Trypanosoma cruzi Bromodomain Proteins: Functional and Structural Studies

JEREZ MB, GÓMEZ BARROSO JA, VILLANOVA V, SERRA E, AGUILAR CF.

San Luis

Congreso; XXVII Reunión Científica Anual de la Sociedad de Biología de Cuyo; 2009

Sociedad de Biología de Cuyo

Trypanosoma cruzi is a protozoan parasite that causes Chagas' disease. The objective of our work is the functional and structural study by X-ray Crystallography and Theoretical Methods of parasite bromodomains proteins: TcBDF1 (bromodomain factor 1) and TcBDF3 (bromodomain factor 3). Bromodomains are conserved structural domains that recognized acetyled-lysine found in many nuclear and no nuclear proteins. The parasite proteins have certain characteristics with respect to their function and location. As preliminary structural  studies and to do comparative studies of both structures we have carried out model-building of the bromo-domain part of our protein on the basis of the  known three dimensional structures of homologous bromo-domains, using the program Modeller. In terms of the experimental studies, TcBDF1 and TcBDF3 have been overexpressed in E. coli as a N-terminal fusion His-tag, solubilized and purified by Ni-Sepharose affinity chromatography. The solubilization of TcBDF1 was performed under different conditions with SDS and Urea. TcBDF3 conformational state was analyzed by Urea-PAGE and Native-PAGE and we have carried out preliminary crystallization screenings of TcBDF3.