title: The beta subunit increases the ginkgolide B sensitivity of inhibitory glycine receptors.
abstract: We investigated the effect of ginkgolide B (GB), a component of the extract from the leaves of the Ginkgo biloba tree, on recombinant glycine receptors (GlyRs) expressed in Xenopus oocytes by using voltage-clamp recording. GB (0.01-10 microM) inhibited glycine-induced currents of homo-oligomeric alpha1, alpha2 and alpha 3 GlyRs, with the highest potency being found at the alpha1 GlyR (IC(50) value=0.61+/-0.1 microM). Coexpression of the alpha subunits with the beta subunit resulted in a shift of the IC(50) value of GB to nanomolar values, indicating selectivity of GB for beta subunit containing GlyRs. We also analyzed the mechanism of GB inhibition and the effect of point mutations introduced into the alpha1 subunit. Our results are consistent with a channel blocking effect, since (i) GB inhibited glycine currents non-competitively, and (ii) a point mutation in the pore forming M2 domain reduced GB potency. In conclusion, GB is a potent blocker of beta subunit containing GlyR channels and hence can be used to discriminate homo- from hetero-oligomeric GlyRs. As hetero-oligomeric GlyRs are known to be synaptically localized, GB represents a channel blocker that may be employed to separate extrasynaptic from synaptic glycine currents.
authors.affiliation: Department of Neurochemistry, Max-Planck-Institute for Brain Research, Deutschordenstrasse 46, 60528 Frankfurt, Germany.
authors.forename: Elena L
authors.initials: EL
authors.lastname: Kondratskaya
authors.forename: Heinrich
authors.initials: H
authors.lastname: Betz
authors.forename: Oleg A
authors.initials: OA
authors.lastname: Krishtal
authors.forename: Bodo
authors.initials: B
authors.lastname: Laube
mesh_terms: D000818:Animals; D004224:Diterpenes / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D004347:Drug Interactions; D004558:Electric Stimulation / Q000379:methods; D005343:Fibrinolytic Agents / Q000494:pharmacology*; D046934:Ginkgolides; D005998:Glycine / Q000493:pharmacokinetics / Q000494:pharmacology; D020128:Inhibitory Concentration 50; D007783:Lactones / Q000494:pharmacology*; D008564:Membrane Potentials / Q000187:drug effects* / Q000502:physiology; D008845:Microinjections / Q000379:methods; D016296:Mutagenesis / Q000502:physiology; D009865:Oocytes; D018408:Patch-Clamp Techniques / Q000379:methods; D021122:Protein Subunits / Q000502:physiology; D018009:Receptors, Glycine / Q000502:physiology*; D011994:Recombinant Proteins; D014981:Xenopus
chemical_list: D004224:Diterpenes; D005343:Fibrinolytic Agents; D046934:Ginkgolides; D007783:Lactones; D021122:Protein Subunits; D018009:Receptors, Glycine; D011994:Recombinant Proteins; C045856:ginkgolide B; D005998:Glycine