Characterization and biochemical and structural analysis of the mitochondrial protein phosphatase PGAM5

• Grant number: 23/16614-0
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: April 01, 2024
• End date: July 31, 2024
• Field of knowledge: Biological Sciences - Biochemistry - Chemistry of Macromolecules
• Principal Investigator: Luciana Elena de Souza Fraga Machado
• Grantee: Laura Santoro de Medeiros
• Host Institution: Instituto de Biociências (IB). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated research grant: 19/02605-3 - Structural and functional characterization of mitochondrial protein phosphatases by redox metabolism and their role in cell biology, AP.JP

Abstract

Mitochondria play an essential role in regulating cellular homeostasis and the stress response, through processes such as fission-fusion, mitophagy and release of apoptotic proteins. Different studies show the importance of protein phosphatases located in subcompartments of the mitochondria. Among mitochondrial protein phosphatases, phosphoglycerate mutase 5 (PGAM5) stands out, a histidine-dependent serine/threonine protein phosphatase, which has been described in the processes of regulating mitochondrial dynamics and programmed cell death. This work aims to characterize biochemically and structurally the mitochondrial protein phosphatase PGAM5 and analyze the interaction interface with the KEAP1 protein. The structural characterization will be analyzed through blue and clear native PAGE, while the interaction of PGAM5 and KEAP1 proteins will be proven with PGAM5 in its monomeric form through pull down and nuclear magnetic resonance experiments. We hope to obtain stable oligomers of PGAM5 and characterize the interaction interface between these two proteins.