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• W2085226739 startingPage "1666" @default.
• W2085226739 abstract "The nucleotide P2Y₁ receptor (P2Y₁R) is expressed in both the endothelial and vascular smooth muscle cells; however, its plasma membrane microregionalization and internalization in human tissues remain unknown. We report on the role of membrane rafts in P2Y₁R signaling by using sodium carbonate or OptiPrep sucrose density gradients, Western blot analysis, reduction of tissue cholesterol content, and vasomotor assays of endothelium-denuded human chorionic arteries. In tissue extracts prepared either in sodium carbonate or OptiPrep, approximately 20 to 30% of the total P2Y₁R mass consistently partitioned into raft fractions and correlated with vasomotor activity. Vessel treatment with methyl β-cyclodextrin reduced the raft partitioning of the P2Y₁R and obliterated the P2Y₁R-mediated contractions but not the vasomotor responses elicited by either serotonin or KCl. Perfusion of chorionic artery segments with 100 nM 2-methylthio ADP or 10 nM [[(1<i>R</i>,2<i>R</i>,3<i>S</i>,4<i>R</i>,5<i>S</i>)-4-[6-amino-2-(methylthio)-9<i>H</i>-purin-9-yl] 2,3dihydroxybicyclo[3.1.0]hex-1-yl]methyl] diphosphoric acid mono ester trisodium salt (MRS 2365), a selective P2Y₁R agonist, not only displaced within 4 min the P2Y₁R localization out of membrane rafts but also induced its subsequent internalization. 2′-Deoxy-<i>N</i>⁶-methyladenosine 3′,5′-bisphosphate tetrasodium salt (MRS 2179), a specific P2Y₁R antagonist, did not cause a similar displacement but blocked the agonist-induced exit from rafts. Neither adenosine nor uridine triphosphate displaced the P2Y₁R from the membrane raft, further evidencing the pharmacodynamics of the receptor-ligand interaction. Vascular reactivity assays showed fading of the ligand-induced vasoconstrictions, a finding that correlated with the P2Y₁R exit from raft domains and internalization. These results demonstrate in intact human vascular smooth muscle the association of the P2Y₁R to membrane rafts, highlighting the role of this microdomain in P2Y₁R signaling." @default.
• W2085226739 created "2016-06-24" @default.
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• W2085226739 date "2008-09-17" @default.
• W2085226739 modified "2023-10-17" @default.
• W2085226739 title "P2Y₁Receptor Activation Elicits Its Partition out of Membrane Rafts and Its Rapid Internalization from Human Blood Vessels: Implications for Receptor Signaling" @default.
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• W2085226739 doi "https://doi.org/10.1124/mol.108.048496" @default.
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