Structural biology of Trypanosoma cruzi energetic metabolism proteins as potential drug targets for Chagas disease.

BARROSO, ARTURO GOMEZ; MIRANDA, MARIANA; BOUVIER, LEON; CANEPA, GASPAR; PEREIRA, CA; AGUILAR, CARLOS

Mar del Plata

Congreso; XLIII Reunión anual Sociedad Argentina de Investigación Bioquímica y Biología Molecular (SAIB); 2007

STRUCTURAL BIOLOGY OF T. CRUZI ENERGETIC METABOLISM PROTEINS AS POTENTIAL DRUG TARGETS FOR CHAGAS DISEASE. Gómez Barroso, J.A Miranda, M.; Bouvier, L.A.; Canepa, G.E.; Pereira, C. and Aguilar, C.F. 1Lab de Biol Mol Estr, U.N.S.L. San Luis; 2Lab de Biol Mol de T.cruzi Inst de Inv Med A Lanari UBA-CONICET, Bs As E-mail: jagomez@unsl.edu.ar Introduction: Trypanosoma cruzi is the etiological agent of Chagas' disease. Proteins involved in T cruzi energetic metabolism are potential targets for drug design. The objective of our work is the resolution by X-ray crystallography of three-dimensional structures of these proteins as a first step for rational drug design. Materials and Methods: T. cruzi proteins under study are: TcAK (arginine kinase), TcNDPK1 (nucleoside diphosphate kinase 1) and TcADK1 (adenylate kinase 1). Proteins have been overexpressed in E. coli and purified by Ni+2-Sepharose affinity chromatography. The crystallization screening has been done by micro-batch technique. Hanging drop and capillary methods have been used for the optimization step. Results and Discussion: TcNDPK1 was overexpressed, purified and crystallized. The initial crystallization condition was found using the microbatch method. Optimal crystals for X-ray diffraction analysis were obtained using capillary-batch and vapor diffusion methods. TcADK1 was overexpressed and purified in the native form. TcADK1 initial steps of crystallization screening are under development. At last, TcAK structure has been refined up to 1.9 Å resolution.