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• W2133993376 abstract "G protein-coupled receptors are known to be organized within different membrane compartments or microdomains of individual cells. Here, we have used a fluorescent A3 adenosine receptor (A3-AR) agonist, ABEA-X-BY630, and the technique of fluorescence correlation spectroscopy (FCS) to investigate the diffusional characteristics of functional agonist-occupied A3-AR complexes in single living cells. In Chinese hamster ovary cells expressing the human A3-AR, the fluorescent A3-AR agonist was able to inhibit forskolin-stimulated [3H]cAMP production (pEC50=8.57), and this was antagonized by the A3-selective antagonist MRS1220 (pK(B)=9.32). The fluorescent ligand also stimulated phosphoinositide hydrolysis (pEC50=7.34). Ligand binding to the A3-AR on the membranes of single cells and subsequent increases in single cell [Ca2+]i were monitored simultaneously in real time using confocal microscopy. FCS measurements in small-membrane microdomains (approximately 0.2 microm2) revealed two agonist-occupied A3-AR components with differing diffusion characteristics (diffusion coefficients=2.65x10(-8) and 1.19x10(-9) cm2/s, respectively). The binding of ligand to these two components was reduced from 5.1 and 14.9 to 2.6 and 3.3 receptors/microm2, respectively, by MRS1220 (100 nM). These data provide direct evidence for at least two populations of agonist-occupied A3-receptor complexes, showing different motilities within the membrane of single living cells." @default.
• W2133993376 created "2016-06-24" @default.
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• W2133993376 date "2007-10-24" @default.
• W2133993376 modified "2023-10-06" @default.
• W2133993376 title "Agonist‐occupied A₃adenosine receptors exist within heterogeneous complexes in membrane microdomains of individual living cells" @default.
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• W2133993376 doi "https://doi.org/10.1096/fj.07-8180com" @default.
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