The pharmacological profile of ELIC, a prokaryotic GABA-gated receptor.
The Erwinia ligand-gated ion channel (ELIC) is a bacterial homologue of vertebrate Cys-loop ligand-gated ion channels. It is activated by GABA, and this property, combined with its structural similarity to GABA(A) and other Cys-loop receptors, makes it potentially an excellent model to probe their structure and function. Here we characterise the pharmacological profile of ELIC, examining the effects of compounds that could activate or inhibit the receptor. We confirm that a range of amino acids and classic GABA(A) receptor agonists do not elicit responses in ELIC, and we show the receptor can be at least partially activated by 5-aminovaleric acid and γ-hydroxybutyric acid, which are weak agonists. A range of GABA(A) receptor non-competitive antagonists inhibit GABA-elicited ELIC responses including α-endosulfan (IC₅₀ = 17 μM), dieldrin (IC₅₀ = 66 μM), and picrotoxinin (IC₅₀ = 96 μM) which were the most potent. Docking suggested possible interactions at the 2' and 6' pore-lining residues, and mutagenesis of these residues supports this hypothesis for α-endosulfan. A selection of compounds that act at Cys-loop and other receptors also showed some efficacy at blocking ELIC responses, but most were of low potency (IC₅₀ > 100 μM). Overall our data show that a number of compounds can inhibit ELIC, but it has limited pharmacological similarity to GLIC and to Cys-loop receptors.
Department of Biochemistry, University of Cambridge, Cambridge CB2 1QW, UK.
Andrew J
AJ
Thompson
Mona
M
Alqazzaz
Chris
C
Ulens
Sarah C R
SC
Lummis
D000595:Amino Acid Sequence; D000818:Animals; D001426:Bacterial Proteins / Q000819:agonists / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000378:metabolism; D002478:Cells, Cultured; D003198:Computer Simulation; D030562:Databases, Protein; D004885:Erwinia / Q000378:metabolism*; D005260:Female; D018755:GABA Agonists / Q000494:pharmacology*; D018756:GABA Antagonists / Q000494:pharmacology*; D007700:Kinetics; D058446:Ligand-Gated Ion Channels / Q000819:agonists / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000378:metabolism; D008024:Ligands; D008969:Molecular Sequence Data; D050505:Mutant Proteins / Q000819:agonists / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism; D018408:Patch-Clamp Techniques; D011485:Protein Binding; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000819:agonists / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism; D016415:Sequence Alignment; D014982:Xenopus laevis; D005680:gamma-Aminobutyric Acid / Q000378:metabolism*
D001426:Bacterial Proteins; D018755:GABA Agonists; D018756:GABA Antagonists; D058446:Ligand-Gated Ion Channels; D008024:Ligands; D050505:Mutant Proteins; D011994:Recombinant Proteins; D005680:gamma-Aminobutyric Acid