Oligomerization of adenosine A2A and dopamine D2 receptors in living cells.
We investigated whether oligomerization of adenosine A(2A) receptor (A(2A)R) and dopamine D(2) receptor (D(2)R) exists in living cells using modified bioluminescence resonance energy transfer (BRET(2)) technology. Fusion of these receptors to a donor, Renilla luciferase (Rluc), and to an acceptor, modified green fluorescent protein (GFP(2)), did not affect the ligand binding affinity, subcellular distribution, and coimmunoprecipitation of the receptors. BRET was detected not only between Myc-D(2)R-Rluc and A(2A)R-GFP(2) but also between HA-tagged A(2A)R-Rluc and A(2A)R-GFP(2). These results indicate A(2A)R, either homomeric or heteromeric with D(2)R, exists as an oligomer in living cells.
Kamiya
Toshio
T
Department of Molecular Cell Signaling, Tokyo Metropolitan Institute for Neuroscience, 2-6 Musashidai, Fuchu, 183-8526, Tokyo, Japan.
Saitoh
Osamu
O
Yoshioka
Kazuaki
K
Nakata
Hiroyasu
H
D002460:Cell Line; D019281:Dimerization; D004735:Energy Transfer; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D006801:Humans; D007700:Kinetics; D008024:Ligands; D008164:Luminescent Proteins / Q000378:metabolism; D008856:Microscopy, Fluorescence; D011233:Precipitin Tests; D011485:Protein Binding; D043705:Receptor, Adenosine A2A; D017448:Receptors, Dopamine D2 / Q000737:chemistry*; D018047:Receptors, Purinergic P1 / Q000737:chemistry*; D013053:Spectrophotometry; D014162:Transfection
D008024:Ligands; D008164:Luminescent Proteins; D043705:Receptor, Adenosine A2A; D017448:Receptors, Dopamine D2; D018047:Receptors, Purinergic P1; D049452:Green Fluorescent Proteins