Abstract
It has not yet been determined whether human liver contains inducible cytochromes P-450 similar to those that catalyze the oxidative metabolism of foreign substances in animals. We carried out immunoblot analyses of liver microsomes isolated from eight patients and found that each contained a cytochrome P-450, termed HLp, that reacted with antibodies directed against P-450p, a rat liver cytochrome that is inducible by the anti-glucocorticoid pregnenolone-16 alpha-carbonitrile, by glucocorticoids, by anti-seizure drugs, and by such macrolide antibiotics as triacetyloleandomycin. In the two patients who received dexamethasone and anti-seizure medications and in the one patient who was given triacetyloleandomycin, the concentrations of immunoreactive HLp and the ability to demethylate erythromycin and/or to convert triacetyloleandomycin to a metabolite that forms a spectral complex with cytochrome P-450 heme (catalytic properties unique to P-450p in rat liver) were significantly higher as compared to the values for patients who received no inducing drugs. We purified HLp to homogeneity and found that it was immunochemically related to P-450p and to its homologue in the rabbit (LM3c), actively demethylated erythromycin in a reconstituted system, exhibited electrophoretic mobility identical to that of P-450p, and shared 57% homology in its NH2-terminal amino acid sequence with that of a pregnenolone-16 alpha-carbonitrile-inducible rat cytochrome P-450. We conclude that HLp is a human representative of the multigene family of the glucocorticoid-inducible cytochromes P-450.
Full text
PDF




Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Alvares A. P., Fischbein A., Anderson K. E., Kappas A. Alterations in drug metabolism in workers exposed to polychlorinated biphenyls. Clin Pharmacol Ther. 1977 Aug;22(2):140–146. doi: 10.1002/cpt1977222140. [DOI] [PubMed] [Google Scholar]
- Elshourbagy N. A., Guzelian P. S. Separation, purification, and characterization of a novel form of hepatic cytochrome P-450 from rats treated with pregnenolone-16 alpha-carbonitrile. J Biol Chem. 1980 Feb 25;255(4):1279–1285. [PubMed] [Google Scholar]
- Fukushima D. K., Bradlow H. L., Hellman L. Effects of o,p'-DDD on cortisol and 6-beta-hydroxycortisol secretion and metabolism in man. J Clin Endocrinol Metab. 1971 Feb;32(2):192–200. doi: 10.1210/jcem-32-2-192. [DOI] [PubMed] [Google Scholar]
- Gonzalez F. J., Nebert D. W., Hardwick J. P., Kasper C. B. Complete cDNA and protein sequence of a pregnenolone 16 alpha-carbonitrile-induced cytochrome P-450. A representative of a new gene family. J Biol Chem. 1985 Jun 25;260(12):7435–7441. [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]
- Guengerich F. P., Wang P., Davidson N. K. Estimation of isozymes of microsomal cytochrome P-450 in rats, rabbits, and humans using immunochemical staining coupled with sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Biochemistry. 1982 Mar 30;21(7):1698–1706. doi: 10.1021/bi00536a035. [DOI] [PubMed] [Google Scholar]
- 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]
- Henry D. A., Sharpe G., Chaplain S., Cartwright S., Kitchingman G., Bell G. D., Langman M. J. The [14C]-aminopyrine breath test. A comparison of different forms of analysis. Br J Clin Pharmacol. 1979 Dec;8(6):539–545. doi: 10.1111/j.1365-2125.1979.tb01041.x. [DOI] [PMC free article] [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]
- Hunter J., Maxwell J. D., Carrella M., Stewart D. A., Williams R. Urinary D-glucaric-acid excretion as a test for hepatic enzyme induction in man. Lancet. 1971 Mar 20;1(7699):572–575. doi: 10.1016/s0140-6736(71)91166-4. [DOI] [PubMed] [Google Scholar]
- Kaminsky L. S., Dunbar D. A., Wang P. P., Beaune P., Larrey D., Guengerich F. P., Schnellmann R. G., Sipes I. G. Human hepatic cytochrome P-450 composition as probed by in vitro microsomal metabolism of warfarin. Drug Metab Dispos. 1984 Jul-Aug;12(4):470–477. [PubMed] [Google Scholar]
- Kellermann G., Luyten-Kellermann M. Phenobarbital-induced drug metabolism in man. Toxicol Appl Pharmacol. 1977 Jan;39(1):97–104. doi: 10.1016/0041-008x(77)90181-8. [DOI] [PubMed] [Google Scholar]
- Kouri R. E., McKinney C. E., Slomiany D. J., Snodgrass D. R., Wray N. P., McLemore T. L. Positive correlation between high aryl hydrocarbon hydroxylase activity and primary lung cancer as analyzed in cryopreserved lymphocytes. Cancer Res. 1982 Dec;42(12):5030–5037. [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]
- Larrey D., Funck-Brentano C., Breil P., Vitaux J., Theodore C., Babany G., Pessayre D. Effects of erythromycin on hepatic drug-metabolizing enzymes in humans. Biochem Pharmacol. 1983 Mar 15;32(6):1063–1068. doi: 10.1016/0006-2952(83)90626-3. [DOI] [PubMed] [Google Scholar]
- Larrey D., Tinel M., Pessayre D. Formation of inactive cytochrome P-450 Fe(II)-metabolite complexes with several erythromycin derivatives but not with josamycin and midecamycin in rats. Biochem Pharmacol. 1983 May 1;32(9):1487–1493. doi: 10.1016/0006-2952(83)90470-7. [DOI] [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]
- 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]
- Ohnhaus E. E., Gerber-Taras E., Park B. K. Enzyme-inducing drug combinations and their effects on liver microsomal enzyme activity in man. Eur J Clin Pharmacol. 1983;24(2):247–250. doi: 10.1007/BF00613826. [DOI] [PubMed] [Google Scholar]
- Ohnhaus E. E., Park B. K. Measurement of urinary 6-beta-hydroxycortisol excretion as an in vivo parameter in the clinical assessment of the microsomal enzyme-inducing capacity of antipyrine, phenobarbitone and rifampicin. Eur J Clin Pharmacol. 1979 Mar 26;15(2):139–145. doi: 10.1007/BF00609878. [DOI] [PubMed] [Google Scholar]
- Pamperl H., Gradner W., Fridrich L., Pointner H., Denk H. Influence of long-term anticonvulsant treatment on liver ultrastructure in man. Liver. 1984 Oct;4(5):294–300. doi: 10.1111/j.1600-0676.1984.tb00941.x. [DOI] [PubMed] [Google Scholar]
- Pessayre D., Descatoire V., Konstantinova-Mitcheva M., Wandscheer J. C., Cobert B., Level R., Benhamou P. J., Jaouen M., Mansuy D. Self-induction by triacetyloleandomycin of its own transformation into a metabolite forming a stable 456 nm-absorbing complex with cytochrome P-450. Biochem Pharmacol. 1981 Mar 15;30(6):553–558. doi: 10.1016/0006-2952(81)90125-8. [DOI] [PubMed] [Google Scholar]
- Pessayre D., Larrey D., Vitaux J., Breil P., Belghiti J., Benhamou J. P. Formation of an inactive cytochrome P-450 Fe(II)-metabolite complex after administration of troleandomycin in humans. Biochem Pharmacol. 1982 May 1;31(9):1699–1704. doi: 10.1016/0006-2952(82)90671-2. [DOI] [PubMed] [Google Scholar]
- Pirttiaho H. I., Sotaniemi E. A., Pelkonen R. O., Pitkänen U. Hepatic blood flow and drug metabolism in patients on enzyme-inducing anticonvulsants. Eur J Clin Pharmacol. 1982;22(5):441–445. doi: 10.1007/BF00542550. [DOI] [PubMed] [Google Scholar]
- Poland A., Smith D., Kuntzman R., Jacobson M., Conney A. H. Effect of intensive occupational exposure to DDT on phenylbutazone and cortisol metabolism in human subjects. Clin Pharmacol Ther. 1970 Sep-Oct;11(5):724–732. doi: 10.1002/cpt1970115724. [DOI] [PubMed] [Google Scholar]
- Remmer H., Schoene B., Fleischmann R. A. Induction of the unspecific microsomal hydroxylase in the human liver. Drug Metab Dispos. 1973 Jan-Feb;1(1):224–230. [PubMed] [Google Scholar]
- Saenger P., Forster E., Kream J. 6 beta-Hydroxycortisol: a noninvasive indicator of enzyme induction. J Clin Endocrinol Metab. 1981 Mar;52(3):381–384. doi: 10.1210/jcem-52-3-381. [DOI] [PubMed] [Google Scholar]
- Schoene B., Fleischmann R. A., Remmer H., von Oldershausen H. F. Determination of drug metabolizing enzymes in needle biopsies of human liver. Eur J Clin Pharmacol. 1972 Mar;4(2):65–73. doi: 10.1007/BF00562499. [DOI] [PubMed] [Google Scholar]
- Schuetz E. G., Wrighton S. A., Barwick J. L., Guzelian P. S. Induction of cytochrome P-450 by glucocorticoids in rat liver. I. Evidence that glucocorticoids and pregnenolone 16 alpha-carbonitrile regulate de novo synthesis of a common form of cytochrome P-450 in cultures of adult rat hepatocytes and in the liver in vivo. J Biol Chem. 1984 Feb 10;259(3):1999–2006. [PubMed] [Google Scholar]
- Simmons D. L., Lalley P. A., Kasper C. B. Chromosomal assignments of genes coding for components of the mixed-function oxidase system in mice. Genetic localization of the cytochrome P-450PCN and P-450PB gene families and the nadph-cytochrome P-450 oxidoreductase and epoxide hydratase genes. J Biol Chem. 1985 Jan 10;260(1):515–521. [PubMed] [Google Scholar]
- 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]
- 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]
- 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]