Expression and purification of the metacase (PbMCA) of Paracoccidioides brasiliensis

Abstract

Cell death is a biological process inherent in all living things. In eukaryotes superior to programmed cell death is mediated by enzymes called caspases, already in lower eukaryotes such as plants, fungi and protozoa, this process is controlled by enzymes called metacaspases. Studies indicate that these proteases have structural homology, however, differ in affinity cleavage with the substrate, while caspases cleave after aspartic acid residues at the P1 position, the metacaspases hydrolyze after arginine or lysine residues at the P1 position. Paracoccidioides brasiliensis (Pb) is the causative agent of paracoccidioidomycosis, an endemic disease in Latin American countries. It is able to encode a single metacaspase (PbMCA), knowing that it may be involved in Pb cell death, studying it is of great interest for the formulation of drugs that may aid in the control and treatment of paracoccidioidomycosis. Therefore, from the PbMCA clone obtained during my first year of scientific initiation (FAPEPS 2017/06526-5), we intend to express and purify the PbMCA by affinity chromatography for further biochemical characterization.