Tandem immunoprecipitation of phosphotyrosine-mass spectrometry (TIPY-MS) indicates C19ORF19 becomes tyrosine-phosphorylated and associated with activated epidermal growth factor receptor.
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.
Tong
Jiefei
J
Program in Molecular Structure and Function, Hospital for Sick Children, Department of Molecular Genetics, University of Toronto, Toronto, Canada.
Taylor
Paul
P
Jovceva
Eleonora
E
St-Germain
Jonathan R
JR
Jin
Lily L
LL
Nikolic
Ana
A
Gu
Xiaoping
X
Li
Zhi Hua
ZH
Trudel
Suzanne
S
Moran
Michael F
MF
D000595:Amino Acid Sequence; D002460:Cell Line; D066246:ErbB Receptors / Q000378:metabolism*; D006801:Humans; D047468:Immunoprecipitation; D013058:Mass Spectrometry / Q000379:methods*; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010766:Phosphorylation; D019000:Phosphotyrosine / Q000737:chemistry*
D019000:Phosphotyrosine; D066246:ErbB Receptors