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Purification and Properties of Na, K-ATPases from the Rectal Gland of Squalus acanthias and the Electric Organ of Electrophorus electricus and Reconstitution of the Na+ - K+Pump from the Purified Enzyme

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Biochemistry of Membrane Transport

Part of the book series: Proceedings in Life Sciences ((LIFE SCIENCES))

Abstract

About twenty years ago Skou (1957) discovered an adenosine triphosphatase in the microsome fraction of the leg nerve of the shore crab which required Mg+, Na+ and K+ for optimal activity. He suggested that this enzyme, which will henceforth be referred to as the Na,K-ATPase, might be involved in the coupled Na+-K+ pump (Baker, 1972), which serves many important physiological functions, such as maintaining osmotic equilibrium in all animal cells, maintaining the gradients of Na+ and K+ in excitable tissues such as nerve and muscle, which permits waves of excitation down their membranes, and effecting transepithelial transport of Na+, and in some cases, K+ in such structures as the intestinal epithelium, the renal tubules, and various glandular structures. It has been estimated that in the mammal about 30% of the ATP which is produced by resting respiration is hydrolyzed via the Na,K-ATPase (Whittam, 1965). Edelman and his coworkers (1974) have demonstrated that thyroid hormone elevates the level of Na,K-ATPase in thyroid-sensitive tissues, and he has suggested that the resultant increased hydrolysis of ATP could be responsible for the calorigenic effect of thyroid hormone.

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References

  • Albers, R.W., Fahn, S., Koval, G.J.: The role of sodium ions in the activation of electrophorus electric organ adenosine triphosphatase. Proc. Natl. Acad. Sci. U.S. 50, 474–481 (1963)

    Article  CAS  Google Scholar 

  • Alexander, D.R., Rodnight, R.: Separation at neutral pH of a 32P-labelled membrane protein associated with the sodium-plus-magnesium ion-activated adenosine triphosphatase from ox brain. Biochem. 119, 44–45 (1970)

    Google Scholar 

  • Avruch, J., Fairbanks, G.: Demonstration of a phosphopeptide intermediate in the Mg++-dependent, Na+- and K+-stimulated adenosine triphosphatase reaction of the erythrocyte membrane. Proc. Natl. Acad. Sci. U.S. 69, 1216–1220 (1972)

    Article  CAS  Google Scholar 

  • Baker, P.F.: Metabolic Pathways. Vol. VI: Metabolic Transport: The Sodium Pump in Animal Tissues and its Role in the Control of Cellular Metabolism and Function. New York: Academic Press 1972, pp. 243–268

    Google Scholar 

  • Baker, P.F., Manil, J.: The rates of action of K+ and ouabain on the sodium pump in squid axons. Biochim. Biophys. Acta 150, 328–330 (1968)

    Article  PubMed  CAS  Google Scholar 

  • Bergh, C.H., Larson, G., Samuelson, B.E.: Fatty acid and aldehyde composition of major phospholipids in salt gland of marine birds and spiny dogfish. Lipids 10, 299–302 (1975)

    Article  PubMed  CAS  Google Scholar 

  • Bonting, S.L.: Studies on sodium-potassium-activated adenosine triphosphatase. XV. The rectal gland of the elasmobranchs. Comp. Biochem. Physiol. 17, 953–966 (1966)

    Article  PubMed  CAS  Google Scholar 

  • Caldwell, P.C., Hodgkin, A.L., Keynes, R.D., Shaw, T.I.: The effects of injecting “energy rich” phosphate compounds on the active transport of ions in the giant axons of LOLIGO. J. Physiol. 17, 561–590 (1960)

    Google Scholar 

  • Collins, R.C., Albers, R.W.: The phosphoryl acceptor protein of Na-K-ATPase from various tissues. J. Neurochem. 19, 1209–1213 (1972)

    Article  PubMed  CAS  Google Scholar 

  • Dahl, J., Hokin, L.E.: The sodium-potassium adenosine triphosphatase. Ann. Rev. Biochem. 43, 327–356 (1974)

    Article  PubMed  CAS  Google Scholar 

  • Dixon, J.F., Hokin, L.E.: Studies on the characterization of the sodium-potassium transport adenosine triphosphatase. Arch. Biochem. Biophys. 163, 749–758 (1974)

    Article  PubMed  CAS  Google Scholar 

  • Edelman, I.S.: Drugs and Transport Processes: Thyroid and Sodium Transport. London: Macmillan 1974, pp. 101–110

    Google Scholar 

  • Gardner, J.D., Conlon, T.P.: The effects of sodium and potassium on ouabain binding by human erythrocytes. J. Gen. Physiol. 60, 609–629 (1972)

    Article  PubMed  CAS  Google Scholar 

  • Gardos, G.: Connection between membrane adenosine triphosphatase activity and potassium transport in erythrocyte ghosts. Experientia 20, 387 (1964)

    Article  PubMed  CAS  Google Scholar 

  • Garrahan, P.J., Glynn, I.M.: Factors affecting the relative magnitude of the sodium-potassium and sodium: sodium exchanges catalysed by the sodium pump. J. Physiol. 192, 189–216 (1967a)

    PubMed  CAS  Google Scholar 

  • Garrahan, P.J., Glynn, I.M.: The stoicheiometry of the sodium pump. J. Physiol. 192, 217–235 (1967b)

    PubMed  CAS  Google Scholar 

  • Glynn, I.M.: Activation of adenosine triphosphatase activity in a cell membrane by external potassium and internal sodium. J. Physiol. 160, 18 (1962)

    Google Scholar 

  • Glynn, I.M., Karlish, S.J.D.: The sodium pump. Ann. Rev. Physiol. 37, 13–55 (1975)

    Article  CAS  Google Scholar 

  • Goldin, S.M., Tong, S.W.: Reconstitution of active transport catalyzed by the purified sodium and potassium ion-stimulated adenosine triphosphatase from canine renal medulla. J. Biol. Chem. 249, 5907–5915 (1974)

    PubMed  CAS  Google Scholar 

  • Hilden, S., Hokin, L.E.: Active potassium transport coupled to active sodium transport in vesicles reconstituted from purified sodium and potassium ion-activated adenosine triphosphatase from the rectal gland of Squalus acanthias. J. Biol. Chem. 250, 6296–6303 (1975)

    PubMed  CAS  Google Scholar 

  • Hilden, S., Hokin, L.E.: Coupled Na+-K+ transport in vesicles containing a purified (NaK)-ATPase and only phosphatidyl choline. Biochem. Biophys. Res. Commun. 69, 521–527 (1976)

    Article  PubMed  CAS  Google Scholar 

  • Hilden, S., Rhee, H.M., Hokin, L.E.: Sodium transport by phospholipid vesicles containing purified sodium and potassium ion-activated adenosine triphosphatase. J. Biol. Chem. 249, 7432–7440 (1974)

    PubMed  CAS  Google Scholar 

  • Hinkle, P.C., Kim, J.J., Racker, E.: Ion transport and respiratory control in vesicles formed from cytochrome oxidase and phospholipids. J. Biol. Chem. 247, 1338–1339 (1972)

    PubMed  CAS  Google Scholar 

  • Hoffman, J.F.: The link between metabolism and the active transport of Na+ in human red cell ghosts. Federation Proc. 19, 127 (1960)

    Google Scholar 

  • Hoffman, J.F.: Organization of Energy-Transducing Membranes: The Molecular Aspects of Na+,K+-pump and Red Blood Cells. Baltimore: University Park Press 1973, pp. 9–21

    Google Scholar 

  • Hokin, L.E.: Purification and properties of the (sodium + potassium)-activated adenosine triphosphatase and reconstitution of sodium transport. Ann. N.Y. Acad. Sci. 242, 12–23 (1974)

    Article  PubMed  CAS  Google Scholar 

  • Hokin, L.E., Dahl, J.D., Deupree, J.D., Dixon, J.F., Hackney, J.F, Perdue, J.R: Studies on the characterization of the sodium-potassium transport adenosine triphosphatase. J. Biol. Chem. 248, 2593–2605 (1973)

    PubMed  CAS  Google Scholar 

  • Jain, M.K., White, F.P., Strickholm, A., Willians, E., Cordes, E.H.: Studies concerning the possible reconstitution of an active cation pump across an artificial membrane. J. Membrane Biol. 8, 363–388 (1972)

    Article  CAS  Google Scholar 

  • Jorgensen, P.L.: Purification and characterization of (Na++K+)-ATPase. III. Purification from the outer medulla of mammalian kidney after selective removal of membrane components by sodium dodecylsulphate. Biochim. Biophys. Acta 356, 36–52 (1974)

    Article  PubMed  CAS  Google Scholar 

  • Jorgensen, P.L.: Isolation and characterization of the components of the sodium pump. Quart. Rev. Biophys. 7, 239–274 (1975)

    Article  Google Scholar 

  • Kagawa, Y., Racker, E.: Partial resolution of the enzymes catalyzing oxidative phosphorylation. XXV. Reconstitution of vesicles catalyzing 32Pi-adenosine triphosphate exchange. J. Biol. Chem. 246, 5477–5487 (1971)

    CAS  Google Scholar 

  • Kepner, G.R., Macy, R.I.: Membrane enzyme systems molecular size determinations by radiation inactivation. Biochim. Biophys. Acta 163, 188–203 (1968)

    Article  PubMed  CAS  Google Scholar 

  • Kyte, J.: Phosphorylation of a purified (Na++K+) adenosine triphosphatase. Biochem. Biophys. Res. Commun. 43, 1259–1265 (1971a)

    Article  PubMed  CAS  Google Scholar 

  • Kyte, J.: Purification of the sodium- and potassium-dependent adenosine triphosphatase from canine renal medulla. J. Biol. Chem. 246, 4156–4165 (1971b)

    Google Scholar 

  • Kyte, J.: Properties of the two polypeptides of sodium- and potassium-dependent adenosine triphosphatase. J. Biol. Chem. 247, 7642–7649 (1972)

    PubMed  CAS  Google Scholar 

  • Laemmle, U.V.: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature (London) 227, 680–685 (1970)

    Article  Google Scholar 

  • Lane, K., Copenhaver, J.H., Lindenmayer, G.E., Schwartz, A.: Purification and characterization of and [3H] ouabain binding to the transport adenosine triphosphatase from outer medulla of canine kidney. J. Biol. Chem. 248, 7197–7200 (1973)

    PubMed  CAS  Google Scholar 

  • Meissner, G., Fleischer, S.: Dissociation and reconstitution of functional sarcoplasmic reticulum vesicles. J. Biol. Chem. 249, 302–309 (1974)

    PubMed  CAS  Google Scholar 

  • Nakao, T., Nakao, M, Mizuno, N., Komatsu, Y., Fujita, M.: Purification and some properties of Na,K-ATPase. I. Solubilization and stability of Lubrol extracts. J. Biochem. 73, 609–619 (1973a)

    PubMed  CAS  Google Scholar 

  • Nakao, T., Nakao, M., Nagai, F., Kawai, K., Fujihara, U., Hara, Y., Fujita, M.: Purification and some properties of Na,K-transport ATPase. II. Preparations with high specific activity obtained using aminoethyl cellulose chromatography. J. Biochem. 73, 781–791 (1973b)

    PubMed  CAS  Google Scholar 

  • Perrone, J.R., Biostein, R.: Asymmetric interaction of inside-out and right-side-out erythrocyte membrane vesicles with ouabain. Biochim. Biophys. Acta 291, 680–689 (1973)

    Article  PubMed  CAS  Google Scholar 

  • Perrone, J.R., Hackney, J.F., Dixon, J.D., Hokin, L.E.: Molecular properties of purified (sodium and potassium)-activated adenosine triphosphatase and their subunits from the rectal gland of Squalus acanthias and the electric organ of Electrophorus electricus. J. Biol. Chem. 250, 4178–4184 (1975)

    PubMed  CAS  Google Scholar 

  • Racker, E.: Reconstitution of a calcium pump with phospholipids and a purified Ca++-adenosine triphosphatase from sarcoplasmic reticulum. J. Biol. Chem. 247, 8198–8220 (1972)

    PubMed  CAS  Google Scholar 

  • Racker, E.: A new procedure for the reconstitution of biologically active phospholipid vesicles, Biochem. Biophys. Res. Commun. 55, 224–230 (1973)

    Article  PubMed  CAS  Google Scholar 

  • Racker, E., Eytan, E.: Reconstitution of an efficient calcium pump without detergents. Biochem. Biophys. Res. Commun. 55, 174–178 (1973)

    Article  PubMed  CAS  Google Scholar 

  • Racker, E., Fisher, L.W.: Reconstitution of an ATP-dependent sodium pump with an ATPase from electric eel and pure phospholipids. Biochem. Biophys. Res. Commun. 67, 1144–1150 (1975)

    Article  PubMed  CAS  Google Scholar 

  • Racker, E., Stoekenius, W.: Reconstitution of purple membrane vesicles catalyzing light-driven proton uptake and adenosine triphosphatase formation. J. Biol. Chem. 249, 662–663 (1974)

    PubMed  CAS  Google Scholar 

  • Repke, K.R.H., Schon, R., Henke, W., Schonfeld, W., Streckenbach, B., Dittrich, F.: Experimental and theoretical examination of the flip-flop model of (Na-K)-ATPase function. Ann. N.Y. Acad. Sci. 242, 203–219 (1974)

    Article  PubMed  CAS  Google Scholar 

  • Rhee, H.M., Hokin, L.E.: Inhibition of the purified sodium-potassium activated adenosine triphosphatase from the rectal gland of Squalus acanthias by antibody against the glycoprotein subunit. Biochem. Biophys. Res. Commun. 63, 1139–1145 (1975)

    Article  PubMed  CAS  Google Scholar 

  • Ruoho, A., Kyte, J.: Photoaffinity labeling of the ouabain-bending site on (Na++K+) adenosine triphosphatase. Proc. Natl. Acad. Sci. U.S. 71, 2352–2356 (1974)

    Article  CAS  Google Scholar 

  • Schwartz, A., Lindenmayer, G.E., Allen, J.C.: The sodium-potassium adenosine triphosphatase: pharmacological, physiological and biochemical aspects. Pharmacol. Rev. 27, 3–134 (1975)

    PubMed  CAS  Google Scholar 

  • Sen, A.K., Post, R.L.: Stoichiometry and localization of adenosine triphosphatase -dependent sodium and potassium transport in the erythrocyte. J. Biol. Chem. 239, 345–352 (1964)

    PubMed  CAS  Google Scholar 

  • Simons, T.J.B.: Potassium:potassium exchange catalysed by the sodium pump in human red cells. J. Physiol. 237, 123–155 (1974)

    PubMed  CAS  Google Scholar 

  • Simpkins, H., Hokin, L.E.: Studies on the characterization of the sodium-potassium transport adenosine triphosphatase. XIII. On the organization and role of phospholipids in the purified enzyme. Arch. Biochem. Biophys. 159, 897–902 (1973)

    Article  CAS  Google Scholar 

  • Skou, J.C.: The influence of some cations on an adenosine triphosphatase from peripheral nerves. Biochim. Biophys. Acta 23, 394–401 (1957)

    Article  PubMed  CAS  Google Scholar 

  • Skou, J.C.: The (Na++K+) activated enzyme system and its relationship to transport of sodium and potassium. Quart. Rev. Biophys. 7, 401–434 (1975)

    Article  Google Scholar 

  • Stein, W.D., Lieb, W.R., Karlish, S.J.D., Eilam, Y.: A model for active transport of sodium and potassium ions as mediated by a tetrameric enzyme. Proc. Natl. Acad. Sci. U.S. 70, 275–278 (1973)

    Article  CAS  Google Scholar 

  • Sweadner, K.J., Goldin, S.M.: Reconstitution of active ion transport by the sodium and potassium ion-stimulated adenosine triphosphatase from canine brain. J. Biol. Chem. 250, 4022–4024 (1975)

    PubMed  CAS  Google Scholar 

  • Uesugi, S., Dulak, M.C., Dixon, J.F., Hexum, T.D., Dahl, J.D., Perdue, J.F., Hokin, L.E.: Studies on the characterization of the sodium-potassium transport adenosine triphosphatase. VI. Large scale partial purification and properties of a Lubrol-solubilized bovine brain enzyme. J. Biol. Chem. 246, 531–543 (1971)

    PubMed  CAS  Google Scholar 

  • Warren, G.B., Toon, P.A., Birdsall, N.J.M., Lee, A.G., Metcalfe, J.C.: Complete control of the lipid environment of membrane-bound proteins: Application to a calcium transport system. FEBS Lett. 41, 122–124 (1974a)

    Article  PubMed  CAS  Google Scholar 

  • Warren, G.B., Toon, P.A., Birdsall, N.J.M., Lee, A.G., Metcalfe, J.C.: Reconstitution of a calcium pump using defined membrane components. Proc. Natl. Acad. Sci. U.S. 71, 622–626 (1974b)

    Article  CAS  Google Scholar 

  • Whittam, R., Ager, M.E.: The connexion between active cation transport and metabolism in erythrocytes. Biochem. J. 97, 214–227 (1965)

    PubMed  CAS  Google Scholar 

  • Whittam, R., Chipperfield, A.R.: The reaction mechanism of the sodium pump. Biochim. Biophys. Acta 415, 149–171 (1975)

    PubMed  CAS  Google Scholar 

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  1. L. E. Hokin
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  1. Laboratorium für Biochemie, Eidgenössische Technische Hochschule Zürich, 8006, Zürich, Switzerland

    Giorgio Semenza  & Ernesto Carafoli  & 

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Hokin, L.E. (1977). Purification and Properties of Na, K-ATPases from the Rectal Gland of Squalus acanthias and the Electric Organ of Electrophorus electricus and Reconstitution of the Na+ - K+Pump from the Purified Enzyme. In: Semenza, G., Carafoli, E. (eds) Biochemistry of Membrane Transport. Proceedings in Life Sciences. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-66564-6_28

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  • DOI: https://doi.org/10.1007/978-3-642-66564-6_28

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