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NADPH-Dependent Cytochrome P450 Reductase

  • Anthony Y. H. Lu
Part of the NATO ASI Series Advanced Science Institutes Series book series (NSSA, volume 202)

Abstract

The pioneering studies of Horecker (1), Phillips and Langdon (2), Williams and Kamin (3) established that the microsomal NADPH-cytochrome c reductase is a flavoprotein capable of reducing various electron acceptors. Subsequent studies by other investigators (4–6) suggested an involvement of this reductase in microsomal hydroxylation since like cytochrome P450, it is inducible by phenobarbital. In addition, microsomal monooxygenase activity can be inhibited by cytochrome c (7) and antibodies against the reductase (8–10). A direct involvement of this enzyme in hydroxylation was established when the liver microsomal monooxygenase system was solubilized, resolved and reconstituted (11, 12). In these studies, the flavoprotein was shown to be an obligatory component in hydroxylation, transferring reducing equivalents from NADPH to cytochrome P450. Since then, the term NADPH-cytochrome P450 reductase has been used to reflect its physiological function.

Keywords

Dynamic Nuclear Polarization P450 Reductase Phospholipid Vesicle Cytochrome P450 Reductase Hydrophilic Domain 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. 1.
    B. L. Horecker, Triphosphopyridine nucleotide-cytochrome c reductase in liver, J. Biol. Chem. 183:593 (1950).Google Scholar
  2. 2.
    A. H. Phillips and R. G. Langdon, Hepatic triphosphopyridine nucleotide-cytochrome c reductase: isolation, characterization and kinetic studies, J. Biol. Chem. 237:2652 (1962).PubMedGoogle Scholar
  3. 3.
    C. H. Williams and H. Kamin, Microsomal triphosphopyridine nucleotide-cytochrome c reductase of liver, J. Biol. Ghem. 237:587 (1962).Google Scholar
  4. 4.
    L. Ernster and S. Orrenius, Substrate-induced synthesis of the hydroxylating enzyme system of liver microsomes, Fed. Proc. 24:1190 (1965).PubMedGoogle Scholar
  5. 5.
    L. Shuster and H. Jick, The turnover of microsomal protein in the livers of phenobarbital-treated mice, J. Biol. Chem. 241:5361 (1966).Google Scholar
  6. 6.
    Y. Kuriyama, T. Omura, P. Siekevitz and G. E. Palade, Effects of phenobarbital on the synthesis and degradation of the protein components of rat liver microsomal membranes, J. Biol. Chem. 244:2017 (1969).PubMedGoogle Scholar
  7. 7.
    J. R. Gillette, B. B. Brodie and B. N. La Du, The oxidation of drugs by liver microsomes: on the role of TPNH and oxygen, J. Pharmacol. Exp. Ther. 119:532 (1957).PubMedGoogle Scholar
  8. 8.
    F. Wada, H. Shibata, M. Goto and Y. Sakamoto, Participation of microsomal electron transport system involving cytochrome P450 in ω-oxidation of fatty acids, Biochim. Biophys. Acta 162:518 (1968).PubMedCrossRefGoogle Scholar
  9. 9.
    R. I. Glazer, J. B. Schenkman and A. C. Sartorrelli, Immunochemical studies on the role of reduced nicotinamide adenine dinucleotide phosphate-cytochrome c reductase in drug oxidation, Mol. Pharmacol. 7:683 (1971).PubMedGoogle Scholar
  10. 10.
    B. S. S. Masters, J. Baron, W. E. Taylor, E. L. Isaacson and L. Spalluto, Immunochemical studies on electron transport chains involving cytochrome P450. I. Effects of antibodies to pig liver microsomal reduced triphosphopyridine nucleotide-cytochrome c reductase and the non-heme iron protein from bovine adrenocorticol mitochondria, J. Biol. Chem. 246:4143 (1971).PubMedGoogle Scholar
  11. 11.
    A. Y. H. Lu and M. J. Coon, Role of hemoprotein P450 in fatty acid ω-hydroxylation in a soluble enzyme system from liver microsomes, J. Biol. Chem. 243:1331 (1968).PubMedGoogle Scholar
  12. 12.
    A. Y. H. Lu, K. W. Junk and M. J. Coon, Resolution of the cytochrome P450-containing ω-hydroxylation system of liver microsomes into three components, J. Biol. Chem. 244:3714 (1969).PubMedGoogle Scholar
  13. 13.
    Y. Yasukochi and B. S. S. Masters, Some properties of a detergent-solubilized NADPH-cytochrome c reductase purified by biospecific affinity chromatography, J. Biol. Chem. 251:5337 (1976).PubMedGoogle Scholar
  14. 14.
    J. D. Dignam and H. W. Strobel, NADPH-cytochrome P450 reductase from rat liver: purification by affinity chromatography and characterization, Biochemistry 16:1116 (1977).PubMedCrossRefGoogle Scholar
  15. 15.
    S. D. Black and M. J. Coon, Structural feature of liver microsomal NADPH-cytochrome P450 reductase: hydrophobic domain, hydrophilic domain and connecting region, J. Biol. Chem. 257:5929 (1982).PubMedGoogle Scholar
  16. 16.
    T. D. Porter and C. B. Kasper, Coding nucleotide sequence of rat NADPH-cytochrome P-450 oxidoreductase cDNA and identification of flavin-binding domains, Proc. Natl. Acad. Sci. USA 82:973 (1985).PubMedCrossRefGoogle Scholar
  17. 17.
    M. Katagiri, H. Murakami, Y. Yabusaki, T. Sugiyama, M. Okamoto, T. Yamano and H. Ohkawa, Molecular cloning and sequence analysis of full-length cDNA for rabbit liver NADPH-cytochrome P450 reductase mRNA, J. Biochem. 100:945 (1986).PubMedGoogle Scholar
  18. 18.
    J. R. Gum and H. W. Strobel, Isolation of the membranebinding peptide of NADPH-cytochrome P450 reductase: characterization of the peptide and its role in the interaction of reductase with cytochrome P450, J. Biol. Chem. 256:7478 (1981).PubMedGoogle Scholar
  19. 19.
    S. D. Black, J. S. French, C. H. Williams and M. J. Coon, Role of a hydrophobic polypeptide in the N-terminal region of NADPH-cytochrome P450 reductase in complex formation with P450LM, Biochem. Biophys. Res. Commun. 91:1528 (1979).PubMedCrossRefGoogle Scholar
  20. 20.
    J. R. Gum and H. W. Strobel, Purified NADPH-cytochrome P450 reductase: interaction with hepatic microsomes and phospholipid vesicles, J. Biol. Chem. 254:4177 (1979).PubMedGoogle Scholar
  21. 21.
    I. Iyanagi and H. S. Mason, Some properties of hepatic reduced nicotinamide adenine dinucleotide phosphate-cytochrome c reductase, Biochemistry 12:2297 (1973).PubMedCrossRefGoogle Scholar
  22. 22.
    T. Iyanagi, H. Makino and H. S. Mason, Redox properties of the reduced nicotinamide adenine dinucleotide phosphate-cytochrome P450 and reduced nicotinamide adenine dinucleotide-cytochrome b5 reductases, Biochemistry 13:1701 (1974).PubMedCrossRefGoogle Scholar
  23. 23.
    J. L. Vermillion and M. J. Coon, Identification of the high and low potential flavins of liver microsomal NADPH-cytochrome P450 reductase, J. Biol. Chem. 253:8812 (1978).Google Scholar
  24. 24.
    J. L. Vermillion and M. J. Coon, Purified liver microsomal NADPH-cytochrome P450 reductase: spectral characterization of oxidation-reduction states, J. Biol. Chem. 253:2694 (1978).Google Scholar
  25. 25.
    G. P. Kurzban and H. W. Strobel, Preparation and characterization of FAD-dependent NADPH-cytochrome P450 reductase, J. Biol. Chem. 261:7824 (1986).PubMedGoogle Scholar
  26. 26.
    Y. Nisimoto and Y. Shibata, Studies on FAD-and FMN-binding domains in NADPH-cytochrome P450 reductase from rabbit liver microsomes, J. Biol. Chem. 257:12532 (1982).PubMedGoogle Scholar
  27. 27.
    M. Haniu, T. Iyanagi, K. Legesse and J. E. Shively, Structural analysis of NADPH-cytochrome P450 reductase from porcine hepatic microsomes: sequences of proteolytic fragments, cysteine-containing peptides and a NADPH-protected cysteine peptide, J. Biol. Chem. 259:13703 (1984).PubMedGoogle Scholar
  28. 28.
    Y. Nisimoto, F. Hayashi, H. Akutsu, Y. Kyogoku and Y. Shibata, photochemically induced dynamic nuclear polarization study on microsomal NADPH-cytochrome P450 reductase, J. Biol. Chem. 259:2480 (1984).PubMedGoogle Scholar
  29. 29.
    A. L. Shen, T. D. Porter, T. E. Wilson and C. B. Kasper, Structural analysis of the FMN binding domain of NADPH-cytochrome P450 oxidoreductase by site-directed mutagenesis, J. Biol. Chem. 264:7584 (1989).PubMedGoogle Scholar
  30. 30.
    H. Inano and B. Tamaoki, Chemical modification of NADPH-cytochrome P450 reductase: presence of a lysine residue in the rat hepatic enzyme as the recognition site of 2-phosphate moiety of the cofactor, Eur. J. Biochem. 155:485 (1986).PubMedCrossRefGoogle Scholar
  31. 31.
    I.A. Slepneva and L. M. Weiner, Affinity modification of NADPH-cytochrome P450 reductase, Biochem. Biophys. Res. Commun. 155:1026 (1988).PubMedCrossRefGoogle Scholar
  32. 32.
    Y. Nisimoto, Localization of cytochrome c binding domain on NADPH-cytochrome P450 reductase, J. Biol. Chem. 261:14232 (1986).PubMedGoogle Scholar
  33. 33.
    S. G. Nadler and H. W. Strobel, Role of electrostatic interactions in the reaction of NADPH-cytochrome P450 reductase with cytochrome P450, Arch. Biochem. Biophys. 261:418 (1988).PubMedCrossRefGoogle Scholar
  34. 34.
    P. P. Tamburini and J. B. Schenkman, Differences in the mechanism of functional interaction between NADPH-cytochrome P450 reductase and its redox partners, Mol. Pharmacol. 30:178 (1986).PubMedGoogle Scholar
  35. 35.
    R. Bernhardt, K. Pommerening and K. Ruckpaul, Modification of carboxyl groups on NADPH-cytochrome P450 reductase involved in binding of cytochrome c and P450 LM2, Biochem. Int. 14:823 (1987).PubMedGoogle Scholar
  36. 36.
    R. E. Ebel, Selective inactivation of NADPH-cytochrome P450 reductase by diazotized 3-aminopyridine adenine dinucleotide phosphate, Arch. Biochem. Biophys. 211:227 (1981).PubMedCrossRefGoogle Scholar
  37. 37.
    G. T. Miwa, S. B. West, M. T. Huang and A. Y. H. Lu, Studies on the association of cytochrome P450 and NADPH-cytochrome c reductase during catalysis in a reconstituted hydroxylating system, J. Biol. Chem. 254:5695 (1979).PubMedGoogle Scholar
  38. 38.
    J. S. French, F. P. Guengerich and M. J. Coon, Interactions of cytochrome P450, NADPH-cytochrome P450 reductase, phospholipid and substrate in the reconstituted liver microsomal enzyme system, J. Biol. Chem. 255:4112 (1980).PubMedGoogle Scholar
  39. 39.
    B. Bosterling and J. R. Trudell, Association of cytochrome b5 and cytochrome P450 reductase with cytochrome P450 in the membrane of reconstituted vesicles, J. Biol. Chem. 257:4783 (1982).PubMedGoogle Scholar
  40. 40.
    Y. Nisimoto, K. Kinosita, A. Ikegami, N. Kawai, I. Ichihara and Y. Shibata, Possible association of NADPH-cytochrome P450 reductase and cytochrome P450 in reconstituted phospholipid vesicles, Biochemistry, 22:3586 (1983).PubMedCrossRefGoogle Scholar
  41. 41.
    J. Gut, C. Richter, R. J. Cherry, K. H. Winterhalter and S. Kawato, Rotation of cytochrome P450. II. Specific interactions of cytochrome P450 with NADPH-cytochrome P450 reductase in phospholipid vesicles, J. Biol. Chem. 257:7030 (1982).PubMedGoogle Scholar
  42. 42.
    R. Bernhardt, A. Makower, G. R. Janig and K. Ruckpaul, Selective chemical modification of a functionally linked lysine in cytochrome P450 LM2, Biochim. Biophys. Acta 785:186 (1984).PubMedCrossRefGoogle Scholar
  43. 43.
    D. R. Nelson and H. W. Strobel, On the membrane topology of vertebrate cytochrome P450 proteins, J. Biol. Chem. 263:6038 (1988).PubMedGoogle Scholar
  44. 44.
    C. A. Brown and S, D. Black, Membrane topology of mammalian cytochromes P450 from liver endoplasmic reticulum: determination by trypsinolysis of phenobarbital-treated microsomes, J. Biol. Chem. 264:4442 (1989).PubMedGoogle Scholar
  45. 45.
    T. D. Porter and C. B. Kasper, NADPH-cytochrome P450 oxidoreductase: flavin mononucleotide domains evolved from different flavoproteins, Biochemistry 25:1682 (1986).PubMedCrossRefGoogle Scholar
  46. 46.
    F. J. Gonzalez and C. B. Kasper, Phenobarbital induction of NADPH-cytochrome P450 oxidoreductase messenger ribonucleic acid, Biochemistry 19:1790 (1980).PubMedCrossRefGoogle Scholar
  47. 47.
    J. P. Hardwick, F. J. Gonzalez and C. B. Kasper, Transcriptional regulation of rat liver epoxide hydratase, NADPH-cytochrome P450 oxidoreductase and cytochrome P450b genes by phenobarbital, J. Biol. Chem. 258:8081 (1983).PubMedGoogle Scholar
  48. 48.
    H. Kappus, Overview of enzyme systems involved in bioreduction of drugs and in redox cycling, Biochem. Pharmacol. 35:1 (1986).PubMedCrossRefGoogle Scholar
  49. 49.
    G. Powis, Free radical formation by antitumor quinones, Free Radical Biol. & Med. 6:63 (1989).CrossRefGoogle Scholar
  50. 50.
    B. K. Sinha, Free radicals in anticancer drug pharmacology, Chem.-Biol. Interactions 69:293 (1989).CrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1991

Authors and Affiliations

  • Anthony Y. H. Lu
    • 1
  1. 1.Merck Sharp & Dohme Research LaboratoriesRahwayUSA

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