title: Stoichiometric analysis of oligomerization of membrane proteins on living cells using coiled-coil labeling and spectral imaging.
abstract: Many membrane proteins are proposed to work as oligomers; however, the conclusion is sometimes controversial, as for β2-adorenergic receptor (β2AR), which is one of the best-studied family A G-protein-coupled receptors. This is due to the lack of methods for easy and precise detection of the oligomeric state of membrane proteins on living cells. Here, we show that a combination of the coiled-coil tag-probe labeling method and spectral imaging enable a stoichiometric analysis of the oligomeric state of membrane proteins on living cells using monomeric, dimeric, and tetrameric standard membrane proteins. Using this method, we found that β2ARs do not form constitutive homooligomers, while they exhibit their functions such as the cyclic adenosine 5'-monophosphate (cAMP) signaling and internalization upon agonist stimulation.
authors: Graduate School of Pharmaceutical Sciences, Kyoto University, Sakyo-ku, Kyoto, Japan.
authors: Kenichi
authors: K
authors: Kawano
authors: Yoshiaki
authors: Y
authors: Yano
authors: Kaoru
authors: K
authors: Omae
authors: Sayaka
authors: S
authors: Matsuzaki
authors: Katsumi
authors: K
authors: Matsuzaki
mesh_terms: D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D057809:HEK293 Cells; D006801:Humans; D008565:Membrane Proteins / Q000032:analysis* / Q000737:chemistry*; D043562:Receptors, G-Protein-Coupled / Q000032:analysis / Q000737:chemistry; D017550:Spectroscopy, Fourier Transform Infrared / Q000379:methods; D013194:Staining and Labeling / Q000379:methods*; D013237:Stereoisomerism
chemical_list: D008565:Membrane Proteins; D043562:Receptors, G-Protein-Coupled