title: Sequential Co-immunoprecipitation and Immunoblot Approach to Determine Oligomerisation of G-Protein-Coupled Receptors.
abstract: G-protein-coupled receptors (GPCRs) play a major role in psychiatric disorders and are the targets of several current therapeutic approaches in this field. A number of studies have now shown that GPCRs can assemble as high molecular weight homo- and hetero-oligomers, which could affect ligand binding, intracellular signalling or trafficking. This information could be critical in design of new drugs to treat neurological and psychiatric disorders. This chapter describes a sequential co-immunoprecipitation and immunoblot protocol for determining oligomerisation of the 5-hydroxytryptamine (HT)1Areceptor with other GPCRs in co-transfected HEK-293 cells.
authors.affiliation: Laboratory of Neuroproteomics, Department of Biochemistry and Tissue Biology, Institute of Biology, University of Campinas (UNICAMP), Cidade Universitária Zeferino Vaz, Rua Monteiro Lobato 255 F/01, 13083-862, Campinas, Brazil. paulcguest@yahoo.com.
authors.forename: Paul C
authors.initials: PC
authors.lastname: Guest
mesh_terms: D015153:Blotting, Western / Q000379:methods*; D002846:Chromatography, Affinity / Q000379:methods*; D057809:HEK293 Cells; D006801:Humans; D047468:Immunoprecipitation / Q000379:methods*; D009842:Oligopeptides; D055503:Protein Multimerization; D016271:Proto-Oncogene Proteins c-myc; D044282:Receptor, Serotonin, 5-HT1A / Q000032:analysis / Q000737:chemistry; D044302:Receptor, Serotonin, 5-HT1B / Q000032:analysis / Q000737:chemistry; D044346:Receptor, Serotonin, 5-HT1D / Q000032:analysis / Q000737:chemistry; D043562:Receptors, G-Protein-Coupled / Q000032:analysis* / Q000737:chemistry; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000737:chemistry; D014162:Transfection
chemical_list: C489427:MYC protein, human; D009842:Oligopeptides; D016271:Proto-Oncogene Proteins c-myc; D044302:Receptor, Serotonin, 5-HT1B; D044346:Receptor, Serotonin, 5-HT1D; D043562:Receptors, G-Protein-Coupled; D011993:Recombinant Fusion Proteins; D044282:Receptor, Serotonin, 5-HT1A; C085706:FLAG peptide
keywords: 5-HT; Drug target; GPCR; Neurotransmitter; Oligomerisation; Psychiatric disease