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Synaptic multiprotein complexes associated with 5‐HT 2C receptors: a proteomic approach
Author(s) -
Bécamel Carine,
Alonso Gérard,
Galéotti Nathalie,
Demey Emmanuelle,
Jouin Patrick,
Ullmer Christoph,
Dumuis Aline,
Bockaert Joël,
Marin Philippe
Publication year - 2002
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.1093/emboj/21.10.2332
Subject(s) - biology , pdz domain , microbiology and biotechnology , postsynaptic density , receptor , rhodopsin like receptors , g protein coupled receptor , scaffold protein , biochemistry , signal transduction , neurotransmission , metabotropic receptor , glutamate receptor
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‐HT 2C ) 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‐HT 2C receptors interact with PSD95 and the Veli3–CASK–Mint1 complex in vivo . Electron microscopy also indicated a synaptic enrichment of Veli3 and 5‐HT 2C receptors and their colocalization in microvilli of choroidal cells. These results indicate that the 5‐HT 2C receptor is associated with protein networks that are important for its synaptic localization and its coupling to the signalling machinery.