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A proteomic approach based on peptide affinity chromatography, 2-dimensional electrophoresis and mass spectrometry to identify multiprotein complexes interacting with membrane-bound receptors
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...
Autores principales: | , , , , , , |
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Formato: | Texto |
Lenguaje: | English |
Publicado: |
Biological Procedures Online
2002
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC145562/ https://www.ncbi.nlm.nih.gov/pubmed/12734563 http://dx.doi.org/10.1251/bpo39 |
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author | Bécamel, Carine Galéotti, Nathalie Poncet, Joël Jouin, Patrick Dumuis, Aline Bockaert, Joël Marin, Philippe |
author_facet | Bécamel, Carine Galéotti, Nathalie Poncet, Joël Jouin, Patrick Dumuis, Aline Bockaert, Joël Marin, Philippe |
author_sort | Bécamel, Carine |
collection | PubMed |
description | 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 (Bécamel et al., EMBO J., 21(10): 2332, 2002). |
format | Text |
id | pubmed-145562 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2002 |
publisher | Biological Procedures Online |
record_format | MEDLINE/PubMed |
spelling | pubmed-1455622003-04-15 A proteomic approach based on peptide affinity chromatography, 2-dimensional electrophoresis and mass spectrometry to identify multiprotein complexes interacting with membrane-bound receptors Bécamel, Carine Galéotti, Nathalie Poncet, Joël Jouin, Patrick Dumuis, Aline Bockaert, Joël Marin, Philippe Biol Proced Online Research Article 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 (Bécamel et al., EMBO J., 21(10): 2332, 2002). Biological Procedures Online 2002-12-09 /pmc/articles/PMC145562/ /pubmed/12734563 http://dx.doi.org/10.1251/bpo39 Text en Copyright © December 12, 2002, C Bécamel et al. Published in Biological Procedures Online under license from the authors. Copying, printing, redistribution and storage permitted. |
spellingShingle | Research Article Bécamel, Carine Galéotti, Nathalie Poncet, Joël Jouin, Patrick Dumuis, Aline Bockaert, Joël Marin, Philippe A proteomic approach based on peptide affinity chromatography, 2-dimensional electrophoresis and mass spectrometry to identify multiprotein complexes interacting with membrane-bound receptors |
title | A proteomic approach based on peptide affinity chromatography, 2-dimensional electrophoresis and mass spectrometry to identify multiprotein complexes interacting with membrane-bound receptors |
title_full | A proteomic approach based on peptide affinity chromatography, 2-dimensional electrophoresis and mass spectrometry to identify multiprotein complexes interacting with membrane-bound receptors |
title_fullStr | A proteomic approach based on peptide affinity chromatography, 2-dimensional electrophoresis and mass spectrometry to identify multiprotein complexes interacting with membrane-bound receptors |
title_full_unstemmed | A proteomic approach based on peptide affinity chromatography, 2-dimensional electrophoresis and mass spectrometry to identify multiprotein complexes interacting with membrane-bound receptors |
title_short | A proteomic approach based on peptide affinity chromatography, 2-dimensional electrophoresis and mass spectrometry to identify multiprotein complexes interacting with membrane-bound receptors |
title_sort | proteomic approach based on peptide affinity chromatography, 2-dimensional electrophoresis and mass spectrometry to identify multiprotein complexes interacting with membrane-bound receptors |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC145562/ https://www.ncbi.nlm.nih.gov/pubmed/12734563 http://dx.doi.org/10.1251/bpo39 |
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