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Accueil > Agenda > Les séminaires Jean Roche > Réseaux multiprotéiques associés aux cibles antidépresseurs et homéostasie de (...)

Réseaux multiprotéiques associés aux cibles antidépresseurs et

Lundi 20 novembre, 11h, salle Lissitzky.


1 : The proteomic analysis of mouse choroid plexus secretome reveals a high protein secretion capacity of choroidal epithelial cells PROTEOMICS Volume 6, Issue 22, Date : No. 22 November 2006, Pages : 5941-5952 Eric Thouvenot, Mireille Lafon-Cazal, Edith Demettre, Patrick Jouin, Joël Bockaert, Philippe Marin Abstract | References | Full Text : PDF (428K)

2 : Mol Biol Cell. 2006 Nov ;17(11):4619-4631. Epub 2006 Aug 16. Related Articles, OMIM (calculated), Cited Articles, Books, LinkOut

Opposite Effects of PSD-95 and MPP3 PDZ Proteins on Serotonin 5-Hydroxytryptamine2C Receptor Desensitization and Membrane Stability.

Gavarini S, Becamel C, Altier C, Lory P, Poncet J, Wijnholds J, Bockaert J, Marin P.

*Centre National de la Recherche Scienti fique Unite Mixte de Recherche 5203, Institut National de la Sante et de la Recherche Medicale, U661, Universite Montpellier I, Universite Montpellier II, and Departement de Neurobiologie, Institut de Genomique Fonctionnelle, F-34094 Montpellier Cedex 5, France ; Faculty of Medicine, University of Calgary, Calgary, Alberta, Canada T2N 1N4.

PSD-95/Disc large/Zonula occludens 1 (PDZ) domain-containing proteins (PDZ proteins) play an important role in the targeting and the trafficking of transmembrane proteins. Our previous studies identified a set of PDZ proteins that interact with the C terminus of the serotonin 5-hydroxytryptamine (5-HT)(2C) receptor. Here, we show that the prototypic scaffolding protein postsynaptic density-95 (PSD-95) and another membrane-associated guanylate kinase, MAGUK p55 subfamily member 3 (MPP3), oppositely regulate desensitization of the receptor response in both heterologous cells and mice cortical neurons in primary culture. PSD-95 increased desensitization of the 5-HT(2C) receptor-mediated Ca(2+) response, whereas MPP3 prevented desensitization of the Ca(2+) response. The effects of the PDZ proteins on the desensitization of the Ca(2+) response were correlated with a differential regulation of cell surface expression of the receptor. Additional experiments were performed to assess how PDZ proteins globally modulate desensitization of the 5-HT(2C) receptor response in neurons, by using a peptidyl mimetic of the 5-HT(2C) receptor C terminus fused to the human immunodeficiency virus type-1 Tat protein transduction domain, which disrupts interaction between the 5-HT(2C) receptor and PDZ proteins. Transduction of this peptide inhibitor into cultured cortical neurons increased the desensitization of the 5-HT(2C) receptor-mediated Ca(2+) response. This indicates that, overall, interaction of 5-HT(2C) receptors with PDZ proteins inhibits receptor desensitization in cortical neurons.

PMID : 16914526 [PubMed - as supplied by publisher]

3 : Cell Tissue Res. 2006 Nov ;326(2):553-72. Epub 2006 Aug 1. Related Articles, OMIM (calculated), Books, LinkOut

Neuronal 5-HT metabotropic receptors : fine-tuning of their structure, signaling, and roles in synaptic modulation.

Bockaert J, Claeysen S, Becamel C, Dumuis A, Marin P.

CNRS, UMR5203, Montpellier, F-34094, France.

Serotonin (5-hydroxytryptamine, 5-HT) is, without doubt, the neurotransmitter for which the number of receptors is the highest. Fifteen genes encoding functional 5-HT receptors have been cloned in mammalian brain. 5-HT(3) receptors are ionotropic receptors, whereas all the others are metabotropic G-protein-coupled receptors (GPCRs). 5-HT receptor diversity is further increased by post-genomic modifications, such as alternative splicing (up to 10 splice variants for the 5-HT(4) receptor) or by mRNA editing in the case of 5-HT(2C) receptors. The cellular and behavioral implications of 5-HT(2C) receptor editing are of great physiological importance. Signaling of 5-HT receptors involves a great variety of pathways, but only some of these have been demonstrated in neurons. The classical view of neurotransmitter receptors localized within the synaptic cleft cannot be applied to 5-HT receptors, which are mostly (but not exclusively) localized at extra-synaptic locations either pre- or post-synaptically. 5-HT receptors are engaged in pre- or post-synaptic complexes composed of many GPCR-interacting proteins. The functions of these proteins are starting to be revealed. These proteins have been implicated in targeting, trafficking to or from the membrane, desensitization, and fine-tuning of signaling.

PMID : 16896947 [PubMed - in process]

4 : J Biol Chem. 2004 May 7 ;279(19):20257-66. Epub 2004 Feb 26. Related Articles, Gene, Gene (GeneRIF), GENSAT, HomoloGene, Nucleotide (RefSeq), OMIM (calculated), Substance via MeSH, Protein (RefSeq), Taxonomy via GenBank, UniGene, Nucleotide, Protein, GEO Profiles, Books, LinkOut

The serotonin 5-HT2A and 5-HT2C receptors interact with specific sets of PDZ proteins.

Becamel C, Gavarini S, Chanrion B, Alonso G, Galeotti N, Dumuis A, Bockaert J, Marin P.

UPR CNRS 2580 and CNRS UMR 5101, 141 rue de la Cardonille, 34094 Montpellier Cedex 5, Fra nce.

The 5-hydroxytryptamine type 2A (5-HT(2A)) receptor and the 5-HT(2C) receptor are closely related members of the G-protein-coupled receptors activated by serotonin that share very similar pharmacological profiles and cellular signaling pathways. These receptors express a canonical class I PDZ ligand (SXV) at their C-terminal extremity. Here, we have identified proteins that interact with the PDZ ligand of the 5-HT(2A) and 5-HT(2C) receptors by a proteomic approach associating affinity chromatography using immobilized synthetic peptides encompassing the PDZ ligand and mass spectrometry. We report that both receptor C termini interact with specific sets of PDZ proteins in vitro. The 5-HT(2C) receptor but not the 5-HT(2A) receptor binds to the Veli-3.CASK.Mint1 ternary complex and to SAP102. In addition, the 5-HT(2C) receptor binds more strongly to PSD-95 and MPP-3 than the 5-HT(2A) receptor. In contrast, a robust interaction between the 5-HT(2A) receptor and the channel-interacting PDZ protein CIPP was found, whereas CIPP did not significantly associate with the 5-HT(2C) receptor. We also show that residues located at the -1 position and upstream the PDZ ligand in the C terminus of the 5-HT(2A) and 5-HT(2C) receptors are major determinants in their interaction with specific PDZ proteins. Immunofluorescence and electron microscopy studies strongly suggested that these specific interactions also take place in living cells and that the 5-HT(2) receptor-PDZ protein complexes occur in intracellular compartments. The interaction of the 5-HT(2A) and the 5-HT(2C) receptor with specific sets of PDZ proteins may contribute to their different signal transduction properties.

PMID : 14988405 [PubMed - indexed for MEDLINE]

5 : Biol Proced Online. 2002 Dec 9 ;4:94-104. Related Articles, Cited Articles, Free in PMC, Cited in PMC, Books, LinkOut

A proteomic approach based on peptide affinity chromatography, 2-dimensional electrophoresis and mass spectrometry to identify multiprotein complexes interacting with membrane-bound receptors.

Becamel C, Galeotti N, Poncet J, Jouin P, Dumuis A, Bockaert J, Marin P.

CNRS UPR 9023. 141, Rue de la Cardonille, 34094 Montpellier Cedex 5. France. marin

There is accumulating evidence that membrane-bound receptors interact with many intracellular proteins. Multiprotein complexes associated with ionotropic receptors have been extensively characterized, but the identification of proteins interacting with G protein-coupled receptors (GPCRs) has so far only been achieved in a piecemeal fashion, focusing on one or two protein species. We describe a method based on peptide affinity chromatography, two-dimensional electrophoresis, mass spectrometry and immunoblotting to identify the components of multiprotein complexes interacting directly or indirectly with intracellular domains of GPCRs or, more generally, any other membrane-bound receptor. Using this global approach, we have characterized multiprotein complexes that bind to the carboxy-terminal tail of the 5-hydroxytryptamine type 2C receptor and are important for its subcellular localization in CNS cells (Becamel et al., EMBO J., 21(10) : 2332, 2002).

PMID : 12734563 [PubMed - as supplied by publisher]

6 : J Biol Chem. 2003 Jul 4 ;278(27):24438-48. Epub 2003 Apr 22. Related Articles, Cited in PMC, Books, LinkOut

Proteomic analysis of astrocytic secretion in the mouse. Comparison with the cerebrospinal fluid proteome.

Lafon-Cazal M, Adjali O, Galeotti N, Poncet J, Jouin P, Homburger V, Bockaert J, Marin P.

Unite Propre CNRS 2580, 141 rue de la Cardonille, 34094 Montpellier cedex 5, France.

Astrocytes, the most abundant cell type in the central nervous system, are intimately associated with synapses. They play a pivotal role in neuronal survival and the brain inflammatory response. Some astrocytic functions are mediated by the secretion of polypeptides. Using a proteomic approach, we have identified more than 30 proteins released by cultured astrocytes. These include proteases and protease inhibitors, carrier proteins, and antioxidant proteins. Exposing astrocytes to brefeldin A, which selectively blocks secretory vesicle assembly, suppressed the release of some of these proteins. This indicates that astrocytes secrete these proteins by a classic vesicular mechanism and others by an alternative pathway. Astrocytes isolated from different brain regions secreted a similar pattern of proteins. However, the secretion of some of them, including metalloproteinase inhibitors and apolipoprotein E, was region-specific. In addition, pro-inflammatory treatments modified the profile of astrocytic protein secretion. Finally, more than two thirds of the proteins identified in the astrocyte-conditioned medium were detectable in the mouse cerebrospinal fluid, suggesting that astrocytes contribute to the cerebrospinal fluid protein content. In conclusion, this study provides the first unbiased characterization of the major proteins released by astrocytes, which may play a crucial role in the modulation of neuronal survival and function.

PMID : 12709418 [PubMed - indexed for MEDLINE]

7 : EMBO J. 2002 May 15 ;21(10):2332-42. Related Articles, Gene, Gene (GeneRIF), Nucleotide (RefSeq), OMIM (calculated), < a CLASS="dblinks" href="">Protein (RefSeq), Cited Articles, Taxonomy via GenBank, UniGene, Nucleotide, Protein, GEO Profiles, Free in PMC, Cited in PMC, Books, LinkOut

Synaptic multiprotein complexes associated with 5-HT(2C) receptors : a proteomic approach.

Becamel C, Alonso G, Galeotti N, Demey E, Jouin P, Ullmer C, Dumuis A, Bockaert J, Marin P.

CNRS UPR9023, CCIPE 141 rue de la Cardonille, F-34094 Montpellier Cedex 05, France.

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.

PMID : 12006486 [PubMed - indexed for MEDLINE]

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