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Structural and kinetic characterization of Mayaro virus C9 protease

Grant number: 20/12198-3
Support type:Scholarships in Brazil - Master
Effective date (Start): August 01, 2021
Effective date (End): September 30, 2022
Field of knowledge:Biological Sciences - Biochemistry - Molecular Biology
Principal researcher:Rafael Elias Marques Pereira Silva
Grantee:Luiza Leme
Home Institution: Centro Nacional de Pesquisa em Energia e Materiais (CNPEM). Ministério da Ciência, Tecnologia e Inovações (Brasil). Campinas , SP, Brazil

Abstract

ABSTRACT Mayaro virus (MAYV) is the causative agent of the Mayaro fever, a febrile syndrome that in severe cases may cause incapacitating arthralgia. Currently, this neglected tropical arbovirus maintains a sylvatic transmission cycle in many countries of Latin America, but evidence shows it has potential to spread to urban areas where it could become a public health crisis, since there is no treatment or vaccine for the Mayaro Fever. The nsP2 (87 kDa) is a non structural protein of MAYV that plays an important role during viral replication by cleaving the viral polyprotein through a cysteine-protease domain on its C-end, this protein is a promising target for the development of antiviral drugs but, unfortunately, its 3D structure is yet to be uncovered. This work aims to achieve the structural and kinetic characterization of the C9 peptidase domain (41 kDa) of nsP2 through x-ray crystallography and enzyme assays. So far we successfully developed a protocol for the expression of the C9 peptidase from recombinant E. coli cells and a purification protocol that allows the obtention of this protein in a pure and stable state, with sufficient yield to perform protein crystallization assays.

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