Summary
The present studies demonstrate that extracellular cations alter ouabain binding by intact human erythrocytes and that this alteration reflects a change in the apparent affinity but not the capacity for ouabain binding. Monovalent cations exert their effect at a single site (the “monovalent cation site”) and each monovalent cation can inhibit competitively the effect of other monovalent cations. Sodium, lithium, and cesium increase while potassium and rubidium decrease the apparent affinity with which ouabain is bound. Divalent cations exert their effect at a single site (the “divalent cation site”) which is functionally distinct from the site at which monovalent cations act and show mutual competitive inhibition. Each divalent cation studied increases the apparent affinity with which oubain binds to the erythrocyte membrane. Magnesium and calcium, but not barium, can also alter the effect of monovalent cations on ouabain binding. To interpret our findings we have proposed that the erythrocyte membrane has a “receptor complex” composed of a monovalent cation site, a divalent cation site and a glycoside-binding site. The number and type of cations occupying the cation sites determine the affinity of the glycoside-binding site for ouabain.
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Baker, P. F., Willis, J. S. 1969. On the number of sodium pumping sites in cell membranes.Biochim. Biophys. Acta 183:646
Baker, P. F., Willis, J. S. 1970. Potassium ions and the binding of cardiac glycosides to mammalian cells.Nature 226:521
Baker, P. F., Willis, J. S. 1972. Binding of the cardiac glycoside ouabain to intact cells.J. Physiol. 224:441
Baker, P. F., Willis, J. S. 1972. Inhibition of the sodium pump in squid giant axons by cardiac glycosides: Dependence on extracellular ions and metabolism.J. Physiol. 224:463
Beaugé, L. A., Adragna, N. 1971. The kinetics of ouabain inhibition and the partition of rubidium influx in human red blood cells.J. Gen. Physiol. 57:576
Davidsohn, I., Wells, B. B. 1963. Clinical Diagnosis by Laboratory Methods. p. 73. W. B. Saunders Co., Philadelphia, Pa.
Draper, N., Smith, H. 1966. Applied Regression Analysis. Chapter 10, p. 263. John Wiley and Sons Inc., New York
Dunham, P. B., Hoffman, J. F. 1971. Active cation transport and ouabain binding in high potassium and low potassium red blood cells of sheep.J. Gen. Physiol. 58:94
Dunham, P. B., Hoffman, J. F. 1971. The number of Na+∶K+ pump sites on red blood cells from HK and LK lambs.Biochim. Biophys. Acta 241:399
Gardner, J. D., Conlon, T. P. 1972. The effects of sodium and potassium on ouabain binding by human erythrocytes.J. Gen. Physiol. 60:609
Gardos, G. 1961. The function of calcium in the regulation of ion transport.In: Membrane Transport and Metabolism. A. Kleinzeller and A. Kotyk, editors. p. 553. Academic Press Inc., New York
Gardos, G. 1968. The function of calcium in the potassium permeability of human erythrocytes.Biochim. Biophys. Acta 30:653
Garrahan, P. J., Glynn, I. M. 1967. The sensitivity of the sodium pump to external sodium.J. Physiol. (London) 192:175
Glynn, I. M. 1957. The action of cardiac glycosides on sodium and potassium movements in human red cells.J. Physiol. 136:148
Hoffman, J. F. 1962. The active transport of sodium by ghosts of human red blood cells.J. Gen. Physiol. 45:837
Hoffman, J. F. 1962. Cation transport and structure of the red-cell plasma membrane.Circulation 26:1201
Hoffman, J. F. 1969. The interaction between tritiated ouabain and the Na−K pump in red blood cells.J. Gen. Physiol. 54:343s
Hoffman, J. F., Ingram, C. J. 1968. Cation transport and the binding of T-ouabain to intact human red blood cells.Proc. First Int. Symp. Metabolism and Membrane Permeability of Erythrocytes and Thrombocytes, p. 420, Vienna
Jarnefelt, J. 1962. Properties and possible mechanism of the Na+ and K+-stimulated microsomal adenosinetriphosphatase.Biochim. Biophys. Acta 59:643
Judah, J. D., Ahmed, K. 1963. Role of phosphoproteins in ion transport: Interactions of sodium with calcium and potassium in liver slices.Biochim. Biophys. Acta 71:34
Judah, J. D., Ahmed, K. 1964. The biochemistry of sodium transport.Biol. Rev. 39:160
Judah, J. D., Ahmed, K., McLean, A. E. M. 1962. Ion transport and phosphoproteins of human red cells.Biochim. Biophys. Acta 65:472
Kahn, J. B., Jr., Acheson, G. H. 1955. Effects of cardiac glycosides and other lactones and of certain other compounds, on cation transfer in human erythrocytes.J. Pharmacol. 115:305
Kregenow, F. M., Hoffman, J. F. 1972. Some kinetic and metabolic characteristics of calcium induced potassium transport in human red cells.J. Gen. Physiol. 60:406
Kyte, J. 1971. Purification of the sodium- and potassium-dependent adenosine triphosphatase from canine renal medulla.J. Biol. Chem. 246:4157
Kyte, J. 1972. The titration of the cardiac glycoside binding site of the (Na++K+)-adenosine triphosphatase.J. Biol. Chem. 247:7634.
Kyte, J. 1972. Properties of the two polypeptides of sodium- and potassium-dependent adenosine triphosphatase.J. Biol. Chem. 247:7642
Matsui, H., Schwartz, A., 1968. Mechanism of cardiac glycoside inhibition of the (Na+−K+)-dependent ATPase from cardiac tissue.Biochim. Biophys. Acta 151:655
Post, R. L., Jolly, P. C. 1957. The linkage of sodium, potassium and ammonium active transport across the human erythrocyte membrane.Biochim. Biophys. Acta 25:118
Priestland, R. N., Whittam, R. 1968. The influence of external sodium ions on the sodium pump in erythrocytes.Biochem. J. 109:369
Sachs, J. R., Welt, L. G. 1967. The concentration dependence of active potassium transport in the human red blood cell.J. Clin. Invest. 46:65
Schwartz, A., Lindenmayer, G. E., Allen, J. C. 1972. The Na+, K+-ATPase membrane transport system: Importance in cellular function.In: Current Topics in Membranes and Transport. F. Bronner and A. Kleinzeller, editors. p. 1. Academic Press Inc., New York
Skou, J. C. 1957. The influence of some cations on an adenosine triphosphatase from peripheral nerves.Biochim. Biophys. Acta 23:394
Skou, J. C. 1960. Further investigations on a Mg+++Na+-activated adenosinetriphosphatase, possibly related to the active, linked transport of Na+ and K+ across the nerve membrane.Biochim. Biophys. Acta 42:6
Skou, J. C. 1965. Enzymatic basis for active transport of Na+ and K+ across cell membrane.Physiol. Rev. 45:596
Skou, J. C., Butler, K. W., Hansen, O. 1971. The effect of magnesium, ATP, Pi and sodium on the inhibition of the (Na++K+)-activated enzyme system byg-strophanthin.Biochim. Biophys. Acta 241:443
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Gardner, J.D., Frantz, C. Effects of cations on ouabain binding by intact human erythrocytes. J. Membrain Biol. 16, 43–64 (1974). https://doi.org/10.1007/BF01872406
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DOI: https://doi.org/10.1007/BF01872406