Skip to main content
Log in

Effects of palytoxin on cation occlusion and phosphorylation of the (Na+,K+)-ATPase

The Journal of Membrane Biology Aims and scope Submit manuscript

Abstract

Palytoxin (PTX) inhibits the (Na+ + K+)-driven pump and simultaneously opens channels that are equally permeable to Na+ and K+ in red cells and other cell membranes. In an effort to understand the mechanism by which PTX induces these fluxes, we have studied the effects of PTX on: 1) K+ and Na+ occlusion by the pump protein; 2) phosphorylation and dephosphorylation of the enzyme when a phosphoenzyme is formed from ATP and from Pi; and 3) p-nitro phenyl phosphatase (p-NPPase) activity associated with the (Na+,K+)-ATPase. We have found that palytoxin 1) increases the rate of deocclusion of K+(Rb+) in a time- and concentration-dependent manner, whereas Na+ occluded in the presence of oligomycin is unaffected by the toxin; 2) makes phosphorylation from Pi insensitive to K+, and 3) stimulates the p-NPPase activity. The results are consistent with the notion that PTX produces a conformation of the Na+,K+-pump that resembles the one observed when ATP is bound to its low-affinity binding site. Further, they suggest that the channels that are formed by PTX might arise as a consequence of a perturbation in the ATPase structure, leading to the loss of control of the outside “gate” of the enzyme and hence to an uncoupling of the ion transport from the catalytic function of the ATPase.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

References

  • Argüello, J.M., Kaplan, J.H. 1991. Evidence for essential carboxyls in the cation-binding domain of the Na,K-ATPase. J. Biol. Chem. 266:14627–14635

    PubMed  Google Scholar 

  • Argüello, J.M., Kaplan, J.H. 1994. Glutamate779, an intramem-brane carboxyl, is essential for monovalent cation binding by the Na,K-ATPase. J. Biol. Chem. 69:6892–6899

    Google Scholar 

  • Askari, A., Huang, W., McCormick, P. 1983. (Na + K)-dependent adenosine triphosphatase. Regulation of inorganic phosphate, magnesium ion and calcium ion interactions with the enzyme by ouabain. J. Biol. Chem. 258:3453–3460

    PubMed  CAS  Google Scholar 

  • Berberian, G., Beauge, L. 1985. Phosphatase activity of the (Na + K)-ATPase. Ligand interactions and related enzyme forms. Biochim. Biophys. Acta 821:17–29

    Article  PubMed  CAS  Google Scholar 

  • Cantley, L.C., Cantley, L.G., Josephson, L. 1978. A characterization of vanadate interactions with the (Na,K)-ATPase. J. Biol. Chem. 20:7361–7368

    Google Scholar 

  • Cornelius, F. 1995. Phosphorylation/dephosphorylation of reconstituted shark Na,K-ATPase: one phosphorylation site per alpha beta protomer. Biochim. Biophys. Acta 1235:197–204

    Article  PubMed  Google Scholar 

  • Cornelius, F., Fedosova, N.U., Klodos, I. 1998. E2P phosphoforms of Na,K-ATPase. II. Interaction of substrate and cation-binding sites in Pi phosphorylation of Na,K-ATPase. Biochem. 37:16686–16696

    Article  CAS  Google Scholar 

  • Esmann, M. 1994. Influence of Na on conformational states in membrane-bound renal Na,K-ATPase. Biochem. 33:8558–8565

    Article  CAS  Google Scholar 

  • Esmann, M., Skou, J.C. 1985. Occlusion of Na by the Na,K-AT-Pase in the presence of oligomycin. Biochim. Biophys. Acta 127:857–863

    CAS  Google Scholar 

  • Fedosova, N.U., Cornelius, F., Klodos, I. 1995. Fluorescent styryl dyes as probes for Na,K-ATPase reaction mechanism: significance of the charge of the hydrophilic moiety of RH dyes. Biochem. 34:16806–16814

    Article  CAS  Google Scholar 

  • Fedosova, N.U., Cornelius, F., Klodos, I. 1998. E2P phosphoforms of Na,K-ATPase. I. Comparison of phosphointermediates formed from ATP and Pi by their reactivity toward hydroxyl-amine and vanadate. Biochem. 37:13634–13642

    Article  CAS  Google Scholar 

  • Forbush III, B. 1987. Rapid release of K and Rb from an occluded state of the Na,K-pump in the presence of ATP or ADP. J. Biol. Chem. 262:11104–11115

    Google Scholar 

  • Glynn, I.M., Hara, Y., Richards, D.E. 1984. The occlusion of sodium ions within the mammalian sodium-potassium pump: its role in sodium transport. J. Physiol. 351:531–547

    PubMed  CAS  Google Scholar 

  • Guennoun, S., Horisberger, J.-D. 2000. Structure of the 5th transmembrane segment of the Na,K-ATPase alpha subunit: a cysteine-scanning mutagenesis study. FEBS Lett. 482:144–148

    Article  PubMed  CAS  Google Scholar 

  • Guennoun, S., Horisberger, J.-D. 2002. Cysteine-scanning mutagenesis study of the sixth transmembrane segment of the Na,K-ATPase alpha subunit. FEBS Lett. 513:277–281

    Article  PubMed  CAS  Google Scholar 

  • Habermann, E. 1989. Palytoxin acts through the Na,K-ATPase. Toxicon 27:1171–1187

    Article  PubMed  CAS  Google Scholar 

  • Habermann, E., Chhatwal, G.S. 1982. Ouabain inhibits the increase due to palytoxin of cation permeability of erythrocytes. Naunyn-Schmiedeberg’s Arch. Pharmac. 319:101–107

    Article  CAS  Google Scholar 

  • Hasenauer, J., Huang, Wu-H., Askari, A. 1993. Allosteric regulation of the access channels to the Rb occlusion sites of (Na + K)-ATPase. J. Biol. Chem. 268:3289–3297

    PubMed  CAS  Google Scholar 

  • Henderson, G.R., Askari, A. 1977. Transport ATPase: further studies on the properties of the thimerosal-treated enzyme. Arch. Biochem. Biophys. 182:221–226

    Article  PubMed  CAS  Google Scholar 

  • Ibrahim, A.R., Shier, W.T. 1987. Palytoxin: mechanism of action of a potent marine toxin. L. Toxicol-Toxin Reviews 6:159–187

    CAS  Google Scholar 

  • Jensen, J., Ottolenghi, P. 1983. ATP binding to solubilized (Na + K)-ATPase. The abolition of subunit-subunit interaction and the maximum weight of the nucleotide-binding unit. J. Biol. Chem. 258:14895–14907

    Google Scholar 

  • Jørgensen, P.L., Nielsen, J.M., Rasmussen, J.H., Pedersen, P.A. 1999. Identification of amino acid residues involved in Na or K (Tl) binding in Na,K-ATPase; contribution to Na/K specificity. In : 9th International Conference on the Na,K-ATPase and related ATPases. K. Taniguchi and S.Kaya, editors, pp. 95–102.Elsevier Science B.V., Sapporo, Japan

    Google Scholar 

  • Kaplan, J.H., Lutsenko, S., Gatto, C, Daoud, S., Kenney, L.J. 1996. Ligand-induced conformational changes in the Na,K-ATPase alpha subunit. In : Na/K-ATPase and Related Transport ATPases. Structure, Mechanism and Regulation. L.A. Beauge, D.C. Gadsby and P.J. Garrahan, editors, pp. 45–55. The New York Academy of Sciences, Mar del Plata, Buenos Aires, Argentina

    Google Scholar 

  • Kaplan, J.H., Mone, M.D. 1985. Modified cation activation of the (Na + K)-ATPase following treatment with thimerosal. Arch. Biochem. Biophys. 237:386–395

    Article  PubMed  CAS  Google Scholar 

  • Lingrel, J.B., Argüello, J.M., van Huysse, J., Kuntzweiler, T.A. 1996. Cation and cardiac glycoside binding sites of the Na,K-ATPase. In : Na/K-ATPase and Related Transport ATPases. Structure, Mechanism and Regulation. Beauge, L.A. Gadsby D.C. and Garrahan, P.J. editors, pp. 194–206. The New York Academy of Sciences, Mar del Plata, Buenos Aires, Argentina

    Google Scholar 

  • Liu, L., Askari, A. 1997. Evidence for the existence of two ATP-sensitive Rb occlusion pockets within the transmembrane domains of Na/K-ATPase. J. Biol. Chem. 272:14380–14386

    Article  PubMed  CAS  Google Scholar 

  • Lutsenko, S., Anderko, R., Kaplan, J.H. 1995. Membrane disposition of the M5-M6 hairpin of Na,K-ATPase alpha subunit is ligand dependent. Proc. Natl. Acad. Sci. USA 92:7936–7940

    Article  PubMed  CAS  Google Scholar 

  • Malo, D. 1951. Hawaiian antiquities. (Translated by Dr.N. B.Emerson). Bishop Museum Press. Honolulu, Hawaii

    Google Scholar 

  • Moore, R.E., Scheuer, P.J. 1971. Palytoxin: a new marine toxin from a coelenterate. Science 172:495–498

    Article  PubMed  CAS  Google Scholar 

  • Ozaki, H., Nagase, H., Urakawa, N. 1985. Interaction of palytoxin and cardiac glycosides on erythrocyte membrane and (Na,K)-ATPase. Eur. J. Biochem. 152:475–80

    Article  PubMed  CAS  Google Scholar 

  • Pitts, B.J., Askari, A. 1971. Stimulation of the phosphatase activity of (Na,K)-ATPase preparation by ouabain. Biochim. Biophys. Acta 225:388–391

    Article  PubMed  CAS  Google Scholar 

  • Post, R.L., Hegyvary, C, Kume, S. 1972. Activation by adenosine triphosphate in the phosphorylation kinetics of sodium and potassium ion transport adenosine triphosphatase. J. Biol. Chem. 247:6530–6540

    PubMed  CAS  Google Scholar 

  • Post, R.L., Taniguchi, K., Toda, G. 1974. Synthesis of adenosine triphosphate by Na,K-ATPase. Ann. New York Acad. Sci. 242:80–91

    Article  CAS  Google Scholar 

  • Robinson, J.D. 1969. Effects of phlorizin on membrane cation-dependent adenosine triphosphatase and p-nitrophenyl phosphatase activities. Mol. Pharmacol. 5:584–592

    PubMed  CAS  Google Scholar 

  • Scheiner-Bobis, G., Heringdorf, D.M.-Z., Christ, M., Habermann, E. 1994. Palytoxin induces K efflux from yeast cells expressing the mammalian sodium pump. Molec. Pharmacol. 45:1132–1138

    CAS  Google Scholar 

  • Skou, J.C. 1974. Effect of ATP on the intermediary steps of the reaction of the (Na + K)-dependent enzyme system. III. Effect on the p-nitrophenylphosphatase activity of the system. Biochim. Biophys. Acta 339:258–273

    Article  CAS  Google Scholar 

  • Tosteson, M.T. 2000. Mechanism of action, pharmacology and toxicology. In : Seafood and freshwater toxins. L.M.Botana, editor, pp. 549 566. Marcel Dekker, New York-Basel

    Google Scholar 

  • Tosteson, M.T., Halperin, J.A., Kishi, Y., Tosteson, D.C. 1991. Palytoxin induces an increase in the cation conductance of red cells. J. Gen. Physiol. 98:969–985

    Article  PubMed  CAS  Google Scholar 

  • Tosteson, M.T., Scriven, D.R.L., Bharadwaj, A., Arnadottir, J., Tosteson, D.C. 1997. Interaction of palytoxin with the Na,K-ATPase. Where are those sites? In : Na/K-ATPase and Related Transport ATPases. Structure, Mechanism and Regulation.

  • L.A. Beauge, D.C. Gadsby and P.J. Garrahan, editors, pp. 424–425. The New York Academy of Sciences, Mar del Plata, Buenos Aires, Argentina

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Tosteson, M.T., Thomas, J., Arnadottir, J. et al. Effects of palytoxin on cation occlusion and phosphorylation of the (Na+,K+)-ATPase. J. Membrane Biol. 192, 181–189 (2003). https://doi.org/10.1007/s00232-002-1074-9

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00232-002-1074-9

Key words

Navigation