Tandem Immunoprecipitation of Phosphotyrosine-Mass Spectrometry (TIPY-MS) Indicates C19ORF19 Becomes Tyrosine-Phosphorylated and Associated with Activated Epidermal Growth Factor Receptor
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- 作者：Jiefei Tong ; Paul Taylor ; Eleonora Jovceva ; Jonathan R. St-Germain ; Lily L. Jin ; Ana Nikolic ; Xiaoping Gu ; Zhi Hua Li ; Suzanne Trudel ; Michael F. Moran
- 刊名：Journal of Proteome Research
- 出版年：2008
- 出版时间：March 2008
- 年：2008
- 卷：7
- 期：3
- 页码：1067 - 1077
- 全文大小：1802K
- 年卷期：v.7,no.3(March 2008)
- ISSN：1535-3907
文摘
To identify phosphotyrosine (pY) sites in the epidermal growth factor receptor (EGFR) network, a tandem immunoprecipitation-mass spectrometry method (TIPY-MS) was applied wherein protease-digested EGFR immune complexes were extracted with anti-pY after Rush et al. (Nat. Biotech. 2005, 23, 94) and analyzed by LC−MS/MS. New pY sites in the pathway were found, including SOS1 Y1065, SOS2 Y1275, CBL-B Y889, and in the EGFR regulatory protein Mig-6 Y458. The novel human C19orf19 gene product was found EGFR-associated and phosphorylated at 5 tyrosines in response to EGFR activation and, therefore, represents a new component of the EGFR signaling network.