Kinetics of P2X7 receptor-operated single channels currents

Biophys J. 2007 Apr 1;92(7):2377-91. doi: 10.1529/biophysj.106.091413. Epub 2006 Dec 22.

Abstract

Human P2X7 receptors were expressed in Xenopus laevis oocytes and single channels were recorded using the patch-clamp technique in the outside-out configuration. ATP4- evoked two types of P2X7 receptor-mediated single channel currents characterized by short-lived and long-lived openings. The short- and long-lasting open states had mean open times of approximately 5 and approximately 20 ms and slope conductances near -60 mV of 9 and 13 pS, respectively. The open probabilities of the short and long openings were strongly [ATP4-]-dependent with EC50 values of approximately 0.3 mM and approximately 0.1 mM ATP4-, respectively. The channel kinetics did not change significantly during sustained P2X7 receptor activation for several minutes, as was also observed in recordings in the cell-attached patch-clamp configuration. Activation and deactivation of the short openings followed exponential time courses with time constants in the range of 20 ms, and displayed a shallow [ATP4-] dependence of the activation process. The kinetics of the short channel openings at negative membrane potentials fitted well to a linear C-C-C-O model with two ATP4- binding steps at equal binding sites with a dissociation constant Kd of 139 microM.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphate / chemistry
  • Adenosine Triphosphate / metabolism*
  • Animals
  • Cells, Cultured
  • Computer Simulation
  • Ion Channel Gating / physiology*
  • Kinetics
  • Models, Biological*
  • Models, Chemical*
  • Oocytes / physiology*
  • Receptors, Purinergic P2 / chemistry*
  • Receptors, Purinergic P2 / physiology*
  • Receptors, Purinergic P2X7
  • Xenopus laevis

Substances

  • P2RX7 protein, human
  • Receptors, Purinergic P2
  • Receptors, Purinergic P2X7
  • Adenosine Triphosphate