Cloning of a novel phosphotyrosine binding domain containing molecule, Odin, involved in signaling by receptor tyrosine kinases

Akhilesh Pandey, Blagoy Blagoev, Irina Kratchmarova, Minerva Fernandez, Mogens Nielsen, Troels Zakarias Kristiansen, Osamu Ohara, Alexandre V. Podtelejnikov, Serge Roche, Harvey F. Lodish, Matthias Mann

Research output: Contribution to journalArticlepeer-review

Abstract

We have used a proteomic approach using mass spectrometry to identify signaling molecules involved in receptor tyrosine kinase signaling pathways. Using affinity purification by anti-phosphotyrosine antibodies to enrich for tyrosine phosphorylated proteins, we have identified a novel signaling molecule in the epidermal growth factor receptor signaling pathway. This molecule, designated Odin, contains several ankyrin repeats, two sterile alpha motifs and a phosphotyrosine binding domain and is ubiquitously expressed. Using antibodies against endogenous Odin, we show that it undergoes tyrosine phosphorylation upon addition of growth factors such as EGF or PDGF but not by cytokines such as IL-3 or erythropoietin. Immunofluorescence experiments as well as Western blot analysis on subcellular fractions demonstrated that Odin is localized to the cytoplasm both before and after growth factor treatment. Deletion analysis showed that the phosphotyrosine binding domain of Odin is not required for its tyrosine phosphorylation. Overexpression of Odin, but not an unrelated adapter protein, Grb2, inhibited EGF-induced activation of c-Fos promoter. Microinjection of wild-type or a mutant version lacking the PTB domain into NIH3T3 fibroblasts inhibited PDGF-induced mitogenesis. Taken together, our results indicate that Odin may play a negative role in growth factor receptor signaling pathways.

Original languageEnglish (US)
Pages (from-to)8029-8036
Number of pages8
JournalOncogene
Volume21
Issue number52
DOIs
StatePublished - Nov 14 2002

Keywords

  • Bioinformatics
  • Mass spectrometry
  • Proteomics

ASJC Scopus subject areas

  • Molecular Biology
  • Cancer Research
  • Genetics

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