title: Molecular basis of Kir6.2 mutations associated with neonatal diabetes or neonatal diabetes plus neurological features.
abstract: Inwardly rectifying potassium channels (Kir channels) control cell membrane K(+) fluxes and electrical signaling in diverse cell types. Heterozygous mutations in the human Kir6.2 gene (KCNJ11), the pore-forming subunit of the ATP-sensitive (K(ATP)) channel, cause permanent neonatal diabetes mellitus (PNDM). For some mutations, PNDM is accompanied by marked developmental delay, muscle weakness, and epilepsy (severe disease). To determine the molecular basis of these different phenotypes, we expressed wild-type or mutant (R201C, Q52R, or V59G) Kir6.2/sulfonylurea receptor 1 channels in Xenopus oocytes. All mutations increased resting whole-cell K(ATP) currents by reducing channel inhibition by ATP, but, in the simulated heterozygous state, mutations causing PNDM alone (R201C) produced smaller K(ATP) currents and less change in ATP sensitivity than mutations associated with severe disease (Q52R and V59G). This finding suggests that increased K(ATP) currents hyperpolarize pancreatic beta cells and impair insulin secretion, whereas larger K(ATP) currents are required to influence extrapancreatic cell function. We found that mutations causing PNDM alone impair ATP sensitivity directly (at the binding site), whereas those associated with severe disease act indirectly by biasing the channel conformation toward the open state. The effect of the mutation on ATP sensitivity in the heterozygous state reflects the different contributions of a single subunit in the Kir6.2 tetramer to ATP inhibition and to the energy of the open state. Our results also show that mutations in the slide helix of Kir6.2 (V59G) influence the channel kinetics, providing evidence that this domain is involved in Kir channel gating, and suggest that the efficacy of sulfonylurea therapy in PNDM may vary with genotype.
authors.affiliation: University Laboratory of Physiology, Oxford University, Oxford OX1 3PT, United Kingdom.
authors.forename: Peter
authors.initials: P
authors.lastname: Proks
authors.forename: Jennifer F
authors.initials: JF
authors.lastname: Antcliff
authors.forename: Jon
authors.initials: J
authors.lastname: Lippiat
authors.forename: Anna L
authors.initials: AL
authors.lastname: Gloyn
authors.forename: Andrew T
authors.initials: AT
authors.lastname: Hattersley
authors.forename: Frances M
authors.initials: FM
authors.lastname: Ashcroft
mesh_terms: D018528:ATP-Binding Cassette Transporters; D000255:Adenosine Triphosphate / Q000378:metabolism / Q000494:pharmacology; D000818:Animals; D048909:Diabetes Complications / Q000235:genetics*; D003920:Diabetes Mellitus / Q000151:congenital* / Q000235:genetics* / Q000378:metabolism / Q000503:physiopathology; D004594:Electrophysiology; D006801:Humans; D007231:Infant, Newborn; D015640:Ion Channel Gating / Q000187:drug effects; D007700:Kinetics; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D009154:Mutation / Q000235:genetics*; D009422:Nervous System Diseases / Q000150:complications* / Q000235:genetics / Q000378:metabolism / Q000503:physiopathology; D009865:Oocytes / Q000187:drug effects / Q000378:metabolism; D024661:Potassium Channels, Inwardly Rectifying / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D051381:Rats; D011955:Receptors, Drug; D013453:Sulfonylurea Compounds / Q000494:pharmacology; D064233:Sulfonylurea Receptors; D014982:Xenopus laevis; D000099161:Kcnj11 Channel
chemical_list: D018528:ATP-Binding Cassette Transporters; D000255:Adenosine Triphosphate; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D024661:Potassium Channels, Inwardly Rectifying; D011955:Receptors, Drug; D013453:Sulfonylurea Compounds; D064233:Sulfonylurea Receptors; D000099161:Kcnj11 Channel