T. cruzi epimastigotes was subjected to affinity chromatography columns using as ligands either S-hexylglutathione or a non-reducible analogue of trypanothione disulphide. A similar protein of 52 kDa was eluted in both cases. Its partial amino acid sequence indicated that it was identical with the protein encoded by the TcAc2 cDNA previously described [Schoneck, Plumas-Marty, Taibi et al. (1994) Biol. Cell 80, 1-10]. This protein showed no significant glutathione transferase activity but surprisingly catalysed the thiol-disulphide exchange between dihydrotrypanothione and glutathione disulphide. The kinetic parameters were in the same range as those determined for trypanothione reductase toward its natural substrate. This trypanothione-glutathione thioltransferase provides a new target for a specific chemotherapy against Chagas' disease and may constitute a link between the glutathione-based metabolism of the host and the trypanothione-based metabolism of the parasite.
Moutiez
M
M
Chimie des Biomolécules, URA CNRS 1309, Institut Pasteur de Lille, France.
Aumercier
M
M
Schöneck
R
R
Meziane-Cherif
D
D
Lucas
V
V
Aumercier
P
P
Ouaissi
A
A
Sergheraert
C
C
Tartar
A
A
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies; D015153:Blotting, Western; D002846:Chromatography, Affinity; D018076:DNA, Complementary; D016054:DNA, Protozoan; D004591:Electrophoresis, Polyacrylamide Gel; D007535:Isomerases / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D007700:Kinetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010455:Peptides / Q000138:chemical synthesis / Q000276:immunology; D019704:Protein Disulfide-Isomerases; D017386:Sequence Homology, Amino Acid; D014349:Trypanosoma cruzi / Q000201:enzymology*
D000906:Antibodies; D018076:DNA, Complementary; D016054:DNA, Protozoan; D010455:Peptides; D007535:Isomerases; D019704:Protein Disulfide-Isomerases