Structural biology of T. cruzi energetic metabolism proteins as potential drug targets for chagas disease

GOMEZ BARROSO JA; MIRANDA MR; BOUVIER LA; CANEPA GE; PEREIRA CA; AGUILAR CF

Mar del Plata, Bs. As.

Congreso; XLIII Reunión anual de la Sociedad Argentina Bioquímica y Biología Molecular; 2007

SB-P06.3-D STRUCTURE OF ENERGETIC METABOLISM PROTEINS AS POTENTIAL DRUG TARGETS FOR CHAGAS DISEASE Gómez Barroso JA , Miranda M , Bouvier LA , Canepa GE , Pereira C , Aguilar CF .Lab de Biol Mol Estr, UNSL, San Luis; Lab de Biol Mol de T.cruzi Introduction: is the etiological agent of Chagas´ disease. Proteins involved in 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: proteins under study are: TcAK (arginine kinase), TcNDPK1 (nucleoside diphosphate kinase 1) and TcADK1(adenylate kinase 1). Proteins have been overexpressed in and purified by Ni -Sepharose affinity chromatography. The crystallization screening has been done by micro-batch technique. Hanging drop and capillary methods have been used for theoptimization 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.Finally, TcAK structure has been refined up to 1.9Åresolution