Expression and purification of Zika virus NS2B-NS3 protease for structural studies

• Grant number: 25/17916-5
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: January 01, 2026
• End date: December 31, 2026
• Field of knowledge: Biological Sciences - Biophysics - Molecular Biophysics
• Principal Investigator: Glaucius Oliva
• Grantee: Lorrana Lara Vieira
• Host Institution: Instituto de Física de São Carlos (IFSC). Universidade de São Paulo (USP). São Carlos , SP, Brazil
• Associated research grant: 13/07600-3 - CIBFar - Center for Innovation in Biodiversity and Drug Discovery, AP.CEPID

Abstract

Zika virus (ZIKV), an arbovirus of the Flavivirus genus, remains a global public health threat due to its association with severe neurological complications and the lack of specific antiviral treatments. The viral NS2B-NS3 protease is essential for processing the ZIKV polyprotein and represents a promising target for the development of new drugs. This project aims to produce the ZIKV NS2B-NS3 protease with high purity and yield, followed by the optimization of crystallization conditions to enable structural studies via X-ray diffraction. The construct used is bicistronic, allowing independent expression of NS2B and NS3 and promoting proper folding of the protein complex. The methodology includes expression in E. coli, purification through affinity and size-exclusion chromatography, quality assessment by SDS-PAGE, and screening of crystallization conditions using the hanging-drop vapor diffusion method. The execution of this project will advance structural knowledge about ZIKV and contribute to the scientific and technical training of the student, involving her in a research line of great relevance to public health in Brazil. (AU)