ABPP-HT - High-Throughput Activity-Based Profiling of Deubiquitylating Enzyme Inhibitors in a Cellular Context
| Autor: | Adan Pinto-Fernandez, Roman Fischer, Raphael Heilig, Hannah B. L. Jones, Benedikt M. Kessler |
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| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
activity-based probes
Chemical biology Context (language use) chemical biology Computational biology Proteomics 01 natural sciences drug discovery lcsh:Chemistry 03 medical and health sciences proteomics ubiquitin Profiling (information science) Throughput (business) deubiquitylating enzymes 030304 developmental biology Original Research Profiling (computer programming) 0303 health sciences Deubiquitylating enzyme 010405 organic chemistry Chemistry Drug discovery Target engagement General Chemistry Small molecule 0104 chemical sciences Protein profiling Workflow lcsh:QD1-999 pharmacology activitomics |
| Zdroj: | Frontiers in Chemistry, Vol 9 (2021) Frontiers in Chemistry |
| ISSN: | 2296-2646 |
| DOI: | 10.3389/fchem.2021.640105/full |
| Popis: | The potency and selectivity of a small molecule inhibitor are key parameters to assess during the early stages of drug discovery. In particular, it is very informative for characterizing compounds in a relevant cellular context in order to reveal potential off-target effects and drug efficacy. Activity-based probes (ABPs) are valuable tools for that purpose, however, obtaining cellular target engagement data in a high-throughput format has been particularly challenging. Here, we describe a new methodology named ABPP-HT (high-throughput-compatible activity-based protein profiling), implementing a semi-automated proteomic sample preparation workflow that increases the throughput capabilities of the classical ABPP workflow approximately ten times while preserving its enzyme profiling characteristics. Using a panel of deubiquitylating enzyme (DUB) inhibitors, we demonstrate the feasibility of ABPP-HT to provide compound selectivity profiles of endogenous DUBs in a cellular context at a fraction of time as compared to previous methodologies. |
| Databáze: | OpenAIRE |
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