Caracterização do complexo proteolitico proteassoma em Strongyloides venezuelensis

AUTOR(ES)
DATA DE PUBLICAÇÃO

2004

RESUMO

Protease is an important biological process in different parasites, especially in enzymatic complex like 26S proteasome. The objective of the present work was to initiate the study of the 26S proteasome in the nematode Strongyloides venezuelensis using biochemical experiments and identifying some components of this multienzymatic complex. This nematode has been used as a model and tool for the studies on strongyloidiasis, a helminthiasis of high prevalence in tropical and subtropical regions. In this study, two development stages of S. venezuelensis, namely infective larvae (third-stage larvae) and parasitic stages, were used for experimental infections in Rattus norvegicus. In biochemical experiments the crude extracts of the both stages were submitted at proteolytic activities (chymotrypsin-like and trypsin-like) using fluorogenic peptides in presence and absence of proteasome inhibitors (MG132 and PSI). Moreover, the crude extracts were submitted at SDS-PAGE and western-blot methods using specifics antibodies. In molecular experiments, RNA of both stages were amplifed with specific oligonucleotides to alfa 6 and beta 6 subunits of the 20S proteasome (proteolytic core) and ATPase 3 and S5a of the regulatory particle (19S). The genomic DNA was amplifed to all subunits, excepting of S5a subunit. Results show the presence of the 26S proteasome in proteolytic activities from S. venezulensis of the chymotrypsin-like and trypsin-like and the presence of the labeled bands by the antibodies, as well as molecular analysis of the subunits. However, new studies will be necessary to define the importance of these multienzymatic complex on the parasite biology

ASSUNTO(S)

biologia molecular strongylidae bioquimica

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