Proteomics of Trypanosoma cruzi: Expression, purification and preliminary structural studies.

BARROSO, ARTURO GOMEZ; MIRANDA, MARIANA; PEREIRA, CA; VAZQUEZ, MARTIN; SERRA, ESTEBAN; AGUILAR, CARLOS

Pinamar

Congreso; XLI Reunión Anual de la Sociedad Argentina de Investigación Bioquímica y Biología Molecular; 2005

Trypanosoma cruzi is the etiologic agent of Chagas' disease. The objective of our work is the resolution by X-ray crystallography of the three-dimensional structure of T. cruzi proteins as a first step for rational drug design based on the structure. The development of new drugs is based in the search for protein targets in metabolic routes absent in the human host and common in trypanosomatids. The proteins studied at the present are: TcAK (arginine kinase), TcNDPK (nucleoside diphosphate kinase), TcFIP1-like (factor interacting with Pap1) and TcCPSF30 (cleavaje polyadenylation specificity factor). The plasmids are provided by the other members of the research network. They are over-expressed as a fusion protein with a N-terminal His-tag. They are purified to be used for crystallization and other structural assays. His-TcNDPK was over-expressed and purified in the native state with a high purity and a high concentration for primary crystallization assays. Work is being done on the over-expression of His-TcFIP1 and TcCPSF30. The structure of TcAK has been solved at 1.8 Å resolution. This work is being done within the framework of the Chagas´ Disease Research Network. The research network has established a common methodological matrix to study protein complexes of different biological functions.