Electrochemical evaluation of chemical selectivity of glutamate receptor ion channel proteins with a multi-channel sensor

Masao Sugawara, Ayumi Hirano, Marián Rehák, Jun Nakanishi, Kunji Kawai, Hitoshi Sato, Yoshio Umezawa

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

A new method for evaluating chemical selectivity of agonists towards receptor ion channel proteins is proposed by using glutamate receptor (GluR) ion channel proteins and their agonists N-methyl-D-aspartic acid (NMDA), L- glutamate, and (2S, 3R, 4S) isomer of 2-(carboxycyclopropyl)glycine (L-CCG- IV). Integrated multi-channel currents, corresponding to the sum of total amount of ions passed through the multiple open channels, were used as a measure of agonists' selectivity to recognize ion channel proteins and induce channel currents. GluRs isolated from rat synaptic plasma membranes were incorporated into planar bilayer lipid membranes (BLMs) formed by the folding method. The empirical factors that affect the selectivity were demonstrated: (i) the number of GluRs incorporated into BLMs varied from one membrane to another; (ii) each BLM contained different subtypes of GluRs (NMDA and/or non-NMDA subtypes); and (iii) the magnitude of multi-channel responses induced by L-glutamate at negative applied potentials was larger than at positive potentials, while those by NMDA and L-CCG-IV were linearly related to applied potentials. The chemical selectivity among NMDA, L- glutamate and L-CCG-IV for NMDA subtype of GluRs was determined with each single BLM in which only NMDA subtype of GluRs was designed to be active by inhibiting the non-NMDA subtypes using a specific antagonist DNQX. The order of selectivity among the relevant agonists for the NMDA receptor subtype was found to be L-CCG-IV>L-glutamate>NMDA, which is consistent with the order of binding affinity of these agonists towards the same NMDA subtypes. The potential use of this approach for evaluating chemical selectivity towards non-NMDA receptor subtypes of GluRs and other receptor ion channel proteins is discussed.

Original languageEnglish
Pages (from-to)425-439
Number of pages15
JournalBiosensors and Bioelectronics
Volume12
Issue number5
DOIs
Publication statusPublished - 1997

Keywords

  • Changes in membrane permeability
  • Folding method
  • Ion channel sensor
  • L- CCG-IV
  • L-glutamate
  • NMDA
  • Planar bilayer lipid membrane
  • Receptor ion channel

ASJC Scopus subject areas

  • Biotechnology
  • Biophysics
  • Biomedical Engineering
  • Electrochemistry

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