Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UNIPROT:P06889 (Mol)
 630,302 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Homologous 1-naphtoxyalcanthiols of the type 1-C10H7O(CH2)nSH (n = 2-7) are used for structural studies of the microsomal cytochrome P450 active centre. It was found that the strongest complex of thiol with P450 is formed for n = 3. Microsomal oxidation of P450 substrates aminopyrine and benz(a) pyrene is inhibited by the 1-naphtoxyalcanthiols studied. A non-monotonous dependence of pI50 on n was found, the compound with a chain length n = 3 appeared to be the most effective inhibitor. The interaction of this thiol (n = 3) with both the heme group of P450 and the hydrophobic substrate zone is supposed and the distance between these points was estimated. It is possible to employ this approach for structural studies on the active centers of different isoforms of P450.
Mol Biol (Mosk)
PMID:[The study of the structure of active center of membrane-bound cytochrome p-450 using 1-naphthoxyalcanthiols]. 318 39

Using highly sensitive probe-substrate analyses, investigations of drug biotransformation in tissues of the rat conceptus during an early stage of organogenesis revealed that three separate tissue components each contained P-450 isozymes capable of catalyzing the monooxygenation of foreign organic chemicals. Tissues of the embryo proper contained constitutive P450(s) that catalyzed readily measurable O-depentylation and O-debenzylation of pentoxyphenoxazone and benzyloxyphenoxazone, respectively, but no measurable O-demethylation of methoxyphenoxazone and barely detectable O-deethylation of ethoxyphenoxazone. Higher specific activities for the O-depentylation and O-debenzylation reactions were measured in preparations of the yolk sac and this organ also appeared to contain constitutive P450(s) for the readily detectable O-deethylation of ethoxyphenoxazone. The O-demethylation of methoxyphenoxazone could not be detected in the yolk sac. Only the O-debenzylation reaction could be detected in tissues of the ectoplacental cone. Treatment of conceptuses in utero with 3-methycholantherene (MC) resulted in significantly increased rates of O-deethylation reactions in preparations of yolk sac and embryo but not ectoplacental cone. Demethylation was not detectable in the same preparations. Treatment with phenobarbital, pregnenolone-16 alpha-carbonitrile, or isosafrole produced no observable effect on any of the reactions studied. Carbon monoxide (CO:O2 = 80:20 versus N2:O2 = 80:20) markedly inhibited all reaction rates and inhibition could be reversed by replacement of CO with N2. Deethylation and debenzylation were inhibited by anti-P450IA1 IgG after MC induction but were not affected by the same IgG fraction in untreated conceptuses. Depentylation reactions were not inhibited by anti-P450IA1 or anti-P450IIB1/2 antibodies under any of the conditions used. Deethylation was strongly inhibited by 1.0 microM 7,8-benzoflavone in tissues from MC-treated but not untreated conceptus. Metyrapone (0.1 mM) failed to significantly inhibit any of the measurable conceptus-catalyzed depentylation reaction. The results indicated the presence of four (or more) functional P450 isozymes in tissues of the conceptus during organogenesis, a constitutive depentylase(s) in the yolk sac and embryo, a constitutive deethylase(s) present in the yolk sac, an MC-inducible deethylase(s) in the embryo and yolk sac, and constitutive debenzylase(s) present in all three tissues. No O-demethylation was detectable in any of the three tissues, even after in utero exposure to inducers.(ABSTRACT TRUNCATED AT 400 WORDS)
Mol Pharmacol 1988 Jul
PMID:Cytochrome P-450-dependent biotransformation of a series of phenoxazone ethers in the rat conceptus during early organogenesis: evidence for multiple P-450 isoenzymes. 339 41

The crystal structure of Pseudomonas putida cytochrome P450cam with its substrate, camphor, bound has been refined to R = 0.19 at a normal resolution of 1.63 A. While the 1.63 A model confirms our initial analysis based on the 2.6 A model, the higher resolution structure has revealed important new details. These include a more precise assignment of sequence to secondary structure, the identification of three cis-proline residues, and a more detailed picture of substrate-protein interactions. In addition, 204 ordered solvent molecules have been found, one of which appears to be a cation. The cation stabilizes an unfavorable polypeptide conformation involved in forming part of the active site pocket, suggesting that the cation may be the metal ion binding site associated with the well-known ability of metal ions to enhance formation of the enzyme-substrate complex. Another unusual polypeptide conformation forms the proposed oxygen-binding pocket. A localized distortion and widening of the distal helix provides a pocket for molecular oxygen. An intricate system of side-chain to backbone hydrogen bonds aids in stabilizing the required local disruption in helical geometry. Sequence homologies strongly suggest a common oxygen-binding pocket in all P450 species. Further sequence comparisons between P450 species indicate common three-dimensional structures with changes focused in a region of the molecule postulated to be associated with the control of substrate specificity.
J Mol Biol 1987 Jun 05
PMID:High-resolution crystal structure of cytochrome P450cam. 365 28

Magnetic circular dichroism spectra (MCD) of reduced cytochromes P450 and P420 from rabbit liver microsomes have been recorded and analyzed for the 350-600 nm spectral region in the temperature interval from 2 to 290 K. The shape, intensity and temperature dependence of the MCD of reduced P450 in the Soret region are quite different from that of other high-spin ferrous hemoproteins, whose heme iron is coordinated to the imidazole of histidine (deoxymyoglobin, deoxyhemoglobin, reduced peroxidase and cytochrome c oxidase). Assuming that in the reduced P450 as well as in its CO-complex the protein-derived ligand is mercaptide (RS-) the differences can be explained by the existence of two electronic transitions in the Soret region: the common for hemoproteins pi----pi porphyrin transition and sulfur to porphyrin charge-transfer transition, p+(Sp)----eg (pi). The unusual spectral characteristics of the CO-complex of P450 have been ascribed earlier to strong configurational interaction of these two transitions. From the similarities of the Soret MCD and their temperature dependences for the reduced P420 and for other high-spin ferrous hemoproteins one can conclude that heme iron of the reduced P420 is high-spin and is coordinated to the imidazole of histidine. The zero-field splitting parameter D of the spin Hamiltonian has been estimated from the MCD temperature dependences. The obtained splitting of approximately 30 cm-1 for P450 and of approximately 10 cm-1 for P420 exceeds that for myoglobin and hemoglobin (approximately 5 cm-1).
Mol Biol (Mosk)
PMID:[Electron structure of the heme of reduced cytochrome P450 and P420 from the data of low-temperature magnetic circular dichroism]. 370 68

Magnetic circular dichroism (MCD) spectra of reduced cytochromes P450 and P420 in equilibrium and non-equilibrium protein conformations are compared at 4.2 K for the 350-800 spectral region. Non-equilibrium forms have been produced by photolysis of CO-complexes at 4.2 K. The differences between MCD spectra of proteins in equilibrium and non-equilibrium conformations, in particular for the visible region, show clearly the structural changes in the heme iron coordination sphere to occur on ligand binding. The comparison of the Soret MCD spectra of reduced proteins in their equilibrium and non-equilibrium forms with those of other high-spin ferrous hemoproteins suggest that mercaptide (RS-) is the protein ligand of the heme iron in reduced P450, as well as in its CO-complex, and that imidazole of histidine is the fifth ligand of the iron both in reduced P420 and its CO-complex. The thermal recombination of the photoproducts with CO have been studied. When temperature rises from 4.2 to 77 K for two hours both proteins have similar temperature characteristics during the recombination processes. The recombination begins at T approximately equal to 10 K and is completed at approximately equal to 50 K. The temperature at which half of the total photolyzed molecules are restored to the CO-form is equal to 25 K. For products of photolysis of CO-complexes of myoglobin and hemoglobin under the same heating conditions these temperatures are equal to 35 and 23 K respectively. Thus, the photoproducts of P450, P420 and hemoglobin have similar parameters of low-temperature recombination and the kinetics of this process is faster than for photodissociated myoglobin.
Mol Biol (Mosk)
PMID:[Comparison of the heme electron state of reduced cytochrome P450 and P420 in equilibrium and non-equilibrium protein conformations. The nature of the protein heme ligand]. 370 69

Cytochrome P450 was partially purified from rat breast tissue from 1-, 2-, 3-, 6-, 9-, and 15-week-old pregnant, lactating, or 3-week postlactation rats. The detergent-solubilized P450 was spectrally quantified, and the P450 isozyme pattern in the different samples was characterized by Western blot analysis with antibodies against cytochromes P450 1A1, 1A2, 2A, 2B, 2D4, 3A, 4A, 2E, and 19. The yield of P450 was 5-60 pmol/g wet weight tissue, with the highest yields in 1- and 2-week-old pups and lactating rats. The cytochromes P450 expressed in the breast can be divided into six main groups on the basis of their pattern of regulation: (a) those present in all samples (4A, 2E1, and 2D4), (b) those highly expressed in 1- to 3-week-old rats (2D4 and 3A), (c) those expressed only after 9 weeks of age [P450 19 (aromatase)], (d) those induced in pregnancy and maintained during lactation (1A1), (e) those induced in pregnancy and not maintained during lactation (2A), and (f) those induced 3 weeks after lactation (2B, 2A, and 3A). Reverse transcription-polymerase reaction amplification was used to confirm the presence of P450 isoforms in the breast. The mRNAs of cytochromes P450 1A1, 2A1, 2B1-3, 2D1, 2D3, 2D4, 2E1, and 4A3 were detected on analysis of total breast RNA. The mRNA of CYP 3A1 was not convincingly detected in untreated rat breast but was inducible by treatment with pregnenolone-16-alpha-carbonitrile. The presence of these various forms of P450 in the breast and their regulation by age and endocrine status may have implications for in situ metabolism of steroids and steroid antagonists and for activation of procarcinogens.
Mol Pharmacol 1995 Oct
PMID:Developmental and endocrine regulation of P450 isoforms in rat breast. 747 88

Growth hormone (GH) secretory patterns regulate the expression of several sex-dependent liver cytochrome P450 (CYP) genes. Studies using the hypophysectomized rat model have established that the intermittent plasma GH secretory pattern associated with adult male rats markedly stimulates liver expression of the male-specific CYP 2C11, a testosterone 2 alpha- and 16 alpha-hydroxylase, but is not required for expression of other male-specific liver enzymes, including CYP 2A2, a testosterone 15 alpha-hydroxylase, and CYP 3A2, a testosterone 6 beta-hydroxylase. In the present study, the effects of intermittent GH treatment on liver CYP expression were studied in adult rats rendered GH deficient by neonatal administration of monosodium glutamate (MSG), which depletes circulating adult GH without the global loss of other pituitary-dependent hormones that is associated with hypophysectomy. Restoration of the normal masculine circulating GH profile of six daily pulses (180-225 ng GH/ml/peak) in MSG-treated male rats by the use of an external pumping apparatus led to a substantial (30-50%) restoration of normal male levels of CYP 2A2 and CYP 3A2 activity, protein, and mRNA. GH pulsation at the nonphysiological frequencies of two or four times per day was less effective unless given at a dose that resulted in supraphysiological plasma GH levels. Although intermittent GH treatment can induce male-specific P450 expression in hypophysectomized female rats, the same hormone treatment did not stimulate CYP 2A2 or CYP 3A2 expression in MSG-treated female rats. Liver GH receptor mRNA levels at adulthood were not significantly altered by neonatal MSG treatment, suggesting that the unresponsiveness of MSG-treated females and the previously reported low responsiveness of MSG-treated males to GH-induced CYP 2C11 expression are not due to the absence of GH receptor. Moreover, normal liver IGF-1 mRNA levels were expressed in the MSG-treated female rats, suggesting that the liver GH receptor is functional in these animals. The present findings establish that the adult male-specific enzymes CYP 2A2 and CYP 3A2 can be positively regulated by intermittent GH pulsation despite their GH-independent expression in hypophysectomized rats. Moreover, neonatal MSG treatment, particularly in female rats, may lead to the loss of factors other than GH that are required for full expression of the pulsatile GH-stimulated CYP 2A2, 3A2, and 2C11 genes.
Mol Pharmacol 1995 Nov
PMID:Growth hormone regulation of male-specific rat liver P450s 2A2 and 3A2: induction by intermittent growth hormone pulses in male but not female rats rendered growth hormone deficient by neonatal monosodium glutamate. 747 8

Adrenal mitochondria metabolize cholesterol at inner membrane (IM) cytochrome P450scc. Exogenous and outer membrane (OM) cholesterol are metabolized more slowly due to a limiting transfer of cholesterol from OM to IM. This process is stimulated by in vivo ACTH treatment and inhibited by cycloheximide (CX)-induced depletion of labile regulatory proteins. In isolated rat adrenal mitochondria, GTP enhances the metabolism of exogenous cholesterol, consistent with enhanced intermembrane cholesterol transfer (Xu et al. (1989) J. Biol Chem. 264, 17674), but metabolism of 20 alpha-hydroxycholesterol, which readily traverses mitochondrial membranes, is not affected. The non-hydrolyzable analog, GTP gamma S, completely inhibits the activation of cholesterol metabolism by GTP, suggesting a requirement for GTP hydrolysis. Low concentrations of Ca2+ (0.4-4 microM) stimulate two independent cholesterol transport processes. For exogenous cholesterol, a Ca(2+)-mediated process can replace GTP since each produces comparable stimulation and the combination produces little additional activity. This Ca2+ stimulation is insensitive to GTP gamma S and also to Ruthenium Red (RR), which prevents Ca2+ entry into the matrix. Ca2+ also enhances availability to P450 scc of endogenous OM cholesterol, which accumulates during in vivo CX-inhibition. This stimulation is, however, distinguished by insensitivity to GTP and complete inhibition by RR. Ca2+, therefore, enhances intermembrane transfer of exogenous cholesterol from OM without entry into the matrix through a process which is independently stimulated by GTP. Ca2+ induces transfer of endogenous OM cholesterol through a completely different mechanism involving RR-inhibited matrix changes.(ABSTRACT TRUNCATED AT 250 WORDS)
Mol Cell Endocrinol 1995 Feb
PMID:Metabolism of exogenous cholesterol by rat adrenal mitochondria is stimulated equally by physiological levels of free Ca2+ and by GTP. 753 86

The carboxyl-terminal 28 amino acids of rabbit cytochrome P450 2C2 are markedly different from those of other rabbit cytochrome P450 2C family members and, substitution of the equivalent amino acids of other cytochrome P450s can confer novel steroid hydroxylase activity to P450 2C2 while the normal lauric acid hydroxylase activity is retained. To determine the basis for the novel steroid hydroxylase activity, amino acids of cytochrome P450 2C1 were substituted for those of cytochrome P450 2C2 and the mutants were expressed in COS-1 cells. There are 13 differences between the sequences of cytochrome P450 2C2 and P450 2C1 in this region, including five nonconservative exchanges of charged and uncharged amino acids. However, only substitution of valine for Ser-473 increased steroid hydroxylase activity to the maximum level expected in a modified cytochrome P450 2C2, which contained additional substitutions in the 368-388 region to maximize progesterone hydroxylase activity. Introduction of this single substitution into cytochrome P450 2C2 resulted in 21-progesterone hydroxylase activity similar to that resulting from substitution of all 28 carboxyl-terminal cytochrome P450 2C1 amino acids. None of the substitutions, with one exception, substantially affected either lauric acid hydroxylase activity or the amount of immunologically reactive cytochrome P450 that was expressed. A glycine substitution for Val-477 reduced activity of both lauric acid hydroxylase and progesterone hydroxylase and altered the regioselectivity of the hydroxylation for both. Homology modeling of cytochrome P450 2C2, based on the cytochrome bacterial P450cam sequence, indicated that the side chains of residue 473 and the other five residues previously shown to affect substrate specificity face the substrate pocket. For four of the six residues, smaller and more hydrophobic residues increased progesterone relative to lauric acid hydroxylation.
Mol Pharmacol 1995 Sep
PMID:Substitution at residue 473 confers progesterone 21-hydroxylase activity to cytochrome P450 2C2. 756 21

The Cyp 2d-9 gene encodes the male-specific steroid 16 alpha-hydroxylase in mouse liver and shares a conserved regulatory element (-100TTCCGGGC-93) with another male-specific Slp promoter. As shown with the Slp promoter (N. Yokomori, R. Moore, and M. Negishi, Proc. Natl. Acad. Sci. USA 92:1302-1306, 1995), the male-preferential demethylation also occurs at CpG/-97 in the Cyp 2d-9 promoter. The transcription factor which specifically binds to the demethylated element has been purified. The peptide sequences reveal that the factor consists of GABP alpha and GABP beta 1 with Ets and Notch motifs, respectively. Both DNase I footprinting and gel shift assays indicate that the bacterially expressed glutathione S-transferase-GABP fusion proteins bind to the regulatory element only when CpG/-97 is demethylated. Moreover, Cyp 2d-9 promoter is trans-activated by coexpression of GABP proteins in HepG2 cells. Given the additional results that CpG/-50 of the female-specific steroid 15 alpha-hydroxylase (Cyp 2a-4) promoter is preferentially demethylated in the females, the sex-specific expressions of the P450 genes correlate very well with DNA demethylation. We also conclude that GABP is a methylation-sensitive transcription factor and is a potential transcription activator of the male-specific Cyp 2d-9 promoter.
Mol Cell Biol 1995 Oct
PMID:A DNA methylation site in the male-specific P450 (Cyp 2d-9) promoter and binding of the heteromeric transcription factor GABP. 756 85


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