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| Subject Categories: Signal Transduction | Neuroscience |
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| The EMBO Journal
(2002) 21, 2332–2342, doi: 10.1093/emboj/21.10.2332 |
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| Synaptic multiprotein complexes associated with 5-HT2C receptors: a proteomic approach |
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| Carine Bécamel1, Gérard Alonso2, Nathalie Galéotti1, Emmanuelle Demey1, Patrick Jouin1, Christoph Ullmer3, Aline Dumuis1, Joël Bockaert1 and Philippe Marin1 |
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1 CNRS UPR9023, CCIPE 141 rue de la Cardonille, F-34094 Montpellier Cedex 05, France 2 CNRS UMR 5101, CCIPE 141 rue de la Cardonille, F-34094 Montpellier Cedex 05, France 3 Biofrontera Pharmaceuticals GmbH, Hemmelratherweg 201, D-51377 Leverkusen, Germany To whom correspondence should be addressed Philippe Marin, marin@montp.inserm.fr Received 21 January 2002; Revised 21 March 2002; Accepted 25 March 2002. |
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| Abstract |
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| Membrane-bound receptors such as tyrosine kinases and ionotropic receptors are associated with large protein networks structured by protein−protein interactions involving multidomain proteins. Although these networks have emerged as a general mechanism of cellular signalling, much less is known about the protein complexes associated with G-protein-coupled receptors (GPCRs). Using a proteomic approach based on peptide affinity chromatography followed by mass spectrometry and immunoblotting, we have identified 15 proteins that interact with the C- terminal tail of the 5-hydroxytryptamine 2C (5-HT2C) receptor, a GPCR. These proteins include several synaptic multidomain proteins containing one or several PDZ domains (PSD95 and the proteins of the tripartite complex Veli3−CASK−Mint1), proteins of the actin/spectrin cytoskeleton and signalling proteins. Coimmunoprecipitation experiments showed that 5-HT2C receptors interact with PSD95 and the Veli3−CASK−Mint1 complex in vivo. Electron microscopy also indicated a synaptic enrichment of Veli3 and 5-HT2C receptors and their colocalization in microvilli of choroidal cells. These results indicate that the 5-HT2C receptor is associated with protein networks that are important for its synaptic localization and its coupling to the signalling machinery. |
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| Keywords: 5-HT2C receptor, PDZ domain, proteomic analysis, scaffold protein |
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