• Brandon C Drafts and Janet L Fisher.
• Department of Pharmacology, Physiology, and Neuroscience, University of South Carolina School of Medicine, Columbia, SC 29208, USA.
• J. Pharmacol. Exp. Ther. 2006 Sep 1;318(3):1094-101.

AbstractBarbiturates act on GABA(A) receptors (GABARs) through three distinct mechanisms, resulting in positive allosteric modulation, direct activation, and inhibition. These effects are observed at different concentrations and are differentially affected by some mutations and by the receptor's subunit composition. Mammalian GABARs can be formed from a combination of 16 different subunit subtypes. Although the effect of barbiturates depends largely on the beta subunit, their agonist activity is substantially influenced by the alpha subunit subtype. Pentobarbital is a more effective agonist than GABA only when receptors contain an alpha6 subunit. Results from chimeric alpha1/alpha6 subunits suggested that structural differences within the extracellular N-terminal domain were responsible for this characteristic. Within this domain, we examined 15 amino acid residues unique to the alpha6 subtype. Each of these sites was individually mutated in the alpha6 subunit to the corresponding residue of the alpha1 subunit. The effect of the mutation on direct activation by pentobarbital was determined with whole-cell electrophysiological recordings. Our results indicate that only one of these mutations, alpha6(T69K), altered pentobarbital efficacy. This single mutation reduced the response to pentobarbital to a level intermediate to the wild-type alpha1beta1gamma2L and alpha6beta1gamma2L isoforms. The mutation did not affect the sensitivity of the receptor to GABA but did reduce the efficacy of etomidate, another i.v. anesthetic with activity similar to pentobarbital. The reverse mutation in the alpha1 subunit (K70T) did not alter the response to pentobarbital. This is the first identification of a structural difference in GABAR alpha subtypes that regulates direct activation by barbiturates.

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