A single amino acid in γ-aminobutyric acid ρ1 receptors affects competitive and noncompetitive components of picrotoxin inhibition

Tian Li Wang, Abigail S Hackam, William B. Guggino, Garry R. Cuttining

Research output: Contribution to journalArticle

86 Citations (Scopus)

Abstract

A class of bicuculline-insensitive γ-aminobutyric acid (GABA) receptors, GABA(C), has been identified in retina. Several lines of evidence indicate that GABA(C) receptors are formed partially or wholly of GABA ρ subunits. These receptors generate a Cl- current in response to GABA but differ from GABA(A) receptors in a number of ways. Picrotoxin, widely accepted as a noncompetitive antagonist of GABA(A) receptors, displays competitive and noncompetitive antagonism of GABA(C) receptors in perch and bovine retina and GABA ρ1 receptors expressed in Xenopus oocytes. The aim of this study was to identify the molecular basis of the two components of picrotoxin inhibition of GABA ρ1 receptors. By using a domain-swapping and mutagenesis strategy, a difference in picrotoxin sensitivity between ρ1 and ρ2 receptors was localized to a single amino acid in the putative second transmembrane domain. Substitution of this amino acid with residues found in the analogous position in highly picrotoxin-sensitive glycine α and GABA(A) subunits increased the sensitivity of ρ1 mutants 10- to 500-fold. Importantly, the competitive component of picrotoxin inhibition of the ρ1 mutant receptors was almost eliminated. These findings demonstrate that an amino acid in the putative channel domain of GABA ρ1 receptors influences picrotoxin sensitivity and mediates agonist binding by an allosteric mechanism.

Original languageEnglish
Pages (from-to)11751-11755
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume92
Issue number25
DOIs
StatePublished - Dec 5 1995
Externally publishedYes

Fingerprint

Aminobutyrates
Picrotoxin
Amino Acids
gamma-Aminobutyric Acid
GABA Receptors
Retina
Perches
GABA-A Receptor Antagonists
Bicuculline
GABA-A Receptors
Amino Acid Substitution
Xenopus
Mutagenesis
Glycine
Oocytes
GABA-C receptor

Keywords

  • chloride channel
  • ligand-gated neuroreceptor
  • mutagenesis
  • Xenopus oocytes

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

A single amino acid in γ-aminobutyric acid ρ1 receptors affects competitive and noncompetitive components of picrotoxin inhibition. / Wang, Tian Li; Hackam, Abigail S; Guggino, William B.; Cuttining, Garry R.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 92, No. 25, 05.12.1995, p. 11751-11755.

Research output: Contribution to journalArticle

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