Interaction studies between the proteins Aha1 and Hsp90 and obtantion of the N- and C-terminal domains of Aha1 of Plasmodium falciparum

Abstract

The molecular chaperones are proteins that assist protein folding and prevent aggregation, among other important intracellular functions. Among the family of chaperones, the Hsp90 is essential for growth of eukaryotic cells and are described as a specialized tool for protein folding. These proteins are capable of interacting with unfolded or partially folded proteins, keeping them in a re-foldable state, and maintaining the conformation, stability and function large number of proteins. The Hsp90 are involved in cellular transcription, cell cycle and apoptosis. A number higher than 150 clients of Hsp90 proteins is already known and they can be classified into three groups: steroid hormone receptors, kinase proteins and unrelated proteins. The function of Hsp90 is driven by dynamic association with their substrates and their co-chaperones, which form large complexes with other molecular chaperones and other cellular proteins. The Hsp90 is essential for cell viability, since its inactivation leads to suppression of the growth of organisms, including protozoaires. Inhibitors of Hsp90 (radicicol and geldanamycin) inhibit the growth of Plasmodium falciparum, Leishmania donovani and Trypanosoma cruzi, and currently have been mentioned as potential drugs for the treatment of some cancers. There are studies that suggest the potential for developing selective inhibitors of Hsp90 against protozoa. The Hsp90 exhibit low ATPase activity, which can be stimulated by client proteins and by the co-chaperone Aha1. The objective of this undergraduate research project is to obtain and characterize the domains N-and C-terminal of the co-chaperone Aha1 of P. falciparum, and to study its interaction with Hsp90 continuing the project already under development by the candidate. It has been shown that Aha1 important role in the functional cycle of Hsp90 in humans and is proposed here to study its interaction with Hsp90, as well as produce and characterize the isolated domains of Aha1 for future studies focusing the mechanism of interaction with Hsp90. The BMB group already has the recombinant Aha1 and M-domain of Hsp90 of P. falciparum produced and purified in its folded, and has the DNA coding for the complete PfHsp90 protein inserted into cloning vector. From the vector containing the DNA coding for the Aha1, the domains N-and C-terminal of this co-chaperone are cloned and the functional tests carried out with recombinant proteins produced.