Characterization of protein kinases involved in the control of metabolism in Trypanosomatdes

• Grant number: 21/12527-0
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: December 01, 2021
• End date: November 30, 2022
• Field of knowledge: Biological Sciences - Biochemistry - Biochemistry of Microorganisms
• Principal Investigator: Sergio Schenkman
• Grantee: Michel Augusto Silva
• Host Institution: Escola Paulista de Medicina (EPM). Universidade Federal de São Paulo (UNIFESP). Campus São Paulo. São Paulo , SP, Brazil
• Associated research grant: 20/07870-4 - Mechanisms of trypanosomatid adaptation to hosts through the control of transcription, protein synthesis and secretion of extracellular vesicles, AP.TEM

Abstract

In our laboratory we obtained parasites expressing subunit alpha1 and alpha 2 as well as the gamma subunit of the AMPK protein kinase of T. cruzi containing the myc peptide sequence using the CRISPR/Cas9 technique. We also found that these subunits are phosphorylated when parasites are incubated with glucose oxidase and glucose to produce hydrogen peroxide. Thus, we intend to immunoprecipitate lysates of these epimastigote forms of the parasite incubated in the presence or absence of the peroxide-inducing system, after separation into polyacrylamide gel containing SDS to identify the associated proteins. We also intend to test the differential protein kinase activity of immunoprecipitated samples and verify if ubiquitination occurs using anti-ubiquitin antibodies by Western blot. (AU)