Production, crystallization, and crystallographic structure determination of dihydrofolate reductase from Cryptococcus neoformans

• Grant number: 25/13004-1
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: August 01, 2025
• End date: July 31, 2026
• Field of knowledge: Biological Sciences - Biophysics - Molecular Biophysics
• Principal Investigator: Marcio Vinicius Bertacine Dias
• Grantee: Luiza Catarina Batista de Alvarenga
• Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated research grant: 24/22643-5 - Structural, functional characterization, and fragment screening for dihydrofolate reductase from invasive pathogenic fungi, AP.R

Abstract

Infectious diseases are currently a significant problem in worldwide public health. Among these diseases, there are those infections caused by invasive pathogenic fungi that cause the death of a large number of patients, particularly those in hospital environments or those who suffer from immunodeficiency. Currently, there are only a few alternatives for treating these infections, and the options are still associated with significant side effects. Additionally, there are still an increasing number of reports of resistant strains to the used antifungal. Therefore, it is crucial to investigate molecular targets and develop new antifungal drugs to manage these diseases, including those caused by Cryptococcus neoformans. This fungus typically causes severe infections in the nervous system in patients with immunosuppression. In this project, our objective is to study the enzyme dihydrofolate reductase (DHFR) from this pathogen. To achieve this, we aim to heterologously produce this enzyme in a bacterial system, purify it, crystallize it, and determine its structure in the presence of the coenzyme NADPH and in the presence of antifolates. At the end of the project, we expect to obtain crucial information about the binding mode of these molecules, which may contribute to the development of new compounds for treating fungal infections. (AU)