Plasma membrane diffusion of G protein-coupled receptor oligomers.
G protein-coupled receptors are known to form homo-and heteromers at the plasma membrane, but the molecular properties of these oligomers are relatively unknown. Here, we show a method that allows the diffusion of G protein-coupled receptors oligomers in the plasma membrane to be monitored in single cells by combining Bimolecular Fluorescence Complementation and Fluorescence Correlation Spectroscopy. With this approach we have measured, for the first time, the membrane diffusional characteristics of adenosine A(1) and A(2A) receptor homo-and heterodimers in Chinese Hamster Ovary cells. Interestingly, both homodimers display similar diffusion co-efficients (D) when expressed in living cells (D=5.0 and 4.8x10(-9) cm(2)/s, respectively) but the heterodimer formed by these receptors exhibit a significantly faster plasma membrane diffusion co-efficent (D=5.6x10(-9) cm(2)/s) when compared to the adenosine A(1) receptor tagged with the full-length yellow fluorescent protein (D=4.0x10(-9) cm(2)/s). Overall, these results demonstrate differences in plasma membrane diffusion between adenosine receptor homo-and heterodimers, providing new insights into the molecular plasticity of G protein-coupled receptor oligomerization.
Briddon
Stephen J
SJ
Institute of Cell Signalling, University of Nottingham, Nottingham, NG7 2UH, UK.
Gandía
Jorge
J
Amaral
Olavo B
OB
Ferré
Sergi
S
Lluís
Carme
C
Franco
Rafael
R
Hill
Stephen J
SJ
Ciruela
Francisco
F
D000818:Animals; D001426:Bacterial Proteins / Q000378:metabolism; D016466:CHO Cells; D002462:Cell Membrane / Q000378:metabolism*; D002478:Cells, Cultured; D006224:Cricetinae; D003412:Cricetulus; D000242:Cyclic AMP / Q000494:pharmacology; D004058:Diffusion; D019281:Dimerization; D031541:Fluorescence Resonance Energy Transfer; D015151:Immunoblotting; D008164:Luminescent Proteins / Q000378:metabolism; D016133:Polymerase Chain Reaction; D021381:Protein Transport; D043682:Receptor, Adenosine A1 / Q000378:metabolism*; D043705:Receptor, Adenosine A2A / Q000378:metabolism*; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism*; D011993:Recombinant Fusion Proteins; D013050:Spectrometry, Fluorescence; D014162:Transfection
D001426:Bacterial Proteins; D008164:Luminescent Proteins; D043682:Receptor, Adenosine A1; D043705:Receptor, Adenosine A2A; D043562:Receptors, G-Protein-Coupled; D011993:Recombinant Fusion Proteins; C054562:yellow fluorescent protein, Bacteria; D000242:Cyclic AMP