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

被引：8
|
作者：
Arya, Rimanshee [1 ,2 ]
Prashar, Vishal [1 ]
Kumar, Mukesh [1 ,2 ]
机构：
[1] Bhabha Atom Res Ctr, Radiat Biol Sr Hlth Sci Div, Prot Crystallog Sect, Mumbai 400085, Maharashtra, India
[2] Homi Bhabha Natl Inst, Mumbai 400094, Maharashtra, India
来源：
MOLECULAR BIOTECHNOLOGY | 2022年 / 64卷 / 01期
关键词：
SARS-CoV-2; COVID-19; Papain-like protease; Enzyme activity; Aggregation; Stability; PAPAIN-LIKE PROTEASE; RESPIRATORY SYNDROME CORONAVIRUS;
D O I：
10.1007/s12033-021-00383-y
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
摘要：
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.
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页码：1 / 8
页数：8
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