Skip to main content
SpringerLink
Log in

Nadph-initiated cytochrome P450-dependent free iron-independent microsomal lipid peroxidation: Specific prevention by ascorbic acid

  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Abstract

In this paper we demonstrate that ascorbic acid specifically prevents NADPH-initiated cytochrome P450 (P450)-mediated microsomal lipid peroxidation in the absence of free iron. Lipid peroxidation has been evidenced by the formations of conjugated dienes, lipid hydroperoxide and malondialdehyde. Other scavengers of reactive oxygen species including superoxide dismutase, catalase, glutathione, α-tocopherol, uric acid, thiourea, mannitol, histidine, β-carotene and probucol are ineffective to prevent the NADPH-initiated P450-mediated free iron-independent microsomal lipid peroxidation. Using a reconstituted system comprised of purified NADPH-P450 reductase, P450 and isolated microsomal lipid or pure L-α-phosphatidylcholine diarachidoyl, a mechanism has been proposed for the iron-independent microsomal lipid peroxidation and its prevention by ascorbic acid. It is proposed that the perferryl moiety P450 Fe3+. O2 initiates lipid peroxidation by abstracting methylene hydrogen from polyunsaturated lipid to form lipid radical, which then combines with oxygen to produce the chain propagating peroxyl radical for subsequent formation of lipid peroxides. Apparently, ascorbic acid prevents initiation of lipid peroxidation by interacting with P450 Fe3+. O2. (Mol Cell Biochem 166: 35-44, 1997)

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

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Ames BN: Dietary carcinogens and anticarcinogens. Science 221: 1256–1264, 1983

    Google Scholar 

  2. Halliwell B, Gutteridge JMC: Free radicals in biology and medicine, 2nd ed. Oxford, Clarendon Press, 1989

    Google Scholar 

  3. Cerutti PA: Prooxidant states and tumor production. Science 227: 375–381, 1985

    Google Scholar 

  4. Chakrabarty S, Nandi A, Mukhopadhyay CK. Chatterjee IB: Protective role of ascorbic acid against lipid peroxidation and myocardial injury. Mol Cell Biochem 111: 41–47, 1992

    Google Scholar 

  5. Mukhopadhyay M, Mukhopadhyay CK, Chatterjee IB: Protective effect of ascorbic acid against lipid peroxidation and oxidative damage in cardiac microsomes. Mol Cell Biochem 126: 69–75, 1993

    Google Scholar 

  6. Ames BN, Shigenaga MK, Hagen TM: Oxidants, antioxidants and the degenerative diseases of aging. Proc Natl Acad Sci USA 90: 7915–7922, 1993

    Google Scholar 

  7. Hochstein P, Ernster L: ADP-activated lipid peroxidation coupled to the TPNH-oxidase system of microsomes. Biochem Biophys Res Commun 12: 388–394, 1963

    Google Scholar 

  8. Aust SD, Roerig DL, Pederson TC: NADPH-dependent lipid peroxidation catalyzed by purified NADPH-cytochrome c reductase from rat liver microsomes. Biochem Biophys Res Commun 47: 1133–1137, 1972

    Google Scholar 

  9. Pederson TC, Aust SD: NADPH-dependent lipid peroxidation catalyzed by purified NADPH-cytochrome c reductase from rat liver microsomes. Biochem Biophys Res Commun 48: 789–795, 1972

    Google Scholar 

  10. Pederson TC, Buege JA, Aust SD: Microsomal electron transport: The role of reduced nicotinamide adenine dinucleotide phosphate-cytochrome c reductase in liver microsomal lipid peroxidation. J Biol Chem 248: 7134–7141, 1973

    Google Scholar 

  11. Pederson TC, Aust SD: The mechanism of liver microsomal lipid peroxidation. Biochem Biophys Acta 385: 232–241, 1975

    Google Scholar 

  12. Buege JA, Aust SD: Microsomal lipid peroxidation. Methods Enzymol 52: 302–310, 1978

    Google Scholar 

  13. Svingen BA, Buege JA, O'Neal FO, Aust SD: The mechanism of NADPH-dependent lipid peroxidation. J Biol Chem 254: 5892–5899, 1979

    Google Scholar 

  14. Ekström G, Ingelman-Sundberg M: Cytochrome P450-dependent lipid peroxidation in reconstituted membrane vesicles. Biochem Pharmacol 33: 2521–2523, 1984

    Google Scholar 

  15. Aust SD, Morehouse LA, Thomas CE: Role of metals in oxygen radical reactions. J Free Radicals Biol and Med 1: 3–25, 1985

    Google Scholar 

  16. Sevanian A, Nordenbrand K, Kim E, Ernster L, Hochstein P: Microsomal lipid peroxidation: The role of NADPH-cytochrome P450 reductase and cytochrome P450. Free Radical Biol Med 8: 145–152, 1990

    Google Scholar 

  17. Ekström G, Ingelman-Sundberg M: Mechanisms of lipid peroxidation dependent upon cytochrome P450 LM2. Eur J Biochem 158: 195–201, 1986

    Google Scholar 

  18. Davies KJA, Goldberg AL: Oxygen radicals stimulate intracellular proteolysis and lipid peroxidation by independent mechanisms in erythrocytes. J Biol Chem 282: 8220–8226, 1987

    Google Scholar 

  19. Mukhopadhyay CK, Chatterjee IB: NADPH-initiated cytochrome P450-mediated free metal ion-independent oxidative damage of microsomal proteins: Exclusive prevention by ascorbic acid. J Biol Chem 269: 13390–13397, 1994

    Google Scholar 

  20. Mukhopadhyay CK, Ghosh MK, Chatterjee IB: Ascorbic acid prevents lipid peroxidation and oxidative damage of proteins in guinea pig extrahepatic tissue microsomes. Mol Cell Biochem 142: 71–78, 1995

    Google Scholar 

  21. Chakrabarty S, Nandi A, Mukhodhyay M, Mukhopadhyay CK, Chatterjee IB: Ascorbate protects guinea pig tissues against lipid peroxidation. Free Radical Biol and Med 16: 417–426, 1993

    Google Scholar 

  22. Van der Hoeven TA, Coon MJ: Preparation and properties of partially purified cytochrome P450 and reduced nicotinamide adenine dinucleotide phosphate-cytochrome P450 reductase from rabbit liver microsomes. J Biol Chem 249: 6302–6310, 1974

    Google Scholar 

  23. Masters BSS, Williams CH Jr, Kamin H: The preparation and properties of microsomal TPNH-cytochrome c reductase from pig liver. Methods Enzymol 10: 565–573, 1967

    Google Scholar 

  24. Omura T, Sato R: The carbon monoxide-binding pigment of liver microsomes. J Biol Chem 239: 2370–2378, 1964

    Google Scholar 

  25. Aikens J, Dix TA: Perhydroxyl radical initiated lipid peroxidation. J Biol Chem 266: 15091–15098, 1991

    Google Scholar 

  26. Flohé L, Günzler WA: Assays of glutathione peroxidase. Methods Enzymol 105: 114–121, 1984

    Google Scholar 

  27. Pryor WA, Castle L: Chemical methods for the detection of lipid hydroperoxides. Methods Enzymol 105: 293–299, 1984

    Google Scholar 

  28. Esterbauer H, Cheeseman KH: Determination of aldehydic lipid peroxidation products: Malonaldehyde and 4-hydroxynonenal. Methods Enzymol 186: part B, 407–421, 1990

    Google Scholar 

  29. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ: Protein measurement with the folin phenol reagent. J Biol Chem 193: 265–275, 1951

    Google Scholar 

  30. Mukhopadhyay CK, Chatterjee IB: Free metal ion-independent oxidative damage of collagen: Protection by ascorbic acid. J Biol Chem 269: 30200–30205, 1994

    Google Scholar 

  31. Chirico S: High-performance liquid chromatography-based thiobarbituric acid tests. Methods Enzymol 233: 314–318, 1994

    Google Scholar 

  32. Buettner GR: The pecking order of free radicals and antioxidants: Lipid peroxidation, α-tocopherol and ascorbate. Arch Biochem Biophys 300: 535–543, 1993

    Google Scholar 

  33. Beyer RE: The role of ascorbate in antioxidant protection of biomembranes: Interaction with vitamin E and coenzyme Q. J Bioenergetics and Biomembranes 26: 349–358, 1994

    Google Scholar 

  34. Yao K, Falick AM, Patel N, Correia MA: Cumene hydroperoxide-mediated inactivation of cytochrome P450 2B1. J Biol Chem 268: 59–65, 1993

    Google Scholar 

  35. Sinclair PR, Gorman N, Walton HS, Bement WJ, Jacobs JM, Sinclair JF: Ascorbic acid inhibition of cytochrome P450 catalyzed uropor phyrin accumulation. Arch Biochem Biophys 304: 464–470, 1993

    Google Scholar 

  36. Basu S, Som S, Deb S, Mukherjee D, Chatterjee IB: Dehydroascorbic acid reduction in human erythrocyte. Biochem Biophys Res Commun 90: 1335–1340, 1979

    Google Scholar 

  37. Som S, Basu S, Deb S, Mukherjee D, Chatterjee IB: Dehydroascorbic acid reduction in guinea pig tissues. Curr Sci 49: 195–196, 1980

    Google Scholar 

  38. Meister A: Glutathione — ascorbic acid antioxidant system in animals. J Biol chem 269: 9397–9400, 1994

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Centre for Genetic Engineering and Biotechnology and Department of Biochemistry, University College of Science, Calcutta, 700 019, India

    Mrinal K. Ghosh, Mau Mukhopadhyay & Indu B. Chatterjee

Authors
  1. Mrinal K. Ghosh
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Mau Mukhopadhyay
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Indu B. Chatterjee
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ghosh, M.K., Mukhopadhyay, M. & Chatterjee, I.B. Nadph-initiated cytochrome P450-dependent free iron-independent microsomal lipid peroxidation: Specific prevention by ascorbic acid. Mol Cell Biochem 166, 35–44 (1997). https://doi.org/10.1023/A:1006841228483

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1006841228483

Search

Navigation