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. 1986 Jun 15;236(3):625–633. doi: 10.1042/bj2360625

Enzymic oxidation of unconjugated bilirubin by rat liver.

R Cardenas-Vazquez, O Yokosuka, B H Billing
PMCID: PMC1146892  PMID: 3790083

Abstract

The presence of the enzyme bilirubin oxidase, which degrades bilirubin in vitro, was demonstrated in the liver. Subcellular-fractionation experiments indicate that bilirubin oxidase is located in both the inner and outer membranes of the mitochondria. The mean rate of the reaction is 1.57 +/- 0.38 (S.D.) nmol of bilirubin degraded/min per mg of mitochondrial protein (munits/mg of protein). With respect to the overall breakdown of bilirubin, the enzyme has a Km' of 136 microM-bilirubin and a Vmax.' of 9.13 munits/mg of protein. Its activity is influenced by the ionic strength of the media and is inhibited by KCN, thiol reagents, NADH and albumin. The enzyme is aerobic, and between 1 and 1.5 mol of O2 are consumed per mol of bilirubin degraded. The products of the reaction include propentdyopents. The hepatic bilirubin oxidase activity of the jaundiced Gunn-rat liver is not significantly different from that of the Sprague-Dawley rat, and it is not induced by beta-naphthoflavone.

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Selected References

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

  1. Benga G., Hodarnau A., Tilinca R., Porutiu D., Dancea S., Pop V., Wrigglesworth J. Fractionation of human liver mitochondria: enzymic and morphological characterization of the inner and outer membranes as compared to rat liver mitochondria. J Cell Sci. 1979 Feb;35:417–429. doi: 10.1242/jcs.35.1.417. [DOI] [PubMed] [Google Scholar]
  2. Blume H. Die Isolierung hochgereinigter Mitochondrienpräparationen für Untersuchungen zur Biotransformation von Arzneistoffen. Arch Pharm (Weinheim) 1979 Jul;312(7):561–572. doi: 10.1002/ardp.19793120702. [DOI] [PubMed] [Google Scholar]
  3. Brodersen R., Bartels P. Enzymatic oxidation of bilirubin. Eur J Biochem. 1969 Oct;10(3):468–473. doi: 10.1111/j.1432-1033.1969.tb00712.x. [DOI] [PubMed] [Google Scholar]
  4. COOPERSTEIN S. J., LAZAROW A. A microspectrophotometric method for the determination of cytochrome oxidase. J Biol Chem. 1951 Apr;189(2):665–670. [PubMed] [Google Scholar]
  5. Chappell J. B. The oxidation of citrate, isocitrate and cis-aconitate by isolated mitochondria. Biochem J. 1964 Feb;90(2):225–237. doi: 10.1042/bj0900225. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. DE DUVE C., PRESSMAN B. C., GIANETTO R., WATTIAUX R., APPELMANS F. Tissue fractionation studies. 6. Intracellular distribution patterns of enzymes in rat-liver tissue. Biochem J. 1955 Aug;60(4):604–617. doi: 10.1042/bj0600604. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Ellis G., Goldberg D. M. Optimal conditions for the kinetic assay of serum glutamate dehydrogenase activity at 37 degrees C. Clin Chem. 1972 Jun;18(6):523–527. [PubMed] [Google Scholar]
  8. GILBERTSEN A. S., LOWRY P. T., HAWKINSON V., WATSON C. J. Studies of the dipyrrylmethene ("fuscin") pigments. I. The anabolic significance of the fecal mesobilifuscin. J Clin Invest. 1959 Jul;38(7):1166–1174. doi: 10.1172/JCI103892. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kapitulnik J., Ostrow J. D. Stimulation of bilirubin catabolism in jaundiced Gunn rats by an induced of microsomal mixed-function monooxygenases. Proc Natl Acad Sci U S A. 1978 Feb;75(2):682–685. doi: 10.1073/pnas.75.2.682. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  11. Leighton F., Poole B., Beaufay H., Baudhuin P., Coffey J. W., Fowler S., De Duve C. The large-scale separation of peroxisomes, mitochondria, and lysosomes from the livers of rats injected with triton WR-1339. Improved isolation procedures, automated analysis, biochemical and morphological properties of fractions. J Cell Biol. 1968 May;37(2):482–513. doi: 10.1083/jcb.37.2.482. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. MICHAUELSSON M., NOSSLIN B., SJOELIN S. PLASMA BILIRUBIN DETERMINATION IN THE NEWBORN INFANT. A METHODOLOGICAL STUDY WITH SPECIAL REFERENCE TO THE INFLUENCE OF HEMOLYSIS. Pediatrics. 1965 Jun;35:925–931. [PubMed] [Google Scholar]
  13. McDonagh A. F., Assisi F. Commercial bilirubin: A trinity of isomers. FEBS Lett. 1971 Nov 1;18(2):315–317. doi: 10.1016/0014-5793(71)80475-1. [DOI] [PubMed] [Google Scholar]
  14. McDonagh A. F., Assisi F. The ready isomerization of bilirubin IX- in aqueous solution. Biochem J. 1972 Sep;129(3):797–800. doi: 10.1042/bj1290797. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. McDonagh A. F., Palma L. A. Preparation and properties of crystalline biliverdin IX alpha. Simple methods for preparing isomerically homogeneous biliverdin and [14C[biliverdin by using 2,3-dichloro-5,6-dicyanobenzoquinone. Biochem J. 1980 Aug 1;189(2):193–208. doi: 10.1042/bj1890193. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Mertens B. B., Van de Vijver M., Heirwegh K. P. Determination of bilirubin in liver homogenate with diazotized p-iodoaniline. Improvement of color recovery by incorporation of antioxidant (2,6-di-tert-butyl-p-cresol) in the reaction system. Anal Biochem. 1972 Dec;50(2):652–655. doi: 10.1016/0003-2697(72)90080-2. [DOI] [PubMed] [Google Scholar]
  17. Nebert D. W., Gelboin H. V. Substrate-inducible microsomal aryl hydroxylase in mammalian cell culture. I. Assay and properties of induced enzyme. J Biol Chem. 1968 Dec 10;243(23):6242–6249. [PubMed] [Google Scholar]
  18. Peters T. J., Seymour C. A. Analytical subcellular fractionation of needle-biopsy specimens from human liver. Biochem J. 1978 Aug 15;174(2):435–446. doi: 10.1042/bj1740435. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. SCHMID R., HAMMAKER L. METABOLISM AND DISPOSITION OF C14-BILIRUBIN IN CONGENITAL NONHEMOLYTIC JAUNDICE. J Clin Invest. 1963 Nov;42:1720–1734. doi: 10.1172/JCI104858. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Seymour C. A., Peters T. J. Enzyme activities in human liver biopsies: assay methods and activities of some lysosomal and membrane-bound enzymes in control tissue and serum. Clin Sci Mol Med. 1977 Mar;52(3):229–239. doi: 10.1042/cs0520229. [DOI] [PubMed] [Google Scholar]
  21. Sottocasa G. L., Kuylenstierna B., Ernster L., Bergstrand A. An electron-transport system associated with the outer membrane of liver mitochondria. A biochemical and morphological study. J Cell Biol. 1967 Feb;32(2):415–438. doi: 10.1083/jcb.32.2.415. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. WATSON C. J. The direct preparation of crystalline urobilin from bilirubin. J Biol Chem. 1953 Feb;200(2):691–696. [PubMed] [Google Scholar]
  23. Zenone E. A., Stoll M. S., Ostrow J. D. The effect of elimination of environmental light on the metabolism of unconjugated bilirubin in the Gunn rat. Dig Dis Sci. 1982 Dec;27(12):1117–1120. doi: 10.1007/BF01391450. [DOI] [PubMed] [Google Scholar]

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