Functional characterization of a recombinant cathepsin from cutting ant Atta sexdens

Abstract

Cathepsins are cysteine proteases widely distributed among living organisms and are linked to several physiological processes operating in protein turnover and as regulatory and signaling molecules. In insects, cathepsins L have been related to tissue remodeling which implies important functions of these enzymes in the development and survival of insects.With the sequencing of the genomes of two cutting ants, Acromyrmex echinatior and Atta cephalotes, three sequences were annotated with identity to cathepsins L. However, although annotation of the genes, our knowledge, no studies reported production and characterization of cathepsins from ants in the literature.So, to contribute to the knowledge of this class of enzymes in cutting ants, this project has the purpose to obtain a recombinant cathepsin from Atta sexdens and characterize characterized it biochemistry and enzymatically. For this, oligonucleotides will be designed based on the sequences of cathepsins from cutting ants deposited in databases and DNA will be amplified by PCR from the RNA. The recombinant enzyme expression system will be the Pichia pastoris, which should allow the enzyme expression in soluble and active form. In the future, with the development of the methodology to obtain the recombinant enzyme, the cathepsin can be used in the search for inhibitors and also be assessed for their possible insecticidal activity.This project is part of the project 'Studies of important metabolic pathways in the control of leaf-cutter ants', coordinated by Prof.. Dulce Helena F. Souza and funded by FAPESP (process 2014 / 12169-2).