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. 1988 Oct;26(4):363–372. doi: 10.1111/j.1365-2125.1988.tb03393.x

A form of cytochrome P450 in man, orthologous to form d in the rat, catalyses the O-deethylation of phenacetin and is inducible by cigarette smoking.

D Sesardic 1, A R Boobis 1, R J Edwards 1, D S Davies 1
PMCID: PMC1386556  PMID: 3190986

Abstract

1. In previous studies (Boobis et al., 1985b) it was shown that a monoclonal antibody (MAb 3/4/2), raised against rat cytochrome P450 form c, reacts with an isoenzyme(s) of cytochrome P450 in human liver. It was predicted that the epitope with which this antibody reacts should be present on both isoenzymes of the P450IA gene sub-family (the orthologues of forms c and d) in man (Edwards et al., 1987). 2. This antibody was used to probe 45 different samples of human liver, by the technique of Western blotting. With one exception, all of the samples contained immunoreactive protein, a single band at Mr 54,000 (orthologous to rat form d), which ranged in content from less than 0.5 to 33.5 pmol mg-1 microsomal protein. The content of the human orthologue of form c was below 0.5 pmol mg-1, the limit of detection of the assay. 3. Thirteen of the samples were from patients of known smoking status. Immunoreactive P450 content was 3.5-fold higher, and phenacetin O-deethylase activity was four-fold higher, in the smokers than in the non-smokers. 4. There was a highly significant correlation between the amount of immunoreactive cytochrome P450 and the high affinity component of phenacetin O-deethylase activity in both smokers and non-smokers. 5. It is concluded that the high affinity component of phenacetin O-deethylase activity in man is catalysed by the orthologue of rat cytochrome P450d, and that this isoenzyme is inducible by cigarette smoking. 6. In a number of previous publications it has been suggested that there is an association between the poor metaboliser (PM) phenotype for debrisoquine and impaired phenacetin O-deethylation. In the present study it was shown that not all subjects PM for debrisoquine are poor metabolisers of phenacetin.

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Selected References

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  1. Adams D. J., Seilman S., Amelizad Z., Oesch F., Wolf C. R. Identification of human cytochromes P-450 analogous to forms induced by phenobarbital and 3-methylcholanthrene in the rat. Biochem J. 1985 Dec 15;232(3):869–876. doi: 10.1042/bj2320869. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Boobis A. R., Brodie M. J., Kahn G. C., Fletcher D. R., Saunders J. H., Davies D. S. Monooxygenase activity of human liver in microsomal fractions of needle biopsy specimens. Br J Clin Pharmacol. 1980 Jan;9(1):11–19. doi: 10.1111/j.1365-2125.1980.tb04790.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Boobis A. R., Hampden C. E., Murray S., Beaune P., Davies D. S. Attempts to phenotype human liver samples in vitro for debrisoquine 4-hydroxylase activity. Br J Clin Pharmacol. 1985 Jun;19(6):721–729. doi: 10.1111/j.1365-2125.1985.tb02706.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Boobis A. R., McQuade J., Sesardic D., Robson R. T., Hayward C., Lock E. A., Elcombe C. R., Rose M. S., Davies D. S. A monoclonal antibody raised to rat liver cytochrome P-448 (form C) which recognises an epitope common to many other forms of cytochrome P-450. Biochem Pharmacol. 1985 May 15;34(10):1671–1681. doi: 10.1016/0006-2952(85)90633-1. [DOI] [PubMed] [Google Scholar]
  5. Conney A. H. Induction of microsomal cytochrome P-450 enzymes: the first Bernard B. Brodie lecture at Pennsylvania State University. Life Sci. 1986 Dec 29;39(26):2493–2518. doi: 10.1016/0024-3205(86)90103-7. [DOI] [PubMed] [Google Scholar]
  6. Conney A. H. Induction of microsomal enzymes by foreign chemicals and carcinogenesis by polycyclic aromatic hydrocarbons: G. H. A. Clowes Memorial Lecture. Cancer Res. 1982 Dec;42(12):4875–4917. [PubMed] [Google Scholar]
  7. Distlerath L. M., Reilly P. E., Martin M. V., Davis G. G., Wilkinson G. R., Guengerich F. P. Purification and characterization of the human liver cytochromes P-450 involved in debrisoquine 4-hydroxylation and phenacetin O-deethylation, two prototypes for genetic polymorphism in oxidative drug metabolism. J Biol Chem. 1985 Jul 25;260(15):9057–9067. [PubMed] [Google Scholar]
  8. Foster J. R., Elcombe C. R., Boobis A. R., Davies D. S., Sesardic D., McQuade J., Robson R. T., Hayward C., Lock E. A. Immunocytochemical localization of cytochrome P-450 in hepatic and extra-hepatic tissues of the rat with a monoclonal antibody against cytochrome P-450 c. Biochem Pharmacol. 1986 Dec 15;35(24):4543–4554. doi: 10.1016/0006-2952(86)90777-x. [DOI] [PubMed] [Google Scholar]
  9. Goldstein J. A., Linko P. Differential induction of two 2,3,7,8-tetrachlorodibenzo-p-dioxin-inducible forms of cytochrome P-450 in extrahepatic versus hepatic tissues. Mol Pharmacol. 1984 Jan;25(1):185–191. [PubMed] [Google Scholar]
  10. Guengerich F. P., Dannan G. A., Wright S. T., Martin M. V., Kaminsky L. S. Purification and characterization of liver microsomal cytochromes p-450: electrophoretic, spectral, catalytic, and immunochemical properties and inducibility of eight isozymes isolated from rats treated with phenobarbital or beta-naphthoflavone. Biochemistry. 1982 Nov 9;21(23):6019–6030. doi: 10.1021/bi00266a045. [DOI] [PubMed] [Google Scholar]
  11. Jaiswal A. K., Nebert D. W., McBride O. W., Gonzalez F. J. Human P(3)450: cDNA and complete protein sequence, repetitive Alu sequences in the 3' nontranslated region, and localization of gene to chromosome 15. J Exp Pathol. 1987 Winter;3(1):1–17. [PubMed] [Google Scholar]
  12. Kahn G. C., Boobis A. R., Brodie M. J., Toverud E. L., Murray S., Davies D. S. Phenacetin O-deethylase: an activity of a cytochrome P-450 showing genetic linkage with that catalysing the 4-hydroxylation of debrisoquine? Br J Clin Pharmacol. 1985 Jul;20(1):67–76. doi: 10.1111/j.1365-2125.1985.tb02800.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kelley M., Hantelle P., Safe S., Levin W., Thomas P. E. Co-induction of cytochrome P-450 isozymes in rat liver by 2,4,5,2',4',5'-hexachlorobiphenyl or 3-methoxy-4-aminoazobenzene. Mol Pharmacol. 1987 Aug;32(1):206–211. [PubMed] [Google Scholar]
  14. Kremers P., Beaune P., Cresteil T., de Graeve J., Columelli S., Leroux J. P., Gielen J. E. Cytochrome P-450 monooxygenase activities in human and rat liver microsomes. Eur J Biochem. 1981 Sep 1;118(3):599–606. doi: 10.1111/j.1432-1033.1981.tb05561.x. [DOI] [PubMed] [Google Scholar]
  15. Kuntzman R., Pantuck E. J., Kaplan S. A., Conney A. H. Phenacetin metabolism: effect of hydrocarbons and cigarette smoking. Clin Pharmacol Ther. 1977 Nov;22(5 Pt 2):757–764. doi: 10.1002/cpt1977225part2757. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  18. Murray S., Boobis A. R. An assay for paracetamol, produced by the O-deethylation of phenacetin in vitro, using gas chromatography/electron capture negative ion chemical ionization mass spectrometry. Biomed Environ Mass Spectrom. 1986 Feb;13(2):91–93. doi: 10.1002/bms.1200130208. [DOI] [PubMed] [Google Scholar]
  19. Nebert D. W., Adesnik M., Coon M. J., Estabrook R. W., Gonzalez F. J., Guengerich F. P., Gunsalus I. C., Johnson E. F., Kemper B., Levin W. The P450 gene superfamily: recommended nomenclature. DNA. 1987 Feb;6(1):1–11. doi: 10.1089/dna.1987.6.1. [DOI] [PubMed] [Google Scholar]
  20. Negishi M., Nebert D. W. Structural gene products of the Ah locus. Genetic and immunochemical evidence for two forms of mouse liver cytochrome P-450 induced by 3-methylcholanthrene. J Biol Chem. 1979 Nov 10;254(21):11015–11023. [PubMed] [Google Scholar]
  21. OMURA T., SATO R. THE CARBON MONOXIDE-BINDING PIGMENT OF LIVER MICROSOMES. I. EVIDENCE FOR ITS HEMOPROTEIN NATURE. J Biol Chem. 1964 Jul;239:2370–2378. [PubMed] [Google Scholar]
  22. Pantuck E. J., Hsiao K. C., Maggio A., Nakamura K., Kuntzman R., Conney A. H. Effect of cigarette smoking on phenacetin metabolism. Clin Pharmacol Ther. 1974 Jan;15(1):9–17. doi: 10.1002/cpt19741519. [DOI] [PubMed] [Google Scholar]
  23. Pantuck E. J., Kuntzman R., Conney A. H. Decreased concentration of phenacetin in plasma of cigarette smokers. Science. 1972 Mar 17;175(4027):1248–1250. doi: 10.1126/science.175.4027.1248. [DOI] [PubMed] [Google Scholar]
  24. Parkinson A., Safe S. H., Robertson L. W., Thomas P. E., Ryan D. E., Reik L. M., Levin W. Immunochemical quantitation of cytochrome P-450 isozymes and epoxide hydrolase in liver microsomes from polychlorinated or polybrominated biphenyl-treated rats. A study of structure-activity relationships. J Biol Chem. 1983 May 10;258(9):5967–5976. [PubMed] [Google Scholar]
  25. Pelkonen O., Pasanen M., Kuha H., Gachalyi B., Kairaluoma M., Sotaniemi E. A., Park S. S., Friedman F. K., Gelboin H. V. The effect of cigarette smoking on 7-ethoxyresorufin O-deethylase and other monooxygenase activities in human liver: analyses with monoclonal antibodies. Br J Clin Pharmacol. 1986 Aug;22(2):125–134. doi: 10.1111/j.1365-2125.1986.tb05239.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Quattrochi L. C., Pendurthi U. R., Okino S. T., Potenza C., Tukey R. H. Human cytochrome P-450 4 mRNA and gene: part of a multigene family that contains Alu sequences in its mRNA. Proc Natl Acad Sci U S A. 1986 Sep;83(18):6731–6735. doi: 10.1073/pnas.83.18.6731. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Reik L. M., Levin W., Ryan D. E., Thomas P. E. Immunochemical relatedness of rat hepatic microsomal cytochromes P-450c and P-450d. J Biol Chem. 1982 Apr 10;257(7):3950–3957. [PubMed] [Google Scholar]
  28. Sesardic D., Boobis A. R., McQuade J., Baker S., Lock E. A., Elcombe C. R., Robson R. T., Hayward C., Davies D. S. Inter-relatedness of some isoenzymes of cytochrome P-450 from rat, rabbit and human, determined with monoclonal antibodies. Biochem J. 1986 Jun 1;236(2):569–577. doi: 10.1042/bj2360569. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Shahidi N. T. Acetophenetidin-induced methemoglobinemia. Ann N Y Acad Sci. 1968 Jul 31;151(2):822–832. doi: 10.1111/j.1749-6632.1968.tb48266.x. [DOI] [PubMed] [Google Scholar]
  30. Thomas P. E., Bandiera S., Maines S. L., Ryan D. E., Levin W. Regulation of cytochrome P-450j, a high-affinity N-nitrosodimethylamine demethylase, in rat hepatic microsomes. Biochemistry. 1987 Apr 21;26(8):2280–2289. doi: 10.1021/bi00382a031. [DOI] [PubMed] [Google Scholar]
  31. Thomas P. E., Reidy J., Reik L. M., Ryan D. E., Koop D. R., Levin W. Use of monoclonal antibody probes against rat hepatic cytochromes P-450c and P-450d to detect immunochemically related isozymes in liver microsomes from different species. Arch Biochem Biophys. 1984 Nov 15;235(1):239–253. doi: 10.1016/0003-9861(84)90273-x. [DOI] [PubMed] [Google Scholar]
  32. Thomas P. E., Reik L. M., Ryan D. E., Levin W. Induction of two immunochemically related rat liver cytochrome P-450 isozymes, cytochromes P-450c and P-450d, by structurally diverse xenobiotics. J Biol Chem. 1983 Apr 10;258(7):4590–4598. [PubMed] [Google Scholar]
  33. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Uotila P., Pelkonen O., Cohen G. M. The effects of cigarette smoke on the metabolism of [3H]benzo(a)pyrene by rat lung microsomes. Cancer Res. 1977 Jul;37(7 Pt 1):2156–2161. [PubMed] [Google Scholar]
  35. Watkins P. B., Wrighton S. A., Maurel P., Schuetz E. G., Mendez-Picon G., Parker G. A., Guzelian P. S. Identification of an inducible form of cytochrome P-450 in human liver. Proc Natl Acad Sci U S A. 1985 Sep;82(18):6310–6314. doi: 10.1073/pnas.82.18.6310. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Wong T. K., Domin B. A., Bent P. E., Blanton T. E., Anderson M. W., Philpot R. M. Correlation of placental microsomal activities with protein detected by antibodies to rabbit cytochrome P-450 isozyme 6 in preparations from humans exposed to polychlorinated biphenyls, quaterphenyls, and dibenzofurans. Cancer Res. 1986 Feb;46(2):999–1004. [PubMed] [Google Scholar]
  37. Wrighton S. A., Campanile C., Thomas P. E., Maines S. L., Watkins P. B., Parker G., Mendez-Picon G., Haniu M., Shively J. E., Levin W. Identification of a human liver cytochrome P-450 homologous to the major isosafrole-inducible cytochrome P-450 in the rat. Mol Pharmacol. 1986 Apr;29(4):405–410. [PubMed] [Google Scholar]
  38. Wrighton S. A., Maurel P., Schuetz E. G., Watkins P. B., Young B., Guzelian P. S. Identification of the cytochrome P-450 induced by macrolide antibiotics in rat liver as the glucocorticoid responsive cytochrome P-450p. Biochemistry. 1985 Apr 23;24(9):2171–2178. doi: 10.1021/bi00330a010. [DOI] [PubMed] [Google Scholar]
  39. Wrighton S. A., Thomas P. E., Molowa D. T., Haniu M., Shively J. E., Maines S. L., Watkins P. B., Parker G., Mendez-Picon G., Levin W. Characterization of ethanol-inducible human liver N-nitrosodimethylamine demethylase. Biochemistry. 1986 Nov 4;25(22):6731–6735. doi: 10.1021/bi00370a001. [DOI] [PubMed] [Google Scholar]
  40. Yeowell H. N., Linko P., Hodgson E., Goldstein J. A. Induction of specific cytochrome P-450 isozymes by methylenedioxyphenyl compounds and antagonism by 3-methylcholanthrene. Arch Biochem Biophys. 1985 Dec;243(2):408–419. doi: 10.1016/0003-9861(85)90517-x. [DOI] [PubMed] [Google Scholar]

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