title: Signaling initiated by overexpression of the fibroblast growth factor receptor-1 investigated by mass spectrometry.
abstract: Overexpression of the fibroblast growth factor receptor-1 (FGFR-1), a prototypic receptor tyrosine kinase, is a feature of several human tumors. In human 293 cells overexpression of the FGFR-1 leads to constitutive activation of the receptor with concomitant sustained high increase in the cellular level of phosphotyrosine-containing proteins. Here we use mass spectrometry to study the tyrosine-phosphorylated proteins induced by overexpression of the FGFR-1. Several well known components of FGFR-1 signaling were identified along with two novel candidates: NS-1-associated protein-1 and target of Myb 1-like protein. We subsequently applied mass spectrometry precursor ion scanning to identify 22 tyrosine phosphorylation sites distributed on six substrate proteins of the FGFR-1 or downstream tyrosine kinases. Novel in vivo tyrosine phosphorylation sites were found in the FGFR-1, phospholipase Cgamma, p90 ribosomal S6 kinase, cortactin, and NS-1-associated protein-1 as a result of sustained FGFR-1 signaling, and we propose these as functional links to downstream molecular and cellular processes.
authors.affiliation: MDS Proteomics A/S, Staermosegaardsvej 6, Odense DK-5230, Denmark. hinsby@plab.ku.dk
authors.forename: Anders M
authors.initials: AM
authors.lastname: Hinsby
authors.forename: Jesper V
authors.initials: JV
authors.lastname: Olsen
authors.forename: Keiryn L
authors.initials: KL
authors.lastname: Bennett
authors.forename: Matthias
authors.initials: M
authors.lastname: Mann
mesh_terms: D015153:Blotting, Western; D002460:Cell Line; D051356:Cortactin; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D034441:Heterogeneous-Nuclear Ribonucleoproteins; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D013058:Mass Spectrometry / Q000379:methods*; D008840:Microfilament Proteins / Q000378:metabolism; D010766:Phosphorylation; D011233:Precipitin Tests; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry; D016601:RNA-Binding Proteins / Q000737:chemistry; D020794:Receptor Protein-Tyrosine Kinases / Q000737:chemistry* / Q000378:metabolism; D051496:Receptor, Fibroblast Growth Factor, Type 1; D017468:Receptors, Fibroblast Growth Factor / Q000737:chemistry* / Q000378:metabolism; D038744:Ribosomal Protein S6 Kinases, 90-kDa / Q000378:metabolism; D015398:Signal Transduction; D014162:Transfection; D014443:Tyrosine / Q000378:metabolism
chemical_list: C495868:CTTN protein, human; D051356:Cortactin; D034441:Heterogeneous-Nuclear Ribonucleoproteins; D047908:Intracellular Signaling Peptides and Proteins; D008840:Microfilament Proteins; D011506:Proteins; D016601:RNA-Binding Proteins; D017468:Receptors, Fibroblast Growth Factor; C425312:SYNCRIP protein, human; C120296:TOM1 protein, human; D014443:Tyrosine; C496348:FGFR1 protein, human; D020794:Receptor Protein-Tyrosine Kinases; D051496:Receptor, Fibroblast Growth Factor, Type 1; D038744:Ribosomal Protein S6 Kinases, 90-kDa