Abstract
Measurements of kinase activity are important for kinase-directed drug development, analysis of inhibitor structure and function, and understanding mechanisms of drug resistance. Sensitive, accurate, and miniaturized assay methods are crucial for these investigations. Here, we describe a label-free, high-throughput mass spectrometry-based assay for studying individual kinase enzymology and drug discovery in a purified system, with a focus on validated drug targets as benchmarks. We demonstrate that this approach can be adapted to many known kinase substrates and highlight the benefits of using mass spectrometry to measure kinase activity in vitro, including increased sensitivity. We speculate that this approach to measuring kinase activity will be generally applicable across most of the kinome, enabling research on understudied kinases and kinase drug discovery.
Original language | English (US) |
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Article number | 101599 |
Journal | Cell Reports Physical Science |
Volume | 4 |
Issue number | 10 |
DOIs | |
State | Published - Oct 18 2023 |
Externally published | Yes |
Keywords
- HTS
- drug discovery
- enzymology
- kinase
- lead optimization
- mass spectroscopy
ASJC Scopus subject areas
- General Chemistry
- General Materials Science
- General Engineering
- General Energy
- General Physics and Astronomy