GABAA receptor needs two homologous domains of the & beta;-subunit for activation by GABA but not by pentobarbital

Abstract

THE predominant inhibitory neurotrasmitter of the brain, GABA (γ-amino butyric acid), activates chloride-selective ion pores integral to the receptor complex. Subunits comprising the pre-sumed hetero-pentameric GABA channel have been cloned1–4, but little information is available on the domains important for activation. Rat wild-type or mutated α1-, & beta;2- and γ2-subunits (designated α, β and γ) were coexpressed in Xenopus oocytes and examined electrophy siologically. We report here the identification of two separate and homologous domains of the β-subunit, each of which contributes a tyrosine and threonine essential for activation by GABA. Conservative substitution of each of these four amino acids dramatically decreased GABA channel sensitivity to activation by GABA and the GABA agonist muscimol. These substitutions, however, did not impair activation by the barbiturate pentobarbital, indicating these two different classes of agonists activate GABA channels through distinct mechanisms. We also present evidence suggesting that the two identified domains of the β-subunit contribute a major component of the GABA receptor.