A single amino acid change within the ion-channel domain of the γ-aminobutyric acid ρ1 receptor accelerates desensitization and increases taurine agonism

Background receptors are part of the ligand-gated ion channel family of receptors that share some functional and structural features: e.g., they have four putative transmembrane domains (TM1–TM4) and the TM2-segment is presumed to form the ion-channel. GABAC receptors open chloride channels and do not desensitize even after long exposures to GABA. These receptors are highly expressed in vertebrate retina, where they may play a unique role due to their unusual biophysical and pharmacologic characteristics. s ermine whether the TM2 domain plays a role in the process of desensitization of GABAC receptors, we used site-directed mutagenesis to produce several permutations within the leucine (L9′) residue of the TM2 domain of the human GABAρ1 subunit. Recombinant receptors were expressed in Xenopus laevis oocytes and their functional and pharmacologic properties were studied by using a two-microelectrode, voltage-clamp. s l amino acid changes led to receptors that did not generate GABA-currents, whereas an Asp for Leu mutation in the well-conserved L9′ position of the ρ1 subunit (L301D-ρ1) generated a fast-desensitizing, bicuculline-resistant receptor that was antagonized by TPMPA, a specific GABAC receptor antagonist. Moreover, in contrast with wild-type ρ1 receptors, which are practically not gated by taurine, L301D-ρ1 mutant receptors generated substantial taurine-currents. sions tution of L9′ residue in the TM2 region of GABAρ1 receptor for an amino acid residue with an acidic lateral chain greatly accelerates its desensitization rate and increases taurine-agonism. This mutant will be useful to study mechanisms involved in gating and desensitization of GABAC receptors in particular, and of neurotransmitter receptors in general.