Production of Carbon Monoxide by Sonication of Medium Containing Glycerol. Effect on Cytochrome P-450 Reconstituted Systems

  • V. Luu-The
  • Ch. Vandresse
  • J. Cumps
  • P. Dumont
Part of the Advances in Experimental Medicine and Biology book series (AEMB)

Abstract

Cytochrome P-450 is well known to be inactivated by carbon monoxide by forming a complex that absorbs at 450 nm in the reduced state. In the preparation of the cytochrome P-450 reconstituted systems, sonication has been used routinely to disperse phasphatidylcholine and to facilitate incorporation of cytochrome P-450 into the phospholipid vesicles. Our studies reveal that such treatment in the presence of dissolved gas and glycerol, which is used to prevent conversion of cytochrome P-450 to cytochrome P-420, produced carbon monoxide that may then inhibit the enzymatic activities of the system. This phenomenon is prevented by degassing the medium completely. A method is proposed to perform the reconstitution of cytochrome P-450 metabolizing systems.

Keywords

Difference Spectrum Phospholipid Vesicle Sonic Treatment Reconstituted System Hantzsch Reaction 
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. Franklin, M.R., 1974, The formation of a 455 nm Complex during Cytochrome P-450-dependent N-Hydroxyamphetamine Metabolism, Mol. Pharmacol., 10: 975.Google Scholar
  2. Luu-The, V., Vandresse,Ch., Cumps, J., and Dumont, P., 1979, Purification of cytochrome P-450 by double chromatography on substituted Sepharose. Effect of type I and type II ligands, Arch. Internat. Physiol. Biochim., 84: 1059.Google Scholar
  3. Nash,T., 1953, The colorimetric estimation of formaldehyde by means of the Hantzsch Reaction, Biochem. J., 55: 416.Google Scholar
  4. Omura,T., and Sato, R., 1964, The Carbon Monoxide-Binding Pigment of liver microsomes. I. Evidence for its hemoprotein nature, J. Biol. Chem. 239: 2370.Google Scholar
  5. Ryan, D.E., Thomas, P.E., Korgeniouski, P., and Levin, W., 1979, Separation and Characterization of Highly Purified Forms of Liver Microsomal Cytochrome P-450 from Rats treated with polychlorinated Biphenyls, Phenobarbital, and 3-Methylcholanthrene J. Biol. Chem., 254: 1365.Google Scholar
  6. Yasukochi,Y., and Masters, B.S.S., 1976, Some properties of a Deter gent-Solubized NADPH-Cytochrome P-450 Reductase Purified by Biospecific Affinity Chromatography, J. Biol. Chem. 251: 5337Google Scholar

Copyright information

© Springer Science+Business Media New York 1982

Authors and Affiliations

  • V. Luu-The
    • 1
  • Ch. Vandresse
    • 1
  • J. Cumps
    • 1
  • P. Dumont
    • 1
  1. 1.Department of Medicinal Chemistry Faculty of MedicineUniversity of LouvainBruxellesBelgium

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