期刊
NATURE CHEMICAL BIOLOGY
卷 12, 期 11, 页码 959-+出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/NCHEMBIO.2178
关键词
-
资金
- China State Key Basic Research Program [2012CB910101, 2016YFA0501402, 2013CB911202]
- National Natural Science Foundation of China [21235006, 21321064, 21535008, 81361128015]
- Canadian Cancer Society
- Canadian Institute of Health Research
- Ontario Research Fund
- National Science Fund of China [21525524]
- Canadian Research Chair in Functional Genomics and Cellular Proteomics
We present a new strategy for systematic identification of phosphotyrosine (pTyr) by affinity purification mass spectrometry (AP-MS) using a Src homology 2 (SH2)-domain-derived pTyr superbinder as the affinity reagent. The superbinder allows for markedly deeper coverage of the Tyr phosphoproteome than anti-pTyr antibodies when an optimal amount is used. We identified similar to 20,000 distinct phosphotyrosyl peptides and >10,000 pTyr sites, of which 36% were 'novel', from nine human cell lines using the superbinder approach. Tyrosine kinases, SH2 domains and phosphotyrosine phosphatases were preferably phosphorylated, suggesting that the toolkit of kinase signaling is subject to intensive regulation by phosphorylation. Cell-type-specific global kinase activation patterns inferred from label-free quantitation of Tyr phosphorylation guided the design of experiments to inhibit cancer cell proliferation by blocking the highly activated tyrosine kinases. Therefore, the superbinder is a highly efficient and cost-effective alternative to conventional antibodies for systematic and quantitative characterization of the tyrosine phosphoproteome under normal or pathological conditions.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据