Structural study of the enzymes P5C, P5CS, P5CR, P5CDH and PRODH of Proline metabolism in Trypanosoma cruzi as targets for the discovery of drugs against Chagas Disease

• Grant number: 23/15602-8
• Support Opportunities: Scholarships in Brazil - Doctorate
• Start date: September 01, 2024
• End date: July 31, 2027
• Field of knowledge: Biological Sciences - Parasitology
• Principal Investigator: Otavio Henrique Thiemann
• Grantee: Maria Eduarda Souza Dias Lino
• Host Institution: Instituto de Física de São Carlos (IFSC). Universidade de São Paulo (USP). São Carlos , SP, Brazil
• Associated research grant: 21/12938-0 - Amino acid metabolism in Trypanosoma cruzi: a toolbox to survive in hostile environments, AP.TEM
• Associated scholarship(s): 25/11958-8 - Structural Characterization of Trypanosoma cruzi P5CS Filaments by Cryo-Electron Microscopy: Insights Into Proline Metabolism and Drug Discovery, BE.EP.DR

Abstract

Chagas disease is caused by the protozoan Trypanosoma cruzi, which resides in the digestive tract of insects such as triatomines and is transmitted by the intestinal contents of this vector. The metabolism of the amino acid Proline in T. cruzi is one of the essential pathways for its survival, being a precursor of Krebs cycle intermediates through the oxidation of proline to ”1-pyrroline-5-carboxylate (P5C) by proline dehydrogenase (PRODH), a FAD-dependent oxidoreductase that donates electrons to the respiratory chain. In mitochondria, P5C is converted to glutamate semialdehyde (GSA), both being in balance. GSA is oxidized to Glutamate by ”1-pyrroline-5-carboxylate dehydrogenase (P5CDH). The reverse reaction occurs by the enzymes ”1-pyrroline-5-carboxylate synthase (P5CS) and ”1-pyrroline-5-carboxylate reductase (P5CR). Thus, the objective is to characterize the atomic structure of the enzymes P5C, P5CS, P5CR, P5CDH and PRODH from Trypanosoma cruzi using different methodologies, i.e., X-ray diffraction of single crystals and Cryo-Electron Microscopy. Furthermore, different biophysical techniques will be used to elucidate the reaction mechanism of enzymes, as well as contribute to the discovery of new inhibitors by searching for compounds in virtual banks and enzymatic assays. This project is part of a collaboration with Prof. Dr. Ariel M. Silber and composes the objectives of the 2021/12938-0 thematic project.