Role of the bile acid transport system in hepatocellular ouabain uptake

  • E. Petzinger
  • K. Fischer
  • H. Fasold


HeLa cells take up 3H-ouabain by internalization together with Na+/K+-ATPase (4). Similar endocytotic uptake was suggested for rat hepatocytes which excrete ouabain into bile (10). On isolated rat hepatocytes carrier mediated uptake of ouabain has been described (5) but any identification of the transport system is still lacking. By using rauwolfia alkaloide cevadine we have proved that ouabain in contrast to HeLa cells is not taken up in rat hepatocytes by Na+/K+-ATPase internalization (17). We speculate additional binding proteins in rat liver cell membrane with properties for glycoside translocation and suggest the bile acid transport system to be such a candidate. Ouabain uptake is inhibited by DIDS, probenecid, furosemide and cevadine which inhibit bile acid uptake concomitantly. Bile acids are inhibitors of ouabain uptake. Cells which lack bile acid uptake, e. g. AS-30 D ascites hepatoma cells and Ehrlich ascites tumor cells do not transport ouabain. Digitoxin, digoxin and cassaine as compared to ouabain are stronger inhibitors of 3H-ouabain uptake. They inhibit cholate uptake markedly.

In order to identify membrane proteins probably involved in ouabain uptake, rat hepatocytes were incubated with benzacidolysin-ouabain for photoaffinity labelling. The label was covalently attached to a 50 kDa protein which was not identified in transport deficient Ehrlich cells. With respect to recent results in the identification of the bile acid transporter from hepatocytes (14, 23) we propose ouabain as substrate for the hepatic bile acid uptake system.


Bile Acid Cardiac Glycoside Ehrlich Ascites Tumor Cell Bile Acid Transport Ascites Hepatoma Cell 
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.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Berry MS, Friend DS (1969) High-yield preparation of isolated rat liver parenchymal cells. J Cell Biol 43: 509–529CrossRefGoogle Scholar
  2. 2.
    Boyer JL, Reno D (1975) Properties of ( Na’ + K’)-activated ATPase in rat liver plasma membranes enriched with bile canaliculi. Biochim Biophys Acta 401: 59–72Google Scholar
  3. 3.
    Cook JS, Will PC, Proctor WR, Brake ET (1976) Turnover of ouabain binding sites and plasma membrane proteins in HeLa cells. In: Cook JS (ed) Biogenesis and Turnover of Membrane Macromolecules. Raven Press, New York, pp 15–36Google Scholar
  4. 4.
    Cook JS, Tate EH, Shaffer C (1982) Uptake of (3H)-ouabain from the cell surface into the lysosomal compartment of HeLa cells. J Cellular Physiol 110: 84–92CrossRefGoogle Scholar
  5. 5.
    Eaton DL, Klaassen CD (1978) Carrier-mediated transport of ouabain in isolated hepatocytes. J Pharmacol Exp Ther 205: 480–488PubMedGoogle Scholar
  6. 6.
    Farah A (1946) On the elimination of G-strophantin in the rat. J Pharmacol Exp Ther 86: 248–257PubMedGoogle Scholar
  7. 7.
    Frimmer M (1980) Organotropism by carrier-mediated transport. Trend Pharmacol Sci 3: 395–397CrossRefGoogle Scholar
  8. 8.
    Frimmer M, Petzinger E, Ziegler K (1980) Protective effect of anionic cholecystographic agents against phalloidin on isolated hepatocytes by competitive inhibition of the phallotoxin uptake. Naunyn Schmiedeberg’s Arch Pharmacol 313: 85–89PubMedCrossRefGoogle Scholar
  9. 9.
    Frimmer M, Ziegler K (1985) Photoaffinity labeling of whole cells by flashed light: A simple apparatus for high-energy ultraviolet flashes. Biochim Biophys Acta (in press)Google Scholar
  10. 10.
    Graf J, Peterlik M (1976) Ouabain-mediated sodium uptake and bile formation by isolated perfused rat liver. Am J Physiol 230: 876–885PubMedGoogle Scholar
  11. 11.
    Grell E, Lewitzki E, Krause G, Richter WJ, Raschdorf F (1984) Mechanism of interaction between cation and ligand binding sites in Na+,K+-ATPase. In: Proceedings of the Third EBEC Conference, Hannover 1984. ICSU Press, MiamiGoogle Scholar
  12. 12.
    Klingenberg M, Pfaff E (1967) Means of terminating reactions. Meth Enzymol 10: 680–684CrossRefGoogle Scholar
  13. 13.
    Kupferberg HJ, Schanker LJ (1968) Biliary secretion of ouabain-3H and its uptake by liver slices in the rat. Am J Physiol 214: 1048–1053PubMedGoogle Scholar
  14. 14.
    Kramer W, Bickel U, Buscher H-P, Gerok W, Kurz G (1982) Bile-salt-binding polypeptides in plasma membranes of hepatocytes revealed by photoaffinity labelling. Eur J Biochem 129: 13–24PubMedCrossRefGoogle Scholar
  15. 15.
    Petzinger E, Joppen C, Frimmer M (1983) Common properties of hepatocellular uptake of cholate, iodipamide and antamanide, as distinct from the uptake of bromsulfophthalein. Naunyn Schmiedeberg’s Arch Pharmacol 322: 174–179PubMedCrossRefGoogle Scholar
  16. 16.
    Petzinger E, Frimmer M (1984) Driving forces in hepatocellular uptake of phalloidin and cholate. Biochim Biophys Acta 778: 539–548PubMedCrossRefGoogle Scholar
  17. 17.
    Petzinger E, Fischer K (1985) Transport functions of the liver. Lack of correlation between hepatocellular ouabain uptake and binding to ( Na’ + K+)-ATPase. Biochim Biophys Acta 815: 334–340Google Scholar
  18. 18.
    Schanker LS (1968) Secretion of organic compounds in the bile. In: Handbook of Physiology Vol 5, Alimentary Canal, p 2433–2449Google Scholar
  19. 19.
    Schwenk M (1980) Transport systems of isolated hepatocytes. Arch Toxicol 44: 113–126PubMedCrossRefGoogle Scholar
  20. 20.
    Schwenk M, Wiedemann T, Remmer H (1981) Uptake, accumulation and release of ouabain by isolated rat hepatocytes. Naunyn Schmiedeberg’s Arch Pharmacol 316: 340–344PubMedCrossRefGoogle Scholar
  21. 21.
    Weber K, Osborn M (1969) The reliability of molecular weight determinations by dodecyl sulfatepolyacrylamide gel electrophoresis. J Biol Chem 244: 4406–4412PubMedGoogle Scholar
  22. 22.
    Ziegler K, Frimmer M, Fasold H (1984) Further characterization of membrane proteins involved in the transport of organic anions in hepatocytes. Comparison of two different affinity labels: 4,4’diisothiocyano-1,2-diphenylethane-2,2’-disulfonic acid and brominated taurodehydrocholic acid. Biochim Biophys Acta 769: 117–129PubMedCrossRefGoogle Scholar
  23. 23.
    Ziegler K, Frimmer M, Müller S, Fasold H (1984) 3’isothiocyanatobenzamido (3H)-cholate, a new affinity label for hepatocellular membrane proteins responsible for the uptake of both bile acids and phalloidin. Biochim Biophys Acta 773: 11–22Google Scholar
  24. 24.
    Ziegler K, Frimmer M (1986) Photoaffinity labeling of whole cells by flashed light: A simple apparatus for high energy ultraviolet flashes. Biochim Biophys Acta (in press)Google Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 1986

Authors and Affiliations

  • E. Petzinger
    • 3
    • 4
  • K. Fischer
    • 1
  • H. Fasold
    • 2
  1. 1.Institute of Pharmacology and Toxicology, Department of Veterinary MedicineJustus-Liebig-University GießenGießenGermany
  2. 2.Institute of Biochemistry, Frankfurt, Department of MedicineJohann Wolfgang Goethe-UniversityFrankfurtWest Germany
  3. 3.Institut für Pharmakologie und Toxikologie im Fachbereich VeterinärmedizinJustus-Liebig-Universität GießenGießenWest Germany
  4. 4.Reprint requests schould be addrssed to E.P.Max Planck Institut für SystemphysiologieDortmundGermany

Personalised recommendations