A Crystallographic Snapshot of SARS-CoV-2 Main Protease Maturation Process

Autor(es):Mostrar menos -

Noske, G. D. ^[1] ; Nakamura, A. M. ^[1] ; Gawriljuk, V. O. ^[1] ; Fernandes, R. S. ^[1] ; Lima, G. M. A. ^{[2, 3]} ; Rosa, H. V. D. ^[1] ; Pereira, H. D. ^[1] ; Zeri, A. C. M. ^[4] ; Nascimento, A. F. Z. ^[4] ; Freire, M. C. L. C. ^[1] ; Fearon, D. ^{[5, 6]} ; Douangamath, A. ^{[5, 6]} ; von Delft, F. ^{[7, 5, 6, 8]} ; Oliva, G. ^[1] ; Godoy, A. S. ^[1]

Afiliação do(s) autor(es)

^[1] Univ Sao Paulo, Inst Phys Sao Carlos, Av Joao Dagnone 1100, BR-13563120 Sao Carlos - Brazil

^[2] Astex Pharmaceut, 436 Cambridge Sci Pk, Milton Rd, Cambridge CB4 0QA - England

^[3] Max IV Lab, BioMAX, Fotongatan 2, S-22484 Lund - Sweden

^[4] Brazilian Ctr Res Energy & Mat CNPEM, Brazilian Synchrotron Light Lab LNLS, BR-13083970 Campinas, SP - Brazil

^[5] Diamond Light Source Ltd, Harwell Sci & Innovat Campus, Didcot OX11 0QX, Oxon - England

^[6] Res Complex & Harwell, Harwell Sci & Innovat Campu, Didcot OX11 0FA, Oxon - England

^[7] Univ Johannesburg, Dept Biochem, ZA-2006 Auckland Pk - South Africa

^[8] Univ Oxford, Ctr Med Discovery, Old Rd Campus, Roosevelt Dr, Headington OX3 7DQ - England

• Tipo de documento: Artigo Científico
• Fonte: Journal of Molecular Biology; v. 433, n. 18 SEP 3 2021.
• Citações Web of Science: 1

Resumo

SARS-CoV-2 is the causative agent of COVID-19. The dimeric form of the viral M-pro is responsible for the cleavage of the viral polyprotein in 11 sites, including its own N and C-terminus. The lack of structural information for intermediary forms of M-pro is a setback for the understanding its self-maturation process. Herein, we used X-ray crystallography combined with biochemical data to characterize multiple forms of SARS-CoV-2 M-pro. For the immature form, we show that extra N-terminal residues caused conformational changes in the positioning of domain-three over the active site, hampering the dimerization and diminishing its activity. We propose that this form preludes the cis and trans-cleavage of N-terminal residues. Using fragment screening, we probe new cavities in this form which can be used to guide therapeutic development. Furthermore, we characterized a serine site-directed mutant of the M-pro bound to its endogenous N and C-terminal residues during dimeric association stage of the maturation process. We suggest this form is a transitional state during the C-terminal trans-cleavage. This data sheds light in the structural modifications of the SARS-CoV-2 main protease during its self-maturation process. (C) 2021 Elsevier Ltd. All rights reserved. (AU)

• Processo FAPESP: 15/16811-3 - EMU concedido no processo 2014/15546-1: robô de crystalização Gryphon
• Beneficiário: Richard Charles Garratt
• Modalidade de apoio: Auxílio à Pesquisa - Programa Equipamentos Multiusuários
• Processo FAPESP: 16/19712-9 - Caracterização estrutural das proteínas do vírus Zika e busca por agentes antivirais
• Beneficiário: Andre Schutzer de Godoy
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 13/07600-3 - CIBFar - Centro de Inovação em Biodiversidade e Fármacos
• Beneficiário: Glaucius Oliva
• Modalidade de apoio: Auxílio à Pesquisa - Centros de Pesquisa, Inovação e Difusão - CEPIDs