Open AccessAgonist-induced closure of constitutively open γ-aminobutyric acid channels with mutated M2 domains
Author(s) -
Zhuo Pan,
Dongxian Zhang,
Xishan Zhang,
Stuart A. Lipton
Publication year - 1997
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.94.12.6490
Subject(s) - agonist , allosteric regulation , gating , biophysics , chemistry , ligand gated ion channel , protein subunit , ion channel , ligand (biochemistry) , aminobutyric acid , receptor , biochemistry , biology , gene
Ligand-gated ion channels display a fundamental property—channels remain virtually closed at rest and open upon agonist binding. Here we show that substituting alanines for either of two amino acid residues (T314 or L317) in the M2 region of the γ-aminobutyric acid (GABA) ρ1 subunit results in spontaneous channel opening in the absence of ligand. Surprisingly, for two single point mutants (T314A or L317A), application of very low concentrations of agonist partially suppressed this spontaneous current, while higher concentrations re-activated the receptors. When both of these sites were mutated (T314A/L317A), GABA nearly completely suppressed the constitutive current and did not re-activate the current even at very high concentrations. This study provides important new insights into the structure–function relationship of ligand-gated ion channels, where modification of the structure of the channel pore region not only alters the allosteric transition of the receptor protein but also reverses the polarity of agonist regulation of channel gating. Our results suggest that the sites where these two residues are located are structurally critical for channel gating.