当前位置: X-MOL 学术ACS Chem. Biol. › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
Differential Kinobeads Profiling for Target Identification of Irreversible Kinase Inhibitors
ACS Chemical Biology ( IF 4 ) Pub Date : 2017-09-12 00:00:00 , DOI: 10.1021/acschembio.7b00617
Lars Dittus 1 , Thilo Werner 1 , Marcel Muelbaier 1 , Marcus Bantscheff 1
Affiliation  

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.

中文翻译:

差分Kinobeads分析用于不可逆激酶抑制剂的靶标鉴定

用Kinobeads对激酶抑制剂进行化学蛋白质组学分析,可以评估细胞系和组织中内源表达的蛋白激酶的抑制剂效能和选择性。使用一小部分靶向共价抑制剂,我们证明了在活细胞中测量共价靶标结合的重要性。我们提出了一种共价激酶抑制剂的不同的kinobeads分析策略,其中将化合物添加到活细胞或细胞提取物中,从而能够全面评估抑制剂对共价和非共价靶标的选择性。我们发现Acalabrutinib,CC-292和Ibrutinib有效和共价结合TEC家族激酶,但只有Ibrutinib也有效结合BLK。由于Cys22的共价修饰,ZAK被鉴定为亚微摩尔亲和力的依鲁替尼脱靶。
更新日期:2017-09-12
down
wechat
bug