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ATPase for the Determination of Cardiac Glycosides

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Part of the Handbook of Experimental Pharmacology book series (HEP,volume 56 / 1)

Abstract

The existence of an Na+, K+ -activated ATPase (Mg2+-dependent, Na+, K+-activated ATP-phosphohydrolase, Ec 3.6.1.3) was discovered by Skou in 1957. Soon afterwards he also found that cardiac glycosides are able to inhibit ATPase activity by binding to the enzyme (Skou, 1960). Extensive work was done by different groups to purify and characterize this enzyme which is specifically associated with active cation transport. Recently the molecular weight of the enzyme, which is composed of two types of polypeptide chain, was found to be 250,000 daltons (Hopkins et al.,1976). The first chain has a molecular weight of 95,000 daltons and contains both an aspartic acid residue that becomes phosphorylated during the transport reaction as well as a binding site for ouabain (Bastide et al., 1973). The second polypeptide has a molecular weight of 50,000 daltons and is characterized as being a glycoprotein (Kyte, 1972). There appear to be one ouabain binding site and either one or two phosphorylation sites per 250,000 daltons molecular weight (Wilson, 1978).

Keywords

  • Cardiac Glycoside
  • Relative Binding Affinity
  • ATPase Assay
  • Displacement Assay
  • Weber Table

These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  • Akera, T., Larsen, F.S., Brody, T.M.: Correlation of cardiac sodium-and potassium-activated adenosine triphosphat activity with ouabain-induced inotropic stimulation. J. Pharmacol. Exp. Ther. 173, 145 (1970)

    PubMed  CAS  Google Scholar 

  • Bastide, F., Meissner, G., Fleischner, S., Post, R.L.: Similarity of the active site of phosphorylation of the adenosine triphosphatase for transport of sodium and potassium ions to that for transport of calcium ions in the sarcoplasmic reticulum of muscle. J. Biol. Chem. 248, 8385 (1973)

    PubMed  CAS  Google Scholar 

  • Belz, G.G., Pflederer, W.: Studies on a plasma cardiac glycoside assay based upon displacement of 3H-ouabain from Na+K+-ATPase. Basic Res. Cardiol. 70, 142 (1975)

    CAS  Google Scholar 

  • Bentley, J.D., Burnett, G.H., Conklin, R.L., Wasserburger, R.H.: Clinical applications of serum digitoxin levels. Circulation 41, 67 (1970)

    PubMed  CAS  Google Scholar 

  • Besch, H.R., Allen, J.C., Glick, G., Schwartz, A.: Correlation between the inotropic action of ouabain and its effect on subcellular enzyme systems from canine myocardium. J. Pharmacol. Exp. Pher. 171, 1 (1970)

    CAS  Google Scholar 

  • Brooker, G., Jelliffe, R.W.: Serum cardiac glycoside assay based upon displacement of 3Houabain from Na-K-ATPase. Circulation 45, 20 (1972)

    PubMed  CAS  Google Scholar 

  • Burnett, G.H., Conklin, R.L.: The enzymatic assay of plasma digitoxin levels. J. Lab. Clin. Med. 71, 1040 (1968)

    PubMed  CAS  Google Scholar 

  • Burnett, G.H., Conklin, R.L.: The enzymatic assay of plasma digoxin. J. Lab. Clin. Med. 78, 779 (1971)

    PubMed  CAS  Google Scholar 

  • Fiske, S., Subbarrow, Y.: A colorimetric method for phosphorus. J. Biol. Chem. 66, 375 (1925)

    CAS  Google Scholar 

  • Friedemann, T.E., Haugen, G.E.: Pyruvic acid. II. The determination of keto acids in blood and urine. J. Biol. Chem. 147, 415 (1943)

    CAS  Google Scholar 

  • Gundert-Remy, U., Thorade, B., Karacsonyi, P., Weber, E.: Inhibition of a NaKMg activated ATPase isolated from ox brain by different cardiac glycosides. Naunyn Schmiedebergs Arch. Pharmacol. 284, R24 (1974)

    Google Scholar 

  • Gundert-Remy, U., Koch, K., Hristka, V.: Chloroform-extractable and polar metabolites examined with different assays. In: Cardiac glycosides. Bodem, G., Dengler, H.J. (eds.), p. 28. Berlin, Heidelberg, New York: Springer 1978

    Google Scholar 

  • Hopkins, B.E., Wagner, J.H., Smith, T.W.: Sodium-and potassium-activated adenosine triphosphatase of the nasal salt gland of the duck, purification, characterization and NH2terminal amino acid sequence of the phosphorylating polypeptide. J. Biol. Chem. 251, 4365 (1976)

    PubMed  CAS  Google Scholar 

  • Kyte, J.: The titration of the cardiac glycoside binding site of the (Na+ + K+)-denosine-triphosphatase. J. Biol. Chem. 247, 7642 (1972)

    PubMed  CAS  Google Scholar 

  • Marcus, F.J., Burkhalter, L., Cuccia, C. Pavlovich, J., Kapadia, G G: Administration of tritiated digoxin with and without loading dose. A metabolic study. Circulation 34, 865 (1966)

    PubMed  CAS  Google Scholar 

  • Marcus, F.I., Ryan, J.N., Stafford, M.G.: The reactivity of digoxin and derivates of digoxin and digitoxin as measured by the Na-K-ATPase displacement assay and by radioimmunoassay. J. Lab. Clin. Med 85, 610 (1975)

    PubMed  CAS  Google Scholar 

  • Mardh, S.: A simple enzymatic assay of cardiac glycosides and its application to analysis of glycoside levels in plasma. Clin. Chim Acta 44, 165 (1973)

    PubMed  CAS  CrossRef  Google Scholar 

  • Marsh, B.B.: The estimation of anorganic phosphate in the presence of adenosine triphosphate. Biochim. Biophys. Acta 32, 357 (1959)

    PubMed  CAS  CrossRef  Google Scholar 

  • Matsui, H., Schwartz, A.: Mechanism of cardiac glycoside inhibition of the (Na+ + K+) -dependent ATPase from cardiac tissue. Biochim. Biophys. Acta 151, 655 (1968)

    PubMed  CAS  Google Scholar 

  • Schoner, W., von Ilberg, C., Kramer R., Seubert, W.: On the mechanism of Na+- and K+ -stimulated hydrolysis of adenosine triphosphate. Eur. J. Biochem. 1, 334 (1967)

    PubMed  CAS  CrossRef  Google Scholar 

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

    PubMed  CAS  CrossRef  Google Scholar 

  • Skou, J.C.: Further investigations on a Mg++-Na+ activated adenosine-tri-phosphatase possibly related to the active, linked transport of Na+ and K+ across the nerve membrane. Biochim. Biophys. Acta 42, 6 (1960)

    CAS  CrossRef  Google Scholar 

  • Skou, J.C., Hilberg C.: The effect of cations, g-strophanthin, and oligomycin on the labeling from (32P)-ATP of the Na+, K+-activated enzyme system and the effect of cations and g-strophanthin on the labeling from (32P)-ITP and 32 Pi. Biochim. Biophys. Acta 185, 198 (1969)

    PubMed  CAS  Google Scholar 

  • Vöhringer, H.F., Rietbrock, N.: Metabolism and excretion of digitoxin in man. Clin. Pharmacol. Ther. 16, 796 (1974)

    PubMed  Google Scholar 

  • Wilson, D.B.: Cellular transport mechanisms. Ann. Rev. Biochem. 47, 933 (1978)

    PubMed  CAS  CrossRef  Google Scholar 

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Gundert-Remy, U., Weber, E. (1981). ATPase for the Determination of Cardiac Glycosides. In: Greeff, K. (eds) Cardiac Glycosides. Handbook of Experimental Pharmacology, vol 56 / 1. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-68163-9_6

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  • DOI: https://doi.org/10.1007/978-3-642-68163-9_6

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