Mass spectrometric assays reveal discrepancies in inhibition profiles for the SARS-CoV-2 papain-like protease
The two SARS-CoV-2 proteases, i.e. the main protease (Mpro) and the papain-like protease (PLpro), which hydrolyze the viral polypeptide chain giving functional non-structural proteins, are essential for viral replication and are medicinal chemistry targets. We report a high-throughput mass spectrome...
| Main Authors: | , , , , , , , , , |
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| Format: | Journal article |
| Language: | English |
| Published: |
Wiley
2022
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| _version_ | 1797107347391250432 |
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| author | Brewitz, L Kamps, JJAG Lukacik, P Strain-Dammerell, C Zhao, Y Tumber, A Malla, T Orville, A Walsh, M Schofield, C |
| author_facet | Brewitz, L Kamps, JJAG Lukacik, P Strain-Dammerell, C Zhao, Y Tumber, A Malla, T Orville, A Walsh, M Schofield, C |
| author_sort | Brewitz, L |
| collection | OXFORD |
| description | The two SARS-CoV-2 proteases, i.e. the main protease (Mpro) and the papain-like protease (PLpro),
which hydrolyze the viral polypeptide chain giving functional non-structural proteins, are essential for
viral replication and are medicinal chemistry targets. We report a high-throughput mass spectrometry
(MS)-based assay which directly monitors PLpro catalysis in vitro. The assay was applied to investigate
the effect of reported small-molecule PLpro inhibitors and selected Mpro inhibitors on PLpro catalysis.
The results reveal that some, but not all, PLpro inhibitor potencies differ substantially from those
obtained using fluorescence-based assays. Some substrate-competing Mpro inhibitors, notably PF07321332 (nirmatrelvir) which is in clinical development, do not inhibit PLpro
. Less selective Mpro
inhibitors, e.g. auranofin, inhibit PLpro
, highlighting the potential for dual PLpro/Mpro inhibition. MSbased PLpro assays, which are orthogonal to widely employed fluorescence-based assays, are of utility
in validating inhibitor potencies, especially for inhibitors operating by non-covalent mechanisms |
| first_indexed | 2024-03-07T07:13:16Z |
| format | Journal article |
| id | oxford-uuid:0ed48f1c-92c3-4491-9f16-5ab311cc999c |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T07:13:16Z |
| publishDate | 2022 |
| publisher | Wiley |
| record_format | dspace |
| spelling | oxford-uuid:0ed48f1c-92c3-4491-9f16-5ab311cc999c2022-07-27T07:43:44ZMass spectrometric assays reveal discrepancies in inhibition profiles for the SARS-CoV-2 papain-like proteaseJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:0ed48f1c-92c3-4491-9f16-5ab311cc999cEnglishSymplectic ElementsWiley2022Brewitz, LKamps, JJAGLukacik, PStrain-Dammerell, CZhao, YTumber, AMalla, TOrville, AWalsh, MSchofield, CThe two SARS-CoV-2 proteases, i.e. the main protease (Mpro) and the papain-like protease (PLpro), which hydrolyze the viral polypeptide chain giving functional non-structural proteins, are essential for viral replication and are medicinal chemistry targets. We report a high-throughput mass spectrometry (MS)-based assay which directly monitors PLpro catalysis in vitro. The assay was applied to investigate the effect of reported small-molecule PLpro inhibitors and selected Mpro inhibitors on PLpro catalysis. The results reveal that some, but not all, PLpro inhibitor potencies differ substantially from those obtained using fluorescence-based assays. Some substrate-competing Mpro inhibitors, notably PF07321332 (nirmatrelvir) which is in clinical development, do not inhibit PLpro . Less selective Mpro inhibitors, e.g. auranofin, inhibit PLpro , highlighting the potential for dual PLpro/Mpro inhibition. MSbased PLpro assays, which are orthogonal to widely employed fluorescence-based assays, are of utility in validating inhibitor potencies, especially for inhibitors operating by non-covalent mechanisms |
| spellingShingle | Brewitz, L Kamps, JJAG Lukacik, P Strain-Dammerell, C Zhao, Y Tumber, A Malla, T Orville, A Walsh, M Schofield, C Mass spectrometric assays reveal discrepancies in inhibition profiles for the SARS-CoV-2 papain-like protease |
| title | Mass spectrometric assays reveal discrepancies in inhibition profiles for the SARS-CoV-2 papain-like protease |
| title_full | Mass spectrometric assays reveal discrepancies in inhibition profiles for the SARS-CoV-2 papain-like protease |
| title_fullStr | Mass spectrometric assays reveal discrepancies in inhibition profiles for the SARS-CoV-2 papain-like protease |
| title_full_unstemmed | Mass spectrometric assays reveal discrepancies in inhibition profiles for the SARS-CoV-2 papain-like protease |
| title_short | Mass spectrometric assays reveal discrepancies in inhibition profiles for the SARS-CoV-2 papain-like protease |
| title_sort | mass spectrometric assays reveal discrepancies in inhibition profiles for the sars cov 2 papain like protease |
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