Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1996 Apr 1;315(Pt 1):171–176. doi: 10.1042/bj3150171

Evidence for an UDP-glucuronic acid/phenol glucuronide antiport in rat liver microsomal vesicles.

G Bánhegyi 1, L Braun 1, P Marcolongo 1, M Csala 1, R Fulceri 1, J Mandl 1, A Benedetti 1
PMCID: PMC1217167  PMID: 8670103

Abstract

The transport of glucuronides synthesized in the luminal compartment of the endoplasmic reticulum by UDP-glucuronosyltransferase isoenzymes was studied in rat liver microsomal vesicles. Microsomal vesicles were loaded with p-nitrophenol glucuronide (5 mM), phenolphthalein glucuronide or UDP-glucuronic acid, by a freeze-thawing method. In was shown that: (i) the loading procedure resulted in millimolar intravesicular concentrations of the different loading compounds; (ii) addition of UDP-glucuronic acid (5 mM) to the vesicles released both intravesicular glucuronides within 1 min; (iii) glucuronides stimulated the release of UDP-glucuronic acid from UDP acid-loaded microsomal vesicles; (iv) trans-stimulation of UDP-glucuronic acid entry by loading of microsomal vesicles with p-nitrophenol glucuronide, phenolphthalein glucuronide, UDP-glucuronic acid and UDP-N-acetyl-glucosamine almost completely abolished the latency of UDP-glucuronosyltransferase, although mannose 6-phosphatase latency remained unaltered; (v) the loading compounds by themselves did not stimulate UDP-glucuronosyltransferase activity. This study indicates that glucuronides synthesized in the lumen of endoplasmic reticulum can leave by an antiport, which concurrently transports USP-glucuronic acid into the lumen of the endoplasmic reticulum.

Full Text

The Full Text of this article is available as a PDF (254.2 KB).

Selected References

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

  1. Akerboom T. P., Narayanaswami V., Kunst M., Sies H. ATP-dependent S-(2,4-dinitrophenyl)glutathione transport in canalicular plasma membrane vesicles from rat liver. J Biol Chem. 1991 Jul 15;266(20):13147–13152. [PubMed] [Google Scholar]
  2. Berg C. L., Radominska A., Lester R., Gollan J. L. Membrane translocation and regulation of uridine diphosphate-glucuronic acid uptake in rat liver microsomal vesicles. Gastroenterology. 1995 Jan;108(1):183–192. doi: 10.1016/0016-5085(95)90023-3. [DOI] [PubMed] [Google Scholar]
  3. Berry C., Hallinan T. Summary of a novel, three-component regulatory model for uridine diphosphate glucuronyltransferase. Biochem Soc Trans. 1976;4(4):650–652. doi: 10.1042/bst0040650. [DOI] [PubMed] [Google Scholar]
  4. Bock K. W., Burchell B., Dutton G. J., Hänninen O., Mulder G. J., Owens I. S., Siest G., Tephly T. R. UDP-glucuronosyltransferase activities. Guidelines for consistent interim terminology and assay conditions. Biochem Pharmacol. 1983 Mar 15;32(6):953–955. doi: 10.1016/0006-2952(83)90610-x. [DOI] [PubMed] [Google Scholar]
  5. Bossuyt X., Blanckaert N. Carrier-mediated transport of intact UDP-glucuronic acid into the lumen of endoplasmic-reticulum-derived vesicles from rat liver. Biochem J. 1994 Aug 15;302(Pt 1):261–269. doi: 10.1042/bj3020261. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bossuyt X., Blanckaert N. Mechanism of stimulation of microsomal UDP-glucuronosyltransferase by UDP-N-acetylglucosamine. Biochem J. 1995 Jan 1;305(Pt 1):321–328. doi: 10.1042/bj3050321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Burchell B., Coughtrie M. W. UDP-glucuronosyltransferases. Pharmacol Ther. 1989;43(2):261–289. doi: 10.1016/0163-7258(89)90122-8. [DOI] [PubMed] [Google Scholar]
  8. Bánhegyi G., Bellomo G., Fulceri R., Mandl J., Benedetti A. Intraluminal calcium of the liver endoplasmic reticulum stimulates the glucuronidation of p-nitrophenol. Biochem J. 1993 May 15;292(Pt 1):99–104. doi: 10.1042/bj2920099. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Bánhegyi G., Garzó T., Fulceri R., Benedetti A., Mandl J. Latency is the major determinant of UDP-glucuronosyltransferase activity in isolated hepatocytes. FEBS Lett. 1993 Aug 9;328(1-2):149–152. doi: 10.1016/0014-5793(93)80983-2. [DOI] [PubMed] [Google Scholar]
  10. Dutton G. J. Commentary: Control of UDP-glucuronyltransferase activity. Biochem Pharmacol. 1975 Oct 15;24(20):1835–1841. doi: 10.1016/0006-2952(75)90399-8. [DOI] [PubMed] [Google Scholar]
  11. Fulceri R., Bellomo G., Gamberucci A., Romani A., Benedetti A. Physiological concentrations of inorganic phosphate affect MgATP-dependent Ca2+ storage and inositol trisphosphate-induced Ca2+ efflux in microsomal vesicles from non-hepatic cells. Biochem J. 1993 Jan 1;289(Pt 1):299–306. doi: 10.1042/bj2890299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fulceri R., Bellomo G., Gamberucci A., Scott H. M., Burchell A., Benedetti A. Permeability of rat liver microsomal membrane to glucose 6-phosphate. Biochem J. 1992 Sep 15;286(Pt 3):813–817. doi: 10.1042/bj2860813. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Fulceri R., Bánhegyi G., Gamberucci A., Giunti R., Mandl J., Benedetti A. Evidence for the intraluminal positioning of p-nitrophenol UDP-glucuronosyltransferase activity in rat liver microsomal vesicles. Arch Biochem Biophys. 1994 Feb 15;309(1):43–46. doi: 10.1006/abbi.1994.1081. [DOI] [PubMed] [Google Scholar]
  14. Guillén E., Hirschberg C. B. Transport of adenosine triphosphate into endoplasmic reticulum proteoliposomes. Biochemistry. 1995 Apr 25;34(16):5472–5476. doi: 10.1021/bi00016a018. [DOI] [PubMed] [Google Scholar]
  15. Hauser S. C., Ziurys J. C., Gollan J. L. A membrane transporter mediates access of uridine 5'-diphosphoglucuronic acid from the cytosol into the endoplasmic reticulum of rat hepatocytes: implications for glucuronidation reactions. Biochim Biophys Acta. 1988 Nov 17;967(2):149–157. doi: 10.1016/0304-4165(88)90004-9. [DOI] [PubMed] [Google Scholar]
  16. Hwang C., Sinskey A. J., Lodish H. F. Oxidized redox state of glutathione in the endoplasmic reticulum. Science. 1992 Sep 11;257(5076):1496–1502. doi: 10.1126/science.1523409. [DOI] [PubMed] [Google Scholar]
  17. Iida S., Mizuma T., Sakuma N., Hayashi M., Awazu S. Transport of acetaminophen conjugates in isolated rat hepatocytes. Drug Metab Dispos. 1989 May-Jun;17(3):341–344. [PubMed] [Google Scholar]
  18. Jansen P. L., Mulder G. J., Burchell B., Bock K. W. New developments in glucuronidation research: report of a workshop on "glucuronidation, its role in health and disease". Hepatology. 1992 Mar;15(3):532–544. doi: 10.1002/hep.1840150328. [DOI] [PubMed] [Google Scholar]
  19. Kobayashi K., Komatsu S., Nishi T., Hara H., Hayashi K. ATP-dependent transport for glucuronides in canalicular plasma membrane vesicles. Biochem Biophys Res Commun. 1991 Apr 30;176(2):622–626. doi: 10.1016/s0006-291x(05)80229-3. [DOI] [PubMed] [Google Scholar]
  20. Matsui M., Nagai F. Assay of uridine diphosphate glucuronosyltransferase by high-pressure liquid chromatography. Anal Biochem. 1980 Jun;105(1):141–146. doi: 10.1016/0003-2697(80)90435-2. [DOI] [PubMed] [Google Scholar]
  21. McPhail M. E., Knowles R. G., Salter M., Dawson J., Burchell B., Pogson C. I. Uptake of acetaminophen (paracetamol) by isolated rat liver cells. Biochem Pharmacol. 1993 Apr 22;45(8):1599–1604. doi: 10.1016/0006-2952(93)90300-l. [DOI] [PubMed] [Google Scholar]
  22. Meissner G. Ionic permeability of isolated muscle sarcoplasmic reticulum and liver endoplasmic reticulum vesicles. Methods Enzymol. 1988;157:417–437. doi: 10.1016/0076-6879(88)57094-5. [DOI] [PubMed] [Google Scholar]
  23. Moldéus P., Vadi H., Berggren M. Oxidative and conjugative metabolism of p-nitroanisole and p-nitrophenol in isolated rat liver cells. Acta Pharmacol Toxicol (Copenh) 1976 Jul;39(1):17–32. doi: 10.1111/j.1600-0773.1976.tb03151.x. [DOI] [PubMed] [Google Scholar]
  24. Nilsson R., Peterson E., Dallner G. Permeability of microsomal membranes isolated from rat liver. J Cell Biol. 1973 Mar;56(3):762–776. doi: 10.1083/jcb.56.3.762. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Norling A., Andersson B., Berggren M., Moldéus P. Uptake of glucuronides into isolated hepatocytes and their effects on glucuronide and sulphate conjugation. Acta Pharmacol Toxicol (Copenh) 1978 Oct;43(4):311–317. doi: 10.1111/j.1600-0773.1978.tb02271.x. [DOI] [PubMed] [Google Scholar]
  26. Ojcius D. M., Young J. D. Cytolytic pore-forming proteins and peptides: is there a common structural motif? Trends Biochem Sci. 1991 Jun;16(6):225–229. doi: 10.1016/0968-0004(91)90090-i. [DOI] [PubMed] [Google Scholar]
  27. Sundheimer D. W., Brendel K. Metabolism of harmol and transport of harmol conjugates in isolated rat hepatocytes. Drug Metab Dispos. 1983 Sep-Oct;11(5):433–440. [PubMed] [Google Scholar]
  28. Vanstapel F., Blanckaert N. Carrier-mediated translocation of uridine diphosphate glucose into the lumen of endoplasmic reticulum-derived vesicles from rat liver. J Clin Invest. 1988 Sep;82(3):1113–1122. doi: 10.1172/JCI113668. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Vanstapel F., Blanckaert N. Endogenous esterification of bilirubin by liver microsomes. Evidence for an intramicrosomal pool of UDP-glucose and lumenal orientation of bilirubin UDP-glycosyltransferase. J Biol Chem. 1987 Apr 5;262(10):4616–4623. [PubMed] [Google Scholar]
  30. Vessey D. A., Zakim D. Stimulation of microsomal uridine diphosphate glucuronyltransferase by glucuronic acid derivatives. Biochem J. 1974 Apr;139(1):243–249. doi: 10.1042/bj1390243. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Waddell I. D., Gibb L., Burchell A. Calcium activates glucose-6-phosphatase in intact rat hepatic microsomes. Biochem J. 1990 Apr 15;267(2):549–551. doi: 10.1042/bj2670549. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. von Bahr C., Borgå O. Uptake, metabolism and excretion of desmethylimipramine and its metabolites in the isolated perfused rat liver. Acta Pharmacol Toxicol (Copenh) 1971;29(4):359–374. doi: 10.1111/j.1600-0773.1971.tb00596.x. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES