title: Phosphotyrosine interactome of the ErbB-receptor kinase family.
abstract: Interactions between short modified peptide motifs and modular protein domains are central events in cell signal-transduction. We determined interaction partners to all cytosolic tyrosine residues of the four members of the ErbB-receptor family in an unbiased fashion by quantitative proteomics using pull-down experiments with pairs of phosphorylated and nonphosphorylated synthetic peptides. Each receptor had characteristic preferences for interacting proteins and most interaction partners had multiple binding sites on each receptor. EGFR and ErbB4 had several docking sites for Grb2, while ErbB3 was characterized by six binding sites for PI3K. We identified STAT5 as a direct binding partner to EGFR and ErbB4 and discovered new recognition motifs for Shc and STAT5. The overall pattern of interaction partners of EGFR and ErbB4 suggests similar roles during signaling through their respective ligands. Phosphorylation kinetics of several tyrosine resides was measured by mass spectrometry and correlated with interaction partner preference. Our results demonstrate that system-wide mapping of peptide-protein interactions sites is possible, and suggest shared and unique roles of ErbB-receptor family members in downstream signaling.
authors.affiliation: Department of Biochemistry and Molecular Biology, Center for Experimental Bioinformatics, University of Southern Denmark, Odense, Denmark.
authors.forename: Waltraud X
authors.initials: WX
authors.lastname: Schulze
authors.forename: Lei
authors.initials: L
authors.lastname: Deng
authors.forename: Matthias
authors.initials: M
authors.lastname: Mann
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000737:chemistry; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D001665:Binding Sites; D066246:ErbB Receptors / Q000737:chemistry*; D051380:GRB2 Adaptor Protein / Q000737:chemistry; D006367:HeLa Cells; D006801:Humans; D047468:Immunoprecipitation; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000737:chemistry; D019869:Phosphatidylinositol 3-Kinases / Q000737:chemistry; D010766:Phosphorylation; D019000:Phosphotyrosine / Q000737:chemistry / Q000276:immunology / Q000502:physiology*; D011485:Protein Binding; D025941:Protein Interaction Mapping; D011499:Protein Processing, Post-Translational; D017434:Protein Structure, Tertiary; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000737:chemistry*; D020893:Receptor, ErbB-3 / Q000737:chemistry*; D066247:Receptor, ErbB-4; D050799:STAT5 Transcription Factor / Q000737:chemistry; D016415:Sequence Alignment; D055765:Shc Signaling Adaptor Proteins; D015398:Signal Transduction / Q000502:physiology*; D000071425:Src Homology 2 Domain-Containing, Transforming Protein 1; D049490:Systems Biology; D025521:Tumor Suppressor Proteins
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D066246:ErbB Receptors; D051380:GRB2 Adaptor Protein; D010446:Peptide Fragments; D019869:Phosphatidylinositol 3-Kinases; D019000:Phosphotyrosine; D018719:Erb-b2 Receptor Tyrosine Kinases; D020893:Receptor, ErbB-3; D066247:Receptor, ErbB-4; D050799:STAT5 Transcription Factor; D055765:Shc Signaling Adaptor Proteins; D000071425:Src Homology 2 Domain-Containing, Transforming Protein 1; D025521:Tumor Suppressor Proteins; C495948:GRB2 protein, human; C528024:SHC1 protein, human; C494092:STAT5A protein, human; C494095:STAT5B protein, human; C587689:ERBB4 protein, human