Benzo Pyrene Hydroxylase Research Articles

Purification and characterization of NADPH-cytochrome P-450 reductase from rat epidermis.

NADPH-cytochrome P-450 oxidoreductase (P-450 red) transfers reducing equivalents from NADPH to cytochrome P-450 (P-450) in the monooxygenase system. Detergent solubilized proteins from the membrane fraction of neonatal rat epidermis were purified by 2',5'-ADP-agarose affinity column chromatography. The purified protein showed an apparent homogeneity on sodium dodecylsulfate-polyacrylamide gel electrophoresis and molecular weight was estimated to be 78 kDa. NADPH-cytochrome c reductase activity increased by 95-fold in the purified enzyme. Epidermal P-450 red in vitro reconstituted benzo(a)pyrene hydroxylase activity in a dose dependent manner with P-450 purified from either rat liver or epidermis. Western blot analysis demonstrated that epidermal P-450 red immunologically cross reacts to liver P-450 red. Immunohistochemical staining showed that the enzyme was predominantly localized in the epidermis. The intensity of immunohistochemical staining of rat skin sections and tissue distribution did not change in the skin treated with beta-naphthoflavone, which results in a substantial increase in P-450 1A1 activity. Quantitative assessment of P-450 red in treated and untreated epidermis also showed no change. These findings indicate that constitutive P-450 red, fully capable of supporting P-450, exists in rat epidermis, and can function in metabolism of endogenous and exogenous compounds.

Differences in inducibility of cytochrome p-4501a1, monooxygenases and glutathione s-transferase in cutaneous and extracutaneous tissues after topical and parenteral administration of β-naphthoflavone to rats

1. 1. The effect of topical and parenteral administration of β-NF to rats on cytochrome P-4501A1, monooxygenases and glutathione S-transferase enzyme activities in various tissues were studied. 2. 2. Topical and I.P. administration of β-NF to rats resulted in variable induction responses (2–28-fold) in various tissues. Benzo(a)pyrene hydroxylase, ethoxyresorufin- O-deethylase, ethoxycoumarin- O-deethylase, and GST activities in liver, kidney and skin were induced to higher extent than in other tissues following both topical and i.p. administration. 3. 3. Western blot analysis of microsomal preparation from β-NF-treated rats using monoclonal antibody to P-4501A1 showed a marked induction of P-4501A1 enzyme protein in skin and liver. 4. 4. These results suggest that both topically as well as parenterally administered xenobiotics may influence drug metabolizing capacity of skin and other tissues.

Effect of intratracheally administered lindane on aldrin and benzo(a)pyrene contents in lungs of rats

Pretreatment with lindane resulted in inhibition of benzo(a)pyrene hydroxylase activity in the lungs of rats. The enzyme activity tended to return to normal 5 days after the administration of lindane. Studies with benzo(a)pyrene and aldrin indicated reduced elimination of these compounds from the lungs of lindane-treated animals, suggesting that lindane may alter the clearance of certain substances or compounds from lungs. The delayed clearance of these compounds from lungs may be an indirect effect of lindane related to inhibition of certain metabolizing enzymes.

Xenobiotic metabolizing enzymes in uninduced and induced rainbow trout ( Oncorhynchus mykiss): Effects of diets and food deprivation

1. Untreated and β-naphtoflavone (BNF)-treated trout ( Oncorhynchus mykiss) were maintained for 15–21 days under 3 different feeding regimens or under starvation. 2. Possible diet effects were studied by measuring the following 13 hepatic parameters: aminopyrine demethylase (APDM), ethoxyresorufin-O-deethylase (EROD), benzo(a)pyrene hydroxylase (AHH), UDP glucuronyltransferase (UDPGT), glutathione reductase (GR), glutathione-S-transferase (GST), glutathione peroxidase (GPx), glucose 6-phosphate dehydrogenase (G6PD) and 6-phosphogluconate dehydrogenase (6PGD) enzyme activities; the content of total cytochrome P-450 (P-450); proteic and non-proteic SH groups (SHP, SHNP) and liver somatic index (LSI). 3. The results show that some feeding regimens affect a large number of these parameters in both uninduced and BNF-treated fish. 4. Hence, caution should be imposed in the use of the biochemical markers as a water-quality monitoring tool. 5. In particular, it is advisable to maintain fish without feeding during short-term experiments, especially because trout exhibit an excellent biochemical response to inducer even after a 21-day starvation period. 6. Conversely, when fish are to be necessarily fed (long-term experiments), diet must be carefully chosen and controlled by preliminary tests because some commercial fish pellets contain both inducers and inhibitors of several microsomal/cytosolic enzymes.

Ascorbic acid deficiency reduces the level of mRNA for cytochrome P-450 on the induction by polychlorinated biphenyls.

Ascorbic acid (AsA) deficiency causes a decrease in hepatic concentration of cytochrome P-450 and a decrease in hepatic activity of drug-metabolizing enzymes in rats unable to synthesize AsA (ODS rats). To study the mechanism of the decrease in hepatic concentration of cytochrome P-450 isozymes by AsA deficiency, we chose the xenobiotics-inducible cytochrome P-450 and performed the experiments indicated below. AsA-deficient rats were fed polychlorinated biphenyls (PCB) which markedly induce both CYP1A subfamily and several isozymes in CYP2B subfamily. First, we assayed the activities of two drug-metabolizing enzymes so that one could be functionally distinguished from another. AsA deficiency significantly reduced the hepatic activity of aminopyrine-N-demethylase in ODS rats with and without dietary PCB, but had no effect on benzo(a)pyrene hydroxylase activity. Secondly, quantitative immunoblot analyses demonstrated that the levels of CYP2B1/2B2 and CYP1A1 in the AsA-deficiency rats fed PCB were approximately 60 and 80% lower than those found in rats fed AsA-supplemented diet. The degree of reduction in CYP2B1/2B2 was greater than CYP1A1. Thirdly, AsA deficiency caused a decrease in hepatic abundance of CYP2B1/2B2 mRNA, whereas it had no effect on the levels of CYP1A1 and 1A2 mRNA. These results indicated that dietary AsA selectively affects the levels of CYP2B1/2B2 mRNA among cytochrome P-450 induced by PCB and plays important roles for optimum induction of drug-inducible cytochrome P-450. We concluded that AsA deficiency decreases specific froms of drug-inducible cytochrome P-450, especially CYP2B1/2B2 and that the reduction of CYP2B1/2B2 mRNA level in AsA-deficient rats caused a decrease in cytochrome P-450 concentration and hepatic activity of drug-metabolizing enzymes.

Anti-P450lpr antiserum inhibits specific monooxygenase activities in LPR house fly microsomes.

Monooxygenase activity in microsomes from the LPR strain of house fly (Musca domestica L.) was inhibited by anti-P450lpr, and antiserum specific for house fly cytochrome P450lpr. Anti-P450lpr did not inhibit house fly cytochrome P450 reductase or rat cytochrome P450 monooxygenase assays, consistent with specific inhibition of P450lpr. Anti-P450lpr inhibited the ability of cytochrome P450 reductase to reduce carbon monoxide treated LPR microsomal cytochrome P450, up to 49% of the total, showing that inhibition of cytochrome P450 reduction is the major mechanism of inhibition. Anti-P450lpr inhibited 98% of methoxyresorufin-O-demethylase activity and all the benzo(a)pyrene hydroxylase activity in LPR microsomes, but none of the pentoxyresorufin-O-dealkylase activity. The antiserum partially inhibited ethoxyresorufin-O-dealkylase and ethoxycoumarin-O-dealkylase activity. These results demonstrate that methoxyresourfin-O-demethylase activity and benzo(a)pyrene hydroxylase activity are characteristic substrates for P450lpr activity in the LPR strain of house fly.

Dose dependent induction of the microsomal monooxygenase aryl hydrocarbon hydroxylase in isolated peripheral blood monocytes: the influence of age.

The induction of the microsomal monooxygenase benzo(alpha)pyrene hydroxylase was investigated in isolated peripheral blood monocytes from young and old donors, after exposure to different concentrations of benzanthracene as an inducing agent. The sensitivity of the cells to the inducing stimulus was not dependent on the age of the donor. The mechanisms underlying impaired monooxygenase induction in the elderly remain to be clarified.

Diminution in kerosene-mediated induction of drug metabolizing enzymes by asbestos in rat lungs.

In order to determine the pulmonary toxicity of kerosene and its ignition product (soot) in asbestos exposed subjects, the activities of phase I and phase II drug metabolizing enzymes in rat lungs after single intratracheal co-exposure to Indian chrysotile asbestos and kerosene or its soot and Indian chrysotile were assayed. Exposure to kerosene or its soot resulted in a significant increase in the level of microsomal cytochrome P-450 and the activity of P-450 dependent monooxygenase, benzo(a)pyrene hydroxylase, as well as in the activities of microsomal epoxide hydrase and cytosolic glutathione-S-transferase (GST). However, in chrysotile exposed animals a reverse pattern in these parameters was recorded. The co-exposure to chrysotile and kerosene or chrysotile and soot led to a significant depletion in cytochrome P-450 level and a decrease in the activities of benzo(a)pyrene hydroxylase, epoxide hydrase and GST when compared to kerosene and soot controls, respectively. These results suggest that asbestos by altering the pulmonary drug metabolizing enzyme system may increase the toxic potential of kerosene and its ignition product in the respiratory system.

Hexachlorocyclohexane (HCH) induced changes in microsomal drug metabolizing enzymes and liver lipids in rats fed different levels of protein

The interaction of HCH (50 mg/kg) and dietary protein levels on microsomal drug metabolizing enzymes system and liver lipids were studied in the rats for 90 days. The results indicated that rats fed a lower protein diet and HCH has a higher rate of mortality, lower rate of growth and an increased liver weight. A significant induction in the hepatic microsomal aminopyrine‐N‐demethylase, p‐nitroanisole‐O‐dealkylase, benzo(a)pyrene hydroxylase and glutathione‐S‐transferase activity was observed in pesticide treated animals as compared to control animals. The pathological changes observed in liver of HCH treated animals consisted mainly of necrosis and fatty degeneration of hepatocytes. HCH also induced the significant accumulation of cholesterol, triglycerides, phospholipid and total lipid in liver in low protein diet animals. Protein accelerates the metabolism of HCH, resulting in a decrease of HCH concentration with the increase of dietary protein level. A close correlation existed between lipid accumulati...

Interaction of mineral fibres with lung cytochrome P-450 system: Impairment of drug metabolizing enzyme activities

In the present study, in vitro effects of carcinogenic (chrysotile, crocidolite, amosite) and non-carcinogenic (anthophyllite) varieties of asbestos and three varieties of Indian wollastonite (kemolit A-60, Kemolit-N, Kemolit ASB-3) boTh on phase I and phase Ii drug metabolizing enzymes and microsomal lipid peroxidatin (LPO) in rat lung were investigated. All the mineral fibres when incubated with lung microsomes adsorbed cytochrome P-450 to different degrees, while heme was released only by chrysotile and crocidolite. Chrysotile, crocidolite, amosite and kemolit A-60 treatments only led to significant depletion in the activities of benzo(a)pyrene hydroxylase, and epoxide hydratase. The carcinogenic varieties of asbestos also decreased the activity of glutathione-S-transferase, while kemolit-N significantly increased the activity. However, kemolit A-60 and kemolit ASB-3 did not alter the activity of this enzyme. All the dusts which impaired the activities of drug metabolizing enzymes also significantly induced LPO in the microsomes. These data indicate the differential behaviour of asbestos and wollastonite, both on the drug metabolizing enzymes and induction of LPO in rat lung and also suggest that these effects may be related to their toxic potentials.

Interactive effects of cadmium and benzo(a)pyrene on cellular structure and biotransformation enzymes of the liver of the European eel Anguilla anguilla

Interactive effects of cadmium and benzo(a)pyrene (BaP) on eel Anguilla anguilla liver were studied at the ultrastructural and biotransformation enzyme levels. Seawater-adapted eels were held for 24 days in either clean or cadmium-containing (5 μg Cd −1) seawater. then given a single intraperitoneal injection of BaP (20 mg kg −1 body weight) and finally sacrificed 24 h later. Cadmium exposure alone caused a hepatic perivascular fibrosis. whereas the short-term BaP intoxication alone produced marked lipid droplet accumulation associated with glycogen depletion. When eels were exposed to both toxicants together, the liver showed a complete disorganisation of the hepatic parenchyma including nuclear degeneration. The existence of elements of a biotransformation enzyme system, i.e. cytochrome P-450 monooxygenase system and glutathione-S-transferase (GST). was demonstrated in the liver. BaP injection alone had no detectable effect on microsomal cytochrome P-450 content and NADPH-cytochrome c reductase activity but markedly increased 7-ethoxyresorufin O-deethylase (EROD) and benzo(a)pyrene hydroxylase (BaPMO) activities (induction factors of 15- and 35-fold, respectively). In contrast, cytosolic GST activity was slightly decreased by this treatment. In cadmium-exposed eels, BaP treatment elicited a higher increase in EROD and BaPMO activities (induction factors of 19- and 71-fold. respectively) and also causes a 1.8-fold increase in cylochrome P-450 microsomal content. The effects of cadmium on biotransformation enzymes are discussed in relation to the ultrastructural effects of both toxicants.

Diminution in phase I and phase II drug metabolizing enzymes of rat lung by asbestos: Anin vitro study

In the present paper studies are presented concerning the effect of three varieties of asbestos namely, chrysotile, crocidolite and amosite, and of titanium dioxide, an inert non-asbestos dust, on the enzymes of phase 1 and phase 2 drug metabolism in isolated rat lung microsomes and post-microsomal fraction. Since 3-methylcholanthrene (3-MC) is known to induce cytochromes P-450 in the IA family and their associated activity, benzo(a)pyrene hydroxylase, therefore, the effect of these mineral fibers on cytochrome P-450 and benzo(a)pyrene hydroxylase in lung microsomes isolated from 3-MC-treated rats was studied.

Effect of Plasmodium berghei infection and chloroquine on the hepatic drug metabolizing system of mice

The hepatic microsomal mixed-function oxidase (MFO) system was markedly impaired during Plasmodium berghei infection in mice. Cytochrome P-450 and other mono-oxygenases, viz. aniline hydroxylase, aminopyrine- N-demethylase and benzo( a)pyrene hydroxylase, were significantly decreased while microsomal heme showed a four-fold increase at peak parasitemia (> 50%). Oral treatment with chloroquine (16 mg kg −1 body wt for 4 days) of P. berghei-infected mice cleared the parasitemia within 72 h and almost normalized the altered levels of MFO indices, a week after cessation of treatment. The findings were further supported by the isoenzymic profile and drug-binding properties of terminal mono-oxygenase, cytochrome P-450.

Administration of testosterone alleviates the constitutive sex difference in rat brain cytochrome P-450

Sex-related difference was observed in the levels of total cytochrome P-450 (P-450) and the mono-oxygenase activity mediated by P-450 (b,e), namely, aminopyrine N-demethylase and morphine N-demethylase activity in rat brain microsomes. Male rat brain had higher activity of the above enzymes as compared to the female rat brain. On the other hand, P-450 (c,d) mediated 7-ethoxycoumarin O-deethylase and benzo(a)pyrene hydroxylase activity showed no sex-related difference in rat brain. Administration of testosterone elevated the levels of total P-450, aminopyrine N-demethylase and morphine N-demethylase in female rat brain to levels comparable with that of the male rat brain. No significant change was observed in the levels of 7-ethoxycoumarin O-deethylase and benzo(a)pyrene hydroxylase and NADPH cytochrome c reductase. All of the above enzyme levels were unaffected in the male rat brain following the treatment with testosterone. These results indicate that testosterone may regulate the forms of cerebral P-450 that are associated with the sex-related difference observed in rat brain.

Effects of acetone administration on cytochrome P-450-dependent monooxygenases in hamster liver, kidney, and lung

The effects of acetone on liver, kidney, and lung monooxygenases were studied using hamsters administered 8% acetone in drinking water. Binding of aniline to liver microsomes induced a type II difference spectrum, and the spectral binding was enhanced in hamsters pretreated with acetone. Administration of acetone caused significant increases of cytochrome P-450 and cytochrome b5 contents in liver microsomes. The increases of the hemeproteins were associated with induction of monooxygenase activities toward test substrates, aniline, N-nitrosodimethylamine, benzphetamine, benzo(a)pyrene, and 7-ethoxycoumarin. In the kidneys, acetone administration increased microsomal contents of the hemeprotein and monooxygenase activities toward aniline. N-nitrosodimethylamine, and 7-ethoxycoumarin, but not benzphetamine or benzo(a)pyrene. In the lungs, acetone pretreatment increased aniline hydroxylase activity without affecting the levels of N-nitrosodimethylamine demethylase, cytochromes P-450 and b5. In marked contrast to the inductive effects in the liver, acetone administration markedly decreased lung microsomal benzo(a)pyrene hydroxylase and 7-ethoxycoumarin O-deethylase activities. Gel electrophoresis of liver and kidney microsomes from control and acetone-treated hamsters revealed that acetone treatment enhanced the intensity of a protein band(s) in the cytochrome P-450 molecular weight region. Immunoblotting of the microsomal proteins showed that the protein band induced by acetone in hamster liver, kidney and lung was cross-reactive with antibody raised against ethanol-inducible human liver cytochrome P-450. These results demonstrate that acetone has the ability to uniformly induce a specific form of cytochrome P-450, designated as IIE1, and to cause differential changes of monooxygenase activities in the hamster tissues.(ABSTRACT TRUNCATED AT 250 WORDS)

The development of drug‐metabolizing enzymes in female sheep livers

The purpose of this investigation was to determine age-related changes of some hepatic drug-metabolizing activities in Lacaune ewes in the foetal, neonatal (1 and 4 weeks), growing (7 months), pregnant (11 months) and adult (6 years) stages. Although microsomal cytochrome P-450 was not detected in 3-month-old foetuses, it increased regularly from 1-week- to 11-month-old animals. Among mixed-function oxidases, the development of aminopyrine and ethylmorphine N-demethylases, benzo(alpha)pyrene hydroxylase and ethoxycoumarin O-deethylase were correlated to that of total cytochrome P-450. Due to their presence in foetal liver or their more rapid evolution, cytochrome b5, NADPH cytochrome c reductase, aniline hydroxylase, benzphetamine N-demethylase and erythromycin N-demethylase did not parallel the ontogenesis of cytochrome P-450. Hepatic transferases showed different developmental patterns from mono-oxygenases, so UDP glucuronyltransferase was detected in the foetus, reached maximum activity in all young ages up to the pregnant stage and subsequently fell in adult ewes. Concerning glutathione S-transferase accepting 1-chloro-2,4-dinitrobenzene as substrate, similar values were obtained in the foetus and all young animals, whereas five- to tenfold higher values were obtained in both pregnant and adult female sheep. N-acetyltransferase using sulphamethazine did not significantly change from foetuses to adults but there were large differences in the capacity of hepatic acetylation between animals belonging to the same group.

Diethylstilbestrol potentiates and testosterone antagonizes the action of 3‐methylcholanthrene on benzo(a) pyrene metabolism in hep G2 cells

We have used the human hepatoma cell line, Hep G2, to examine the ability of hormones and xenobiotics to modulate the hepatic induction of benzo(a)pyrene hydroxylase and epoxide hydrolase. Hep G2 cells were cultured in Eagle's Minimum Essential Medium supplemented with 10% fetal calf serum. 3-Methylcholanthrene, diethylstilbestrol, testosterone propionate, and combinations of 3-methylcholanthrene, and each of the hormones were added directly to the culture media. We subsequently studied the metabolism of benzo(a)pyrene using cell lysates of the Hep G2 cells. Metabolites were quantitated by high-performance liquid chromatography (HPLC) using fluorodetection. Exposure to 3-methylcholanthrene alone resulted in an eightfold increase in total benzo(a)pyrene metabolites with a change of the predominant metabolite from the 3-hydroxybenzo(a)pyrene to the carcinogenic pathway of the benzo(a)pyrene-7,8-diol. Diethylstilbestrol and testosterone propionate resulted in small, but significant, decreases in metabolism of benzo(a)pyrene. When exposed in combination with 3-methylcholanthrene, testosterone propionate antagonized and diethylstilbestrol potentiated the metabolism of benzo(a)pyrene. 3-Methylcholanthrene, diethylstilbestrol, and combinations of 3-methylcholanthrene and diethylstilbestrol or testosterone propionate resulted in increased epoxide hydrolase activity as compared to controls. These results, carried out in a human hepatoma cell line, lend support to a concern for potentiated toxicity and carcinogenicity following exposure to complex chemical mixtures.

Presence of functional cytochrome P-450 on isolated rat hepatocyte plasma membrane

Antibodies against cytochrome P-450 are found in some children with autoimmune hepatitis (antiliver/kidney microsome 1) and in patients with ticrynafen hepatitis (antiliver/kidney microsome 2). For an immune reaction against cytochrome P-450 to possibly destroy the hepatocytes, one must assume that cytochrome P-450 is present on the plasma membrane surface of hepatocytes. In a first series of experiments, plasma membranes were prepared with a technique based on the electrostatic attachment of isolated hepatocytes to polyethyleneimine-coated beads. After vortexing, beads were coated with a very pure plasma membrane fraction. Microsomal contamination, judged from the specific activities of glucose-6-phosphatase or NADH-cytochrome c reductase, was less than 1%. Nevertheless, the specific content (per milligram of protein) of CO-binding cytochrome P-450 was 20% of that in microsomes; the specific benzo(a)pyrene hydroxylase activity was 25%, and ethoxycoumarin deethylase 11%. Immunoblots showed the presence of cytochromes P-450 UT-A, UT-H, PB-B, ISF-G and PCN-E, the last three isoenzymes being inducible by, respectively, phenobarbital, 3-methylcholanthrene and dexamethasone. In a second series of experiments, nonpermeabilized isolated hepatocytes from untreated rats were incubated with anticytochrome P-450 antibodies. Immunofluorescence and immunoperoxidase staining confirmed the presence of cytochromes P-450 UT-A, PB-B and ISF-G on the membrane. In a last series of experiments, human antiliver-kidney microsomal 1 antibodies were found to react specifically with rat liver plasma membrane cytochrome P-450 UT-H (IID subfamily). We conclude that several cytochrome P-450 isoenzymes are present, active and inducible on the plasma membrane surface of hepatocytes. It is therefore conceivable that immunization against plasma membrane cytochrome P-450 might lead to the immunological destruction of hepatocytes in some patients.

Regional and subcellular distribution of cytochrome P-450-dependent drug metabolism in monkey brain: The olfactory bulb and the mitochondrial fraction have high levels of activity

In brain of female monkey ( M. fascicularis ) the content of cytochrome P-450 and benzo(a)pyrene hydroxylase activity in the mitochondrial fraction exceeded that of the microsomes by more than 4-fold. The mitochondrial drug metabolism activity exhibited substrate specificity and, unlike the microsomes, did not catalyze 7-ethoxyresorufin O-deethylase reaction. Moreover, the rate of benzo(a)pyrene hydroxylase activity by both the mitochondrial and the microsomal fractions displayed regional variation with the olfactory bulb displaying the highest hydroxylase activity. In contrast, the microsomal 7-ethoxyresorufin O-deethylase activity was uniformally distributed in all brain regions.

Presence of Functional Cytochrome P–450 on Isolated Rat Hepatocyte Plasma Membrane

Antibodies against cytochrome P-450 are found in some children with autoimmune hepatitis (antiliver/kidney microsome 1) and in patients with ticrynafen hepatitis (antiliver/kidney microsome 2). For an immune reaction against cytochrome P-450 to possibly destroy the hepatocytes, one must assume that cytochrome P-450 is present on the plasma membrane surface of hepatocytes. In a first series of experiments, plasma membranes were prepared with a technique based on the electrostatic attachment of isolated hepatocytes to polyethyleneimine-coated beads. After vortexing, beads were coated with a very pure plasma membrane fraction. Microsomal contamination, judged from the specific activities of glucose-6-phosphatase or NADH-cytochrome c reductase, was less than 1%. Nevertheless, the specific content (per milligram of protein) of CO-binding cytochrome P-450 was 20% of that in microsomes; the specific benzo(a)pyrene hydroxylase activity was 25%, and ethoxycoumarin deethylase 11%. Immunoblots showed the presence of cytochromes P-450 UT-A, UT-H, PB-B, ISF-G and PCN-E, the last three isoenzymes being inducible by, respectively, phenobarbital, 3-methylcholanthrene and dexamethasone. In a second series of experiments, nonpermeabilized isolated hepatocytes from untreated rats were incubated with anticytochrome P-450 antibodies. Immunofluorescence and immunoperoxidase staining confirmed the presence of cytochromes P-450 UT-A, PB-B and ISF-G on the membrane. In a last series of experiments, human antiliver-kidney microsomal 1 antibodies were found to react specifically with rat liver plasma membrane cytochrome P-450 UT-H (IID subfamily). We conclude that several cytochrome P-450 isoenzymes are present, active and inducible on the plasma membrane surface of hepatocytes. It is therefore conceivable that immunization against plasma membrane cytochrome P-450 might lead to the immunological destruction of hepatocytes in some patients.