Abstract
The properties of five structurally related forms of cytochrome P-450 (PB1a, PB1b, PB2a, PB2b and PB2d) isolated from rats treated with phenobarbital have been compared with two forms isolated previously now termed 'PB1c' and 'PB2c'. These enzymes were characterized by their marginal inducibility by phenobarbital and are clearly distinguishable from the major phenobarbital-inducible proteins. PB1a and PB1b differed in Mr (52,700 and 52,900), absorption spectra and papain-proteolysis fragments. However, they had identical N-terminal sequences. PB2a, PB2b and PB2d had apparent Mr values of 52,900, 52,900 and 50,800. PB2a and PB2b had different N-terminal sequences and, after digestion with papain, gave different papain-proteolysis fragments. The N-terminal sequence of PB2b was similar to, but not identical with, that of pregnenolone-16 alpha-carbonitrile-inducible P-450 species, and PB2b was the protein most closely related to PB2c. The extent of immunocross-reactivity among the forms was stronger within, than between, the PB1 and PB2 groups. Even structurally similar forms were functionally diverse, exhibiting large differences in metabolic specificity in the dealkylation of a series of alkoxyresorufins.
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- 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]
- 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]
- Buchmann A., Kuhlmann W., Schwarz M., Kunz W., Wolf C. R., Moll E., Friedberg T., Oesch F. Regulation and expression of four cytochrome P-450 isoenzymes, NADPH-cytochrome P-450 reductase, the glutathione transferases B and C and microsomal epoxide hydrolase in preneoplastic and neoplastic lesions in rat liver. Carcinogenesis. 1985 Apr;6(4):513–521. doi: 10.1093/carcin/6.4.513. [DOI] [PubMed] [Google Scholar]
- Burke M. D., Mayer R. T. Differential effects of phenobarbitone and 3-methylcholanthrene induction on the hepatic microsomal metabolism and cytochrome P-450-binding of phenoxazone and a homologous series of its n-alkyl ethers (alkoxyresorufins). Chem Biol Interact. 1983 Jul 15;45(2):243–258. doi: 10.1016/0009-2797(83)90072-8. [DOI] [PubMed] [Google Scholar]
- Burke M. D., Mayer R. T. Ethoxyresorufin: direct fluorimetric assay of a microsomal O-dealkylation which is preferentially inducible by 3-methylcholanthrene. Drug Metab Dispos. 1974 Nov-Dec;2(6):583–588. [PubMed] [Google Scholar]
- Burke M. D., Thompson S., Elcombe C. R., Halpert J., Haaparanta T., Mayer R. T. Ethoxy-, pentoxy- and benzyloxyphenoxazones and homologues: a series of substrates to distinguish between different induced cytochromes P-450. Biochem Pharmacol. 1985 Sep 15;34(18):3337–3345. doi: 10.1016/0006-2952(85)90355-7. [DOI] [PubMed] [Google Scholar]
- Cheng K. C., Schenkman J. B. Purification and characterization of two constitutive forms of rat liver microsomal cytochrome P-450. J Biol Chem. 1982 Mar 10;257(5):2378–2385. [PubMed] [Google Scholar]
- Cleveland D. W., Fischer S. G., Kirschner M. W., Laemmli U. K. Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis. J Biol Chem. 1977 Feb 10;252(3):1102–1106. [PubMed] [Google Scholar]
- Gonzalez F. J., Song B. J., Hardwick J. P. Pregnenolone 16 alpha-carbonitrile-inducible P-450 gene family: gene conversion and differential regulation. Mol Cell Biol. 1986 Aug;6(8):2969–2976. doi: 10.1128/mcb.6.8.2969. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- Heuman D. M., Gallagher E. J., Barwick J. L., Elshourbagy N. A., Guzelian P. S. Immunochemical evidence for induction of a common form of hepatic cytochrome P-450 in rats treated with pregnenolone-16 alpha-carbonitrile or other steroidal or non-steroidal agents. Mol Pharmacol. 1982 May;21(3):753–760. [PubMed] [Google Scholar]
- 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]
- Kumar A., Raphael C., Adesnik M. Cloned cytochrome P-450 cDNA. Nucleotide sequence and homology to multiple phenobarbital-induced mRNA species. J Biol Chem. 1983 Sep 25;258(18):11280–11284. [PubMed] [Google Scholar]
- 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]
- 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]
- Lu A. Y., West S. B. Multiplicity of mammalian microsomal cytochromes P-45. Pharmacol Rev. 1979 Dec;31(4):277–295. [PubMed] [Google Scholar]
- NASH T. The colorimetric estimation of formaldehyde by means of the Hantzsch reaction. Biochem J. 1953 Oct;55(3):416–421. doi: 10.1042/bj0550416. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- 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]
- Slaughter S. R., Wolf C. R., Marciniszyn J. P., Philpot R. M. The rabbit pulmonary monooxygenase system. partial structural characterization of the cytochrome P-450 components and comparison to the hepatic cytochrome P-450. J Biol Chem. 1981 Mar 10;256(5):2499–2503. [PubMed] [Google Scholar]
- Suwa Y., Mizukami Y., Sogawa K., Fujii-Kuriyama Y. Gene structure of a major form of phenobarbital-inducible cytochrome P-450 in rat liver. J Biol Chem. 1985 Jul 5;260(13):7980–7984. [PubMed] [Google Scholar]
- Ullrich V., Weber P. The O-dealkylation of 7-ethoxycoumarin by liver microsomes. A direct fluorometric test. Hoppe Seylers Z Physiol Chem. 1972 Jul;353(7):1171–1177. doi: 10.1515/bchm2.1972.353.2.1171. [DOI] [PubMed] [Google Scholar]
- 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]
- Waxman D. J., Walsh C. Cytochrome P-450 isozyme 1 from phenobarbital-induced rat liver: purification, characterization, and interactions with metyrapone and cytochrome b5. Biochemistry. 1983 Sep 27;22(20):4846–4855. doi: 10.1021/bi00289a035. [DOI] [PubMed] [Google Scholar]
- Wolf C. R., Moll E., Friedberg T., Oesch F., Buchmann A., Kuhlmann W. D., Kunz H. W. Characterization, localization and regulation of a novel phenobarbital-inducible form of cytochrome P450, compared with three further P450-isoenzymes, NADPH P450-reductase, glutathione transferases and microsomal epoxide hydrolase. Carcinogenesis. 1984 Aug;5(8):993–1001. doi: 10.1093/carcin/5.8.993. [DOI] [PubMed] [Google Scholar]
- Wolf C. R., Oesch F. Isolation of a high spin form of cytochrome P-450 induced in rat liver by 3-methylcholanthrene. Biochem Biophys Res Commun. 1983 Mar 16;111(2):504–511. doi: 10.1016/0006-291x(83)90335-2. [DOI] [PubMed] [Google Scholar]
- Wolf C. R., Smith B. R., Ball L. M., Serabjit-Singh C., Bend J. R., Philpot R. M. The rabbit pulmonary monooxygenase system. Catalytic differences between two purified forms of cytochrome P-450 in the metabolism of benzo(a)pyrene. J Biol Chem. 1979 May 10;254(9):3658–3663. [PubMed] [Google Scholar]