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Query: UNIPROT:P06889 (Mol)
 630,302 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Pulmonary microsomes, highly purified pulmonary flavin-containing monooxygenase, and highly purified pulmonary cytochrome P-450IIB-4 from pregnant female rabbits catalyze the NADPH-dependent S-oxygenation of a series of 2-aryl-1,3-dithiolanes. The S-oxide is the only detectable product formed during the short time period of the enzymatic reactions. Studies on the biochemical mechanism for S-oxygenation of 2-aryl-1,3-dithiolanes suggest that this reaction is catalyzed preferentially by the flavin-containing monooxygenase, although cytochromes P-450 also contribute to S-oxygenation. This conclusion is based on the effects of a cytochrome P-450 inhibitor, aminobenzotriazole, as well as on studies of the stereoselectivity of the reaction. Although both purified rabbit pulmonary cytochrome P-450IIB-4 and purified flavin-containing monooxygenase have identical diastereoselectivity, producing the (trans)-S-oxide, these monooxygenases possess opposite S-oxygenation enantioselectivity. Pulmonary cytochrome P-450IIB-4 S-oxygenates 2-aryl-1,3-dithiolanes almost exclusively at the pro-S-sulfur atom, whereas pulmonary flavin-containing monooxygenase S-oxygenates 2-aryl-1,3-dithiolanes exclusively at the pro-R-sulfur atom. 2-Aryl-1,3-dithiolane S-oxides are S-oxygenated a second time on the S'-sulfide sulfur atom but only by rabbit lung microsomes and pulmonary flavin-containing monooxygenase and not by cytochrome P-450IIB-4. That pulmonary flavin-containing monooxygenase only catalyzes formation of (trans)- and not (cis)-2-aryl-1,3-dithiolane S-oxide formation suggests that the active site of pulmonary flavin-containing monooxygenase exerts great steric limitations on 2-aryl-1,3-dithiolane S-oxygenation.
Mol Pharmacol 1990 Feb
PMID:Enantioselective S-oxygenation of 2-aryl-1,3-dithiolanes by rabbit lung enzyme preparations. 230 56

The cytochrome P-450-mediated activation of phenacetin (PHEN) to reactive intermediates by two hypothetical mechanisms has been studied by use of SV 6-31G ab initio energy and spin distribution calculations. In our calculations, the cytochrome P-450 enzyme system has been substituted by a singlet oxygen atom in order to reduce the computational efforts and to fulfill the requirements as to spin conservation. Both mechanisms are based on the currently increasingly accepted view that radical intermediates, formed via sequential one-electron steps, play a crucial role in the metabolic activation of substrates by cytochrome P-450. The first pathway is proposed to involve an initial abstraction of an electron and a proton from the alpha-methylene carbon atom in the ethoxy side chain and can explain the O-deethylation products paracetamol and acetaldehyde. In the second pathway, an initial abstraction of an electron and a proton from the nitrogen atom in the acetylamino side chain is proposed. The calculated spin densities of the formed nitrogen radical indicate that the unpaired electron is primarily localized at the nitrogen atom and to a smaller extent at the ortho- and paracarbon atoms relative to the acetylamino group. Radical recombination reactions between a hydroxyl radical and the spin delocalization-radicalized reactive centers of the nitrogen radical can explain the formation of the metabolites N-hydroxy-PHEN, 2-hydroxy-PHEN, and the arylating metabolite N-acetyl-p-benzoquinone imine (NAPQI), which forms a 3-(S-glutathionyl)paracetamol conjugate in the presence of glutathione. NAPQI is proposed to be formed via intermediate formation of a hemiketal. Proposals are made for the decomposition of this hemiketal into NAPQI that are consistent with currently available experimental data on 14C- and 18O-labeled PHEN.
Mol Pharmacol 1990 Mar
PMID:Mechanisms of activation of phenacetin to reactive metabolites by cytochrome P-450: a theoretical study involving radical intermediates. 231 92

Complementary DNA clones encoding the male-specific rat liver cytochrome P-450 g have been isolated by cross-hybridization with sequences from the female-specific rat liver cytochrome P-450 15 beta. Tissue distribution analysis indicates the liver as the organ with major expression of this cytochrome P-450 gene. Minimal P-450 g expression was also detected in prostate, kidney, heart, and brain. A developmental analysis reveals liver expression in the 8-week-old male and to a lesser extent in the 4-week-old male, but no detectable expression is seen in females of these ages or in 1- and 2-week-old rats from both sexes. Hypophysectomy of female rats dramatically increases hepatic expression of P-450 g, whereas continuous GH administration represses hepatic expression in male or female hypophysectomized rats. In similarity to P-450 15 beta and P-450 16 alpha, therefore, the cytochrome P-450 g gene in liver is GH regulated.
Mol Endocrinol 1990 Jan
PMID:Structural and regulatory analysis of the male-specific rat liver cytochrome P-450 g: repression by continuous growth hormone administration. 232 68

The cytochrome P-450-dependent aromatase pathway utilizes the androgens testosterone (T) and androstenedione, as substrates for estrogen formation. In addition, androgens have been shown to influence the level of aromatase activity in various tissues. In cultured human skin fibroblasts, incubation with T for 14 h resulted in a dose-dependent decline in aromatase activity, the concentration of T producing a half-maximal decline being 6 nM. In the presence of T (50 nM), aromatase activity declined in a time-dependent fashion with maximal reduction occurring by 9 h. When aromatase kinetics were determined after preincubation of cells with T, there was a significant decline in the calculated Vmax with no significant change in the apparent Km, suggesting that incubation of cells with T reduced the number of active enzyme sites. Aromatase activity was unaffected by preincubation of cells with the synthetic androgen methyltrienolone. In addition, the decline in aromatase activity following preincubation with T was observed in cells derived from patients with complete androgen insensitivity demonstrating that the effect of T was not mediated by androgen receptors. Furthermore, new protein synthesis was not necessary for the T-mediated effect as the presence of cycloheximide (50 micrograms/ml) did not prevent it. When cells were incubated at low oxygen tension, the inhibition of aromatase activity by T was diminished. Testosterone is rapidly metabolized in genital skin fibroblasts to dihydrotestosterone, androstanedione, androsterone, 3 alpha-androstanediol, 3 beta-androstanediol and estradiol. To determine if a metabolite of T might be responsible for the repression of aromatase activity, aromatase activity was determined in cells following preincubation with various metabolites of T. Preincubation of cells with androstenedione, androstanedione or 3 alpha-androstanediol produced a small but significant decline in aromatase activity, whereas preincubation of cells with dihydrotestosterone, androsterone, or 3 beta-androstanediol did not have a significant effect. Aromatase activity was also unaffected by preincubation of cells with estradiol or diethylstilbestrol. When aromatase activity was assayed in microsomal preparations from cells preincubated with T, activity was reduced. Although cells preincubated with 50 nM [3H]T contained between 0.25 and 0.51 pmol of residual steroid/mg microsomal protein, the amount of [1-3H]androstenedione and T was insufficient to account for the observed decline in aromatase activity on the basis of competitive inhibition.(ABSTRACT TRUNCATED AT 400 WORDS)
Mol Cell Endocrinol 1990 Mar 05
PMID:Testosterone lowers aromatase activity in cultured human genital skin fibroblasts. 232 27

The purpose of this study was to evaluate the hypothesis that NADPH supply in intact cells is regulated by oxygen tension. This was accomplished by studying monooxygenation in perfused livers from Ah locus-responsive C57BL/6J mice, where rates of monooxygenation are high. Elevation of flow rate decreases the hepatic O2 gradient and increases O2 delivery to the organ. Under these conditions, rates of p-nitroanisole O-demethylation were 2-3 times higher in perfused livers from fed or fasted mice at high (10 ml/min) compared with normal (5 ml/min) flow rates. Rates of monooxygenation were directly proportional to oxygen tension (half-maximal rates occurred with approximately 400 microM O2). On the other hand, rates were independent of oxygen concentration in isolated microsomes where NADPH was supplied in excess. The decrease in rate due to diminished O2 concentration in the intact organ could not be attributed to hypoxia, because O2 tension in the effluent perfusate exceeded 50 microM even when influent perfusate was saturated with 25% O2 and ATP/ADP ratios were in the normal range. Thus, monooxygenation of p-nitroanisole in perfused mouse liver is dependent on oxygen tension. Similarly, glucuronidation of p-nitrophenol was oxygen dependent in the intact organ but not in isolated microsomes supplemented with UDP-glucuronic acid. Taken together, these data support the hypothesis that, at high oxygen tensions (e.g., in periportal regions of the liver lobule), mitochondrial activity is increased, which in turn enhances NADPH and UDP-glucuronic acid turnover, leading to accelerated rates of monooxygenation and glucuronidation in intact cells. In support of this idea, NH4Cl, which utilizes NADPH for urea synthesis, inhibited monooxygenation in the perfused mouse liver at high but not low flow rates. Thus, important phase I and II detoxification reactions are regulated indirectly by the hepatic oxygen gradient, via mechanisms involving cofactor supply, when cytochrome P-450 is not limiting.
Mol Pharmacol 1990 Jul
PMID:Unique role of oxygen in regulation of hepatic monooxygenation and glucuronidation. 237 Aug 51

The effects of growth hormone (GH) on two constitutively expressed rat liver cytochrome P-450 (P-450) enzymes, namely P-450f (IIC7) and P-450PB1 (IIC6), have been investigated. Hypophysectomy of both male and female rats results in low expression of P-450f. Single daily injections of GH cannot restore expression of P-450f but further repress it. Continuous administration of GH, on the other hand, increases expression of P-450f to levels comparable to those of the normal male and female rat. P-450PB1 is minimally affected by hypophysectomy or GH treatment, although a weak but significant repression by continuous administration of GH can be detected. Run-on analysis shows that continuous GH treatment increases transcription of P-450f. In addition, the sequence of the 5' flanking region of this gene reveals DNA segments that might have a putative role in the transcriptional regulation of P-450f by GH.
Mol Pharmacol 1990 Aug
PMID:Growth hormone regulation of the cytochrome P-450IIC subfamily in the rat: inductive, repressive, and transcriptional effects on P-450f (IIC7) and P-450PB1 (IIC6) gene expression. 238 31

In mouse hepatoma cells, the environmental contaminant 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) increases the transcription rate of the CYP1A1 gene, which encodes a cytochrome P-450 enzyme. In this study, we analyzed the DNA region immediately upstream of the CYP1A1 gene. A domain that extends upstream to nucleotide--166 was found to function as a transcriptional promoter. The promoter was silent when uncoupled from the dioxin-responsive enhancer located farther upstream. DNase footprinting experiments indicated that nuclear proteins interact with distinct domains of the promoter in a TCDD-independent fashion. Mutational analyses indicated that the CYP1A1 promoter contains at least three functional domains, including a TATAAA sequence, a CCAAT box transcription factor/nuclear factor I-like recognition motif, and a guanine-rich G box.
Mol Cell Biol 1990 Oct
PMID:Functional analysis of the transcriptional promoter for the CYP1A1 gene. 239 86

Various 4-alkyl analogues of 3,5-diethoxycarbonyl-1,4-dihydro-2,4,6-trimethylpyridine (DDC) cause mechanism-based inactivation of cytochrome P-450 (P-450) via heme destruction. We have examined the time course of effects of DDC analogues on the catalytic activities and apoproteins of the major beta-naphthoflavone-, dexamethasone-, and phenobarbital-inducible isozymes of rat liver P-450 following in vivo administration. In beta-naphthoflavone-treated rats, all DDC analogues examined caused loss of the P-450 chromophore and dramatic loss of 7-ethoxyresorufin O-deethylase activity, a catalytic marker for P-450c. The isopropyl, hexyl, and isobutyl analogues caused the most pronounced loss/alteration of P-450c apoprotein levels, as revealed by two monoclonal antibodies (MAbs), 1-31-2 and 1-7-1. The apoprotein of P-450d was not altered. In dexamethasone-treated rats, all analogues except 4-hexyl-DDC caused loss of the P-450 chromophore and erythromycin N-demethylase activity, a catalytic marker for P-450p-related isozymes. Only 4-isopropyl-DDC caused significant loss/alteration of the apoprotein of P-450p-related forms, as revealed by MAb 2-13-1. In phenobarbital-treated rats, all analogues reduced the level of the P-450 chromophore, whereas only 4-hexyl-DDC and 4-isopropyl-DDC lowered 7-pentoxyresorufin O-dealkylase activity, a catalytic marker for P-450b. MAbs 2-66-3 and 2-8-1 revealed no change in the level of phenobarbital-inducible apoproteins recognized by these probes. In agreement with our previous in vitro studies [Mol. Pharmacol. 35;626-634 (1989)], P-450 c and p are targets for mechanism-based inactivation by DDC analogues. However, unlike the situation in vitro, loss of enzyme activity in vivo is, at least in some instances, accompanied by loss/alteration of the corresponding P-450 apoprotein.
Mol Pharmacol 1990 Jan
PMID:Effects of 4-alkyl analogues of 3,5-diethoxycarbonyl-1,4-dihydro-2,4,6-trimethylpyridine on hepatic cytochrome P-450 heme, apoproteins, and catalytic activities following in vivo administration to rats. 240 48

Monoclonal antibodies specific for cytochromes P-450 induced by 3-methylcholanthrene (Mab 1-7-1) and phenobarbital (Mab 2-66-3) have been used in an unlabeled peroxidase-antiperoxidase immunohistochemical procedure to investigate the intralobular distribution and induction sites of the hemoproteins within the livers of CD-1, C57BL/6, and DBA/2 mice. 3-Methylcholanthrene-specific cytochromes P-450 were localized predominantly in centrilobular hepatocytes of control mice from all strains and were present at higher levels in CD-1 and C57BL/6 mice than in DBA/2 mice. Treatment with either 3-methylcholanthrene or beta-naphthoflavone produced striking increases of 3-methylcholanthrene-specific cytochromes P-450 in hepatocytes from all regions of the hepatic lobule in CD-1 and C57BL/6 mice, but not in DBA/2 mice. Phenobarbital-specific cytochromes P-450 were localized in hepatocytes throughout all segments of the lobule in control mice, with slightly greater hemoprotein content in centrilobular hepatocytes. Treatment with phenobarbital resulted in enhancement of cytochrome P-450 that was visualized in hepatocytes in all regions of the lobule. Strain-related differences were not observed for phenobarbital-specific cytochromes P-450. These results demonstrate that constitutive levels of 3-methylcholanthrene- and phenobarbital-specific cytochromes P-450 are localized predominantly in centrilobular hepatocytes of murine livers, and induction of the hemoproteins is manifested to the greatest extent in periportal hepatocytes, resulting in a more uniform distribution throughout the hepatic lobule.
Mol Pharmacol 1988 Dec
PMID:Distribution and induction sites of phenobarbital- and 3-methylcholanthrene-inducible cytochromes P-450 in murine liver: immunohistochemical localization with monoclonal antibodies. 246 60

Vasoactive intestinal peptide (VIP), a neuropeptide present in ovarian nerves, has been previously shown to induce synthesis of the side-chain cleavage cytochrome P-450 enzyme which catalyzes the conversion of cholesterol to pregnenolone (the rate-limiting step in progesterone synthesis). In the present study we demonstrate, by means of a bovine 3'-specific P-450scc cDNA probe, that this VIP effect is exerted at least partially at the level of gene expression in cultured granulosa cells that were isolated from estrogen-primed, immature rats. The size and level of the 2.0 kilobase P-450scc mRNA species was assessed by Northern blot analysis, while the translatability of this mRNA was assayed by immunoisolation of the 35S-labeled P-450scc precursor protein translated from total RNA of control and stimulated granulosa cells. FSH was much more effective than VIP at increasing P-450scc mRNA concentrations in cultured granulosa cells, whereas secretin treatment was ineffective. The results suggest that, like FSH, the stimulatory effect of the neuropeptide VIP on ovarian progesterone secretion involves regulation of P-450scc gene expression during functional maturation of the prepubertal ovary.
Mol Endocrinol 1987 Jul
PMID:Vasoactive intestinal peptide regulates cholesterol side-chain cleavage cytochrome P-450 (P-450scc) gene expression in granulosa cells from immature rat ovaries. 248 21


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