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. 1991 Sep 15;278(Pt 3):637–641. doi: 10.1042/bj2780637

Hepatocanalicular organic-anion transport is regulated by protein kinase C.

H Roelofsen 1, R Ottenhoff 1, R P Oude Elferink 1, P L Jansen 1
PMCID: PMC1151394  PMID: 1716882

Abstract

In order to investigate the regulation of canalicular organic-anion transport, we used a hepatocyte transport assay in which canalicular secretion of a model organic anion, dinitrophenyl-glutathione (GS-DNP), was measured in the presence of stimulators and inhibitors of the Ca2+/protein kinase C (PKC) second-messenger system and of the cyclic AMP (cAMP) second-messenger system. Vasopressin (24 nM) and the phorbol ester phorbol 12-myristate 13-acetate (1 microgram/ml), both stimulators of PKC, stimulated GS-DNP efflux by 65 +/- 36% and 55 +/- 28% respectively, whereas staurosporine (10 microM), an inhibitor of PKC, inhibited efflux by 53 +/- 13%. Glucagon and forskolin, both stimulators of the cAMP second-messenger system, as well as the cAMP analogue dibutyryl cAMP and the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine, did not significantly influence the GS-DNP efflux. It can be concluded that canalicular organic-anion transport in hepatocytes is either directly or indirectly regulated by PKC.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Akerboom T. P., Bilzer M., Sies H. Competition between transport of glutathione disulfide (GSSG) and glutathione S-conjugates from perfused rat liver into bile. FEBS Lett. 1982 Apr 5;140(1):73–76. doi: 10.1016/0014-5793(82)80523-1. [DOI] [PubMed] [Google Scholar]
  2. Ashendel C. L. The phorbol ester receptor: a phospholipid-regulated protein kinase. Biochim Biophys Acta. 1985 Sep 9;822(2):219–242. doi: 10.1016/0304-4157(85)90009-7. [DOI] [PubMed] [Google Scholar]
  3. Awasthi Y. C. The interrelationship between P-glycoprotein and glutathionyl S-conjugate transporter(s) Trends Biochem Sci. 1990 Oct;15(10):376–377. doi: 10.1016/0968-0004(90)90233-2. [DOI] [PubMed] [Google Scholar]
  4. Ballatori N., Truong A. T. Cholestasis, altered junctional permeability, and inverse changes in sinusoidal and biliary glutathione release by vasopressin and epinephrine. Mol Pharmacol. 1990 Jul;38(1):64–71. [PubMed] [Google Scholar]
  5. Berry M. N., Friend D. S. High-yield preparation of isolated rat liver parenchymal cells: a biochemical and fine structural study. J Cell Biol. 1969 Dec;43(3):506–520. doi: 10.1083/jcb.43.3.506. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Boyer J. L. New concepts of mechanisms of hepatocyte bile formation. Physiol Rev. 1980 Apr;60(2):303–326. doi: 10.1152/physrev.1980.60.2.303. [DOI] [PubMed] [Google Scholar]
  7. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  8. Chambers T. C., Chalikonda I., Eilon G. Correlation of protein kinase C translocation, P-glycoprotein phosphorylation and reduced drug accumulation in multidrug resistant human KB cells. Biochem Biophys Res Commun. 1990 May 31;169(1):253–259. doi: 10.1016/0006-291x(90)91461-z. [DOI] [PubMed] [Google Scholar]
  9. Chambers T. C., McAvoy E. M., Jacobs J. W., Eilon G. Protein kinase C phosphorylates P-glycoprotein in multidrug resistant human KB carcinoma cells. J Biol Chem. 1990 May 5;265(13):7679–7686. [PubMed] [Google Scholar]
  10. Coleman R. Biochemistry of bile secretion. Biochem J. 1987 Jun 1;244(2):249–261. doi: 10.1042/bj2440249. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Corasanti J. G., Smith N. D., Gordon E. R., Boyer J. L. Protein kinase C agonists inhibit bile secretion independently of effects on the microcirculation in the isolated perfused rat liver. Hepatology. 1989 Jul;10(1):8–13. doi: 10.1002/hep.1840100103. [DOI] [PubMed] [Google Scholar]
  12. De Bruijn M. H. Substrate specificity and the mdr pump. Trends Biochem Sci. 1990 Jun;15(6):218–219. doi: 10.1016/0968-0004(90)90031-6. [DOI] [PubMed] [Google Scholar]
  13. Dewald B., Thelen M., Wymann M. P., Baggiolini M. Staurosporine inhibits the respiratory burst and induces exocytosis in human neutrophils. Biochem J. 1989 Dec 15;264(3):879–884. doi: 10.1042/bj2640879. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Díaz-Guerra M. J., Boscá L. Lack of translocation of protein kinase C from the cytosol to the membranes in vasopressin-stimulated hepatocytes. Biochem J. 1990 Jul 1;269(1):163–168. doi: 10.1042/bj2690163. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Elferink R. P., Ottenhoff R., Liefting W., de Haan J., Jansen P. L. Hepatobiliary transport of glutathione and glutathione conjugate in rats with hereditary hyperbilirubinemia. J Clin Invest. 1989 Aug;84(2):476–483. doi: 10.1172/JCI114189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Garrison J. C., Johnsen D. E., Campanile C. P. Evidence for the role of phosphorylase kinase, protein kinase C, and other Ca2+-sensitive protein kinases in the response of hepatocytes to angiotensin II and vasopressin. J Biol Chem. 1984 Mar 10;259(5):3283–3292. [PubMed] [Google Scholar]
  17. Huber M., Guhlmann A., Jansen P. L., Keppler D. Hereditary defect of hepatobiliary cysteinyl leukotriene elimination in mutant rats with defective hepatic anion excretion. Hepatology. 1987 Mar-Apr;7(2):224–228. doi: 10.1002/hep.1840070204. [DOI] [PubMed] [Google Scholar]
  18. Hyde S. C., Emsley P., Hartshorn M. J., Mimmack M. M., Gileadi U., Pearce S. R., Gallagher M. P., Gill D. R., Hubbard R. E., Higgins C. F. Structural model of ATP-binding proteins associated with cystic fibrosis, multidrug resistance and bacterial transport. Nature. 1990 Jul 26;346(6282):362–365. doi: 10.1038/346362a0. [DOI] [PubMed] [Google Scholar]
  19. Ishikawa T. Is the glutathione S-conjugate carrier an mdr1 gene product? Trends Biochem Sci. 1990 Jun;15(6):219–220. doi: 10.1016/0968-0004(90)90032-7. [DOI] [PubMed] [Google Scholar]
  20. Ishikawa T., Müller M., Klünemann C., Schaub T., Keppler D. ATP-dependent primary active transport of cysteinyl leukotrienes across liver canalicular membrane. Role of the ATP-dependent transport system for glutathione S-conjugates. J Biol Chem. 1990 Nov 5;265(31):19279–19286. [PubMed] [Google Scholar]
  21. Jakobson I., Warholm M., Mannervik B. The binding of substrates and a product of the enzymatic reaction to glutathione S-transferase A. J Biol Chem. 1979 Aug 10;254(15):7085–7089. [PubMed] [Google Scholar]
  22. Jansen P. L., Groothuis G. M., Peters W. H., Meijer D. F. Selective hepatobiliary transport defect for organic anions and neutral steroids in mutant rats with hereditary-conjugated hyperbilirubinemia. Hepatology. 1987 Jan-Feb;7(1):71–76. doi: 10.1002/hep.1840070116. [DOI] [PubMed] [Google Scholar]
  23. Jansen P. L., Oude Elferink R. P. Hereditary hyperbilirubinemias: a molecular and mechanistic approach. Semin Liver Dis. 1988 May;8(2):168–178. doi: 10.1055/s-2008-1040537. [DOI] [PubMed] [Google Scholar]
  24. Jansen P. L., Peters W. H., Lamers W. H. Hereditary chronic conjugated hyperbilirubinemia in mutant rats caused by defective hepatic anion transport. Hepatology. 1985 Jul-Aug;5(4):573–579. doi: 10.1002/hep.1840050408. [DOI] [PubMed] [Google Scholar]
  25. Juranka P. F., Zastawny R. L., Ling V. P-glycoprotein: multidrug-resistance and a superfamily of membrane-associated transport proteins. FASEB J. 1989 Dec;3(14):2583–2592. doi: 10.1096/fasebj.3.14.2574119. [DOI] [PubMed] [Google Scholar]
  26. Kamimoto Y., Gatmaitan Z., Hsu J., Arias I. M. The function of Gp170, the multidrug resistance gene product, in rat liver canalicular membrane vesicles. J Biol Chem. 1989 Jul 15;264(20):11693–11698. [PubMed] [Google Scholar]
  27. Kitamura T., Jansen P., Hardenbrook C., Kamimoto Y., Gatmaitan Z., Arias I. M. Defective ATP-dependent bile canalicular transport of organic anions in mutant (TR-) rats with conjugated hyperbilirubinemia. Proc Natl Acad Sci U S A. 1990 May;87(9):3557–3561. doi: 10.1073/pnas.87.9.3557. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Klaassen C. D., Watkins J. B., 3rd Mechanisms of bile formation, hepatic uptake, and biliary excretion. Pharmacol Rev. 1984 Mar;36(1):1–67. [PubMed] [Google Scholar]
  29. Oude Elferink R. P., Ottenhoff R., Liefting W. G., Schoemaker B., Groen A. K., Jansen P. L. ATP-dependent efflux of GSSG and GS-conjugate from isolated rat hepatocytes. Am J Physiol. 1990 May;258(5 Pt 1):G699–G706. doi: 10.1152/ajpgi.1990.258.5.G699. [DOI] [PubMed] [Google Scholar]
  30. Oude Elferink R. P., de Haan J., Lambert K. J., Hagey L. R., Hofmann A. F., Jansen P. L. Selective hepatobiliary transport of nordeoxycholate side chain conjugates in mutant rats with a canalicular transport defect. Hepatology. 1989 Jun;9(6):861–865. doi: 10.1002/hep.1840090612. [DOI] [PubMed] [Google Scholar]
  31. Ringe D., Petsko G. A. Cystic fibrosis. A transport problem? Nature. 1990 Jul 26;346(6282):312–313. doi: 10.1038/346312a0. [DOI] [PubMed] [Google Scholar]
  32. Rüegg U. T., Burgess G. M. Staurosporine, K-252 and UCN-01: potent but nonspecific inhibitors of protein kinases. Trends Pharmacol Sci. 1989 Jun;10(6):218–220. doi: 10.1016/0165-6147(89)90263-0. [DOI] [PubMed] [Google Scholar]
  33. Sato W., Yusa K., Naito M., Tsuruo T. Staurosporine, a potent inhibitor of C-kinase, enhances drug accumulation in multidrug-resistant cells. Biochem Biophys Res Commun. 1990 Dec 31;173(3):1252–1257. doi: 10.1016/s0006-291x(05)80921-0. [DOI] [PubMed] [Google Scholar]
  34. Taniguchi H., Pyerin W. Glutathione S-transferase is an in vitro substrate of Ca++-phospholipid-dependent protein kinase (protein kinase C). Biochem Biophys Res Commun. 1989 Aug 15;162(3):903–907. doi: 10.1016/0006-291x(89)90757-2. [DOI] [PubMed] [Google Scholar]
  35. Thiebaut F., Tsuruo T., Hamada H., Gottesman M. M., Pastan I., Willingham M. C. Cellular localization of the multidrug-resistance gene product P-glycoprotein in normal human tissues. Proc Natl Acad Sci U S A. 1987 Nov;84(21):7735–7738. doi: 10.1073/pnas.84.21.7735. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Vegesna R. V., Wu H. L., Mong S., Crooke S. T. Staurosporine inhibits protein kinase C and prevents phorbol ester-mediated leukotriene D4 receptor desensitization in RBL-1 cells. Mol Pharmacol. 1988 May;33(5):537–542. [PubMed] [Google Scholar]
  37. West I. C. What determines the substrate specificity of the multi-drug-resistance pump? Trends Biochem Sci. 1990 Feb;15(2):42–46. doi: 10.1016/0968-0004(90)90171-7. [DOI] [PubMed] [Google Scholar]
  38. Wolf M., Baggiolini M. The protein kinase inhibitor staurosporine, like phorbol esters, induces the association of protein kinase C with membranes. Biochem Biophys Res Commun. 1988 Aug 15;154(3):1273–1279. doi: 10.1016/0006-291x(88)90277-x. [DOI] [PubMed] [Google Scholar]

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