MassIVE MSV000079489

Partial Public PXD003563

Ultra-deep tyrosine phosphoproteome analysis enabled by an SH2 domain-derived phosphotyrosine superbinder

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Description

We present a new strategy for systematic identification of phosphotyrosine (pTyr) by AP-MS using an SH2 domain-derived pTyr-superbinder as the affinity reagent. The superbinder allows for markedly deeper and broader coverage of the Tyr phosphoproteome than conventional anti-phosphotyrosine antibodies when an optimal amount is used. Indeed, we identified approximately 20,000 distinct phosphotyrosyl peptides and more than 10,000 pTyr sites, of which 36% are novel, from 9 cell lines using the superbinder approach. Intriguingly, tyrosine kinases, SH2 domains and phosphotyrosine phosphatases were found preferably phosphorylated, suggesting that the toolkit of kinase signaling is subjected 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 analysis of the tyrosine phosphoproteome under normal or pathological conditions. [dataset license: CC0 1.0 Universal (CC0 1.0)]

Keywords: tyrosine phosphorylation ; phosphotyrosine enrichment

Contact

Principal Investigators:
(in alphabetical order) 		hanfa Zou; Shawn Li
Submitting User: lilei
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