Kinetic and structural characterization of Mayaro virus nsP2 protease domain

Author(s)

Luiza Leme

• Document type: Master's Dissertation
• Press: Limeira, SP.
• Institution: Universidade Estadual de Campinas (UNICAMP). Instituto de Biologia
• Defense date: 2023-02-27

Examining board members

Rafael Elias Marques; Clarice Weis Arns; Gustavo Fernando Mercaldi

• Advisor: Rafael Elias Marques

Abstract

Mayaro virus (MAYV) belongs to the alphavirus genus and is the causative agent of Mayaro fever, a febrile syndrome that may cause incapacitating arthralgia in severe cases. Currently, this neglected tropical arbovirus maintains a sylvatic transmission cycle in many countries of Latin America, but evidence shows MAYV has the potential to spread to urban areas where it could become a public health crisis. There is no treatment or vaccine available against Mayaro Fever. The non-structural protein 2 (nsP2) is an 87 kDa nonstructural protein conserved among alphaviruses that plays an essential role during viral replication. nsP2 cleaves the viral polyprotein through a cysteine-protease domain on its C-terminal (nsP2pro), generating the non-structural proteins in their mature and functional state. The nsP2pro is considered a promising target for the development of antiviral drugs but, unfortunately, the literature about nsP2pro functionality and 3D structure is incomplete, and these insights have yet to be uncovered. Thus, we aimed at elucidating the tridimensional structure of nsP2pro (41 kDa) using X-rays crystallography and at characterizing nsP2 proteolytic activity using an enzymatic assay. We successfully developed a protocol for the expression of the nsP2pro, also named C9 protease, from recombinant E. coli cells and a chromatographic purification protocol that allows the obtention of this protein in a pure, monomeric, structured, stable and active form (AU)

• FAPESP's process: 20/12198-3 - Structural and kinetic characterization of Mayaro virus C9 protease
• Grantee: Luiza Leme
• Support Opportunities: Scholarships in Brazil - Master