Structural and functional characterization of proteins envolved in the virulence of the protozoan parasite Leishmania major

Abstract

The study of the structure and function of Leishmania proteins presents an opportunity to understand the molecular basis of the host/parasite interaction and discovery new targets for development of therapies against the parasite. The poposed project has the objective to investigate the structural and functional aspects of Leishmania major proteins that were previously identified in a subproteomic study of total extract of vesicles of Leishmania species. The strategy is based on the convergence of structural biology and functional analysis for the elucidation of the functional relevance of these proteins to the biology of the parasite, with particular enphasis on pathogenic factors. Two targets (NDK, Nucleoside Diphosphate Kinase and MIF, Macrophage Migration Inhibitory Factor) have been selected from the subproteome study wich present immunomodulatory functions in other pathogenic organisms. These two selected targets for this project have not been characterized in trypanosomatid parasites. The project will be developed by means of two approachs. The first is directed forwards structural and enzymatic characterization of the two targets , spectroscopy studies and resolution of the 3D structure by X-ray. The second approach will be directed forwords the functional characterization of proteins in the L. major (virulent lines) by immunolocalization, interaction of proteins with macrophages, constitutive superexpression of the genes in the promastigote forms of L. major and virulence analysis of the parasite transfected forms.The approval of this project will allow the establishement of the Laboratory of Biochemistry of Parasite Proteins in the FFCLRP-USP, which will be the first laboratory to focus on parasite proteins within the FFCLRP-USP, and will strengthen a network of biochemistry and parasitology laboratories both within the department and between other units on the campus of USP-Ribeirão Preto. (AU)