Voltage- and [ATP]-dependent gating of the P2X(2) ATP receptor channel.

The Journal of General Physiology
Yuichiro FujiwaraYoshihiro Kubo

Abstract

P2X receptors are ligand-gated cation channels activated by extracellular adenosine triphosphate (ATP). Nonetheless, P2X(2) channel currents observed during the steady-state after ATP application are known to exhibit voltage dependence; there is a gradual increase in the inward current upon hyperpolarization. We used a Xenopus oocyte expression system and two-electrode voltage clamp to analyze this "activation" phase quantitatively. We characterized the conductance-voltage relationship in the presence of various [ATP], and observed that it shifted toward more depolarized potentials with increases in [ATP]. By analyzing the rate constants for the channel's transition between a closed and an open state, we showed that the gating of P2X(2) is determined in a complex way that involves both membrane voltage and ATP binding. The activation phase was similarly recorded in HEK293 cells expressing P2X(2) even by inside-out patch clamp after intensive perfusion, excluding a possibility that the gating is due to block/unblock by endogenous blocker(s) of oocytes. We investigated its structural basis by substituting a glycine residue (G344) in the second transmembrane (TM) helix, which may provide a kink that could mediate "gating." We foun...Continue Reading

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Citations

Sep 2, 2010·Purinergic Signalling·Yan Zhu, Hong-Bo Zhao
Jan 23, 2010·The Journal of Biological Chemistry·Sebastian KracunBaljit S Khakh
Oct 12, 2012·Proceedings. Biological Sciences·Minoru UchiyamaJean M P Joss
Nov 13, 2009·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Lishuang CaoR Alan North
May 28, 2014·The Journal of General Physiology·Batu Keceli, Yoshihiro Kubo
Sep 16, 2009·The Journal of Physiology·Yoshihiro KuboKoichi Nakajo
Apr 14, 2016·Proceedings of the National Academy of Sciences of the United States of America·Yuichi MinatoIchio Shimada
Mar 24, 2017·Scientific Reports·Go KasuyaOsamu Nureki
Oct 23, 2019·Proceedings of the National Academy of Sciences of the United States of America·Benjamin GeorgeMufeng Li

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Methods Mentioned

BETA
atomic force microscopy
transfection

Software Mentioned

KyPlot
pClamp9
Igor
Clampfit
Igor Pro
pCLAMP

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