Interaction of allosteric ligands with GABAA receptors containing one, two, or three different subunits.
The presence of allosteric binding sites on recombinant GABAA receptors formed after transfection of human embryonic kidney (HEK) 293 cells with alpha 1-, beta 3-, or gamma 2-subunits, or with various combinations of these subunits, was systematically investigated. From all possible subunit combinations, high affinity [3H]muscimol binding sites were induced in cells transfected with alpha 1 beta 3- or alpha 1 beta 3 gamma 2-subunits only. GABAA receptor associated [3H]flunitrazepam binding sites were induced in cells after transfection with alpha 1 gamma 2- or alpha 1 beta 3, gamma 2-subunits, and [35S]r-butylbicyclophosphorothionate (TBPS) binding sites were found in cells transfected with beta 3-, beta 3 gamma 2-, alpha 1 beta 3-, or alpha 1 beta 3 gamma 2-subunits. Binding of [35S]TBPS could be inhibited by pentobarbital, etazolate, (+)-etomidate, alphaxalone, propofol, chlormethiazole, and 4'-chlorodiazepam (Ro 5-4864) with a potency which differed in cells transfected with beta 3-, beta 3 gamma 2-, alpha 1 beta 3-, or alpha 1 beta 3 gamma 2-subunits. Results obtained indicate that receptors with different subunit composition actually can be formed in HEK cells and exhibit distinct pharmacological properties.
Zezula
J
J
Department of Biochemical Psychiatry, University Clinic for Psychiatry, Vienna, Austria.
Slany
A
A
Sieghart
W
W
D000495:Allosteric Site; D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000493:pharmacokinetics; D002460:Cell Line; D003292:Convulsants / Q000493:pharmacokinetics; D005445:Flunitrazepam / Q000493:pharmacokinetics; D018755:GABA Agonists / Q000493:pharmacokinetics; D018757:GABA Modulators / Q000493:pharmacokinetics; D058787:GABA-A Receptor Antagonists; D006801:Humans; D007668:Kidney / Q000187:drug effects / Q000378:metabolism*; D008566:Membranes / Q000187:drug effects / Q000378:metabolism; D009118:Muscimol / Q000493:pharmacokinetics; D011869:Radioligand Assay; D011963:Receptors, GABA-A / Q000187:drug effects / Q000378:metabolism*
D019086:Bridged Bicyclo Compounds, Heterocyclic; D003292:Convulsants; D018755:GABA Agonists; D018757:GABA Modulators; D058787:GABA-A Receptor Antagonists; D011963:Receptors, GABA-A; D009118:Muscimol; D005445:Flunitrazepam; C037476:tert-butylbicyclophosphorothionate