Functional characterization of P2X3 receptors fused with fluorescent proteins

Alexander Grote, Zsolt Boldogkoi, Andreas Zimmer, Christian Steinhäuser, Ronald Jabs

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

P2X receptor function in the CNS is poorly understood, and currently available data are partly inconsistent. In the presented study, we investigated P2X3 receptors stably expressed in HEK293 cells. Non-stationary noise analysis of whole cell currents and rapid ATP application through flash photolysis allowed for assessing the single channel conductance (6.6 pS) and the fast activation kinetics of the receptor (20 ms). The characteristics of channel desensitization and pharmacological properties matched previous findings. The properties of wild type receptors were compared with P2X 3 constructs carrying a fluorescent tag (ECFP or DsRed2) at the C-terminus. These fluorescently labeled subunits formed functional receptors, with neither the affinity of the ligand binding site nor channel properties (ion selectivity, gating kinetics, single channel conductance) differing from wild type. We conclude that both fusion proteins tested here are suitable for generating transgenic mice, which can be expected to promote understanding of the physiological role of P2X3 receptors in CNS signaling.

Original languageEnglish
Pages (from-to)497-506
Number of pages10
JournalMolecular Membrane Biology
Volume22
Issue number6
DOIs
Publication statusPublished - Nov 1 2005

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Keywords

  • DsRed
  • ECFP
  • Fusion proteins
  • Gating kinetics
  • P2X receptor

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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