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

Arya, Rimanshee; Prashar, Vishal; Kumar, Mukesh

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.