title: Mechanism of block of hEag1 K+ channels by imipramine and astemizole.
abstract: Ether à go-go (Eag; KV10.1) voltage-gated K+ channels have been detected in cancer cell lines of diverse origin and shown to influence their rate of proliferation. The tricyclic antidepressant imipramine and the antihistamine astemizole inhibit the current through Eag1 channels and reduce the proliferation of cancer cells. Here we describe the mechanism by which both drugs block human Eag1 (hEag1) channels. Even if both drugs differ in their affinity for hEag1 channels (IC50s are approximately 2 microM for imipramine and approximately 200 nM for astemizole) and in their blocking kinetics, both drugs permeate the membrane and inhibit the hEag1 current by selectively binding to open channels. Furthermore, both drugs are weak bases and the IC50s depend on both internal an external pH, suggesting that both substances cross the membrane in their uncharged form and act from inside the cell in their charged forms. Accordingly, the block by imipramine is voltage dependent and antagonized by intracellular TEA, consistent with imipramine binding in its charged form to a site located close to the inner end of the selectivity filter. Using inside- and outside-out patch recordings, we found that a permanently charged, quaternary derivative of imipramine (N-methyl-imipramine) only blocks channels from the intracellular side of the membrane. In contrast, the block by astemizole is voltage independent. However, as astemizole competes with imipramine and intracellular TEA for binding to the channel, it is proposed to interact with an overlapping intracellular binding site. The significance of these findings, in the context of structure-function of channels of the eag family is discussed.
authors.affiliation: Abteilung Molekulare Biologie Neuronaler Signale, Max-Planck Institut für Experimentelle Medizin, 37075 Göttingen, Germany. rgarcia@gwdg.de
authors.forename: Rafael E
authors.initials: RE
authors.lastname: García-Ferreiro
authors.forename: Daniel
authors.initials: D
authors.lastname: Kerschensteiner
authors.forename: Felix
authors.initials: F
authors.lastname: Major
authors.forename: Francisco
authors.initials: F
authors.lastname: Monje
authors.forename: Walter
authors.initials: W
authors.lastname: Stühmer
authors.forename: Luis A
authors.initials: LA
authors.lastname: Pardo
mesh_terms: D016589:Astemizole / Q000494:pharmacology*; D002462:Cell Membrane / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004347:Drug Interactions; D000072237:ERG1 Potassium Channel; D051638:Ether-A-Go-Go Potassium Channels; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007099:Imipramine / Q000494:pharmacology*; D015640:Ion Channel Gating / Q000187:drug effects* / Q000502:physiology*; D007668:Kidney / Q000187:drug effects* / Q000502:physiology*; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D008954:Models, Biological; D024642:Potassium Channels, Voltage-Gated / Q000187:drug effects* / Q000502:physiology*; D011994:Recombinant Proteins / Q000378:metabolism; D013329:Structure-Activity Relationship
chemical_list: D000072237:ERG1 Potassium Channel; D051638:Ether-A-Go-Go Potassium Channels; D024642:Potassium Channels, Voltage-Gated; D011994:Recombinant Proteins; D016589:Astemizole; D007099:Imipramine