On the mechanism of nucleotide diphosphate activation of the ATP-sensitive K+ channel in ventricular cell of guinea-pig.
1. Effects of intracellular nucleotide diphosphates (NDPs) on the ATP-sensitive K+ channel (K+ATP channel) were examined in ventricular cells of guinea-pig heart, using the inside-out patch clamp technique. On formation of inside-out patches in the ATP-free internal solution, the K+ATP channel appeared and then ran down spontaneously. This run-down of the K+ATP channel activity was probably due to dephosphorylation. 2. Millimolar concentrations of various NDPs, e.g. UDP (uridine diphosphate), IDP (inosine diphosphate), CDP (cytidine diphosphate) and GDP (guanosine diphosphate), applied to the internal side of the patch membrane, induced openings of the K+ATP channel after run-down, i.e. in the dephosphorylated state. ADP opened the channel weakly at low concentrations (100 microM) but inhibited it at higher concentrations (1-10 mM). 3. NDP-induced openings of the channel were Mg2+ dependent and inhibited by ATP (100 microM) and glibenclamide (1 microM). None of nucleosides, nucleotide monophosphates nor nucleotide triphosphates induced openings of the channel. Thus, the K+ATP channel may have a Mg(2+)-dependent NDP-binding site, which induces openings of the dephosphorylated channel in ATP-free solution, in addition to the Mg(2+)-independent ATP-binding inactivation site and phosphorylation site. 4. In inside-out patches, pinacidil (a K+ATP channel opener) activated the K+ATP channel in the phosphorylated state but not in the dephosphorylated state. In the presence of NDPs (UDP, IDP, CDP, GDP), however, pinacidil (30 microM) enhanced openings of the dephosphorylated K+ATP channel prominently. 5. From the above results, we concluded that NDP-binding to the specific site has similar effects to channel phosphorylation, i.e. it keeps the K+ATP channel in an operative state in ATP-free solution and enhances the pinacidil-induced channel openings.
Tung
R T
RT
Department of Internal Medicine, Mayo Clinic, Rochester, MN 55905.
Kurachi
Y
Y
D000244:Adenosine Diphosphate / Q000502:physiology; D000255:Adenosine Triphosphate / Q000502:physiology*; D000818:Animals; D002478:Cells, Cultured; D003565:Cytidine Diphosphate / Q000502:physiology; D006153:Guanosine Diphosphate / Q000502:physiology; D006168:Guinea Pigs; D006352:Heart Ventricles / Q000378:metabolism; D007290:Inosine Diphosphate / Q000502:physiology; D009206:Myocardium / Q000378:metabolism*; D009711:Nucleotides / Q000502:physiology*; D011188:Potassium / Q000378:metabolism; D015221:Potassium Channels / Q000502:physiology*; D014530:Uridine Diphosphate / Q000502:physiology
D009711:Nucleotides; D015221:Potassium Channels; D006153:Guanosine Diphosphate; D014530:Uridine Diphosphate; D000244:Adenosine Diphosphate; D003565:Cytidine Diphosphate; D007290:Inosine Diphosphate; D000255:Adenosine Triphosphate; D011188:Potassium