Open Access

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

  • Carine Bécamel1,
  • Nathalie Galéotti1,
  • Joël Poncet1,
  • Patrick Jouin1,
  • Aline Dumuis1,
  • Joël Bockaert1 and
  • Philippe Marin1Email author
Biological Procedures Online4:41094

https://doi.org/10.1251/bpo39

Received: 17 September 2002

Accepted: 15 November 2002

Abstract

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).

Indexing terms

proteomics spectrum analysis, mass

Notes

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