The N-terminal domain of gamma-aminobutyric Acid(B) receptors is sufficient to specify agonist and antagonist binding

Mol Pharmacol. 1999 Aug;56(2):448-54. doi: 10.1124/mol.56.2.448.

Abstract

The recently identified gamma-aminobutyric acid type B receptors (GABA(B)Rs) share low sequence similarity with the metabotropic glutamate (mGlu) receptors. Like the mGlu receptors, the N-terminal extracellular domain (NTED) of GABA(B)Rs is proposed to be related to bacterial periplasmic binding proteins (PBPs). However, in contrast to the mGlu receptors, the GABA(B)Rs lack a cysteine-rich region that links the PBP-like domain to the first transmembrane domain. This cysteine-rich region is necessary for the PBP-like domain of mGlu receptors to bind glutamate. To delimit the ligand-binding domain of GABA(B)Rs, we constructed a series of chimeric GABA(B)R1/mGluR1 and truncated GABA(B)R1 receptor mutants. We provide evidence that despite the lack of a cysteine-rich region, the NTED of GABA(B)Rs contains all of the structural information that is necessary and sufficient for ligand binding. Moreover, a soluble protein corresponding to the NTED of GABA(B)Rs reproduces the binding pharmacology of wild-type receptors. This demonstrates that the ligand-binding domain of the GABA(B)Rs can correctly fold when dissociated from the transmembrane domains.

MeSH terms

  • Animals
  • Antibodies / immunology
  • COS Cells
  • Cell Membrane / metabolism
  • Cells, Cultured
  • Epitopes / immunology
  • GABA-B Receptor Agonists
  • GABA-B Receptor Antagonists
  • Ligands
  • Mammals
  • Protein Conformation
  • Receptors, GABA-B / metabolism*
  • Receptors, Glutamate / genetics
  • Receptors, Glutamate / metabolism
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Spodoptera
  • Transfection

Substances

  • Antibodies
  • Epitopes
  • GABA-B Receptor Agonists
  • GABA-B Receptor Antagonists
  • Ligands
  • Receptors, GABA-B
  • Receptors, Glutamate
  • Recombinant Fusion Proteins