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Oligomerization of adenosine A2A and dopamine D2 receptors in living cells
Authors:Kamiya Toshio  Saitoh Osamu  Yoshioka Kazuaki  Nakata Hiroyasu
Affiliation:Department of Molecular Cell Signaling, Tokyo Metropolitan Institute for Neuroscience, 2-6 Musashidai, Fuchu, 183-8526, Tokyo, Japan.
Abstract:
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.
Keywords:Oligomerization   Adenosine A2A receptor   Dopamine D2 receptor   Bioluminescence resonance energy transfer   Purinergic   Striatum
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