title: Deciphering activation of olfactory receptors using heterologous expression in Saccharomyces cerevisiae and bioluminescence resonance energy transfer.
abstract: Hetero- and homo-oligomerization of G protein-coupled receptors (GPCRs) has been addressed in the past years using various approaches such as co-immunoprecipitation, fluorescence resonance energy transfer and bioluminescence resonance energy transfer (BRET). Here, we report the methodological details from a previously published study to investigate the relationships between oligomerization and activation states of olfactory receptors (ORs). This methodology combines heterologous expression of ORs in Saccharomyces cerevisiae and BRET assays on membrane fractions, in particular, upon odorant stimulation. We have demonstrated that ORs constitutively homodimerize at the plasma membrane and that high odorant concentrations promote a conformational change of the dimer, which becomes inactive. We proposed a model in which one odorant molecule binding the dimer would induce activation, while two odorant molecules, each binding one protomer of the dimer, would blunt signaling.
authors.affiliation: Unité de Neurobiologie de l'Olfaction et Modélisation en Imagerie & Equipe Biologie de l'Olfaction et Biosenseurs, INRA, Jouy-en-Josas, France.
authors.forename: Guenhaël
authors.initials: G
authors.lastname: Sanz
authors.forename: Edith
authors.initials: E
authors.lastname: Pajot-Augy
mesh_terms: D060807:Bioluminescence Resonance Energy Transfer Techniques / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D015870:Gene Expression; D005818:Genetic Engineering / Q000379:methods*; D006801:Humans; D015151:Immunoblotting; D008024:Ligands; D008567:Membranes, Artificial; D010957:Plasmids / Q000235:genetics; D055503:Protein Multimerization; D020836:Protein Structure, Quaternary; D018035:Receptors, Odorant / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000254:growth & development; D014170:Transformation, Genetic
chemical_list: D008024:Ligands; D008567:Membranes, Artificial; D018035:Receptors, Odorant