Authors
Carine Bécamel, Nathalie Galéotti, Joël Poncet, Patrick Jouin, Aline Dumuis, Joël Bockaert, Philippe Marin
Summary
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).Further details
The protocol was imported from Springer Protocols. To see the entire protocol, click on the source link, or download the pdf here.Advertisement
Stats
- Recommendations n/a n/a positive of 0 vote(s)
- Views 262
- Comments 0