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. 1987 Oct;80(4):1017–1022. doi: 10.1172/JCI113154

Purification of a human liver cytochrome P-450 immunochemically related to several cytochromes P-450 purified from untreated rats.

S A Wrighton 1, P E Thomas 1, P Willis 1, S L Maines 1, P B Watkins 1, W Levin 1, P S Guzelian 1
PMCID: PMC442340  PMID: 3654967

Abstract

Among characterized forms of liver microsomal cytochromes P-450 in rats are four related isozymes (P-450f-i) notable for their lack of inducibility. Immunoblot analyses demonstrated that human livers microsomes contained several proteins related to these rat P-450s. A human liver P-450, termed HLx, was purified and found by immunochemical assays to resemble rat P-450g. Analysis of the NH2-terminal amino acid sequence of HLx indicates that it is related to rat P-450s f-i and human liver P-450MP. A monoclonal antibody was used to measure the amounts of HLx in 21 human liver specimens. No correlation between the levels of HLx protein in these specimens and the patients' environmental histories was observed. However, statistical analysis of the data suggests that the distribution of HLx is at least bimodal. We conclude that HLx is a member of a family of human liver P-450s that resembles in its structure, and possibly in its distribution, several liver P-450s found in other animals.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Adesnik M., Atchison M. Genes for cytochrome P-450 and their regulation. CRC Crit Rev Biochem. 1986;19(3):247–305. doi: 10.3109/10409238609084657. [DOI] [PubMed] [Google Scholar]
  2. Bandiera S., Ryan D. E., Levin W., Thomas P. E. Age- and sex-related expression of cytochromes p450f and P450g in rat liver. Arch Biochem Biophys. 1986 Aug 1;248(2):658–676. doi: 10.1016/0003-9861(86)90521-7. [DOI] [PubMed] [Google Scholar]
  3. Conney A. H. Pharmacological implications of microsomal enzyme induction. Pharmacol Rev. 1967 Sep;19(3):317–366. [PubMed] [Google Scholar]
  4. Dieter H. H., Muller-Eberhard U., Johnson E. F. Rabbit hepatic progesterone 21-hydroxylase exhibits a bimodal distribution of activity. Science. 1982 Aug 20;217(4561):741–743. doi: 10.1126/science.6808664. [DOI] [PubMed] [Google Scholar]
  5. Gonzalez F. J., Kimura S., Song B. J., Pastewka J., Gelboin H. V., Hardwick J. P. Sequence of two related P-450 mRNAs transcriptionally increased during rat development. An R.dre.1 sequence occupies the complete 3' untranslated region of a liver mRNA. J Biol Chem. 1986 Aug 15;261(23):10667–10672. [PubMed] [Google Scholar]
  6. Guengerich F. P., Distlerath L. M., Reilly P. E., Wolff T., Shimada T., Umbenhauer D. R., Martin M. V. Human-liver cytochromes P-450 involved in polymorphisms of drug oxidation. Xenobiotica. 1986 May;16(5):367–378. doi: 10.3109/00498258609050245. [DOI] [PubMed] [Google Scholar]
  7. Haniu M., Ryan D. E., Iida S., Lieber C. S., Levin W., Shively J. E. NH2-terminal sequence analyses of four rat hepatic microsomal cytochromes P-450. Arch Biochem Biophys. 1984 Dec;235(2):304–311. doi: 10.1016/0003-9861(84)90202-9. [DOI] [PubMed] [Google Scholar]
  8. Johnson E. F., Schwab G. E. Constitutive forms of rabbit-liver microsomal cytochrome P-450: enzymatic diversity, polymorphism and allosteric regulation. Xenobiotica. 1984 Jan-Feb;14(1-2):3–18. doi: 10.3109/00498258409151395. [DOI] [PubMed] [Google Scholar]
  9. Kalow W. Genetics of drug transformation. Clin Biochem. 1986 Apr;19(2):76–82. doi: 10.1016/s0009-9120(86)80052-2. [DOI] [PubMed] [Google Scholar]
  10. Kamataki T., Maeda K., Yamazoe Y., Nagai T., Kato R. Sex difference of cytochrome P-450 in the rat: purification, characterization, and quantitation of constitutive forms of cytochrome P-450 from liver microsomes of male and female rats. Arch Biochem Biophys. 1983 Sep;225(2):758–770. doi: 10.1016/0003-9861(83)90087-5. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. 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]
  13. McClellan-Green P., Waxman D. J., Caveness M., Goldstein J. A. Phenotypic differences in expression of cytochrome P-450g but not its mRNA in outbred male Sprague-Dawley rats. Arch Biochem Biophys. 1987 Feb 15;253(1):13–25. doi: 10.1016/0003-9861(87)90632-1. [DOI] [PubMed] [Google Scholar]
  14. Nebert D. W. Multiple forms of inducible drug-metabolizing enzymes: a reasonable mechanism by which any organism can cope with adversity. Mol Cell Biochem. 1979 Sep 28;27(1):27–46. doi: 10.1007/BF00849277. [DOI] [PubMed] [Google Scholar]
  15. Okey A. B., Roberts E. A., Harper P. A., Denison M. S. Induction of drug-metabolizing enzymes: mechanisms and consequences. Clin Biochem. 1986 Apr;19(2):132–141. doi: 10.1016/s0009-9120(86)80060-1. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Reubi I., Griffin K. J., Raucy J. L., Johnson E. F. Three monoclonal antibodies to rabbit microsomal cytochrome P-450 1 recognize distinct epitopes that are shared to different degrees among other electrophoretic types of cytochrome P-450. J Biol Chem. 1984 May 10;259(9):5887–5892. [PubMed] [Google Scholar]
  18. Ryan D. E., Iida S., Wood A. W., Thomas P. E., Lieber C. S., Levin W. Characterization of three highly purified cytochromes P-450 from hepatic microsomes of adult male rats. J Biol Chem. 1984 Jan 25;259(2):1239–1250. [PubMed] [Google Scholar]
  19. Ryan D. E., Thomas P. E., Levin W. Purification of characterization of a minor form of hepatic microsomal cytochrome P-450 from rats treated with polychlorinated biphenyls. Arch Biochem Biophys. 1982 Jun;216(1):272–288. doi: 10.1016/0003-9861(82)90212-0. [DOI] [PubMed] [Google Scholar]
  20. Ryan D. E., Thomas P. E., Reik L. M., Levin W. Purification, characterization and regulation of five rat hepatic microsomal cytochrome P-450 isozymes. Xenobiotica. 1982 Nov;12(11):727–744. doi: 10.3109/00498258209038947. [DOI] [PubMed] [Google Scholar]
  21. Shimada T., Guengerich F. P. Participation of a rat liver cytochrome P-450 induced by pregnenolone 16 alpha-carbonitrile and other compounds in the 4-hydroxylation of mephenytoin. Mol Pharmacol. 1985 Aug;28(2):215–219. [PubMed] [Google Scholar]
  22. Shimada T., Misono K. S., Guengerich F. P. Human liver microsomal cytochrome P-450 mephenytoin 4-hydroxylase, a prototype of genetic polymorphism in oxidative drug metabolism. Purification and characterization of two similar forms involved in the reaction. J Biol Chem. 1986 Jan 15;261(2):909–921. [PubMed] [Google Scholar]
  23. Thomas P. E., Reik L. M., Ryan D. E., Levin W. Regulation of three forms of cytochrome P-450 and epoxide hydrolase in rat liver microsomes. Effects of age, sex, and induction. J Biol Chem. 1981 Jan 25;256(2):1044–1052. [PubMed] [Google Scholar]
  24. Tukey R. H., Okino S., Barnes H., Griffin K. J., Johnson E. F. Multiple gene-like sequences related to the rabbit hepatic progesterone 21-hydroxylase cytochrome P-450 1. J Biol Chem. 1985 Oct 25;260(24):13347–13354. [PubMed] [Google Scholar]
  25. Umbenhauer D. R., Martin M. V., Lloyd R. S., Guengerich F. P. Cloning and sequence determination of a complementary DNA related to human liver microsomal cytochrome P-450 S-mephenytoin 4-hydroxylase. Biochemistry. 1987 Feb 24;26(4):1094–1099. doi: 10.1021/bi00378a016. [DOI] [PubMed] [Google Scholar]
  26. Wang P. P., Beaune P., Kaminsky L. S., Dannan G. A., Kadlubar F. F., Larrey D., Guengerich F. P. Purification and characterization of six cytochrome P-450 isozymes from human liver microsomes. Biochemistry. 1983 Nov 8;22(23):5375–5383. doi: 10.1021/bi00292a019. [DOI] [PubMed] [Google Scholar]
  27. 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]
  28. Waxman D. J. Rat hepatic cytochrome P-450 isoenzyme 2c. Identification as a male-specific, developmentally induced steroid 16 alpha-hydroxylase and comparison to a female-specific cytochrome P-450 isoenzyme. J Biol Chem. 1984 Dec 25;259(24):15481–15490. [PubMed] [Google Scholar]
  29. Whitlock J. P., Jr The regulation of cytochrome P-450 gene expression. Annu Rev Pharmacol Toxicol. 1986;26:333–369. doi: 10.1146/annurev.pa.26.040186.002001. [DOI] [PubMed] [Google Scholar]
  30. Winkel C. A., Simpson E. R., Milewich L., MacDonald P. C. Deoxycorticosterone biosynthesis in human kidney: potential for formation of a potent mineralocorticosteroid in its site of action. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7069–7073. doi: 10.1073/pnas.77.12.7069. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. 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]
  32. 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]
  33. 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]
  34. van der Hoeven T. A., Coon M. J. Preparation and properties of partially purified cytochrome P-450 and reduced nicotinamide adenine dinucleotide phosphate-cytochrome P-450 reductase from rabbit liver microsomes. J Biol Chem. 1974 Oct 10;249(19):6302–6310. [PubMed] [Google Scholar]

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