Comparison of the Na+ Pump and the Ouabain-Resistant K+ Transport System with Other Metal Ion Transport ATPases

  • Leigh English
  • Benjamin White
  • Lewis Cantley
Part of the New Horizons in Therapeutics book series (NHTH)


Digitalis has long been of medical importance and was mentioned in herbal treatment as early as 1250 (Schery, 1972). This drug entered into accepted medical practice in 1785 following the experimental observations of William Withering (Withering, 1785). Today it is commonly used as a cardiac drug to strengthen heart muscle contraction. In the 1950s the receptor for this drug was shown to be the plasma membrane ATPase, which actively pumps Na+ out of the cell and K+ into the cell to maintain cytoplasmic ion concentrations (Skou, 1965). The ability of ouabain, a member of the digitalis family of cardiac glycosides, to inhibit the Na+-K+ ATPase has been widely used to characterize this important enzyme. Recently, a gene was cloned that, when transfected into green monkey fibroblasts, rescued cells from ouabain toxicity (Levenson, 1984). Characterization of the resistant cells indicated the presence of a new ouabain-resistant potassium transport system with characteristics similar to but distinct from the native Na+-K+ ATPase (English et al., 1985a,b). In this chapter we discuss some of the structural and kinetic properties of the Na+-K+ ATPase and a family of related cation transport systems. The nature of the transport system induced by the ouabain resistance gene is discussed in relation to this family of proteins.


Sarcoplasmic Reticulum Catalytic Subunit Cardiac Glycoside Adenosine Triphosphatase Proteolytic Site 
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  1. Allen, G., Trinnaman, B. J., and Green, N.M., 1980, The primary structure of the calcium ion-transporting adenosine triphosphatase protein of rabbit skeletal sarcoplasmic reticulum, Biochem. J. 187:591–616.PubMedGoogle Scholar
  2. Avruch, J., and Fairbanks, G., 1972, Demonstration of a phosphopeptide intermediate in Mg2+-dependent Na+- and K+-stimulated adenosine triphosphatase reaction of the erythrocyte membrane, Proc. Natl. Acad. Sci. U.S.A. 69:1216–1220.PubMedCrossRefGoogle Scholar
  3. Bastide, F., Meissner, G., Fleischer, S., and Post, R. L., 1973, Similarity of the active site of phosphorylation of the adenosine triphosphatase for transport of sodium and potassium ions in kidney to that for transport of calcium ions in the sarcoplasmic reticulum of muscle, J. Biol. Chem. 248:8385–8391.PubMedGoogle Scholar
  4. Brunette, D. M. and Till, J. E., 1971, A rapid method for the isolation of L cell surface membranes using an aqueous two-phase polymer system, J. Membr. Biol. 5:215–224.CrossRefGoogle Scholar
  5. Cantley, L. C., 1981, Structure and mechanism of the (Na+, K+) ATPase, Curr. Top. Bioenerg. 11:201–237.Google Scholar
  6. Carilli, C. T., Farley, R. A., Perlman, D. M., and Cantley, L. C., 1982, The active site structure of Na+- and K+ -stimulated ATPase, J. Biol. Chem. 257:5601–5606.PubMedGoogle Scholar
  7. Castro, J., and Farley, R. A., 1979, Proteolytic fragmentation of the catalytic subunit of the sodium and potassium adenosine triphosphatase, J. Biol. Chem. 254:2221–2228.PubMedGoogle Scholar
  8. Chin, G., and Forgac, M., 1984, Purification and proteolysis of vesicles containing inside- out and rightside-out oriented reconstituted ATPase, J. Biol. Chem. 259:5255–5263.PubMedGoogle Scholar
  9. Chipman, D. M., and Lev, A., 1983, Modification of the conformational equilibria in the sodium and potassium dependent adenosinetriphosphatase with glutaraldehyde, Biochemistry 22:4450–4459.PubMedCrossRefGoogle Scholar
  10. de Meis, L., and Vianna, A. L., 1979, Energy interconversion by the Ca2+-dependent ATPase of the sarcoplasmic reticulum, Annu. Rev. Biochem. 48:275–292.PubMedCrossRefGoogle Scholar
  11. English, L. H., and Cantley, L. C., 1984, Characterization of monovalent ion transport in an insect cell line (Manduca sexta embryonic cell line CHE), J. Cell. Physiol. 121:125–132.PubMedCrossRefGoogle Scholar
  12. English, L. H. and Cantley, L. C., 1985, Delta endotoxin inhibits K+-uptake, lowers cytoplasmic pH, and inhibits a K+ ATPase in the Manduca sexta CHE cell, J. Membr. Biol. 85:199–204.PubMedCrossRefGoogle Scholar
  13. English, L. H., Epstein, J., Cantley, L., Housman, D., and Levenson, R., 1985a, Expression of an ouabain resistance gene in transfected cells, J. Biol. Chem. 260:1114–1119.PubMedGoogle Scholar
  14. English, L. H., Epstein, J., Cantley, L., Housman, D., and Levenson, R., 1985b, Ouabain treatment induces an amiloride-sensitive K+ transport system in cells transfected with the ouabain resistance gene, in: The Sodium Pump (L M. Glynn and J. C. Ellory, eds.). The Company of Biologists Ltd., London, pp. 193–196.Google Scholar
  15. Farley, R. A., and Faller, L. D., 1985, The amino acid sequence of a fluorescein-labeled peptide from the ATP-binding site of gastric H,K-ATPase, Fed. Proc. 44:2527.Google Scholar
  16. Farley, R. A., Goldman, D., and Bayley, H., 1980, Identification of the regions of the catalytic subunit of (Na-K)-ATPase embedded within the cell membrane, J. Biol. Chem. 255:860–864.PubMedGoogle Scholar
  17. Farley, R. A., Tran, C. M., Carilli, C. T., Hawke, D., and Shively, J. E., 1984, The amino acid sequence of a fluorescein-labeled peptide from the active site of (Na,K)-ATPase, J. Biol. Chem. 259:9532–9535.PubMedGoogle Scholar
  18. Hall, C., and Ruoho, A., 1980, Ouabain-binding-site photoaffmity probes that label both subunits of Na+, K+ -ATPase, Proc. Natl. Acad. Sci. U.S.A. 77:4529–4533.PubMedCrossRefGoogle Scholar
  19. Hesse, J. E., Wieczorek, L., Altendorf, K., Reicin, A. S., Doms, E., and Epstein, W., 1984, Sequence homology between two membrane transport ATPases, the Kdp-ATPase ofEscherichia coli and the Ca2+-ATPase of sarcoplasm reticulum, Proc. Natl. Acad. Sci. Sci. U.S.A. 81:4746–4750.CrossRefGoogle Scholar
  20. Jorgensen, P. L., 1982, Mechanism of the Na+, K+ pump: Protein structure and conformations of the pure (Na+, K+)ATPase, Biochim. Biophys. Acta 694:27–68.PubMedGoogle Scholar
  21. Laimins, L. A., Rhoads, D. B., Altendorf, K., and Epstein, W., 1978, Identification of the structural proteins of an ATP-driven potassium transport system in Escherichia coli, Proc. Natl. Acad. Sci. U.S.A. 75:3216–3219.CrossRefGoogle Scholar
  22. Levenson, R., Racaniello, V., Albritton, L., and Housman, D., 1984, Molecular cloning of the mouse ouabain-resistance gene, Proc. Natl. Acad. Sci. U.S.A. 81:1489–1493.PubMedCrossRefGoogle Scholar
  23. Lytton, J., Lin, J. C., and Guidotti, G., 1985, Identification of two molecular forms of (Na+ K+)-ATPase in rat adipocytes, J. Biol. Chem. 260:1177–1184.PubMedGoogle Scholar
  24. MacLennan, D. H., Seeman, P., Iles, G. H., and Yip, C. C., 1971, Membrane formation by the adenosine triphosphatase of sarcoplasmic reticulum, J. Biol. Chem. 246:2702–2710.PubMedGoogle Scholar
  25. MacLennan, D. H., Reithmeier, R. A. F., Shoshan, V., Campbell, K. P., and LeBel, D., 1980, Ion pathways in protein of the sarcoplasmic reticulum, Ann. N.Y. Acad. Sci. 358:138–148.PubMedCrossRefGoogle Scholar
  26. Mitchinson, C., Wilderspin, A. F., Trinnaman, B. J., and Green, N. M., 1982, Identification of a labelled peptide after stoichiometric reaction of fluorescein isothiocyanate with Ca2+-dependent adenosine triphosphatase of sarcoplasmic reticulum, FEBS Lett. 146:87–92.PubMedCrossRefGoogle Scholar
  27. Montecucco, C., Bisson, R., Gach, C., and Johansson, A., 1981, Labelling of the hydrophobic domain of the Na+, K+ -ATPase, FEBS Lett. 128:17–21.PubMedCrossRefGoogle Scholar
  28. Robinson, J. D., 1976, Substrate sites of the (Na+ +K+)-dependent ATPase, Biochim. Biophys. Acta 429:1006–1019.PubMedGoogle Scholar
  29. Rogers, T. B., and Lazdunski, M., 1979, Photoaffmity labeling of the digitalis receptor in the (sodium + potassium)-activated adenosine triphosphatase, Biochemistry 18:135–140.PubMedCrossRefGoogle Scholar
  30. Ruoho, A., and Kyte, J., 1974, Photoaffmity labeling of the ouabain-binding site on (Na++K+) adenosinetriphosphatase, Proc. Natl. Acad. Sci. U.S.A. 71:2352–2356.PubMedCrossRefGoogle Scholar
  31. Sachs, G., Chang, H. H., Rabon, E., Shackman, R., Lewin, M., and Saccomani, G., 1976, A nonelectrogenic H+ pump in plasma membranes of hog stomach, J. Biol. Chem. 251:7690–7698.PubMedGoogle Scholar
  32. Schery, R. W., 1972, Plants For Man, Prentice-Hall, Englewood Cliffs, NJ, p. 312.Google Scholar
  33. Skou, J. C., 1965, Enzymatic basis for active transport of Na+ and K+ across cell membrane, Physiol. Rev. 45:596–617.PubMedGoogle Scholar
  34. Sweadner, K. J., 1979, Two molecular forms of (Na+ +K+)-stimulated ATPase in Brain, J. Biol. Chem. 254:6060–6067.PubMedGoogle Scholar
  35. Thorley-Lawson, D. A., and Green, N. M., 1975, Separation and characterization of tryptic fragments from the adenosine triphosphatase of sarcoplasmic reticulum, Eur. J. Biochem. 59:193–200.PubMedCrossRefGoogle Scholar
  36. Withering, W., 1785, An account of the foxglove and some of its medical uses: with practical remarks on dropsy and other diseases, Swiney, Birmingham.CrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1986

Authors and Affiliations

  • Leigh English
    • 1
  • Benjamin White
    • 1
  • Lewis Cantley
    • 1
  1. 1.Department of Biochemistry and Molecular BiologyHarvard UniversityCambridgeUSA

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