Evaluating Stability and Activity of SARS-CoV-2 PLpro for High-throughput Screening of Inhibitors.
Mol Biotechnol
; 64(1): 1-8, 2022 Jan.
Article
in English
| MEDLINE | ID: covidwho-1366408
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.
Keywords
Full text:
Available
Collection:
International databases
Database:
MEDLINE
Main subject:
High-Throughput Screening Assays
/
Coronavirus Papain-Like Proteases
Type of study:
Experimental Studies
Language:
English
Journal:
Mol Biotechnol
Journal subject:
Molecular Biology
/
Biotechnology
Year:
2022
Document Type:
Article
Affiliation country:
S12033-021-00383-y
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