Obtaining metacaspase (ScMCA-IA) from Saccharomyces cerevisiae containing site-directed alterations of the calcium interaction site

• Grant number: 24/05526-5
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: October 01, 2024
• End date: September 30, 2025
• Field of knowledge: Biological Sciences - Biochemistry - Enzymology
• Principal Investigator: Mauricio Ferreira Marcondes Machado
• Grantee: Emilly Rolim Leme
• Host Institution: Pró-Reitoria Acadêmica. Universidade de Mogi das Cruzes (UMC). Campus da Sede Mogi das Cruzes. Mogi das Cruzes , SP, Brazil
• Associated research grant: 22/06394-0 - Obtaining and biochemical characterization of recombinant protozoan metacaspases of clinical interest, AP.R

Abstract

1.338 / 5.000Resultados de traduçãoResultado da traduçãoThe present research project aims to produce metacaspase from Saccharomyces cerevisiae (ScMCA-IA) containing site-directed mutations in the aspartate residues responsible for the interaction of calcium with this enzyme. The ScMCA-IA metacaspase undergoes self-processing to become catalytically active, and requires calcium ion as an enzymatic cofactor both for its self-processing and to carry out its enzymatic activity. Previous studies carried out by our group showed that the mutation of a single residue demonstrated that only 3 of the 4 aspartate residues demonstrated in the 3D structure of ScMCA-IA affect the interaction of Ca2+ with this enzyme, but none is crucial for the binding of this cofactor. For this reason, we intend to produce this enzyme containing site-directed mutations in the four aspartate residues, replacing them with alanine, but we will produce 2 double mutants (D253/283A and D236/283A), in addition to producing a mutant of the four aspartate residues. (ScMCA-IA Asp-full), to try to elucidate the importance of calcium in the self-processing and regulation of the activity of this protease, such a study could contribute to the rational design of molecules that can inhibit and/or activate ScMCA-IA being of great importance for the development of possible new drugs.