Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1985 Feb;82(4):983–987. doi: 10.1073/pnas.82.4.983

Isolation and sequence of a human cytochrome P-450 cDNA clone.

I R Phillips, E A Shephard, A Ashworth, B R Rabin
PMCID: PMC397177  PMID: 3856261

Abstract

A previously reported cDNA clone [pP450(1)] coding for a phenobarbital-inducible cytochrome P-450 variant of rat liver microsomal membranes, designated P-450e(U.C.), was used as a specific hybridization probe to screen a human liver cDNA library. Restriction mapping showed that two of the colonies isolated contained plasmids coding for overlapping regions of the same cDNA sequence. The clone [pHP450(1)] having the longer cDNA insert (1.25 kilobase pairs) was sequenced. The homology between the rat and human cDNAs is 62% in their coding regions but is only random (24%) in the 3'-noncoding nucleotides. The amino acid sequence deduced from the human cDNA is 50% identical to that of P-450e(U.C.). The homology increases to 72% if conservative changes in amino acid residues are permitted. The hydropathy profile of the polypeptide encoded by pHP450(1) is almost identical to that of P-450e(U.C.). Regions known to be highly conserved in cytochrome P-450 isozymes isolated from rat, rabbit, and mouse were found to be conserved in the amino acid sequence derived from pHP450(1). Analysis by Southern blotting indicated that the human cytochrome P-450 encoded by pHP450(1) is part of a multigene family.

Full text

PDF
983

Images in this article

984Image
on p.984
984Image
on p.984
984Image
on p.984
987Image
on p.987

Selected References

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

  1. Atchison M., Adesnik M. A cytochrome P-450 multigene family. Characterization of a gene activated by phenobarbital administration. J Biol Chem. 1983 Sep 25;258(18):11285–11295. [PubMed] [Google Scholar]
  2. Bell G. I., Sanchez-Pescador R., Laybourn P. J., Najarian R. C. Exon duplication and divergence in the human preproglucagon gene. 1983 Jul 28-Aug 3Nature. 304(5924):368–371. doi: 10.1038/304368a0. [DOI] [PubMed] [Google Scholar]
  3. Biggin M. D., Gibson T. J., Hong G. F. Buffer gradient gels and 35S label as an aid to rapid DNA sequence determination. Proc Natl Acad Sci U S A. 1983 Jul;80(13):3963–3965. doi: 10.1073/pnas.80.13.3963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Blin N., Stafford D. W. A general method for isolation of high molecular weight DNA from eukaryotes. Nucleic Acids Res. 1976 Sep;3(9):2303–2308. doi: 10.1093/nar/3.9.2303. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. 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]
  6. Denhardt D. T. A membrane-filter technique for the detection of complementary DNA. Biochem Biophys Res Commun. 1966 Jun 13;23(5):641–646. doi: 10.1016/0006-291x(66)90447-5. [DOI] [PubMed] [Google Scholar]
  7. Fujii-Kuriyama Y., Mizukami Y., Kawajiri K., Sogawa K., Muramatsu M. Primary structure of a cytochrome P-450: coding nucleotide sequence of phenobarbital-inducible cytochrome P-450 cDNA from rat liver. Proc Natl Acad Sci U S A. 1982 May;79(9):2793–2797. doi: 10.1073/pnas.79.9.2793. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Grosveld F. G., Dahl H. H., de Boer E., Flavell R. A. Isolation of beta-globin-related genes from a human cosmid library. Gene. 1981 Apr;13(3):227–237. doi: 10.1016/0378-1119(81)90028-7. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Hanahan D., Meselson M. Plasmid screening at high colony density. Gene. 1980 Jun;10(1):63–67. doi: 10.1016/0378-1119(80)90144-4. [DOI] [PubMed] [Google Scholar]
  11. Haniu M., Armes L. G., Yasunobu K. T., Shastry B. A., Gunsalus I. C. Amino acid sequence of the Pseudomonas putida cytochrome P-450. II. Cyanogen bromide peptides, acid cleavage peptides, and the complete sequence. J Biol Chem. 1982 Nov 10;257(21):12664–12671. [PubMed] [Google Scholar]
  12. Hardwick J. P., Gonzalez F. J., Kasper C. B. Cloning of DNA complementary to cytochrome P-450 induced by pregnenolone-16 alpha-carbonitrile. Characterization of its mRNA, gene, and induction response. J Biol Chem. 1983 Aug 25;258(16):10182–10186. [PubMed] [Google Scholar]
  13. Idle J. R., Smith R. L. Polymorphisms of oxidation at carbon centers of drugs and their clinical significance. Drug Metab Rev. 1979;9(2):301–317. doi: 10.3109/03602537908993896. [DOI] [PubMed] [Google Scholar]
  14. Ish-Horowicz D., Burke J. F. Rapid and efficient cosmid cloning. Nucleic Acids Res. 1981 Jul 10;9(13):2989–2998. doi: 10.1093/nar/9.13.2989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kato R. Characteristics and differences in the hepatic mixed function oxidases of different species. Pharmacol Ther. 1979;6(1):41–98. doi: 10.1016/0163-7258(79)90056-1. [DOI] [PubMed] [Google Scholar]
  16. Kawajiri K., Gotoh O., Sogawa K., Tagashira Y., Muramatsu M., Fujii-Kuriyama Y. Coding nucleotide sequence of 3-methylcholanthrene-inducible cytochrome P-450d cDNA from rat liver. Proc Natl Acad Sci U S A. 1984 Mar;81(6):1649–1653. doi: 10.1073/pnas.81.6.1649. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kimura S., Gonzalez F. J., Nebert D. W. Mouse cytochrome P3-450: complete cDNA and amino acid sequence. Nucleic Acids Res. 1984 Mar 26;12(6):2917–2928. doi: 10.1093/nar/12.6.2917. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Leighton J. K., DeBrunner-Vossbrinck B. A., Kemper B. Isolation and sequence analysis of three cloned cDNAs for rabbit liver proteins that are related to rabbit cytochrome P-450 (form 2), the major phenobarbital-inducible form. Biochemistry. 1984 Jan 17;23(2):204–210. doi: 10.1021/bi00297a005. [DOI] [PubMed] [Google Scholar]
  19. Messing J., Vieira J. A new pair of M13 vectors for selecting either DNA strand of double-digest restriction fragments. Gene. 1982 Oct;19(3):269–276. doi: 10.1016/0378-1119(82)90016-6. [DOI] [PubMed] [Google Scholar]
  20. Mizukami Y., Fujii-Kuriyama Y., Muramatsu M. Multiplicity of deoxyribonucleic acid sequences with homology to a cloned complementary deoxyribonucleic acid coding for rat phenobarbital-inducible cytochrome P-450. Biochemistry. 1983 Mar 1;22(5):1223–1229. doi: 10.1021/bi00274a036. [DOI] [PubMed] [Google Scholar]
  21. Nebert D. W., Negishi M. Multiple forms of cytochrome P-450 and the importance of molecular biology and evolution. Biochem Pharmacol. 1982 Jul 15;31(14):2311–2317. doi: 10.1016/0006-2952(82)90523-8. [DOI] [PubMed] [Google Scholar]
  22. Negishi M., Swan D. C., Enquist L. W., Nebert D. W. Isolation and characterization of a cloned DNA sequence associated with the murine Ah locus and a 3-methylcholanthrene-induced form of cytochrome P-450. Proc Natl Acad Sci U S A. 1981 Feb;78(2):800–804. doi: 10.1073/pnas.78.2.800. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Ozols J., Heinemann F. S., Johnson E. F. Amino acid sequence of an analogous peptide from two forms of cytochrome P-450. J Biol Chem. 1981 Nov 25;256(22):11405–11408. [PubMed] [Google Scholar]
  24. Penno M. B., Dvorchik B. H., Vesell E. S. Genetic variation in rates of antipyrine metabolite formation: a study in uninduced twins. Proc Natl Acad Sci U S A. 1981 Aug;78(8):5193–5196. doi: 10.1073/pnas.78.8.5193. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Perler F., Efstratiadis A., Lomedico P., Gilbert W., Kolodner R., Dodgson J. The evolution of genes: the chicken preproinsulin gene. Cell. 1980 Jun;20(2):555–566. doi: 10.1016/0092-8674(80)90641-8. [DOI] [PubMed] [Google Scholar]
  26. Phillips I. R., Shephard E. A., Ashworth A., Rabin B. R. Cloning and sequence analysis of a rat liver cDNA coding for a phenobarbital-inducible microheterogenous cytochrome P-450 variant: regulation of its messenger level by xenobiotics. Gene. 1983 Dec;26(1):41–52. doi: 10.1016/0378-1119(83)90034-3. [DOI] [PubMed] [Google Scholar]
  27. Phillips I. R., Shephard E. A., Rabin B. R., Bayney R. M., Pike S. F., Ashworth A., Estall M. R. Factors controlling the expression of genes coding for drug-metabolizing enzymes. Biochem Soc Trans. 1983 Aug;11(4):460–463. doi: 10.1042/bst0110460. [DOI] [PubMed] [Google Scholar]
  28. 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]
  29. Sanger F., Coulson A. R., Barrell B. G., Smith A. J., Roe B. A. Cloning in single-stranded bacteriophage as an aid to rapid DNA sequencing. J Mol Biol. 1980 Oct 25;143(2):161–178. doi: 10.1016/0022-2836(80)90196-5. [DOI] [PubMed] [Google Scholar]
  30. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  31. Tarr G. E., Black S. D., Fujita V. S., Coon M. J. Complete amino acid sequence and predicted membrane topology of phenobarbital-induced cytochrome P-450 (isozyme 2) from rabbit liver microsomes. Proc Natl Acad Sci U S A. 1983 Nov;80(21):6552–6556. doi: 10.1073/pnas.80.21.6552. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. 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]
  33. White P. C., New M. I., Dupont B. Cloning and expression of cDNA encoding a bovine adrenal cytochrome P-450 specific for steroid 21-hydroxylation. Proc Natl Acad Sci U S A. 1984 Apr;81(7):1986–1990. doi: 10.1073/pnas.81.7.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Woods D. E., Markham A. F., Ricker A. T., Goldberger G., Colten H. R. Isolation of cDNA clones for the human complement protein factor B, a class III major histocompatibility complex gene product. Proc Natl Acad Sci U S A. 1982 Sep;79(18):5661–5665. doi: 10.1073/pnas.79.18.5661. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Yabusaki Y., Shimizu M., Murakami H., Nakamura K., Oeda K., Ohkawa H. Nucleotide sequence of a full-length cDNA coding for 3-methylcholanthrene-induced rat liver cytochrome P-450MC. Nucleic Acids Res. 1984 Mar 26;12(6):2929–2938. doi: 10.1093/nar/12.6.2929. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Yuan P. M., Ryan D. E., Levin W., Shively J. E. Identification and localization of amino acid substitutions between two phenobarbital-inducible rat hepatic microsomal cytochromes P-450 by micro sequence analyses. Proc Natl Acad Sci U S A. 1983 Mar;80(5):1169–1173. doi: 10.1073/pnas.80.5.1169. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES