Fluorogenic in vitro activity assay for the main protease Mpro from SARS-CoV-2 and its adaptation to the identification of inhibitors
Summary: This protocol describes an in vitro fluorogenic assay to measure the proteolytic activity and identify inhibitors of Mpro, the main protease produced by SARS-CoV-2 (Severe acute respiratory syndrome coronavirus 2). Studies to identify potential inhibitors of Mpro mainly rely on in silico mo...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2021-09-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166721004998 |
| _version_ | 1818663599808184320 |
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| author | Julien Ihssen Greta Faccio Chunyan Yao Teja Sirec Urs Spitz |
| author_facet | Julien Ihssen Greta Faccio Chunyan Yao Teja Sirec Urs Spitz |
| author_sort | Julien Ihssen |
| collection | DOAJ |
| description | Summary: This protocol describes an in vitro fluorogenic assay to measure the proteolytic activity and identify inhibitors of Mpro, the main protease produced by SARS-CoV-2 (Severe acute respiratory syndrome coronavirus 2). Studies to identify potential inhibitors of Mpro 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 Mpro inhibitors. |
| first_indexed | 2024-12-17T05:19:25Z |
| format | Article |
| id | doaj.art-f2e04d742fb448069d8aa011c6784a0c |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-12-17T05:19:25Z |
| publishDate | 2021-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-f2e04d742fb448069d8aa011c6784a0c2022-12-21T22:02:01ZengElsevierSTAR Protocols2666-16672021-09-0123100793Fluorogenic in vitro activity assay for the main protease Mpro from SARS-CoV-2 and its adaptation to the identification of inhibitorsJulien Ihssen0Greta Faccio1Chunyan Yao2Teja Sirec3Urs Spitz4Biosynth Carbosynth, Rietlistrasse 4, 9422 Staad, Switzerland; Corresponding authorBiosynth Carbosynth, Rietlistrasse 4, 9422 Staad, Switzerland; Corresponding authorBiosynth Carbosynth, Rietlistrasse 4, 9422 Staad, SwitzerlandBiosynth Carbosynth, Rietlistrasse 4, 9422 Staad, SwitzerlandBiosynth Carbosynth, Rietlistrasse 4, 9422 Staad, SwitzerlandSummary: This protocol describes an in vitro fluorogenic assay to measure the proteolytic activity and identify inhibitors of Mpro, the main protease produced by SARS-CoV-2 (Severe acute respiratory syndrome coronavirus 2). Studies to identify potential inhibitors of Mpro 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 Mpro inhibitors.http://www.sciencedirect.com/science/article/pii/S2666166721004998Biotechnology and bioengineeringChemistryClinical ProtocolHealth SciencesHigh Throughput ScreeningMolecular/Chemical Probes |
| spellingShingle | Julien Ihssen Greta Faccio Chunyan Yao Teja Sirec Urs Spitz Fluorogenic in vitro activity assay for the main protease Mpro from SARS-CoV-2 and its adaptation to the identification of inhibitors STAR Protocols Biotechnology and bioengineering Chemistry Clinical Protocol Health Sciences High Throughput Screening Molecular/Chemical Probes |
| title | Fluorogenic in vitro activity assay for the main protease Mpro from SARS-CoV-2 and its adaptation to the identification of inhibitors |
| title_full | Fluorogenic in vitro activity assay for the main protease Mpro from SARS-CoV-2 and its adaptation to the identification of inhibitors |
| title_fullStr | Fluorogenic in vitro activity assay for the main protease Mpro from SARS-CoV-2 and its adaptation to the identification of inhibitors |
| title_full_unstemmed | Fluorogenic in vitro activity assay for the main protease Mpro from SARS-CoV-2 and its adaptation to the identification of inhibitors |
| title_short | Fluorogenic in vitro activity assay for the main protease Mpro from SARS-CoV-2 and its adaptation to the identification of inhibitors |
| title_sort | fluorogenic in vitro activity assay for the main protease mpro from sars cov 2 and its adaptation to the identification of inhibitors |
| topic | Biotechnology and bioengineering Chemistry Clinical Protocol Health Sciences High Throughput Screening Molecular/Chemical Probes |
| url | http://www.sciencedirect.com/science/article/pii/S2666166721004998 |
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