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Title: Kinetics of conformational changes revealed by voltage-clamp fluorometry give insight to desensitization at ATP-gated human P2X1 receptors.
Authors: Fryatt, Alistair G.
Evans, Richard J.
First Published: 8-Oct-2014
Publisher: American Society for Pharmacology and Experimental Therapeutics (ASPET)
Citation: Mol Pharmacol, 2014, 86 (6), pp. 707-715
Abstract: ATP acts as an extracellular signaling molecule at cell-surface P2X receptors, mediating a variety of important physiologic and pathophysiologic roles. Homomeric P2X1 receptors open on binding ATP and then transition to an ATP-bound closed, desensitized state that requires an agonist-free washout period to recover. Voltage-clamp fluorometry was used to record ion channel activity and conformational changes simultaneously at defined positions in the extracellular loop of the human P2X1 receptor during not only agonist binding and desensitization but also during recovery. ATP evoked distinct conformational changes adjacent to the agonist binding pocket in response to channel activation and desensitization. The speed of recovery of the conformational change on agonist washout was state-dependent, with a faster time constant from the open (5 seconds) compared with the desensitized (75 seconds) form of the channel. The ability of ATP to evoke channel activity on washout after desensitization was not dependent on the degree of conformational rearrangement in the extracellular loop, and desensitization was faster from the partially recovered state. An intracellular mutation in the carboxyl terminus that slowed recovery of P2X1 receptor currents (7-fold less recovery at 30 seconds) had no effect on the time course of the extracellular conformational rearrangements. This study highlights that the intracellular portion of the receptor can regulate recovery and shows for the first time that this is by a mechanism independent of changes in the extracellular domain, suggesting the existence of a distinct desensitization gate in this novel class of ligand gated ion channels.
DOI Link: 10.1124/mol.114.095307
ISSN: 0026-895X
eISSN: 1521-0111
Embargo on file until: 1-Jan-10000
Version: Post-print
Status: Peer-reviewed
Type: Journal Article
Rights: Dark archived with reference to SHERPA/RoMEO and publisher website. Copyright © 2014 by The American Society for Pharmacology and Experimental Therapeutics.
Description: This article has supplemental material available at
Appears in Collections:Published Articles, Dept. of Cell Physiology and Pharmacology

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