Subproteômica de trypanosoma cruzi : proteínas ácidas e fração enriquecida em organelas de alta densidade

AUTOR(ES)
DATA DE PUBLICAÇÃO

2009

RESUMO

The flagellate parasite Trypanosoma cruzi (T. cruzi), the etiological agent of Chagas disease, possesses an alternate life cycle between triatomine vectors and mammalian hosts. The fact that T. cruzi gene expression regulation is posttranscriptional prevents the direct correlation between mRNA and protein levels and makes the proteomic approach very attractive. The present work aimed at contributing to the parasite proteomic studies following two different approaches, the comparative proteomics of T. cruzi life stages emphasizing the acidic proteins and the subproteome characterization of a T. cruzi fraction enriched in high density organelles. Thus, narrow pH range (pH 3-5.3 and pH 5.3-6.5) were optimized for protein extracts of epimastigote, tripomastigote and amastigote cells. The resulting tripomastigote and amastigote 2-DE profiles, that were rather complex, displaying more than 1,000 spots/gel, were computationally analyzed in order to determine stage-specific an differentially expressed spots.The 2-DE spots of interest were identified by mass spectrometry (MS) methods. Concerning the T. cruzi subproteome we isolated a fraction from epimastigote cells that was enriched in high density organelles including nuclei as confirmed by transmission electron microscopy, and immunoblotting. The spots of the 2-DE gels in the pH ranges 3- 10, 4-7 and 6-11 were identified by MS displaying nuclear, mitochondrial, flagellar, glycosomal as well as hypothetical proteins.

ASSUNTO(S)

tripanossoma cruzi helminto doença de chagas chagas transporte biológico biologia molecular

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