Developing Chemical Biology Tools to Advance the Study and Drug Discovery for Mycobacterium tuberculosis Serine/Threonine Kinases

• Grant number: 24/22005-9
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Start date: September 01, 2025
• End date: August 31, 2028
• Field of knowledge: Health Sciences - Pharmacy
• Principal Investigator: Katlin Brauer Massirer
• Grantee: Daiane Laise da Silva
• Host Institution: Centro de Biologia Molecular e Engenharia Genética (CBMEG). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil

Abstract

Tuberculosis (TB), caused by Mycobacterium tuberculosis (Mtb), remains a leading infectious disease, claiming 1.3 million lives annually. The rise of multidrug-resistant (MDR) and rifampicin-resistant (RR) TB underscores the urgent need for innovative therapeutic strategies. Mtb's ability to survive within host macrophages depends on signaling networks, including 11 serine/threonine protein kinases (STPKs), which regulate critical processes such as cell wall biosynthesis, metabolic adaptation, immune evasion, and stress responses. Among these, PknA and PknB are essential for bacterial survival, coordinating cell division, cell envelope integrity, and metabolic regulation. Despite their importance, the roles of most STPKs and their signaling pathways remain poorly understood. This proposal seeks to address these knowledge gaps by developing tools and resources for comprehensively studying Mtb STPKs. Leveraging our group's expertise in kinase biochemistry and structural biology, the project aims to: (1) produce recombinant proteins for all 11 Mtb STPKs, focusing on their cytoplasmic kinase domains; (2) develop robust biochemical assays to measure kinase activity and screen inhibitors; (3) identify and characterize inhibitors from diverse compound libraries using thermal shift and activity-based biochemical assays; and (4) determine the co-crystal structures of STPKs bound to identified inhibitors via X-ray crystallography. We anticipate that our findings will provide essential tools and data to advance understanding of Mtb kinase biology and lay the foundation for developing novel therapeutics to combat tuberculosis, particularly MDR strains. (AU)