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Differential Kinobeads Profiling for Target Identification of Irreversible Kinase Inhibitors
- Source :
- ACS chemical biology. 12(10)
- Publication Year :
- 2017
-
Abstract
- Chemoproteomics profiling of kinase inhibitors with kinobeads enables the assessment of inhibitor potency and selectivity for endogenously expressed protein kinases in cell lines and tissues. Using a small panel of targeted covalent inhibitors, we demonstrate the importance of measuring covalent target binding in live cells. We present a differential kinobeads profiling strategy for covalent kinase inhibitors where a compound is added either to live cells or to a cell extract that enables the comprehensive assessment of inhibitor selectivity for covalent and noncovalent targets. We found that Acalabrutinib, CC-292, and Ibrutinib potently and covalently bind TEC family kinases, but only Ibrutinib also potently binds to BLK. ZAK was identified as a submicromolar affinity Ibrutinib off-target due to covalent modification of Cys22. In contrast to Ibrutinib, 5Z-7-Oxozeaenol reacted with Cys150 next to the DFG loop, demonstrating an alternative route to covalent inactivation of this kinase, e.g., to inhibit canonical TGF-β dependent processes.
- Subjects :
- 0301 basic medicine
Proteomics
Cell
Plasma protein binding
Biochemistry
Cell Line
03 medical and health sciences
chemistry.chemical_compound
Piperidines
medicine
Humans
Chemoproteomics
Protein Kinase Inhibitors
Acrylamides
B-Lymphocytes
Kinase
Adenine
General Medicine
Protein-Tyrosine Kinases
Combinatorial chemistry
030104 developmental biology
medicine.anatomical_structure
Pyrimidines
chemistry
Cell culture
Covalent bond
Ibrutinib
Pyrazines
Benzamides
Molecular Medicine
Acalabrutinib
Pyrazoles
Protein Binding
Subjects
Details
- ISSN :
- 15548937
- Volume :
- 12
- Issue :
- 10
- Database :
- OpenAIRE
- Journal :
- ACS chemical biology
- Accession number :
- edsair.doi.dedup.....2fb3fe43c1c934a226b95e10f0cd507e