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Establishing expression systems for the biophysical analysis of integral membrane proteins of pathogenic kinetoplastids


Leishmaniasis and Chagas disease are insect vector borne diseases caused by microscopic protozoan parasites. Whilst recent progress against many NTDs has been promising, with an increasing number of cases in an ever wider geographical area, coupled with a limited number of drugs and rising antimicrobial resistance, the battle against both diseases has been described as a losing one. In this environment new drugs as urgently required, and these demand new validated drug targets. Development of these targets towards drug discovery requires assay systems, and functional and biophysical studies (e.g. crystallography). Despite the progress in the study of different kind of proteins, it is noteworthy that the development of information is unbalanced against membrane proteins. More than 97% (>126,000) of these structures are soluble proteins which are relatively easy to express, purify, functionally characterize and crystallize. However, more than 1 in 4 of eukaryotic proteins are lipid-bound, transmembrane (TM) macromolecules. In the context of a partnership between USP and Durham we are initiating a systematic analysis of membrane integral proteins of both, Leishmania spp. and Trypanosoma cruzi to express them in conditions of making a biophysical characterization for obtaining information to analyze their perspectives for developing new targets. The goal of this proposal is to establish in São Paulo common protocols for the expression of such membrane-associated proteins. In addition, we plan to expand the impact of the visit by offering a post-graduation short intensive course, and at least three seminars in research Institutions in the State of São Paulo. (AU)

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