Evidence for the co-existence of glutathione reductase and trypanothione reductase in the non-trypanosomatid Euglenozoa: Euglena gracilis Z.

Two NADPH-dependent disulfide reductases, glutathione reductase and trypanothione reductase, were shown to be present in Euglena gracilis, purified to homogeneity and characterized. The glutathione reductase (Mr 50 kDa) displays a high specificity towards glutathione disulfide with a KM of 54 microM. The amino acid sequences of two peptides derived from the trypanothione reductase (Mr 54 kDa) show a high level of identity (81% and 64%) with sequences of trypanothione reductases from trypanosomatids. The trypanothione reductase is able to efficiently reduce trypanothione disulfide (KM 30.5 microM) and glutathionylspermidine disulfide (KM 90.6 microM) but not glutathione disulfide, nor Escherichia coli thioredoxin disulfide, nor 5,5'-dithiobis(2-nitrobenzoate) (DTNB). These results demonstrate for the first time (i) the existence of trypanothione reductase in a non-trypanosomatid organism and (ii) the coexistence of trypanothione reductase and glutathione reductase in E. gracilis.