Estudo de interação bioquímica da metaloproteases de Leishmania spp. e Trypanosoma cruzi com substratos proteicos

Abstract

Trypanosomatids parasites are responsible for neglected diseases in humans and with animal importance. Despite numerous studies, Leishmaniasis and Chagas' disease represent an important public health problem, especially in Brazil. Currently, there are no vaccines for the control of human parasitic diseases of these diseases and there is still a great need for new drugs or new drug formulations, which are safe, more effective, and less toxic. Metalloproteinases are surface molecules, present in trypanosomatids and are fundamental for parasite-host interaction, changing the homeostasis of the extracellular matrix. Therefore, the aim of this study is to characterize the proteolytic activity of metalloproteases of trypanosomatids of medical importance, causing diseases, such as Chagas' disease and leishmaniasis against substrates. From the axenic cultivation, the total extracts of the parasites were chosen, from which the quantification was made, extracts with the best ones were selected. Protein profiles, as well as enzymatic activity and enzymatic activity in vitro of different crude extracts of the Trypanosomatids, were evaluated. In all assays performed by SDS-PAGE, a clear distinction was observed between the parasite profiles. Through zymography tests, the proteolytic profile of the proteins in the crude extract of Leishmania spp. on the gelatin and albumin substrate. Regarding Trypanosoma cruzi, a proteolytic pattern similar to that described in the literature was found for cruzipain. These proteases were subjected to degradation tests against serum albumin substrates, at different time intervals, using the SDS- PAGE method, where it was possible to verify the affinity of the proteolytic activity profile of the species selected for this study.