title: Identification of signalling components in tyrosine kinase cascades using phosphopeptide affinity chromatography.
abstract: Various methods are now available to identify the molecular partners of the component of a signal transduction pathway. Some interactions, however, can be technically difficult to detect because they depend upon transient tyrosine phosphorylation. Here, we present a simple affinity chromatography approach based on synthetic phosphopeptides to purify potential partners of phosphotyrosine-containing proteins. With this approach, we confirm the previously characterized interaction between Grb2 and the EGF receptor, and we identify novel partners of the IGF-1 receptor and of the JAK proteins. Methenyltetrahydrofolate synthetase (MTHFS) was identified as a potential mediator of IGF-1R dependent transformation. P85alpha, the regulatory subunit of PI3 kinase, was identified as one of four proteins recruited by a phosphopeptide mimicking a motif conserved in all JAK family members.
authors.affiliation: Unité INSERM 276, Institut Pasteur, Paris, France. gdumenil@Pasteur.fr
authors.forename: G
authors.initials: G
authors.lastname: Duménil
authors.forename: M
authors.initials: M
authors.lastname: Rubini
authors.forename: G
authors.initials: G
authors.lastname: Dubois
authors.forename: R
authors.initials: R
authors.lastname: Baserga
authors.forename: M
authors.initials: M
authors.lastname: Fellous
authors.forename: S
authors.initials: S
authors.lastname: Pellegrini
mesh_terms: D048868:Adaptor Proteins, Signal Transducing; D000595:Amino Acid Sequence; D000818:Animals; D002846:Chromatography, Affinity / Q000379:methods*; D066246:ErbB Receptors / Q000378:metabolism; D051380:GRB2 Adaptor Protein; D018922:HL-60 Cells; D006801:Humans; D008969:Molecular Sequence Data; D010748:Phosphopeptides / Q000737:chemistry*; D010766:Phosphorylation; D011485:Protein Binding; D011505:Protein-Tyrosine Kinases / Q000737:chemistry / Q000378:metabolism*; D011506:Proteins / Q000378:metabolism; D017526:Receptor, IGF Type 1 / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D015398:Signal Transduction
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D051380:GRB2 Adaptor Protein; C495948:GRB2 protein, human; D010748:Phosphopeptides; D011506:Proteins; D066246:ErbB Receptors; D011505:Protein-Tyrosine Kinases; D017526:Receptor, IGF Type 1