The NADP+ linked glutamate dehydrogenase from Trypanosoma cruzi : sequence, genomic organization and expression

摘要: NADP-linked glutamate dehydrogenase (NADP+-GluDH, EC 1.4.1.4) has been purified to homogeneity from epimastigotes of Trypanosoma cruzi by an improved procedure, and the amino acid sequences 11 internal peptides obtained digestion with trypsin, endopeptidase Lys-C, Arg-C or CNBr have obtained. Using oligonucleotide primers synthesized according sequence N-terminus mature enzyme nucleotide a clone corresponding C-terminus, immunological screening expression library, two complete open reading frames (TcGluDH1 TcGluDH2) were isolated sequenced. The are most similar that NADP+-GluDH Escherichia coli (70-72% identity), less (50-56%) those lower eukaryotes. TcGluDH1 as probe, evidence for presence several genes developmental regulation in different parasite stages was encodes enzymically active protein, since its E. resulted production GluDH activity kinetic parameters natural enzyme.