title: Allosteric modulation and accelerated resensitization of human P2X(3) receptors by cibacron blue.
abstract: The activity of ATP as a fast neurotransmitter is mediated by the P2X family of ligand-gated ion channels. P2X receptor subtypes are subject to functional modulation by a diverse set of factors, including pH, divalent cations, and temperature. The human P2X(3) (hP2X(3)) receptor subunit is expressed primarily in sensory ganglia where it exists as either a homomultimeric receptor or, in combination with P2X(2), as a heteromultimeric receptor. This article describes the allosteric modulatory effect of the putative P2X receptor antagonist cibacron blue on the activity of recombinant hP2X(3) receptors. In 1321N1 cells expressing the hP2X(3) receptor, cibacron blue mediated a 3- to 7-fold increase in both the magnitude and the potency of ATP-activated Ca(2+) influx and transmembrane currents. The half-maximal concentration of cibacron blue required to mediate maximal potentiation (EC(50) = 1.4 microM) was independent of the agonist used to activate the hP2X(3) receptor. The nonselective P2 receptor antagonist PPADS (pyridoxal-5-phosphate-6-azophenyl-2',4'-disulfonic acid) caused a rightward shift of the cibacron blue concentration-effect curve, whereas increasing concentrations of cibacron blue attenuated PPADS antagonism. In addition to potentiating the effects of ATP at the hP2X(3) receptor, cibacron blue also produced a 6-fold increase in the rate of hP2X(3) receptor recovery from desensitization (from T(1/2) = 15.9 to 2.6 min), as evidenced by its ability to restore ATP responsiveness to acutely desensitized receptors. Consistent with the properties of other ligand-gated ion channels, these results suggest that hP2X(3) receptor activity can be allosterically modulated by a ligand distinct from the endogenous agonist.
authors.affiliation: Neurological and Urological Diseases Research, Abbott Laboratories, Abbott Park, Illinois 60064-3500, USA.
authors.forename: K
authors.initials: K
authors.lastname: Alexander
authors.forename: W
authors.initials: W
authors.lastname: Niforatos
authors.forename: B
authors.initials: B
authors.lastname: Bianchi
authors.forename: E C
authors.initials: EC
authors.lastname: Burgard
authors.forename: K J
authors.initials: KJ
authors.lastname: Lynch
authors.forename: E A
authors.initials: EA
authors.lastname: Kowaluk
authors.forename: M F
authors.initials: MF
authors.lastname: Jarvis
authors.forename: T
authors.initials: T
authors.lastname: van Biesen
mesh_terms: D000255:Adenosine Triphosphate / Q000502:physiology; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D004594:Electrophysiology; D006801:Humans; D007700:Kinetics; D009865:Oocytes / Q000187:drug effects / Q000378:metabolism; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology*; D058910:Purinergic P2 Receptor Agonists; D058919:Purinergic P2 Receptor Antagonists; D011732:Pyridoxal Phosphate / Q000031:analogs & derivatives / Q000494:pharmacology; D018048:Receptors, Purinergic P2 / Q000378:metabolism*; D058477:Receptors, Purinergic P2X3; D011994:Recombinant Proteins / Q000187:drug effects / Q000378:metabolism; D014227:Triazines / Q000494:pharmacology*; D014982:Xenopus laevis
chemical_list: C546407:P2RX3 protein, human; D011500:Protein Synthesis Inhibitors; D058910:Purinergic P2 Receptor Agonists; D058919:Purinergic P2 Receptor Antagonists; D018048:Receptors, Purinergic P2; D058477:Receptors, Purinergic P2X3; D011994:Recombinant Proteins; D014227:Triazines; C077792:pyridoxal phosphate-6-azophenyl-2',4'-disulfonic acid; C012842:Cibacron Blue F 3GA; D011732:Pyridoxal Phosphate; D000255:Adenosine Triphosphate; D002118:Calcium