Screening for EphB signaling effectors using SILAC with a linear ion trap-orbitrap mass spectrometer.
Erythropoietin-producing hepatocellular carcinoma (Eph) receptors play important roles in development, neural plasticity, and cancer. We used an Orbitrap mass spectrometer and stable isotope labeling by amino acids in cell culture (SILAC) to identify and quantify 204 proteins with significantly changed abundance in antiphosphotyrosine immunoprecipitates after ephrinB1-Fc stimulation. More than half of all known effectors downstream of EphB receptors were identified in this study, as well as numerous novel candidates for EphB signaling.
Zhang
Guoan
G
Department of Pharmacology and Kimmel Center for Biology and Medicine at the Skirball Institute, New York University School of Medicine, New York, New York 10016, USA.
Fenyö
David
D
Neubert
Thomas A
TA
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000378:metabolism*; D000818:Animals; D045744:Cell Line, Tumor; D005910:Glioma / Q000473:pathology; D006822:Hybrid Cells / Q000378:metabolism; D007553:Isotope Labeling; D013058:Mass Spectrometry; D051379:Mice; D008954:Models, Biological; D008969:Molecular Sequence Data; D009447:Neuroblastoma / Q000473:pathology; D040901:Proteomics / Q000379:methods; D051381:Rats; D036081:Receptors, Eph Family / Q000378:metabolism / Q000502:physiology*; D015398:Signal Transduction
D000596:Amino Acids; D036081:Receptors, Eph Family