Fluorogenic in vitro activity assay for the main protease M(pro) from SARS-CoV-2 and its adaptation to the identification of inhibitors

This protocol describes an in vitro fluorogenic assay to measure the proteolytic activity and identify inhibitors of M(pro), the main protease produced by SARS-CoV-2 (Severe acute respiratory syndrome coronavirus 2). Studies to identify potential inhibitors of M(pro) mainly rely on in silico molecul...

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Autores principales: Ihssen, Julien, Faccio, Greta, Yao, Chunyan, Sirec, Teja, Spitz, Urs
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8367757/
https://www.ncbi.nlm.nih.gov/pubmed/34423318
http://dx.doi.org/10.1016/j.xpro.2021.100793
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author Ihssen, Julien
Faccio, Greta
Yao, Chunyan
Sirec, Teja
Spitz, Urs
author_facet Ihssen, Julien
Faccio, Greta
Yao, Chunyan
Sirec, Teja
Spitz, Urs
author_sort Ihssen, Julien
collection PubMed
description This protocol describes an in vitro fluorogenic assay to measure the proteolytic activity and identify inhibitors of M(pro), the main protease produced by SARS-CoV-2 (Severe acute respiratory syndrome coronavirus 2). Studies to identify potential inhibitors of M(pro) mainly rely on in silico molecular dynamics simulations or on FRET (Fluorescence Resonance Energy Transfer) substrates. The protocol is based on an aminomethyl coumarin substrate. High sensitivity, specificity, and an easily detectable fluorescent read-out are the advantages offered by this rapid assay, which allows high throughput screening of new M(pro) inhibitors.
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spelling pubmed-83677572021-08-17 Fluorogenic in vitro activity assay for the main protease M(pro) from SARS-CoV-2 and its adaptation to the identification of inhibitors Ihssen, Julien Faccio, Greta Yao, Chunyan Sirec, Teja Spitz, Urs STAR Protoc Protocol This protocol describes an in vitro fluorogenic assay to measure the proteolytic activity and identify inhibitors of M(pro), the main protease produced by SARS-CoV-2 (Severe acute respiratory syndrome coronavirus 2). Studies to identify potential inhibitors of M(pro) mainly rely on in silico molecular dynamics simulations or on FRET (Fluorescence Resonance Energy Transfer) substrates. The protocol is based on an aminomethyl coumarin substrate. High sensitivity, specificity, and an easily detectable fluorescent read-out are the advantages offered by this rapid assay, which allows high throughput screening of new M(pro) inhibitors. Elsevier 2021-08-17 /pmc/articles/PMC8367757/ /pubmed/34423318 http://dx.doi.org/10.1016/j.xpro.2021.100793 Text en © 2021 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Ihssen, Julien
Faccio, Greta
Yao, Chunyan
Sirec, Teja
Spitz, Urs
Fluorogenic in vitro activity assay for the main protease M(pro) from SARS-CoV-2 and its adaptation to the identification of inhibitors
title Fluorogenic in vitro activity assay for the main protease M(pro) from SARS-CoV-2 and its adaptation to the identification of inhibitors
title_full Fluorogenic in vitro activity assay for the main protease M(pro) from SARS-CoV-2 and its adaptation to the identification of inhibitors
title_fullStr Fluorogenic in vitro activity assay for the main protease M(pro) from SARS-CoV-2 and its adaptation to the identification of inhibitors
title_full_unstemmed Fluorogenic in vitro activity assay for the main protease M(pro) from SARS-CoV-2 and its adaptation to the identification of inhibitors
title_short Fluorogenic in vitro activity assay for the main protease M(pro) from SARS-CoV-2 and its adaptation to the identification of inhibitors
title_sort fluorogenic in vitro activity assay for the main protease m(pro) from sars-cov-2 and its adaptation to the identification of inhibitors
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8367757/
https://www.ncbi.nlm.nih.gov/pubmed/34423318
http://dx.doi.org/10.1016/j.xpro.2021.100793
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