title: Differential agonist-induced desensitization of P2Y2 nucleotide receptors by ATP and UTP.
abstract: The equal potency and efficacy of the agonists, ATP and UTP, pharmacologically distinguish the P2Y2 receptor from other nucleotide receptors. Investigation of the desensitization of the P2Y2 receptors is complicated by the simultaneous expression of different P2 nucleotide receptor subtypes. The co-expression of multiple P2 receptor subtypes in mammalian cells may have led to contradictory reports on the efficacy of the natural agonists of the P2Y2 receptor to induce desensitization. We decided to investigate the desensitization of human and murine isoforms of the P2Y2 receptor, and to rigorously examine their signaling and desensitization properties. For these purposes, we used 1321N1 astrocytoma cells stably transfected with the human or murine P2Y2 receptor cDNA, as well as human A431 cells that endogenously express the receptor. The mobilization of intracellular calcium by extracellular nucleotides was used as a functional assay for the P2Y2 receptors. While ATP and UTP activated the murine and human P2Y2 receptors with similar potencies (EC50 values were 1.5-5.8 microM), ATP was approximately 10-fold less potent (IC50 = 9.1-21.2 microM) than UTP (IC50 = 0.7-2.9 microM) inducing homologous receptor desensitization in the cell systems examined. Individual cell analyses of the rate and dose dependency of agonist-induced desensitization demonstrated that the murine receptor was slightly more resistant to desensitization than its human counterpart. To our knowledge, this is the first individual cell study that has compared the cellular heterogeneity of the desensitized states of recombinant and endogenously expressed receptors. This comparison demonstrated that the recombinant system conserved the cellular regulatory elements needed to attenuate receptor signaling by desensitization.
authors.affiliation: Department of Chemistry, University of Puerto Rico, San Juan 00931-3346, USA.
authors.forename: B
authors.initials: B
authors.lastname: Velázquez
authors.forename: R C
authors.initials: RC
authors.lastname: Garrad
authors.forename: G A
authors.initials: GA
authors.lastname: Weisman
authors.forename: F A
authors.initials: FA
authors.lastname: González
mesh_terms: D000255:Adenosine Triphosphate / Q000819:agonists / Q000494:pharmacology*; D000818:Animals; D001254:Astrocytoma / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D015870:Gene Expression; D006801:Humans; D008856:Microscopy, Fluorescence; D018048:Receptors, Purinergic P2 / Q000235:genetics / Q000378:metabolism*; D058473:Receptors, Purinergic P2Y2; D011994:Recombinant Proteins; D015398:Signal Transduction; D013997:Time Factors; D014162:Transfection; D014407:Tumor Cells, Cultured / Q000187:drug effects / Q000378:metabolism; D014544:Uridine Triphosphate / Q000819:agonists / Q000494:pharmacology*
chemical_list: C546385:P2RY2 protein, human; D018048:Receptors, Purinergic P2; D058473:Receptors, Purinergic P2Y2; D011994:Recombinant Proteins; D000255:Adenosine Triphosphate; D002118:Calcium; D014544:Uridine Triphosphate