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Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1974 Nov;71(11):4293–4297. doi: 10.1073/pnas.71.11.4293

Cobalt Induction of Hepatic Heme Oxygenase; with Evidence That Cytochrome P-450 Is Not Essential for This Enzyme Activity

M D Maines 1, Attallah Kappas 1
PMCID: PMC433868  PMID: 4530983

Abstract

Treatment of rats in vivo with cobalt chloride stimulated heme oxidation by hepatic microsomes to levels up to 800% above controls. This treatment also caused increases in liver weight and in total microsomal protein; in contrast, marked decreases were produced in microsomal oxidation of ethylmorphine (80%), and in cytochrome P-450 (60-70%) and heme (30-50%) contents. Cobalt chloride treatment did not affect heme oxidation by the spleen heme oxygenase system.

The rate of heme oxidation by hepatic microsomal enzymes and the microsomal content of cytochrome P-450 were found to be unrelated. This conclusion was reached from studies in which microsomal heme oxygenase activity from cobalt-treated animals could be increased by 900% above control levels in the same microsomal preparation in which cytochrome P-450 content was decreased to spectrally unmeasurable amounts after incubation with 4 M urea. The same treatment eliminated ehtylmorphine demethylation and decreased microsomal NADPH-cytochrome c reductase (EC 1.6.2.4) activity by 75%.

It is concluded that (i) the hepatic microsomal enzyme system that oxidizes heme compounds is not the same as that which metabolizes drugs, (ii) cytochrome P-450 is not essential for the oxidation of heme by liver cells, (iii) there is no direct relationship between the rate of heme oxidation and the level of NADPH-cytochrome c reductase activity, and (iv) the oxidation of heme is protein-dependent and the active proteins are inducible, but are different from those involved in drug metabolism.

Keywords: heme oxidation, enzyme induction, mixed-function oxidation

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Selected References

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

  1. Correia M. A., Mannering G. J. Reduced diphosphopyridine nucleotide synergism of the reduced triphosphopyridine nucleotide-dependent mixed-function oxidase system of hepatic microsomes. I. Effects of activation and inhibition of the fatty acyl coenzyme A desaturation system. Mol Pharmacol. 1973 Jul;9(4):455–469. [PubMed] [Google Scholar]
  2. 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]
  3. Lathe G. H. The degradation of haem by mammals and its excretion as conjugated bilirubin. Essays Biochem. 1972;8:107–148. [PubMed] [Google Scholar]
  4. Maines M. D., Anders M. W., Muller-Eberhard U. Studies on heme transfer from microsomal hemoproteins to heme-binding plasma proteins. Mol Pharmacol. 1974 Mar;10(2):204–213. [PubMed] [Google Scholar]
  5. Mason H. S., North J. C., Vanneste M. Microsomal mixed-function oxidations: the metabolism of xenobiotics. Fed Proc. 1965 Sep-Oct;24(5):1172–1180. [PubMed] [Google Scholar]
  6. NASH T. The colorimetric estimation of formaldehyde by means of the Hantzsch reaction. Biochem J. 1953 Oct;55(3):416–421. doi: 10.1042/bj0550416. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Nichol A. W. The formation of biliverdin by chicken macrophages in tissue culture. Observations on the effect of inhibitors. Biochim Biophys Acta. 1970 Oct 27;222(1):28–40. doi: 10.1016/0304-4165(70)90348-x. [DOI] [PubMed] [Google Scholar]
  8. OMURA T., SATO R. THE CARBON MONOXIDE-BINDING PIGMENT OF LIVER MICROSOMES. II. SOLUBILIZATION, PURIFICATION, AND PROPERTIES. J Biol Chem. 1964 Jul;239:2379–2385. [PubMed] [Google Scholar]
  9. OSTROW J. D., HAMMAKER L., SCHMID R. The preparation of crystalline bilirubin-C14. J Clin Invest. 1961 Aug;40:1442–1452. doi: 10.1172/JCI104375. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Schacter B. A., Mason J. I. The effect of phenobarbital, 3-methylcholanthrene, 3,4-benzpyrene, and pregnenolone-16 alpha-carbonitrile on microsomal heme oxygenase and splenic cytochrome P-450. Arch Biochem Biophys. 1974 Jan;160(1):274–278. doi: 10.1016/s0003-9861(74)80034-2. [DOI] [PubMed] [Google Scholar]
  11. Schacter B. A., Nelson E. B., Marver H. S., Masters B. S. Immunochemical evidence for an association of heme oxygenase with the microsomal electron transport system. J Biol Chem. 1972 Jun 10;247(11):3601–3607. [PubMed] [Google Scholar]
  12. Tenhunen R., Marver H. S., Schmid R. Microsomal heme oxygenase. Characterization of the enzyme. J Biol Chem. 1969 Dec 10;244(23):6388–6394. [PubMed] [Google Scholar]
  13. Tenhunen R., Marver H. S., Schmid R. The enzymatic catabolism of hemoglobin: stimulation of microsomal heme oxygenase by hemin. J Lab Clin Med. 1970 Mar;75(3):410–421. [PubMed] [Google Scholar]
  14. Tenhunen R., Marver H. S., Schmid R. The enzymatic conversion of heme to bilirubin by microsomal heme oxygenase. Proc Natl Acad Sci U S A. 1968 Oct;61(2):748–755. doi: 10.1073/pnas.61.2.748. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Tenhunen R., Marver H. S., Schmid R. The enzymatic conversion of hemoglobin to bilirubin. Trans Assoc Am Physicians. 1969;82:363–371. [PubMed] [Google Scholar]
  16. Tenhunen R., Marver H., Pimstone N. R., Trager W. F., Cooper D. Y., Schmid R. Enzymatic degradation of heme. Oxygenative cleavage requiring cytochrome P-450. Biochemistry. 1972 Apr 25;11(9):1716–1720. doi: 10.1021/bi00759a029. [DOI] [PubMed] [Google Scholar]
  17. Tephly T. R., Hibbeln P. The effect of cobalt chloride administration on the synthesis of hepatic microsomal cytochrome P-450. Biochem Biophys Res Commun. 1971 Feb 19;42(4):589–595. doi: 10.1016/0006-291x(71)90528-6. [DOI] [PubMed] [Google Scholar]
  18. WILLIAMS C. H., Jr, KAMIN H. Microsomal triphosphopyridine nucleotide-cytochrome c reductase of liver. J Biol Chem. 1962 Feb;237:587–595. [PubMed] [Google Scholar]

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