Functional characterization of stage-specific aminotransferases from trypanosomatids.
Mol Biochem Parasitol
; 166(2): 172-82, 2009 Aug.
Article
em En
| MEDLINE
| ID: mdl-19443056
ABSTRACT
As part of a study on aminotransferases, genes coding for putative enzymes from Trypanosoma brucei and Leishmania major (alanine aminotransferases ALATs, Tb927.1.3950 and LmjF12.0630; kynurenine aminotransferase KAT, Tb10.389.1810; and tyrosine aminotransferase TAT, LmjF36.2360) were cloned and functionally expressed in Escherichia coli. The putative T. brucei KAT, in fact coded for a glutamine aminotransferase (GlnAT), which exhibited a notably high affinity (in the micromolar range) towards glutamine and cysteine; in addition, like bacterial GlnATs and mammalian KATs, it was able to utilize different 2-oxoacids as amino acceptors. L. major TAT resembled T. cruzi TAT in substrate specificity, although the leishmanial enzyme did not exhibit ALAT activity. On the other hand, T. brucei ALAT, shortened by the first 65 amino acids assigned in the data bases, was functional and actively transaminated the substrate pair l-alanine and 2-oxoglutarate. Moreover in Western blots, the molecular size of the protein detected in crude extracts of T. brucei procyclics was identical to the value of the recombinant enzyme. Like T. brucei and T. cruzi orthologues, L. major ALAT displayed narrow substrate specificity. The leishmanial ALAT, like the T. cruzi enzyme, exhibited a dual subcellular localization, in the cytosol and in the mitochondrion. In line with the findings of comparative proteomic analyses of insect and mammalian stages of T. brucei and Leishmania parasites, our results also showed that T. cruzi ALAT is constitutively expressed, with remarkably higher levels being detected in amastigotes than in epimastigotes. ALATs are expressed in the clinically important stages of TriTryps, probably fulfilling an essential role, which deserves further studies.
Texto completo:
1
Coleções:
01-internacional
Contexto em Saúde:
3_ND
Base de dados:
MEDLINE
Assunto principal:
Trypanosoma brucei brucei
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Tripanossomíase Africana
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Proteínas de Protozoários
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Leishmaniose Cutânea
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Leishmania major
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Transaminases
Limite:
Animals
/
Humans
Idioma:
En
Revista:
Mol Biochem Parasitol
Ano de publicação:
2009
Tipo de documento:
Article