Advertisement

Involvement of the Na,K-ATPase in the induction of ion channels by palytoxin

  • Soung Y. Kim
  • Kenneth A. Marx
  • Chau H. Wu
Original Article

Abstract

The effects of ouabain, ATP, and vanadate on palytoxin induction of ion channels were examined with the aim of elucidating the role of Na,K-ATPase in palytoxin action. Palytoxin-induced membrane depolarization of crayfish giant axons and single channel currents of frog erythrocytes and mouse neuroblastoma N1E-115 cells were examined using the intracellular microelectrode and patch-clamp techniques. External application of palytoxin in nanomolar concentrations induced depolarization in the crayfish giant axons, and the depolarization was inhibited by pretreatment of the axon with ouabain (10 μM). Internally perfused axons were less sensitive to palytoxin unless ATP (6 mM) was added internally. In patch-clamp experiments, picomolar palytoxin in the patch electrode induced single channels in both cell-attached and inside-out patches of erythrocytes and neuroblastoma cells. The induced channels had a conductance of about 10 pS, reversed near 0 mV in physiological saline solution, and was permeable to Na+, K+, Cs+, and NH inf4 sup+ , but not to choline. Single channel activities induced by palytoxin were inhibited by ouabain (10 μM) and vanadate (1 mM), but promoted by ATP (1 mM). The modulating effects of ouabain, vanadate, and ATP on palytoxin action suggest that the Na,K-ATPase is involved in the induction of single channels by palytoxin. Palytoxin-induced and ouabain-inhibitable single channels were observed in planar lipid bilayer incorporated with purified Na,K-ATPase. The results indicate that an interaction between palytoxin and Na, K-ATPase leads to opening of a 10-pS ion channel. They further raise the possibility that a channel structure may exist in the sodium pump which is uncovered by the action of palytoxin.

Key words

Na,K-ATPase Palytoxin Ion channels Ouabain ATP Vanadate 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Akera T (1984) Methods for studying digitalis receptors, Na,KATPase, and sodium pump activity in heart membranes and myocardium. In: Dhalla NS (ed) Methods in studying cardiac membranes, vol 11, CRC Press, Inc., Boca Raton, FL, pp 163–180Google Scholar
  2. Béress L (1983) Carribean palytoxin — a new tool in membrane research. In: Hucho F, Ovchinnikov YA (eds) Toxins as tools in neurochemistry. de Gruyter., Berlin, pp 83–89Google Scholar
  3. Böttinger H, Béress L, Habermann E (1986) Involvement of (Na+ + K+)-ATPase in binding and actions of palytoxin on human erythrocytes. Biochim Biophys Acta 861:164–176Google Scholar
  4. Cantley LC, Resh M, Guidotti G (1978) Vanadate inhibits the red cell (Na+,K+)-ATPase from the cytoplasmic side. Nature 272:552–554Google Scholar
  5. Castle NA, Strichartz GR (1988) Palytoxin induces a relatively non-selective cation permeability in frog sciatic nerve which can be inhibited by cardiac glycosides. Toxicon 26:941–951Google Scholar
  6. Chhatwal GS, Hessler H-J, Habermann E (1983) The action of palytoxin on erythrocytes and resealed ghosts. Naunyn-Schmiedeberg's Arch Pharmacol 323:261–268Google Scholar
  7. Ecault E, Sauviat M-P (1991) Characterization of the palytoxininduced sodium conductance in frog skeletal muscle. Br J Pharmacol 102:523–529Google Scholar
  8. Floreani M, Tessari M, Debetto P, Luciani S, Carpenedo F (1987) Effects of N-chlorobenzyl analogues of amiloride on myocardial contractility, Na-Ca exchange carrier and other cardiac enzymatic activities. Naunyn-Schmiedeberg's Arch Pharmacol 336:661–669Google Scholar
  9. Frelin C, Vigne P, Breittmayer J-P (1990a) Palytoxin acidifies chick cardiac cells and activates the Na+/H+ antiporter. FEBS Lett 264:63–66Google Scholar
  10. Frelin C, Vigne P, Breittmayer J-P (1990b) Mechanism of the cardiotoxic action of palytoxin. Mol Pharmacol 38:904–909Google Scholar
  11. Gadsby DC, Rakowski RF, De Weer P (1993) Extracellular access to the Na,K pump: Pathway similar to ion channel. Science 260:100–103PubMedGoogle Scholar
  12. Garritsen A, Ijzerman AP, Tulp MT, Cragoe EJ, Jr, Soudijn W (1991) Receptor binding profiles of amiloride analogues provide no evidence for a link between receptors and the Na+/H+ exchanger, but indicate a common structure on receptor proteins. J Receptor Res 11:891–907Google Scholar
  13. Glynn IM (1985) The Na+,K+-transporting adenosine triphosphatase. In: Martonosi AN (ed) The enzymes of biological membranes, 2nd edn, vol 3, Plenum Press, New York, pp 35–114Google Scholar
  14. Gresser MJ, Tracey AS (1990) Vanadates as phosphate analogs in biochemistry. In: Chasteen ND (ed) Vanadium in biological systems, Kluwer, Dordrecht, pp 63–79Google Scholar
  15. Habermann E (1989) Palytoxin acts through Na+,K+-ATPase. Toxicon 27:1171–1187Google Scholar
  16. Habermann E, Chhatwal GS (1982) Ouabain inhibits the increase due to palytoxin of cation permeability of erythrocytes. NaunynSchmiedeberg's Arch Pharmacol 319:101–107Google Scholar
  17. Halliwell JV, Whitaker MJ (1987) Using microelectrodes. In: Standen NB, Gray PTA, Whitaker MJ (eds) Microelectrode techniques. The Company of Biologists Limited, Cambridge, pp 1–12Google Scholar
  18. Hamill OP, Marty A, Neher E, Sakmann B, Sigworth SF (1981) Improved patch-clamp techniques for high resolution current recording from cell and cell-free membrane patches. Pflügers Arch 391:85–100Google Scholar
  19. Hilgemann DW (1994) Channel-like function of the Na,K pump probed at microsecond resolution in giant membrane patches. Science 263:1429–1432Google Scholar
  20. Huang JMC, Wu CH, Baden DG (1984) Depolarizing action of a red-tide dinoflagellate brevetoxin on axonal membranes. J Pharmacol Exp Ther 229:615–621Google Scholar
  21. Huang W-H, Kakar SS, Periyasamy SM, Askari A (1988) Use of cross-linking reagents for detection of subunit interactions of membrane-bound Na+,K+-ATPase. Meth Enzymol 156: 345–350Google Scholar
  22. Ibrahim AR, Shier WT (1987) Palytoxin: Mechanism of action of a potent marine toxin. J Toxicol — Toxin Rev 6:159–187Google Scholar
  23. Ikeda M, Mitani K, Ito (1988) Palytoxin induces a nonselective cation channel in single ventricular cells of rat. NaunynSchmiedeberg's Arch Pharmacol 337:591–593Google Scholar
  24. Ishida Y, Kajiwara A, Takagi K, Ohizumi Y, Shibata S (1985a) Dual effect of ouabain on the palytoxin-induced contraction and norepinephrine release in the guinea-pig vas deferens. J Pharmacol Exp Ther 232:551–556Google Scholar
  25. Ishida Y, Satake N, Habon H, Kitano H, Shibata A (1985b) Inhibitory effect of ouabain on the palytoxin-induced contraction of human umbilical artery. J Pharmacol Exp Ther 232:557–560Google Scholar
  26. Ito K, Karaki H, Urakawa N (1979) Effects of palytoxin on mechanical and electrical activities of guinea pig papillary muscle. Jpn J Pharmacol 29:467–476Google Scholar
  27. Karlish SJD, Beauge L, Glynn IM (1979) Vanadate inhibits (Na+ + K+)ATPase by blocking a conformational change of the unphosphorylated form. Nature 282:333–335Google Scholar
  28. Kim SY, Wu CH, Beress L (1991) Palytoxin forms ion channels through Na,K-ATPase. In: De Weer P, Kaplan JH (eds) The sodium pump: Recent developments. Rockefeller University Press, New York, pp 505–508Google Scholar
  29. Kinoshita K, Ikeda M, Ito K (1991) Properties of palytoxin-induced whole cell current in single rat ventricular myocytes. NaunynSchmiedeberg's Arch Pharmacol 344:247–251Google Scholar
  30. Kleyman TR, Cragoe EJ, Jr (1988) Amiloride and its analogs as tools in the study of ion transport. J Membr Biol 105:1–21Google Scholar
  31. Knauf H, Simon B, Wais U (1976) Non-specific inhibition of membrane-ATPase by amiloride. Naunyn-Schmiedeberg's Arch Pharmacol 292:189–192Google Scholar
  32. Kumazawa N, Tsujimoto T, Fukushima Y (1986) Influence of voltage and ATP on ion-channel of (Na,K)ATPase incorporated into solvent-free phospholipid planar bilayers. Biochem Biophys Res Commun 136:767–772Google Scholar
  33. Last TA, Gantzer ML, Tyler CD (1983) Ion-gated channel induced in planar bilayers by incorporation of (Na+,K+)-ATPase. J Biol Chem 258:2399–2404Google Scholar
  34. Lauffer L, Stengelin L, Beress L, Hucho F (1985) Palytoxin induced permeability changes in excitable membranes. Biochim Biophys Acta 818:55–60Google Scholar
  35. Läuger P (1979) A channel mechanism for electrogenic ion pumps. Biochim Biophys Acta 552:143–161Google Scholar
  36. Läuger P (1991) Electrogenic ion pumps. Sinauer Associates, Sunderland, MA, USAGoogle Scholar
  37. Levitt DG (1980) The mechanism of the sodium pump. Biochim Biophys Acta 604:321–345Google Scholar
  38. Mironova GD, Bocharnikova NI, Mirsalikhova N, Mironov GP (1986) Ion-transporting properties and ATPase activity of (Na+ + K+)-ATPase large subunit incorporated into bilayer lipid membranes. Biochim Biophys Acta 861:224–236Google Scholar
  39. Mironova GD, Zolotarjova NI (1991) Properties of the ion channel formed in bilayer lipid membranes by α-subunits of (Na+ + K+)ATPase. In: De Weer P, Kaplan JH (eds) The sodium pump: Recent developments. Rockefeller University Press, New York, pp 509–514Google Scholar
  40. Mueller P, Rudin DO (1969) Bimolecular lipid membranes: Techniques of formation, study of electrical properties, and induction of ionic gating phenomena. In: Passow H, Stämpfli R (eds) Laboratory techniques in membrane biophysics. Springer Berlin Heidelburg, New York, pp 141–156Google Scholar
  41. Muramatsu I, Uemura D, Fujiwara M, Narahashi T (1984) Characteristics of palytoxin-induced depolarization in squid axon. J Pharmacol Exp Ther 231:488–494Google Scholar
  42. Muramatsu I, Nishio M, Kigoshi S, Uemura D (1988) Single-ion channels induced by palytoxin in guinea-pig ventricular myocytes. Br J Pharmacol 93:811–816Google Scholar
  43. Ozaki H, Nagase H, Urakawa N (1984a) Involvement of the sugar moiety in the inhibitory action of the cardiac glycosides on the palytoxin-induced responses in vascular smooth muscles. J Pharmacol Exp Ther 231:153–158Google Scholar
  44. Ozaki H, Nagase H, Urakawa (1984b) Sugar moiety of cardiac glycosides is essential for the inhibitory action on the palytoxin-induced K+ release from red blood cells. FEBS Lett 173:196–198Google Scholar
  45. Ozaki H, Nagase H, Ito K, Urakawa N (1984c) Effects of palytoxin on Na, K and ATP contents of vascular smooth muscle of rabbit aorta. Jpn J Pharmacol 34:57–66Google Scholar
  46. Ozaki H, Nagase H, Urakawa N (1985) Interaction of palytoxin and cardiac glycosides on erythrocyte membrane and Na,K-ATPase. Eur J Biochem 152:475–480Google Scholar
  47. Pichon Y (1982) Effects of palytoxin on sodium and potassium permeabilities in unmyelinated axons. Toxicon 20:41–47Google Scholar
  48. Reinhardt R, Lindmann B, Anner BM (1984) Leakage-channel conductance of single (Na+ + K+)-ATPase molecules incorporated into planar bilayers by fusion of liposomes. Biochim Biophys Acta 774:147–150Google Scholar
  49. Rouzaire-Dubois B, Dubois J-M (1990) Characterization of palytoxin-induced channels in mouse neuroblastoma cells. Toxicon 28:1147–1158Google Scholar
  50. Sagar A, Rakowski RF (1994) Access channel model for the voltage dependence of the forward-running Na+/K+ pump. J Gen ] Physiol 103:869–894Google Scholar
  51. Sauviat M-P, Pater C, Berton J (1987) Does palytoxin open a sodium-sensitive channel in cardiac muscle? Toxicon 25:695–704Google Scholar
  52. Scheiner-Bobis G, Meyer zu Heringdorf D, Christ M, Habermann E (1994) Palytoxin induces K+ efflux from yeast cells expressing the mammalian sodium pump. Mol Pharmacol 45:1132–1136Google Scholar
  53. Schwartz A, Whitmer K, Grupp G, Grupp I, Adams RJ, Lee S-W (1982) Mechanism of action of digitalis: Is the Na,K-ATPase the pharmacological receptor? Ann NY Acad Sci 402:253–271Google Scholar
  54. Shamoo AE, Myers M (1974) Na+-Dependent ionophore as part of the small polypeptide of the (Na+ + K+)-ATPase from eel electroplax membrane. J Membr Biol 19, 163–178Google Scholar
  55. Soltoff SP, Mandel LJ (1983) Amiloride directly inhibits the Na,K-ATPase activity of rabbit kidney proximal tubules. Science 220:957–959Google Scholar
  56. Stengelin S, Béress L, Lauffer L, Hucho F (1983) Palytoxin — A cation ionophore? In: Hucho F, Ovchinnikov YA (eds) Toxins as tools in neurochemistry, de Gruyter, Berlin, pp 102–112Google Scholar
  57. Tatsumi M, Takahashi M, Ohizumi Y (1984) Mechanism of palytoxin-induced [3H]norepinephrine release from a rat pheochromocytoma cell line. Mol Pharmacol 25:379–383Google Scholar
  58. Tosteson MT, Halperin JA, Kishi Y, Tosteson DC (1991) Palytoxin induces an increase in the cation conductance of red cells. J Gen Physiol 98:969–985Google Scholar
  59. Van Renterghem C, Frelin C (1993) 3,4-Dichlorobenzamil-sensitive, monovalent cation channel induced by palytoxin in cultured aortic myocytes. Br J Pharmacol 109:859–865Google Scholar
  60. Weidmann S (1977) Effects of palytoxin on the electrical activity of dog and rabbit heart. Experiencia 33:1487–1489Google Scholar
  61. Wu CH, Marx KA (1987) Mechanism of the depolarizing action of palytoxin on axonal membranes. Biophys J 51:387aGoogle Scholar
  62. Yoshizumi M, Houchi H, Ishimura Y, Masuda Y, Morita K, Oka M (1991) Mechanism of palytoxin-induced Na+ influx into cultured bovine adrenal chromaffin cells: possible involvement of Na+/H+ exchange system. Neurosci Lett 130:103–106Google Scholar
  63. Zolotarjova N, Ivkova LV, Mironova GD, Mirsalikhova NM, Sokolova SF (1991) Ouabain, amiloride, and gossypol inhibit the channels formed by α-subunit of Na+,K+-ATPase in bilayer lipid membranes. In: De Weer P, Kaplan JH (eds) The sodium pump: Recent developments. Rockefeller University Press, New York, pp 519–523Google Scholar

Copyright information

© Springer-Verlag 1995

Authors and Affiliations

  • Soung Y. Kim
    • 1
  • Kenneth A. Marx
    • 1
  • Chau H. Wu
    • 1
  1. 1.Department of Molecular Pharmacology and Biological ChemistryNorthwestern University Medical SchoolChicagoUSA

Personalised recommendations