Skip to main content
SpringerLink
Log in

Biochemical Characterization of Highly Purified Cytochrome P-450 and Other Components of the Mixed Function Oxidase System of Liver Microsomal Membranes

  • Chapter
Cytochromes P-450 and b5

Part of the book series: Advances in Experimental Medicine and Biology ((AEMB,volume 58))

Abstract

The cytochrome P-450-containing enzyme system of liver microsomes is of particular interest to biochemists, pharmacologists, and toxicologists because of its ability to hydroxylate or otherwise modify not only steroids and fatty acids, but also drugs, insecticides, carcinogens, and a variety of other foreign compounds. Our laboratory accomplished the solubilization, resolution, and reconstitution of this enzyme some years ago (Lu and Coon, 1968; Coon and Lu, 1969) and has subsequently been concerned with the purification and biochemical characterization of the individual components.

This research was supported by Grant GB-30491X from the National Science Foundation and Grant AM-10339 from the United States Public Health Services.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Chapter
USD 29.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 39.99
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 54.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  • Autor, A. P., Kaschnitz, R. M., Heidema, J. K., van der Hoeven, T. A., Duppel, W., and Coon, M. J. 1973a. Role of phospholipid in the reconstituted liver microsomal mixed function oxidase system containing cytochrome P-450 and NADPH-cyto-chrome P-450 reductase. Drug Metab. Disp. 1:156–161.

    CAS  Google Scholar 

  • Autor, A. P., Kaschnitz, R. M., Heidema, J. K., and Coon, M. J. 1973b. Sedimentation and other properties of the reconstituted liver microsomal enzyme system containing cytochrome P-450, reduced triphosphopyridine nucleotide-cytochrome P-450 reductase, and phosphatidylcholine. Mol. Pharmacol. 9:93–104.

    PubMed  CAS  Google Scholar 

  • Ballou, D. P., Veeger, C., van der Hoeven, T. A., and Coon, M. J. 1974. Properties of partially purified liver microsomal cytochrome P-450: acceptance of two electrons during anaerobic titration. FEBS Letters 38:337–340.

    Article  PubMed  CAS  Google Scholar 

  • Bligh, E. G., and Dyer, W. J. 1959. A rapid method of total lipid extraction and purification. Can. J. Biochem. Physiol. 37:911–917.

    Article  PubMed  CAS  Google Scholar 

  • Burleigh, B. D., Jr., Foust, G. P., and Williams, C. H., Jr. 1969. A method for titrating oxygen-sensitive organic redox systems with reducing agents in solution. Anal. Biochem. 27:536–544.

    Article  PubMed  CAS  Google Scholar 

  • Coon, M. J., and Lu, A. Y. H. 1969. Fatty acid ω-oxidation in a soluble microsomal enzyme system containing P-450, pp. 151–166. In J. R. Gillette et al. (ed.). Microsomes and Drug Oxidations. Academic Press, New York.

    Google Scholar 

  • Coon, M. J., van der Hoeven, T. A., Kaschnitz, R. M., and Strobel, H. W. 1973a. Biochemical studies on cytochrome P-450 solubilized from liver microsomes: partial purification and mechanism of catalysis. Ann. N. Y. Acad. Sci. 212:449–457.

    Article  PubMed  CAS  Google Scholar 

  • Coon, M. J., Strobel, H. W., and Boyer, R. F. 1973b. On the mechanism of hydroxylation reactions catalyzed by cytochrome P-450. Drug Metab. Disp. 1:92–97.

    CAS  Google Scholar 

  • Duppel, W., Lebeault, J. M., and Coon, M. J. 1973. Properties of a yeast cytochrome P-450-containing enzyme system which catalyzes the hydroxylation of fatty acids, alkanes and drugs. Eur. J. Biochem. 36:583–592.

    Article  PubMed  CAS  Google Scholar 

  • Fujita, T., Shoeman, D. W., and Mannering, G. J. 1973. Differences in P-450 cytochromes from livers of rats treated with phenobarbital and with 3-methylcholanthrene. J. Biol. Chem. 248:2192–2201.

    Google Scholar 

  • Iyanagi, T., and Mason, H. S. 1973. Some properties of hepatic reduced nicotinamide adenine dinucleotide phosphate-cyto-chrome c reductase. Biochemistry 12:2297–2308.

    Article  PubMed  CAS  Google Scholar 

  • Kaschnitz, R. M., and Coon, M. J. 1972. Solubilized human liver cytochrome P-450: phospholipid requirement in hydroxylation reactions. Abstracts, Fifth International Congress on Pharmacology, San Francisco, p. 120.

    Google Scholar 

  • Kaschnitz, R. M., and Coon, M. J. 1974. Drug, fatty acid, and hydrocarbon hydroxylation by solubilized human liver cytochrome P-450: phospholipid requirement. Biochem. Pharmacol. in press.

    Google Scholar 

  • King, E. J., 1932. The colorimetrie determination of phosphorus. Biochem. J. 26:292–297.

    PubMed  CAS  Google Scholar 

  • Kusunose, M., Kusunose, E., and Coon, M. J. 1964. Enzymatic ω-oxidation of fatty acids. I. Products of octanoate, decanoate and laurate oxidation. J. Biol. Chem. 239:1374–1380.

    PubMed  CAS  Google Scholar 

  • Laemmli, U. K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685.

    Article  PubMed  CAS  Google Scholar 

  • Lebeault, J. M., Lode, E. T., and Coon, M. J. 1971. Fatty acid and hydrocarbon hydroxylation in yeast: role of cytochrome P-450 in Candida tropicalis. Biochem. Biophys. Res. Commun.42: 413–419.

    Article  PubMed  CAS  Google Scholar 

  • Levin, W., Lu, A. Y. H., Ryan, D., West, S., Kuntzman, R. and Conney, A. H. 1972. Partial purification and properties of cytochromes P-450 and P-448 from rat liver microsomes. Arch. Biochem. Biophys. 153:543–553.

    Article  PubMed  CAS  Google Scholar 

  • Levin, W., Ryan, D., West, S., and Lu, A. Y. H. 1974. Preparation of partially purified, lipid depleted cytochrome P-450 and reduced nicotinamide adenine dinucleotide phosphate-cytochrome c reductase from rat liver microsomes. J. Biol. Chem. 249: 1747–1754.

    PubMed  CAS  Google Scholar 

  • Lowry, O., Rosebrough, N. J., Farr, A. L., and Randall, R. J. 1951. Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193:265–275.

    PubMed  CAS  Google Scholar 

  • Lu, A. Y. H., and Coon, M. J. 1968. Role of hemoprotein P-450 in fatty acid ω-hydroxylation in a soluble enzyme system from liver microsomes. J. Biol. Chem. 243:1331–1332.

    PubMed  CAS  Google Scholar 

  • Lu, A. Y. H., Junk, K. W., and Coon, M. J. 1969a. Resolution of the cytochrome P-450-containing ω-hydroxylation system of liver microsomes into three components. J. Biol. Chem. 244:3714–3721.

    PubMed  CAS  Google Scholar 

  • Lu, A. Y. H., Strobel, H. W., and Coon, M. J. 1969b. Hydroxylation of benzphetamine and other drugs by a solubilized form of cytochrome P-450 from liver microsomes: lipid requirement for drug demethylation. Biochem. Biophys. Res. Commun. 36:545–551.

    Article  PubMed  CAS  Google Scholar 

  • Lu, A. Y. H., Strobel, H. W., and Coon, M. J. 1970. Properties of a solubilized form of the cytochrome P-450-containing mixed function oxidase of liver microsomes. Mol. Pharmacol. 6:213–220.

    PubMed  CAS  Google Scholar 

  • Lu, A. Y. H., Kuntzman, R., West, S., Jacobson, M., and Conney, A. H. 1972. Reconstituted liver microsomal enzyme system that hydroxylates drugs, other foreign compounds, and endogenous substrates. II. Role of cytochrome P-450 and P-448 fractions in drug and steroid hydroxylations. J. Biol. Chem. 247:1727–1734.

    PubMed  CAS  Google Scholar 

  • Nebert, D. W., Heidema, J. K., Strobel, H. W. and Coon, M. J. 1973. Genetic expression of aryl hydrocarbon hydroxylase induction. Genetic specificity resides in the fraction containing cytochrome P448 and P450. J. Biol. Chem. 248:7631–7636.

    PubMed  CAS  Google Scholar 

  • Omura, T., and Sato, R. 1964. The carbon monoxide-binding pigment of liver microsomes. I. Evidence for its hemeprotein nature. J. Biol. Chem. 239:2370–2378.

    Google Scholar 

  • Poland, A. P., and Nebert, D. W. 1973. A sensitive radiometric assay of aminopyrine N-demethylation. J. Pharmacol. Exp. Ther. 184:269–277.

    PubMed  CAS  Google Scholar 

  • Sato, R., Satake, H., and Imai, Y. 1973. Partial purification and some spectral properties of hepatic microsomal cytochrome P-450. Drug Metab. Disp. 1:6–13.

    CAS  Google Scholar 

  • Shoeman, D. W., White, J. G., and Mannering, G. J. 1970. Role of lipid induced aggregation in the function of “soluble” cytochrome P-450. Pharmacologist 12:260.

    Google Scholar 

  • Spatz, L., and Strittmatter, P. 1971. A form of cytochrome 5 that contains an additional hydrophobic sequence of 40 amino acid residues. Proc. Nat. Acad. Sci. U. S. 68:1042–1046.

    Article  CAS  Google Scholar 

  • Strobel, H. W., Lu, A. Y. H., Heidema, J., and Coon, M. J. 1970. Phosphatidylcholine requirement in the enzymatic reduction of hemoprotein P-450 and in fatty acid, hydrocarbon, and drug hydroxylation. J. Biol. Chem. 245:4851–4854.

    PubMed  CAS  Google Scholar 

  • Strobel, H. W., and Coon, M. J. 1971. Effect of superoxide generation and dismutation on hydroxylation reactions catalyzed by liver microsomal cytochrome P-450. J. Biol. Chem. 246:7826–7829.

    PubMed  CAS  Google Scholar 

  • van der Hoeven, T. A., and Coon, M. J. 1974. Preparation and properties of partially purified cytochrome P-450 and NADPH-cytochrome P-450 reductase from rabbit liver microsomes. J. Biol. Chem. 249: in press.

    Google Scholar 

  • van der Hoeven, T. A., Haugen, D. A., and Coon, M. J. 1974. Separation and characterization of multiple forms of liver microsomal cytochrome P-450. Pharmacologist 16: in press.

    Google Scholar 

  • Wiliams, C. H., Jr., and Kamin, H. 1962. Microsomal triphospho-pyridine nucleotide-cytochrome c reductase of liver. J. Biol. Chem. 237:587–595.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Biological Chemistry, Medical School, The University of Michigan, Ann Arbor, Michigan, 48104, USA

    Minor J. Coon, Theodore A. van der Hoeven, David A. Haugen (Predoctoral Trainee), F. Peter Guengerich, Janice L. Vermilion (Postdoctoral Fellow) & David P. Ballou

  2. United States Public Health Service, USA

    Janice L. Vermilion (Postdoctoral Fellow)

  3. United States Public Health Service, Grant, GM-00187, USA

    David A. Haugen (Predoctoral Trainee)

Authors
  1. Minor J. Coon
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Theodore A. van der Hoeven
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. David A. Haugen
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. F. Peter Guengerich
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Janice L. Vermilion
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. David P. Ballou
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. Harrison Department of Surgical Research, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania, USA

    David Y. Cooper  & Otto Rosenthal  & 

  2. Department of Pharmacology, Thomas Jefferson University, Philadelphia, Pennsylvania, USA

    Robert Snyder  & Charlotte Witmer  & 

Rights and permissions

Reprints and permissions

Copyright information

© 1975 Plenum Press, New York

About this chapter

Cite this chapter

Coon, M.J., van der Hoeven, T.A., Haugen, D.A., Guengerich, F.P., Vermilion, J.L., Ballou, D.P. (1975). Biochemical Characterization of Highly Purified Cytochrome P-450 and Other Components of the Mixed Function Oxidase System of Liver Microsomal Membranes. In: Cooper, D.Y., Rosenthal, O., Snyder, R., Witmer, C. (eds) Cytochromes P-450 and b5 . Advances in Experimental Medicine and Biology, vol 58. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-9026-2_2

Download citation

  • DOI: https://doi.org/10.1007/978-1-4615-9026-2_2

  • Publisher Name: Springer, Boston, MA

  • Print ISBN: 978-1-4615-9028-6

  • Online ISBN: 978-1-4615-9026-2

  • eBook Packages: Springer Book Archive

Publish with us

Policies and ethics

Search

Navigation