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
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author Arya, Rimanshee
Prashar, Vishal
Kumar, Mukesh
author_facet Arya, Rimanshee
Prashar, Vishal
Kumar, Mukesh
author_sort Arya, Rimanshee
collection PubMed
description 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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spelling pubmed-83804142021-08-23 Evaluating Stability and Activity of SARS-CoV-2 PLpro for High-throughput Screening of Inhibitors Arya, Rimanshee Prashar, Vishal Kumar, Mukesh Mol Biotechnol Original Paper 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. Springer US 2021-08-22 2022 /pmc/articles/PMC8380414/ /pubmed/34420183 http://dx.doi.org/10.1007/s12033-021-00383-y Text en © The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature 2021 This article is made available via the PMC Open Access Subset for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic.
spellingShingle Original Paper
Arya, Rimanshee
Prashar, Vishal
Kumar, Mukesh
Evaluating Stability and Activity of SARS-CoV-2 PLpro for High-throughput Screening of Inhibitors
title Evaluating Stability and Activity of SARS-CoV-2 PLpro for High-throughput Screening of Inhibitors
title_full Evaluating Stability and Activity of SARS-CoV-2 PLpro for High-throughput Screening of Inhibitors
title_fullStr Evaluating Stability and Activity of SARS-CoV-2 PLpro for High-throughput Screening of Inhibitors
title_full_unstemmed Evaluating Stability and Activity of SARS-CoV-2 PLpro for High-throughput Screening of Inhibitors
title_short Evaluating Stability and Activity of SARS-CoV-2 PLpro for High-throughput Screening of Inhibitors
title_sort evaluating stability and activity of sars-cov-2 plpro for high-throughput screening of inhibitors
topic Original Paper
url 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
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