Competition NMR for Detection of Hit/Lead Inhibitors of Protein–Protein Interactions
Screening for small-molecule fragments that can lead to potent inhibitors of protein–protein interactions (PPIs) is often a laborious step as the fragments cannot dissociate the targeted PPI due to their low μM–mM affinities. Here, we describe an NMR competition assay called w-AIDA-NMR (weak-antagon...
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| Format: | Article |
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MDPI AG
2020-07-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/25/13/3017 |
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| author | Bogdan Musielak Weronika Janczyk Ismael Rodriguez Jacek Plewka Dominik Sala Katarzyna Magiera-Mularz Tad Holak |
| author_facet | Bogdan Musielak Weronika Janczyk Ismael Rodriguez Jacek Plewka Dominik Sala Katarzyna Magiera-Mularz Tad Holak |
| author_sort | Bogdan Musielak |
| collection | DOAJ |
| description | Screening for small-molecule fragments that can lead to potent inhibitors of protein–protein interactions (PPIs) is often a laborious step as the fragments cannot dissociate the targeted PPI due to their low μM–mM affinities. Here, we describe an NMR competition assay called w-AIDA-NMR (weak-antagonist induced dissociation assay-NMR), which is sensitive to weak μM–mM ligand–protein interactions and which can be used in initial fragment screening campaigns. By introducing point mutations in the complex’s protein that is not targeted by the inhibitor, we lower the effective affinity of the complex, allowing for short fragments to dissociate the complex. We illustrate the method with the compounds that block the Mdm2/X-p53 and PD-1/PD-L1 oncogenic interactions. Targeting the PD-/PD-L1 PPI has profoundly advanced the treatment of different types of cancers. |
| first_indexed | 2024-03-10T18:44:24Z |
| format | Article |
| id | doaj.art-45da7299931d40908422397680daff30 |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-03-10T18:44:24Z |
| publishDate | 2020-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-45da7299931d40908422397680daff302023-11-20T05:35:49ZengMDPI AGMolecules1420-30492020-07-012513301710.3390/molecules25133017Competition NMR for Detection of Hit/Lead Inhibitors of Protein–Protein InteractionsBogdan Musielak0Weronika Janczyk1Ismael Rodriguez2Jacek Plewka3Dominik Sala4Katarzyna Magiera-Mularz5Tad Holak6Department of Organic Chemistry, Faculty of Chemistry, Jagiellonian University, Gronostajowa 2, 30-387 Krakow, PolandMax Planck Institute for Biochemistry, Am Klopferspitz 18, 82152 Martinsried, GermanyDepartment of Organic Chemistry, Faculty of Chemistry, Jagiellonian University, Gronostajowa 2, 30-387 Krakow, PolandDepartment of Organic Chemistry, Faculty of Chemistry, Jagiellonian University, Gronostajowa 2, 30-387 Krakow, PolandDepartment of Organic Chemistry, Faculty of Chemistry, Jagiellonian University, Gronostajowa 2, 30-387 Krakow, PolandDepartment of Organic Chemistry, Faculty of Chemistry, Jagiellonian University, Gronostajowa 2, 30-387 Krakow, PolandDepartment of Organic Chemistry, Faculty of Chemistry, Jagiellonian University, Gronostajowa 2, 30-387 Krakow, PolandScreening for small-molecule fragments that can lead to potent inhibitors of protein–protein interactions (PPIs) is often a laborious step as the fragments cannot dissociate the targeted PPI due to their low μM–mM affinities. Here, we describe an NMR competition assay called w-AIDA-NMR (weak-antagonist induced dissociation assay-NMR), which is sensitive to weak μM–mM ligand–protein interactions and which can be used in initial fragment screening campaigns. By introducing point mutations in the complex’s protein that is not targeted by the inhibitor, we lower the effective affinity of the complex, allowing for short fragments to dissociate the complex. We illustrate the method with the compounds that block the Mdm2/X-p53 and PD-1/PD-L1 oncogenic interactions. Targeting the PD-/PD-L1 PPI has profoundly advanced the treatment of different types of cancers.https://www.mdpi.com/1420-3049/25/13/3017NMRMdm2/p53PD-1/PD-L1protein–protein interactionsmall molecule |
| spellingShingle | Bogdan Musielak Weronika Janczyk Ismael Rodriguez Jacek Plewka Dominik Sala Katarzyna Magiera-Mularz Tad Holak Competition NMR for Detection of Hit/Lead Inhibitors of Protein–Protein Interactions Molecules NMR Mdm2/p53 PD-1/PD-L1 protein–protein interaction small molecule |
| title | Competition NMR for Detection of Hit/Lead Inhibitors of Protein–Protein Interactions |
| title_full | Competition NMR for Detection of Hit/Lead Inhibitors of Protein–Protein Interactions |
| title_fullStr | Competition NMR for Detection of Hit/Lead Inhibitors of Protein–Protein Interactions |
| title_full_unstemmed | Competition NMR for Detection of Hit/Lead Inhibitors of Protein–Protein Interactions |
| title_short | Competition NMR for Detection of Hit/Lead Inhibitors of Protein–Protein Interactions |
| title_sort | competition nmr for detection of hit lead inhibitors of protein protein interactions |
| topic | NMR Mdm2/p53 PD-1/PD-L1 protein–protein interaction small molecule |
| url | https://www.mdpi.com/1420-3049/25/13/3017 |
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