Assessing reversible and irreversible binding effects of kinase covalent inhibitors through ADP-Glo assays
Summary: Typical enzymatic inhibition assays often demonstrate improved potency for kinase covalent inhibitors compared to reversible inhibitors. This can primarily be attributed to the irreversible mode of action and could affect the evaluations of the ATP-competitive nature of covalent inhibitors,...
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2021-09-01
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doaj-2681d2892e6c4a1984cfc58599de1c012021-09-19T05:00:44ZengElsevierSTAR Protocols2666-16672021-09-0123100717Assessing reversible and irreversible binding effects of kinase covalent inhibitors through ADP-Glo assaysMartin Schröder0Apirat Chaikuad1Institute of Pharmaceutical Chemistry, Goethe-University Frankfurt, 60438 Frankfurt, Germany; Structural Genomics Consortium, BMLS, Goethe-University Frankfurt, 60438 Frankfurt, Germany; Corresponding authorInstitute of Pharmaceutical Chemistry, Goethe-University Frankfurt, 60438 Frankfurt, Germany; Structural Genomics Consortium, BMLS, Goethe-University Frankfurt, 60438 Frankfurt, Germany; Corresponding authorSummary: Typical enzymatic inhibition assays often demonstrate improved potency for kinase covalent inhibitors compared to reversible inhibitors. This can primarily be attributed to the irreversible mode of action and could affect the evaluations of the ATP-competitive nature of covalent inhibitors, hindering optimization of these compounds. Here, we describe a version of ADP-Glo assay, in which modification of inhibitor incubation time in the presence or absence of ATP enables a quick assessment of relative reversible and irreversible effects of kinase covalent inhibitors.For complete details on the use and execution of this protocol, please refer to Schröder et al. (2020).http://www.sciencedirect.com/science/article/pii/S266616672100424XSingle-molecule AssaysHigh Throughput ScreeningMolecular/Chemical ProbesProtein BiochemistryChemistry |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Martin Schröder Apirat Chaikuad |
spellingShingle |
Martin Schröder Apirat Chaikuad Assessing reversible and irreversible binding effects of kinase covalent inhibitors through ADP-Glo assays STAR Protocols Single-molecule Assays High Throughput Screening Molecular/Chemical Probes Protein Biochemistry Chemistry |
author_facet |
Martin Schröder Apirat Chaikuad |
author_sort |
Martin Schröder |
title |
Assessing reversible and irreversible binding effects of kinase covalent inhibitors through ADP-Glo assays |
title_short |
Assessing reversible and irreversible binding effects of kinase covalent inhibitors through ADP-Glo assays |
title_full |
Assessing reversible and irreversible binding effects of kinase covalent inhibitors through ADP-Glo assays |
title_fullStr |
Assessing reversible and irreversible binding effects of kinase covalent inhibitors through ADP-Glo assays |
title_full_unstemmed |
Assessing reversible and irreversible binding effects of kinase covalent inhibitors through ADP-Glo assays |
title_sort |
assessing reversible and irreversible binding effects of kinase covalent inhibitors through adp-glo assays |
publisher |
Elsevier |
series |
STAR Protocols |
issn |
2666-1667 |
publishDate |
2021-09-01 |
description |
Summary: Typical enzymatic inhibition assays often demonstrate improved potency for kinase covalent inhibitors compared to reversible inhibitors. This can primarily be attributed to the irreversible mode of action and could affect the evaluations of the ATP-competitive nature of covalent inhibitors, hindering optimization of these compounds. Here, we describe a version of ADP-Glo assay, in which modification of inhibitor incubation time in the presence or absence of ATP enables a quick assessment of relative reversible and irreversible effects of kinase covalent inhibitors.For complete details on the use and execution of this protocol, please refer to Schröder et al. (2020). |
topic |
Single-molecule Assays High Throughput Screening Molecular/Chemical Probes Protein Biochemistry Chemistry |
url |
http://www.sciencedirect.com/science/article/pii/S266616672100424X |
work_keys_str_mv |
AT martinschroder assessingreversibleandirreversiblebindingeffectsofkinasecovalentinhibitorsthroughadpgloassays AT apiratchaikuad assessingreversibleandirreversiblebindingeffectsofkinasecovalentinhibitorsthroughadpgloassays |
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1717376105240854528 |