Evaluating Stability and Activity of SARS-CoV-2 PLpro for High-throughput Screening of Inhibitors.
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ABSTRACT: 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.
SUBMITTER: Arya R
PROVIDER: S-EPMC8380414 | biostudies-literature |
REPOSITORIES: biostudies-literature
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