Biosynthesis of trypanothione in Trypanosoma brucei brucei.
Trypanothione [T(SH)2], the major redox mediator in pathogenic trypanosomatids, is synthetized stepwise by two distinct enzymes in Crithidia fasciculata, while in Trypanosoma cruzi a single enzyme catalyzes both steps. A full-length reading frame presumed to encode trypanothione synthetase (TryS) was obtained by PCR using DNA of T. brucei as template and primers based on fragments of putative TryS genes. The recombinant protein produced by E. coli Origami (DE3) was purified to homogeneity by chelate and ion exchange chromatography. The enzyme catalyzed both reactions of T(SH)2 biosynthesis. Thus, T(SH)2 synthesis appears to be similar in African (T. brucei) and New World (T. cruzi) trypanosomes but distinct from that of Crithidia.
Department of Biochemistry, Technical University of Braunschweig, Mascheroder Weg 1, D-38124 Braunschweig, Germany.
Marcelo
M
Comini
Ulrich
U
Menge
Leopold
L
Flohé
D000595:Amino Acid Sequence; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D016831:Crithidia fasciculata / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism; D005978:Glutathione / Q000031:analogs & derivatives* / Q000096:biosynthesis*; D006868:Hydrolysis; D008969:Molecular Sequence Data; D008970:Molecular Weight; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013095:Spermidine / Q000031:analogs & derivatives* / Q000096:biosynthesis*; D014346:Trypanosoma brucei brucei / Q000378:metabolism*; D014349:Trypanosoma cruzi / Q000378:metabolism; D014357:Trypsin
C044809:trypanothione; D014357:Trypsin; D005978:Glutathione; D013095:Spermidine