title: Transgenic mouse proteomics identifies new 14-3-3-associated proteins involved in cytoskeletal rearrangements and cell signaling.
abstract: Identification of protein-protein interactions is crucial for unraveling cellular processes and biochemical mechanisms of signal transduction. Here we describe, for the first time, the application of the tandem affinity purification (TAP) and LC-MS method to the characterization of protein complexes from transgenic mice. The TAP strategy developed in transgenic mice allows the emplacement of complexes in their physiological environment in contact with proteins that might only be specifically expressed in certain tissues while simultaneously ensuring the right stoichiometry of the TAP protein versus their binding partners and represents a novelty in proteomics approaches used so far. Mouse lines expressing TAP-tagged 14-3-3zeta protein were generated, and protein interactions were determined. 14-3-3 proteins are general regulators of cell signaling and represent up to 1% of the total brain protein. This study allowed the identification of almost 40 novel 14-3-3zeta-binding proteins. Biochemical and functional characterization of some of these interactions revealed new mechanisms of action of 14-3-3zeta in several signaling pathways, such as glutamate receptor signaling via binding to homer homolog 3 (Homer 3) and in cytoskeletal rearrangements and spine morphogenesis by binding and regulating the activity of the signaling complex formed by G protein-coupled receptor kinase-interactor 1 (GIT1) and p21-activated kinase-interacting exchange factor beta (betaPIX).
authors.affiliation: Cellzome AG and the European Molecular Biology Laboratory, Meyerhofstrasse 1, D-69117 Heidelberg, Germany. pierre-olivier.angrand@ibl.fr
authors.forename: Pierre-Olivier
authors.initials: PO
authors.lastname: Angrand
authors.forename: Inmaculada
authors.initials: I
authors.lastname: Segura
authors.forename: Pamela
authors.initials: P
authors.lastname: Völkel
authors.forename: Sonja
authors.initials: S
authors.lastname: Ghidelli
authors.forename: Rebecca
authors.initials: R
authors.lastname: Terry
authors.forename: Miro
authors.initials: M
authors.lastname: Brajenovic
authors.forename: Kristina
authors.initials: K
authors.lastname: Vintersten
authors.forename: Rüdiger
authors.initials: R
authors.lastname: Klein
authors.forename: Giulio
authors.initials: G
authors.lastname: Superti-Furga
authors.forename: Gerard
authors.initials: G
authors.lastname: Drewes
authors.forename: Bernhard
authors.initials: B
authors.lastname: Kuster
authors.forename: Tewis
authors.initials: T
authors.lastname: Bouwmeester
authors.forename: Amparo
authors.initials: A
authors.lastname: Acker-Palmer
mesh_terms: D048948:14-3-3 Proteins / Q000378:metabolism*; D000818:Animals; D002352:Carrier Proteins / Q000378:metabolism; D018797:Cell Cycle Proteins / Q000378:metabolism; D002478:Cells, Cultured; D003599:Cytoskeleton / Q000378:metabolism*; D006367:HeLa Cells; D000072233:Homer Scaffolding Proteins; D006801:Humans; D047468:Immunoprecipitation; D051379:Mice; D008815:Mice, Inbred Strains; D008822:Mice, Transgenic; D008954:Models, Biological; D046912:Multiprotein Complexes / Q000302:isolation & purification; D009419:Nerve Tissue Proteins / Q000302:isolation & purification / Q000378:metabolism*; D011485:Protein Binding; D020033:Protein Isoforms / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D015398:Signal Transduction
chemical_list: D048948:14-3-3 Proteins; D002352:Carrier Proteins; D018797:Cell Cycle Proteins; D000072233:Homer Scaffolding Proteins; D046912:Multiprotein Complexes; D009419:Nerve Tissue Proteins; D020033:Protein Isoforms; C111833:MARK3 protein, human; C122043:Mark2 protein, mouse; D017346:Protein Serine-Threonine Kinases