Structural biology of potential protein drug design targets from Trypanosoma cruzi.

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

Rosario

Congreso; XLII Reunión anual Sociedad Argentina de Investigación Bioquímica y Biología Molecular (SAIB); 2006

Introduction: 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 main objective of the Chagas´ Disease Research Network is the search for protein targets in metabolic routes absent in the human host and common in trypanosomatids Materials and Methods: The T. cruzi proteins studied at the present are: TcAK (arginine kinase), TcNDPK (nucleoside diphosphate kinase), TcALArac (alanine racemace), TcTAF9 (tata binding factor 9), TcFIP1-like (factor interacting with Pap1) and TcCPSF30 (cleavaje polyadenylation specificity factor). They are expressed in E. coli as a fusion protein with N-terminal His-tag. We have overexpressed and purified these proteins for crystallization and other structural assays. We have carried out activity assays. Results and Discussion: His-TcNDPK was overexpressed, purified and crystallized in differents conditions. It has shown biological activity. We are overexpressed and purified TcTAF9, His-TcFIP1 and TcCPSF30. His-TcAlarac are overexpressed.