Evaluating Stability and Activity of SARS-CoV-2 PLpro for High-throughput Screening of Inhibitors

Because of the essential roles of SARS-CoV-2 papain-like protease (PLpro) in the viral polyprotein processing and suppression of host immune responses, it is a crucial target for drug discovery against COVID-19. To develop robust biochemical methodologies for inhibitor screening against PLpro, exten...

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Detalles Bibliográficos
Autores principales: Arya, Rimanshee, Prashar, Vishal, Kumar, Mukesh
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer US 2021
Materias:
Original Paper
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8380414/
https://www.ncbi.nlm.nih.gov/pubmed/34420183
http://dx.doi.org/10.1007/s12033-021-00383-y
Descripción
Sumario:Because of the essential roles of SARS-CoV-2 papain-like protease (PLpro) in the viral polyprotein processing and suppression of host immune responses, it is a crucial target for drug discovery against COVID-19. To develop robust biochemical methodologies for inhibitor screening against PLpro, extensive characterization of recombinant protein is important. Here we report cloning, expression, and purification of the recombinant SARS-CoV-2 PLpro, and explore various parameters affecting its stability and the catalytic activity. We also report the optimum conditions which should be used for high-throughput inhibitor screening using a fluorogenic tetrapeptide substrate. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12033-021-00383-y.

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