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 fulllength 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.
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