title: The phosphatidylinositol 3-kinase inhibitor LY294002 potently blocks K(V) currents via a direct mechanism.
abstract: Voltage-dependent K+ (Kv) channels negatively regulate Ca2+ entry into pancreatic beta-cells by repolarizing glucose-stimulated action potentials. A role for phosphatidylinositol 3-kinase (PI3K) modulation of Kv channel function was investigated using the PI3K inhibitors wortmannin and LY294002, and LY303511, a negative control compound with respect to PI3K activity. In MIN6 insulinoma cells, wortmannin (100 nM) had no effect on whole-cell outward K+ currents, but LY294002 and LY303511 reversibly blocked currents in a dose-dependent manner (IC50=9.0+/-0.7 microM and 64.6+/-9.1 microM, respectively). Western blotting confirmed the specific inhibitory effects of LY294002 and wortmannin on insulin-stimulated PI3K activity. Kv currents in rat beta-cells at near physiological temperatures were inhibited 92% by 25 microM LY294002. Kv2.1 and Kv1.4 are highly expressed in beta-cells, and in Kv2.1-transfected tsA201 cells, 50 microM LY294002 and 100 microM LY303511 reversibly inhibited currents by 99% and 41%, respectively. In Kv1.4-transfected tsA201 cells, 50 microM LY294002 reduced the inactivation time constant from 73 to 18 ms. The insulinotropic properties of LY294002 and its effects in other excitable cells may be caused by inhibition of Kv currents rather than PI3K antagonism. Furthermore, LY294002 may represent a novel structure from which future Kv channel blockers may be developed.
authors.affiliation: Department of Medicine, University of Toronto, Ontario, Canada.
authors.forename: Wasim
authors.initials: W
authors.lastname: El-Kholy
authors.forename: Patrick E
authors.initials: PE
authors.lastname: Macdonald
authors.forename: Jia-Hui
authors.initials: JH
authors.lastname: Lin
authors.forename: Jing
authors.initials: J
authors.lastname: Wang
authors.forename: Jocelyn Manning
authors.initials: JM
authors.lastname: Fox
authors.forename: Peter E
authors.initials: PE
authors.lastname: Light
authors.forename: Qinghua
authors.initials: Q
authors.lastname: Wang
authors.forename: Robert G
authors.initials: RG
authors.lastname: Tsushima
authors.forename: Michael B
authors.initials: MB
authors.lastname: Wheeler
mesh_terms: D000730:Androstadienes / Q000494:pharmacology; D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D002867:Chromones / Q000494:pharmacology*; D004004:Dichlororibofuranosylbenzimidazole / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005947:Glucose / Q000494:pharmacology; D006801:Humans; D007328:Insulin / Q000378:metabolism; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000187:drug effects / Q000378:metabolism / Q000502:physiology; D008564:Membrane Potentials / Q000187:drug effects; D009025:Morpholines / Q000494:pharmacology*; D018408:Patch-Clamp Techniques; D000081082:Phosphoinositide-3 Kinase Inhibitors; D010879:Piperazines / Q000494:pharmacology; D024642:Potassium Channels, Voltage-Gated / Q000235:genetics / Q000502:physiology*; D051381:Rats; D015398:Signal Transduction / Q000187:drug effects; D014162:Transfection; D000077191:Wortmannin
chemical_list: D000730:Androstadienes; D002867:Chromones; D004791:Enzyme Inhibitors; D007328:Insulin; D009025:Morpholines; D000081082:Phosphoinositide-3 Kinase Inhibitors; D010879:Piperazines; D024642:Potassium Channels, Voltage-Gated; C093555:LY 303511; C085911:2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one; D004004:Dichlororibofuranosylbenzimidazole; D005947:Glucose; D000077191:Wortmannin