Structural studies for drug discovery against human infectious diseases

• Grant number: 17/24901-8
• Support type: Scholarships in Brazil - Doctorate (Direct)
• Effective date (Start): February 01, 2018
• Effective date (End): January 31, 2022
• Field of knowledge: Biological Sciences - Biophysics - Molecular Biophysics
• Principal researcher: Alessandro Silva Nascimento
• Grantee: Lívia Oliveira Dantas Clementino
• Home Institution: Instituto de Física de São Carlos (IFSC). Universidade de São Paulo (USP). São Carlos , SP, Brazil
• Associated research grant: 15/26722-8 - Drug discovery against human infectious diseases, AP.TEM

Abstract

In the context of the thematic project entitled "Drug discovery against human infectious diseases", the enzymes involved in the biosynthesis of pyridoxal phosphate (PLP, vitamin B6) and thiamine pyrophosphate (TPP, vitamin B1) for some pathogenic microorganisms will be studied. More specifically, the enzymes from these pathways will be evaluated for Staphylococcus aureus and Plasmodium falciparum, looking for candidate targets for the discovery of new agents for the treatment of infection diseases in humans. In this DD proposal, we will carry out structural studies involving the cloning, expression, crystallization and structural determination of the enzymes of the biosynthesis pathway of vitamin B6 in P. falciparum. To this end, synthetic genes are already being purchased with the coding sequences for the enzymes optimized for expression in E. coli. These genes will be amplified and cloned into pET-TRx1A expression vector employing the LIC system for the heterologous expression of the enzymes. Then the genes will be expressed and the products will be purified for crystallization. This stage will be carried out through robotic tests. The crystals, if and when obtained, will be used to collect diffraction data in domestic source or LNLS. In addition, other promising molecular targets in S. aureus and other pathogenic microorganisms may be explored in order to identify compounds with potential inhibitors based on crystallographic structures in later stages. (AU)