| Abstract [eng] |
In this master’s research project thermodynamic and crystallographic investigation of Hsp90 proteins from different organisms was performed. The goal was to gain insight into protein-ligand binding interactions that would be beneficial for the design of new anti-parasitic drugs. In this study the DNA construct encoding TcHsp90N was constructed, successfully expressed in E. coli BL(DE3) strain and then the protein was purified. Fluorescence thermal shift assay was carried out to determine the stability of recombinant protein and binding parameters of select Hsp90 inhibitors ICPD26, ICPD47 and 17-AAG. In addition, optimal crystallization condition screens were performed for several Hsp90N proteins of interest: LmHsp90N, LdHsp90N, PfHsp90N, TgHsp90N, TbHsp90N, TsHsp90N and Hsp90αN. The aim was to obtain crystals that are suitable for structural X-ray analysis. Optimal conditions for Hsp90αN crystallization were achieved and 3D crystals of this protein in complex with inhibitors ICPD26, Pk-11, Pk-16 and ABIT-07 were grown. The data of high resolution (1,6-3,1 Å) X-ray diffraction was acquired for these protein-ligand complexes. |
