Differences in agonist/antagonist binding affinity and receptor transduction using recombinant human gamma-aminobutyric acid type A receptors.
Using human gamma-aminobutyric acid type A (GABAA) receptor subunit combinations, expressed in cell lines and Xenopus laevis oocytes, the pharmacology of a number of ligands interacting directly with the GABA recognition site has been studied in [3H]muscimol binding and electrophysiologically. The binding affinity of GABAA agonist and antagonist ligands showed small but statistically significant dependence on the subunit composition of receptors that include gamma 2 and different alpha and beta subunits. The potency of antagonist ligands was largely independent of receptor subunit composition, whereas the composition of receptors expressed in oocytes strongly influenced the EC50 value of agonists. An apparent reciprocal correlation between subunits favoring agonist binding and antagonist binding, respectively, was observed. Whereas antagonists showed comparable potencies in binding and functional studies, the potency of agonists in binding studies was generally two to three orders of magnitude higher than the agonist potencies measured electrophysiologically. 5-(4-Piperidyl)isothiazol-3-ol, which behaves as a low efficacy partial agonist at GABAA receptors in cultured cortical neurons, showed no efficacy in oocytes, but produced pure antagonist effects with a binding/functional affinity ratio between those observed for the agonists and antagonists. It is concluded that the GABAA receptor mechanisms transducing binding into physiological response, but not the binding per se, is dependent on the receptor subunit composition.
Ebert
B
B
Department of Pharmacology, Merck Sharp and Dohme Research Laboratories, Harlow, Essex, England. bjarke@medchem.dfh.dk
Thompson
S A
SA
Saounatsou
K
K
McKernan
R
R
Krogsgaard-Larsen
P
P
Wafford
K A
KA
D000818:Animals; D001667:Binding, Competitive; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D004594:Electrophysiology; D018755:GABA Agonists / Q000378:metabolism* / Q000494:pharmacology; D018756:GABA Antagonists / Q000378:metabolism* / Q000494:pharmacology; D006801:Humans; D007555:Isoxazoles / Q000378:metabolism / Q000494:pharmacology; D007700:Kinetics; D046911:Macromolecular Substances; D009118:Muscimol / Q000378:metabolism / Q000494:pharmacology; D009865:Oocytes / Q000378:metabolism / Q000648:ultrastructure; D010880:Piperidines / Q000494:pharmacology; D011963:Receptors, GABA-A / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D014161:Transduction, Genetic; D014162:Transfection; D014316:Tritium; D014982:Xenopus laevis
D018076:DNA, Complementary; D018755:GABA Agonists; D018756:GABA Antagonists; D007555:Isoxazoles; D046911:Macromolecular Substances; D010880:Piperidines; D011963:Receptors, GABA-A; D014316:Tritium; D009118:Muscimol; C023402:piperidine-4-sulfonic acid; C015542:gaboxadol