Identification of a domain in the delta subunit (S238-V264) of the alpha4beta3delta GABAA receptor that confers high agonist sensitivity.
We have expressed the alpha4beta3delta and alpha4beta3gamma2L subtypes of the rat GABAA receptor in Xenopus oocytes and have investigated their agonist activation properties. GABA was a more potent agonist of the alpha4beta3delta receptor (EC50 approximately 1.4 micromol/L) than of the alpha4beta3gamma2L subtype (EC50 approximately 27.6 micromol/L). Other GABAA receptor agonists (muscimol, 4,5,6,7-tetrahydroisoxazolo[5,4-c]pyridin-3-ol, imidazole-4-amino acid) displayed similar subtype selectivity. The structural determinants underlying these differences have been investigated by co-expressing chimeric delta/gamma2L subunits with alpha4 and beta3 subunits. A stretch of amino acids in the delta subunit, S238-V264, is shown to play an important role in determining both agonist potency and the efficacies of full or partial agonists. This segment includes transmembrane domain 1 and the short intracellular loop that leads to the second transmembrane domain. The effects of the competitive antagonists, bicuculline and SR95531, and the channel blocker, picrotoxin, were not significantly affected by the incorporation of chimeric subunits. As the delta and gamma2L subunits have not been previously implicated directly in agonist binding, we suggest that the effects are likely to arise from changes in the transduction mechanisms that link agonist binding to channel activation.
You
Haitao
H
Department of Pharmacology, University of Alberta, Edmonton, Alberta, Canada.
Dunn
Susan M J
SM
D000595:Amino Acid Sequence / Q000235:genetics; D000818:Animals; D001667:Binding, Competitive / Q000187:drug effects / Q000235:genetics; D002462:Cell Membrane / Q000737:chemistry / Q000187:drug effects; D018755:GABA Agonists / Q000494:pharmacology*; D018756:GABA Antagonists / Q000494:pharmacology; D015640:Ion Channel Gating / Q000187:drug effects / Q000235:genetics; D008024:Ligands; D009865:Oocytes; D017434:Protein Structure, Tertiary / Q000187:drug effects / Q000235:genetics; D021122:Protein Subunits / Q000737:chemistry / Q000187:drug effects / Q000235:genetics; D051381:Rats; D011963:Receptors, GABA-A / Q000737:chemistry* / Q000187:drug effects* / Q000235:genetics; D014982:Xenopus laevis; D005680:gamma-Aminobutyric Acid / Q000378:metabolism / Q000494:pharmacology
D018755:GABA Agonists; D018756:GABA Antagonists; D008024:Ligands; D021122:Protein Subunits; D011963:Receptors, GABA-A; D005680:gamma-Aminobutyric Acid