TY - JOUR. T1 - Determination of the redox properties of human NADPH-cytochrome P450 reductase. AU - Munro, Andrew W. AU - Noble, Michael A. AU - Robledo, Laura AU - Daff, Simon N. AU - Chapman, Stephen K. PY - 2001/2/20. Y1 - 2001/2/20. N2 - Midpoint reduction potentials for the flavin cofactors in human NADPH-cytochrome P450 oxidoreductase were determined by anaerobic redox titration of the diflavin (FAD and FMN) enzyme and by separate titrations of its isolated FAD/NADPH and FMN domains. Flavin reduction potentials are similar in the isolated domains (FAD domain El [oxidized/semiquinone] = -286 +/- 6 mV, E-2 [semiquinone/ reduced] = -371 +/- 7 mV; FMN domain E-1 = -43 +/- 7 mV, E-2 = -280 +/- 8 mV) and the soluble diflavin reductase (E-1 [FMN] = -66 +/- 8 mV, E-2 [FMN] = -269 +/- 10 mV; E1 [FAD] = -283 +/- 5 mV, E2 [FAD] = -382 +/- 8 mV). The lack of perturbation of the individual flavin potentials in the FAD and FMN domains indicates that the flavins are located in discrete environments and ...

Diflavin reductases are essential proteins capable of splitting the two-electron flux from reduced pyridine nucleotides to a variety of one electron acceptors. The primary sequence of diflavin reductases shows a conserved domain organization harboring two catalytic domains bound to the FAD and FMN flavins sandwiched by one or several non-catalytic domains. The catalytic domains are analogous to existing globular proteins: the FMN domain is analogous to flavodoxins while the FAD domain resembles ferredoxin reductases. The first structural determination of one member of the diflavin reductases family raised some questions about the architecture of the enzyme during catalysis: both FMN and FAD were in perfect position for interflavin transfers but the steric hindrance of the FAD domain rapidly prompted more complex hypotheses on the possible mechanisms for the electron transfer from FMN to external acceptors. Hypotheses of domain reorganization during catalysis in the context of the different members of

TY - JOUR. T1 - Possible association of NADPH-cytochrome P-450 reductase and cytochrome P-450 in reconstituted phospholipid vesicles. AU - Nisimoto, Yukio. AU - Kinosita, Kazuhiko. AU - Ikegami, Akira. AU - Kawai, Norio. AU - Ichihara, Ichiro. AU - Shibata, Yukio. PY - 1983. Y1 - 1983. N2 - A fluorescent probe, N-(1-anilinonaphth-4-yl)-maleimide (ANM), was specifically labeled to SH group(s) in the hydrophilic moiety of NADPH-cytochrome P-450 reductase at a ratio of 1 ± 0.1 ANM/mol of protein. The ANM-labeled reductase and P-450 were reconstituted in phosphatidylcholine-phosphatidylethanolamine-phosphatidylserine vesicles in which all of the enzymes were functionally active. The reconstitution of the mixed-function oxidase system was found to be strongly dependent on both the lipid to protein molar ratio and phospholipid composition. The interactions of ANM-labeled reductase with P-450 in proteoliposomes were investigated by perturbation of the fluorescence of ANM. Upon incorporation of P-450 ...

Cytochrome P450 enzymes (P450s or CYPs) catalyze an enormous variety of oxidative reactions in organisms from all major domains of life. Their monooxygenase activity relies on the reductive scission of molecular oxygen (O2) bound to P450 heme iron, and thus on the delivery of two electrons to the heme iron at discrete points in the catalytic cycle. Early studies suggested that P450 redox partner machinery fell into only two major classes: either the eukaryotic diflavin enzyme NADPH-cytochrome P450 oxidoreductase, or bacterial/mitochondrial NAD(P)H-ferredoxin reductase and ferredoxin partners. However, more recent studies, aided by genome sequence data, reveal a much more complex scenario. Several new types of P450 redox partner systems have now been characterized, including P450s naturally linked to their redox partners, or to a component protein of their P450 electron delivery system. Other P450s have evolved to bypass requirements for redox partners, and instead react directly with hydrogen ...

The crystal structure of NADPH-cytochrome P450 reductase (CYPOR) implies that a large domain movement is essential for electron transfer from NADPH via FAD and FMN to its redox partners. To test this hypothesis, a disulfide bond was engineered between residues Asp(147) and Arg(514) in the FMN and FAD domains, respectively. The cross-linked form of this mutant protein, designated 147CC514, exhibited a significant decrease in the rate of interflavin electron transfer and large (,/=90%) decreases in rates of electron transfer to its redox partners, cytochrome c and cytochrome P450 2B4. Reduction of the disulfide bond restored the ability of the mutant to reduce its redox partners, demonstrating that a conformational change is essential for CYPOR function. The crystal structures of the mutant without and with NADP(+) revealed that the two flavin domains are joined by a disulfide linkage and that the relative orientations of the two flavin rings are twisted approximately 20 degrees compared with the ...

Cytochromes P450 (P450s) belong to a superfamily of heme-containing monooxygenases and are the predominant enzyme responsible for phase I metabolism of clinically relevant drugs (Wang and Tompkins, 2008). Through the incorporation of a single oxygen atom, P450s generate products that are more water-soluble and are either readily excreted in the urine or more amenable substrates for phase II conjugation. Previous studies have demonstrated that many of these enzymes are highly flexible (Domanski and Halpert, 2001; Zhao and Halpert, 2006), allowing them to accommodate a wide range of substrates, including numerous steroids, pharmaceuticals, and environmental pollutants (Johnson and Stout, 2005).. P450 2B enzymes were among the first mammalian P450s to be purified and cloned and have served as a prototype for biochemical and biophysical experiments, as well as studies of substrate specificity and of interactions with the redox partners NADPH-cytochrome P450 reductase and cytochrome b5 (Zhao and ...

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TY - GEN. T1 - Antley-Bixler Syndrome-Related Mutants of Human NADPH-Cytochrome P450 Reductase: Effects on Drug Metabolism and Mutagenesis. AU - Duarte, Maria Paula Amaro de Castilho. AU - Kranendonk, Michel. AU - Rueff, José. PY - 2007/1/1. Y1 - 2007/1/1. KW - Antley-Bixler Syndrome. KW - Cytochrome P450 Reductase. M3 - Conference contribution. VL - na. SP - 78. BT - NA. Y2 - 1 January 2007. ER - ...

Steroids are the most widely marketed products by the pharmaceutical industry after antibiotics. Steroid hydroxylation is one of the most important functionalizations because their derivatives enable a higher biological activity compared to their less polar non-hydroxylated analogs. Bacterial cytochrome P450s constitute promising biocatalysts for steroid hydroxylation due to their high expression level in common workhorses like Escherichia coli. However, they often suffer from wrong or insufficient regio- and/or stereoselectivity, low activity, narrow substrate range as well as insufficient thermostability, which hampers their industrial application. Fortunately, these problems can be generally solved by protein engineering based on directed evolution and rational design. In this work, an overview of recent developments on the engineering of bacterial cytochrome P450s for steroid hydroxylation is presented.

In this study we present evidence that brain cells consume NO by a membrane-localized process that involves NADPH oxidation by CYPOR, a microsomal protein that transfers electrons from NADPH to an acceptor, classically a haem-containing cytochrome P450 [27]. Cytochrome P450 inhibitors decreased NO consumption by brain membranes, suggesting that reduction of these proteins by CYPOR underlies NO consumption.. Cytochrome P450s perform hydroxylation reactions which, in the brain, are involved in diverse functions including steroid hormone synthesis, cholesterol homoeostasis and vitamin, eicosanoid and xenobiotic metabolism [28,29]. Both CYPOR and several members of the cytochrome P450 family are expressed in brain, though at much lower levels than in the liver (1-10%; [30]). Interestingly, NO has been shown to bind and inhibit several cytochrome P450s [31,32], making them intriguing candidates for NO consumption by brain membranes. Indeed, the reductase domain of NOS is very similar to that of CYPOR ...

TY - JOUR. T1 - Synthetic anticonvulsants, antihypoxics, and liver monooxygenase system inducers based on amides and urea. XI. Synthesis of alkyl- and arylalkylureas and their effects on the liver monooxygenase system. AU - Bakibaev, S. S.. AU - Akhmedzhanov, R. R.. AU - Filimonov, V. D.. AU - Novocheeva, T. P.. AU - Saratikov, A. S.. AU - Tignibidina, L. G.. AU - Pustovoitov, A. V.. PY - 1993/9. Y1 - 1993/9. UR - http://www.scopus.com/inward/record.url?scp=14744281917&partnerID=8YFLogxK. UR - http://www.scopus.com/inward/citedby.url?scp=14744281917&partnerID=8YFLogxK. U2 - 10.1007/BF00780583. DO - 10.1007/BF00780583. M3 - Article. AN - SCOPUS:14744281917. VL - 27. SP - 631. EP - 634. JO - Pharmaceutical Chemistry Journal. JF - Pharmaceutical Chemistry Journal. SN - 0091-150X. IS - 9. ER - ...

A number of studies, including the original from Arlt et al. (2004) detailing the discovery of the exon 5 POR 541T→G mutation, have shown the catalytic incompetence of Y181D-substituted CYPOR using various assays and means of recombinant protein expression (Huang et al., 2005; Wang et al., 2007; Agrawal et al., 2008). Before the discovery of POR deficiency as a cause of human disease, Shen et al. (1989) showed that the orthologous Y178D variant of rat CYPOR, recombinantly expressed and purified, was unable to catalyze NCR activity because of loss of FMN. Since their report, the crystal structure of the rat CYPOR has shown the role of Y178 (corresponding to human Y181) as a stacking residue in the binding of FMN (Wang et al., 1997). Here, both purified and membrane-associated Y181D models were used to confirm and analyze, in more detail, compromised FMN binding as the specific molecular defect leading to multiple microsomal P450 deficiencies.. Data reported here show that FMN binding by human ...

Joyce, M.G., Ekanem, I.S., Roitel, O., Dunford, A.J., Neeli, R., Girvan, H.M., Baker, G.J., Curtis, R.A., Munro, A.W. and Leys, D. (2012) The crystal structure of the FAD/NADPH-binding domain of flavocytochrome P450 BM3. FEBS J. 279, 1694-1706 ...

Carcinogen-induced lesions were identified by use of the known markers of hepatocarcinogenesis adenosintriphosphatase and ã-glutamyl transpeptidase. While the GSTs and mEHb were increased in all preneoplastic and neoplastic lesions, the levels of the individual cyt. P-450 isoenzymes were characteristically different from each other. In many of the early ATPase deficient islets PB1 was elevated, whereas the content of the other cyt P-450 forms and NADPH-cytochrome P-450 reductase was either unchanged or slightly lowered. At later stages of hepatocarcinogenesis PB1 returned to the levels of the surrounding tissue, while the other cyt. P-450 isoenzymes were decreased, the most prominent reduction being found in MC1. In neoplastic nodules all cyt. P-450s and NADPH-cyt. P-450 reductase were diminuished, some of them dramatically ...

A. Pastorek, J. Hrnčířová, L. Jankovič, L. Nejdl, S. Civiš, O. Ivanek, V. Shestivska, A. Knížek, P. Kubelík, J. Šponer, L. Petera, A. Křivková, G. Cassone, M. Vaculovičová, J.E. Šponer, M. Ferus ...

Upon incubation of detergent-solubilized NADPH-cytochrome P-450 reductase and either cytochrome b5 or cytochrome c in the presence of a water-soluble carbodiimide, a 1-ethyl-3-[3-(dimethylamino)propyl]carbodiimide (EDC), covalently cross-linked compl

Hepatic NADPH-cytochrome P450 oxidoreductase null (HRN™) mice exhibit no functional expression of hepatic cytochrome P450 (P450) when compared to wild type (WT) mice, but have normal hepatic and extrahepatic expression of other biotransformation enzymes. We have assessed the utility of HRN™ mice for investigation of the role of metabolic bioactivation in liver toxicity caused by the nonsteroidal anti-inflammatory drug (NSAID) fenclozic acid. In vitro studies revealed significant NADPH-dependent (i.e. P450-mediated) covalent binding of [14C]-fenclozic acid to liver microsomes from WT mice and HRN™ mice, whereas no in vitro covalent binding was observed in the presence of the UDP-glucuronyltransferase cofactor UDPGA. Oral fenclozic acid administration did not alter the liver histopathology or elevate the plasma liver enzyme activities of WT mice, or affect their hepatic miRNA contents. Livers from HRN™ mice exhibited abnormal liver histopathology (enhanced lipid accumulation, bile duct ...

Expression and monooxygenase activity of various cytochrome P450 (CYP) enzymes along with constitutive androstane (CAR) and the pregnane X (PXR) receptors were investigated in the brain of control and phenobarbital-treated rabbits (80 mg/kg for 4 days). RT-PCR analysis, using specific primers, demonstrated that in control rabbits mRNAs of CYP 2A10, 2B4/5 and 3A6 were expressed, though to a different extent, in the liver, as well as in brain cortex, midbrain, cerebellum, striatum, hippocampus and hypothalamus, whilst CYP2A11 and 4B1 were not expressed in the hypothalamus. CAR was expressed in liver and all the brain regions examined, whereas the PXR was expressed only in liver and cortex. Real time RT-PCR analysis demonstrated that in vivo treatment with phenobarbital, in contrast with what happened in liver, did not induce the expression of CYP 2B4/5 mRNA in cortex, midbrain and cerebellum. NADPH cytochrome c reductase and some other enzymatic activities markers of CYP 2A, 2B, 3A and 4B ...

The pharmaceutical industry is committed to market safer drugs with fewer side effects, predictable pharmacokinetic properties and quantifiable drug-drug interactions. There is an increasing need to develop robust, enhanced-throughput in vitro assays, which accurately extrapolate to humans. The major drug metabolizing human hepatic cytochrome P450s (CYPs; CYP1A2, 2C9, 2C19, 2D6 and 3A4) have been co-expressed functionally in Escherichia coli with human NADPH-cytochrome P450 reductase and validated as surrogates to their counterparts in human liver microsomes (HLM) with respect to their kinetic and inhibition properties. Using these recombinant enzymes, fully automated in vitro assays to assess CYP inhibition and determine the enzymology of drug oxidation have been developed and validated. IC50 values determined for a series of test compounds in HLM and recombinant CYPs were similar (r2 = 0.9, P , 0.001). There was a good correlation between the sum of individual CYP intrinsic clearance (Clint) ...

The present investigation was undertaken to more precisely establish where xenobiotics can be oxidatively metabolized and bioactivated within the lung. To accomplish this, antibodies raised against NADPH-cytochrome P-450 reductase (EC 1.6.2.4) and cytochromes P-450 BNF-B, PB-B, and PCN-E (the major forms of cytochrome P-450 induced by beta-naphthoflavone, phenobarbital, and pregnenolone-16 alpha-carbonitrile, respectively) that had been purified to apparent homogeneity from rat liver microsomes were used to determine the localizations and distributions of these enzymes immunohistochemically at the light microscopic level within lungs of untreated rats. Additionally, the intrapulmonary sites at which benzo(alpha)pyrene undergoes hydroxylation were identified in situ by means of fluorescence histochemistry. Immunohistochemical staining for NADPH-cytochrome P-450 reductase and cytochromes P-450 BNF-B, PB-B, and PCN-E was detected in bronchial epithelial cells, both ciliated and nonciliated (Clara) ...

NDOR1 - NDOR1 (Myc-DDK-tagged)-Human NADPH dependent diflavin oxidoreductase 1 (NDOR1), transcript variant 2 available for purchase from OriGene - Your Gene Company.

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Kyselina acetylsalicylová, kombinácie s výnimkou psycholeptík, Granulát na perorálnu suspenziu, ATC N02BA51, SPC (Súhrn údajov o prípravku) Terapeutické indikácie: Symptomatická liečba upchatého nosa/nosných dutín (rinosinusitídy) s bolesťou a horúčkou, ktoré sa spájajú s nachladnutím, a/alebo príznakmi podobnými chrípke. ASPIRIN COMPLEX je indikovaný dospelým a dospievajúcim vo veku od 16 rokov.

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TY - JOUR. T1 - Effect of in utero and postnatal exposure to environmental tobacco smoke on the developmental expression of pulmonary cytochrome P450 monooxygenases. AU - Lee, Chanhung Z.. AU - Royce, Fred H.. AU - Denison, Michael S.. AU - Pinkerton, Kent E. PY - 2000. Y1 - 2000. N2 - Pulmonary cytochrome P450 monooxygenases metabolize xenobiotic chemicals, including those found in environmental tobacco smoke (ETS). Exposure to ETS beginning at birth has been shown to induce the P450 CYP1A1 by seven days of life. The effects of perinatal exposure to ETS of the rat lung on the expression of CYP1A1,1B1, 2B1, and NADPH cytochrome P450 reductase were measured using semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR). Timed pregnant dams and their pups were exposed to aged and diluted sidestream cigarette smoke (ADSS) as a surrogate for ETS for four hours/ day from gestational day 5 through postnatal day 21. For all genes analyzed, mRNA could be detected in the fetal lung ...

The phagocyte NADPH oxidase catalyzes the reduction of O-2 to reactive oxygen species with microbicidal activity. It is composed of two membrane-spanning subunits, gp91-phox and p22-phox (encoded by CYBB and CYBA, respectively), and three cytoplasmic subunits, p40-phox, p47-phox, and p67-phox (encoded by NCF4, NCF1, and NCF2, respectively). Mutations in any of these genes can result in chronic granulomatous disease, a primary immunodeficiency characterized by recurrent infections. Using evolutionary mapping, we determined that episodes of adaptive natural selection have shaped the extracellular portion of gp91-phox during the evolution of mammals, which suggests that this region may have a function in host-pathogen interactions. On the basis of a resequencing analysis of approximately 35 kb of CYBB, CYBA, NCF2, and NCF4 in 102 ethnically diverse individuals (24 of African ancestry, 31 of European ancestry, 24 of Asian/Oceanians, and 23 US Hispanics), we show that the pattern of CYBA diversity is ...

This enzyme is required for electron transfer from NADP to cytochrome P450 in microsomes. It can also provide electron transfer to heme oxygenase and cytochrome B5.

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EPAs official web site is www.epa.gov. Some links on this page may redirect users from the EPA website to specific content on a non-EPA, third-party site. In doing so, EPA is directing you only to the specific content referenced at the time of publication, not to any other content that may appear on the same webpage or elsewhere on the third-party site, or be added at a later date ...

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This is a case from the Mediastinal Vascular Cases section of Chest Imaging Cases. The specific content of each case and its diagnosis are necessarily hidden from this abstract. Each case contains case history, followed immediately by the diagnosis, which is supported with findings, differential diagnosis, teaching points, management, and further reading suggestions.

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Corrección del intercambio gaseoso utilizando ventilación no invasiva guiada por manometría esofágica en paciente con Síndrome de Obesidad e Hipoventilación. Reporte de un caso.

Corrección del intercambio gaseoso utilizando ventilación no invasiva guiada por manometría esofágica en paciente con Síndrome de Obesidad e Hipoventilación. Reporte de un caso.

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A 20-year-old woman presented 6 months after an initial injury to her left elbow with pain and restricted movements. She was diagnosed with a type I malunited (Hahn-Steinthal) type of capitellum fracture through radiographic studies. Classically, the treatment has been excision of the fragment, which carries a risk of valgus instability of the elbow and late osteoarthrosis. We report a case of malunited type I capitellum fracture, for which corrective osteotomy through fracture site, open reduction and internal fixation was done 6 months following missed trauma. At 24 months follow-up the capitellum fracture had united and the patient has a stable elbow and excellent range of motion. Our case demonstrates that for type I malunited capitellum fractures corrective osteotomy through fracture site and internal fixation rather than excision of the fragment in young can result in successful union and stable elbow.. ...

Withania somnifera (L.) Dunal, a highly reputed medicinal plant, synthesizes a large array of steroidal lactone triterpenoids called withanolides. Although its chemical profile and pharmacological activities have been studied extensively during the last two decades, limited attempts have been made to decipher the biosynthetic route and identification of key regulatory genes involved in withanolide biosynthesis. Cytochrome P450 reductase is the most imperative redox partner of multiple P450s involved in primary and secondary metabolite biosynthesis. We describe here the cloning and characterization of two paralogs of cytochrome P450 reductase from W. somnifera. The full length paralogs of WsCPR1 and WsCPR2 have open reading frames of 2058 and 2142 bp encoding 685 and 713 amino acid residues, respectively. Phylogenetic analysis demonstrated that grouping of dual CPRs was in accordance with class I and class II of eudicotyledon CPRs. The corresponding coding sequences were expressed in Escherichia coli as

1. The intracellular distribution of nitrogen, DPNH cytochrome c reductase, succinic dehydrogenase, and cytochrome c oxidase has been studied in fractions derived by differential centrifugation from rat and guinea pig spleen homogenates.. 2. In the spleens of each species, the nuclear fraction accounted for 40 to 50 per cent of the total nitrogen content of the homogenate, and the mitochondrial, microsome, and supernatant fractions contained about 8, 12, and 30 per cent of the total nitrogen, respectively.. 3. Per mg. of nitrogen, DPNH cytochrome c reductase was concentrated in the mitochondria and microsomes of both rat and guinea pig spleens. Seventy per cent of the total DPNH cytochrome c reductase activity was recovered in these two fractions. The reductase activity associated with the nuclear fraction was lowered markedly by isolating nuclei from rat spleens with the sucrose-CaCl2 layering technique. The lowered activity was accompanied by the recovery of about 90 per cent of the homogenate ...

Definition of cytochrome c reductase. Provided by Stedmans medical dictionary and Drugs.com. Includes medical terms and definitions.

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Enzyme Activity Calculations by Curvilinear Regression Analysis. A Program for a Computing Desk Top Calculator, Used in the Study of Cytochrome c Reductase and Hyaluronidase.. Hultin, Eskil; Liljeqvist, Gisela; Lundblad, Gunnar; Pal us, Sven; St hl, Gunnar ...

Estradiol p.o. 1 mg, Filmom obalená tableta, ATC G03CA03, SPC (Súhrn údajov o prípravku) Terapeutické indikácie: Hormonálna substitučná liečba (HSL) príznakov nedostatku estrogénov u žien v postmenopauze. Na prevenciu osteoporózy u žien v postmenopauze, so zvýšeným rizikom vzniku fraktúr, ktoré netolerujú, alebo sú pre ne kontraindikované iné lieky schválené na prevenciu osteoporózy. Estrofem sa hlavne indikuje u žien po hysterektómii, ktoré z tohto dôvodu nevyžadujú kombinovanú liečbu estrog...

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These cases are from the Liver section of Gastrointestinal Imaging Cases. The specific content of each case and its diagnosis are necessarily concealed in this abstract. Each case contains case history, followed immediately by the diagnosis, which is supported with findings, differential diagnosis, teaching points, management, and further reading suggestions.

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