Cloning, expression and purification of paracaspase Dictyostelium discoideum

Abstract

The cell cycle is determined primarily by the processes of division, differentiation and death, in organisms such as mammals it is described that cell death is linked to the action of enzymes known as caspases. In recent years enzymes of the caspase family have been described that are involved in programmed cell death in simple eukaryotic organisms such as fungi and protozoa (metacaspases) and in metazoa and amebóide Dictyostelium sp (paracaspases). The biochemical study of metacaspases and paracaspases can aid in the understanding of programmed cell death in unicellular organisms such as fungi and protozoa, in addition to being possible targets for inhibitors and/or activators for the development of drugs for neglected diseases such as Chagas disease, Leishmaniasis, among others. The cell model of Dictyostelium discoideum is evolutionarily closer to animals and fungi than plants and several signals of transduction are conserved between Dictyostelium and mammals. Dictyostelium has a differentiated life cycle, which consists of a unicellular and a multicellular phase. Dictyostelium is a unique model for studying the differences and similarities in the regulation of programmed cell death and cell cycle control in uni- and multicellular organisms. The present research project aims to obtain the paracaspase of recombinant Dictyostelium discoideum that will be used to carry out the biochemical characterization of this protease.