Biochemical characterization of beta2-adrenergic receptor dimers and oligomers.
G Protein-coupled receptor dimerization/oligomerization has been well established during the last several years. Studies have demonstrated the existence of dimers/digomers both in vitro and in living cells. However, a thorough characterization of the biochemical nature of receptor dimers and oligomers as well as their occurrence at the cell surface has not been properly addressed. In this study, we show that both beta2-adrenergic receptor (beta2AR) dimers and oligomers exist at the plasma membrane and that the detection of such species, following receptor solubilization and resolution by denaturing polyacrylamide gel electrophoresis (SDS-PAGE), does not result from the formation of spurious disulfide bonds during cell lysis. Moreover, our results indicate that the biochemical nature of beta2AR dimers is different from that of the oligomers. Although both complexes are partially resistant to SDS denaturation, disulfide bonding is absolutely required for the stability of beta2AR oligomers but not dimers in SDS-PAGE. Indeed, dimeric species can be detected even in the presence of high concentrations of reducing and alkylating agents. Although the different biochemical nature of the dimers and oligomers may be indicative of distinct biological roles in cells, additional studies will be required to further elucidate the biosynthesis and function of these receptor forms.
Salahpour
Ali
A
Department of Biochemistry and Groupe de Recherche sur le Système Nerveux Autonome, Université de Montréal, 2900 Edouard Montpetit, P.O. Box 6128, Succ. Centre-Ville, Montréal H3C 3J7, Canada.
Bonin
Hélène
H
Bhalla
Suparna
S
Petäjä-Repo
Ulla
U
Bouvier
Michel
M
D015153:Blotting, Western; D002460:Cell Line; D002846:Chromatography, Affinity; D003432:Cross-Linking Reagents; D004220:Disulfides / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D019204:GTP-Binding Proteins / Q000378:metabolism; D006801:Humans; D007120:Immunochemistry; D008566:Membranes / Q000378:metabolism; D018343:Receptors, Adrenergic, beta-2 / Q000737:chemistry / Q000378:metabolism*; D014462:Ultrafiltration
D003432:Cross-Linking Reagents; D004220:Disulfides; D018343:Receptors, Adrenergic, beta-2; D019204:GTP-Binding Proteins