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cytochrome P450 reductase

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https://www.readbyqxmd.com/read/28812125/recombinant-expression-and-biochemical-characterization-of-mycobacterium-tuberculosis-3fe-4s-ferredoxin-rv1786
#1
Yun Lu, Feng Qiao, Yue Li, Xiao-Hong Sang, Cong-Ran Li, Jian-Dong Jiang, Xin-Yi Yang, Xue-Fu You
Ferredoxins are iron-sulfur protein that mediate electron transfer in cytochrome P450 mono-oxygenase (CYP)-related catalytic reactions in a wide variety of organisms. Rv1786 is a putative ferredoxin, encoded by a gene located downstream of the gene encoding CYP143A1 in the Mycobacterium tuberculosis genome. However, the structure and function of Rv1786 have remained unclear. Here, the recombinant Mtb Rv1786 was expressed, purified as a His-tagged form and characterized with [3Fe-4S] clusters as its cofactors using a series of measurements including SDS-PAGE, western blot, UV/Visible, MALDI-TOF/TOF-MS, and electron paramagnetic resonance spectroscopic analysis...
August 15, 2017: Applied Microbiology and Biotechnology
https://www.readbyqxmd.com/read/28803762/apigenin-protects-against-alcohol-induced-liver-injury-in-mice-by-regulating-hepatic-cyp2e1-mediated-oxidative-stress-and-ppar%C3%AE-mediated-lipogenic-gene-expression
#2
Feng Wang, Jin-Cheng Liu, Rui-Jun Zhou, Xi Zhao, Mei Liu, Hua Ye, Mei-Lin Xie
Alcohol is a major cause of liver injury, and there are currently no ideal pharmacological reagents that can prevent or reverse this disease. Apigenin is one of the most common flavonoids present in numerous plants and has many beneficial effects. But whether or not apigenin may protect against alcohol-induced liver injury remains unknown. Our aim was to examine the effect and potential mechanisms. The experimental mice were given 56% erguotou wine or simultaneously given apigenin 150-300 mg/kg by gavage for 30 days...
August 10, 2017: Chemico-biological Interactions
https://www.readbyqxmd.com/read/28800917/thyrotoxic-rubber-antioxidants-2-mercaptobenzimidazole-and-its-methyl-derivatives-cause-both-inhibition-and-induction-of-drug-metabolizing-activity-in-rat-liver-microsomes-after-repeated-oral-administration
#3
Atsuko Miyajima, Kazue Sakemi-Hoshikawa, Makoto Usami, Katsuyoshi Mitsunaga, Tomohiko Irie, Yasuo Ohno, Momoko Sunouchi
We examined the effects of thyrotoxic rubber antioxidants, 2-mercaptobenzimidazole (MBI, 0.3 mmol/kg/day) and its methyl derivatives, methyl-MBIs [4-methyl-MBI (4-MeMBI, 0.6 mmol/kg/day), 5-methyl-MBI (5-MeMBI, 0.6 mmol/kg/day), and 4(or 5)-methyl-MBI (4(5)-MeMBI, 0.6 or 1.2 mmol/kg/day)], on the drug-metabolizing activity in male rat liver microsomes by 8-day repeated oral administration. The weight of liver and thyroid were increased by all the test chemicals; MBI was most potent, and there was no additive or synergistic effect between 4-MeMBI and 5-MeMBI...
August 8, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/28799023/expression-of-cholesterol-hydroxylase-lyase-system-proteins-in-yeast-s-cerevisiae-cells-as-a-self-processing-polyprotein
#4
Vera S Efimova, Ludmila V Isaeva, Desislava S Makeeva, Mikhail A Rubtsov, Ludmila A Novikova
2A peptide discovered in Picornaviridae is capable of self-cleavage providing an opportunity to carry out synthesis of several proteins using one transcript. Dissociation in the 2A sequence during translation leads to the individual proteins formation. We constructed cDNA including genes of the bovine cholesterol hydroxylase/lyase (CHL) system proteins-cytochrome P450scc (CYP11A1), adrenodoxin (Adx) and adrenodoxin reductase (AdR), that are fused into a single ORF using FMDV 2A nucleotide sequences. The constructed vectors direct the expression of cDNA encoding polyprotein P450scc-2A-Adx-2A-AdR (CHL-2A) in Escherichia coli and Saccharomyces cerevisiae...
August 10, 2017: Molecular Biotechnology
https://www.readbyqxmd.com/read/28794165/steroidogenesis-during-postnatal-testicular-development-of-galea-spixii
#5
Paulo Ramos da Silva Santos, Francilieusa Delys Oliveira, Maria Angélica Machado Arroyo, Moacir Franco Oliveira, Patricia Castelucci, Alan Conley, Antônio Chaves de Assis Neto
The androgen/estrogen balance is essential for normal sexual development and reproduction in mammals. Studies performed herein investigated the potential for estrogen synthesis in cells of the testes of a hystricomorph rodent, Galea spixii. The study characterized the expression of the key enzymes responsible for estrogen and androgen synthesis, cytochromes P450 aromatase (P450arom), 17α-hydroxylase/17,20-lyase (P450c17) respectively, as well as the redox partner NADPH cytochrome P450 oxido-reductase (CPR) required to support electron transfer and catalysis of these P450s, by immunohistochemistry (IHC) and quantitative polymerase chain reaction (qPCR) analysis, throughout postnatal sexual development...
August 8, 2017: Reproduction: the Official Journal of the Society for the Study of Fertility
https://www.readbyqxmd.com/read/28792090/distinct-activation-mechanisms-trigger-the-trypanocidal-activity-of-dna-damaging-prodrugs
#6
Emma Louise Meredith, Ambika Kumar, Aya Konno, Joanna Szular, Sam Alsford, Karin Seifert, David Horn, Shane R Wilkinson
Quinone-based compounds have been exploited to treat infectious diseases and cancer, with such chemicals often functioning as inhibitors of key metabolic pathways or as prodrugs. Here, we screened an aziridinyl-1,4-benzoquinone (ABQ) library against the causative agents of trypanosomiasis, and cutaneous leishmaniasis, identifying several potent structures that exhibited EC50 values of <100 nM. However, these compounds also displayed significant toxicity towards mammalian cells indicating that they are not suitable therapies for systemic infections...
August 9, 2017: Molecular Microbiology
https://www.readbyqxmd.com/read/28769808/metabolism-of-f18-a-derivative-of-calanolide-a-in-human-liver-microsomes-and-cytosol
#7
Xiangmeng Wu, Qinghao Zhang, Jiamei Guo, Yufei Jia, Ziqian Zhang, Manman Zhao, Yakun Yang, Baolian Wang, Jinping Hu, Li Sheng, Yan Li
10-Chloromethyl-11-demethyl-12-oxo-calanolide (F18), an analog of calanolide A, is a novel potent nonnucleoside reverse transcriptase inhibitor against HIV-1. Here, we report the metabolic profile and the results of associated biochemical studies of F18 in vitro and in vivo. The metabolites of F18 were identified based on liquid chromatography-electrospray ionization mass spectrometry and/or nuclear magnetic resonance. Twenty-three metabolites of F18 were observed in liver microsomes in vitro. The metabolism of F18 involved 4-propyl chain oxidation, 10-chloromethyl oxidative dechlorination and 12-carbonyl reduction...
2017: Frontiers in Pharmacology
https://www.readbyqxmd.com/read/28758225/metabolism-of-7-ethoxycoumarin-flavanone-and-steroids-by-cytochrome-p450-2c9-variants
#8
Tomohide Uno, Ryosuke Nakano, Kengo Kanamaru, Shinji Takenaka, Yuichi Uno, Hiromasa Imaishi
CYP2C9 is a human microsomal cytochrome P450c (CYP). Much variation in CYP2C9 levels and activity can be attributed to polymorphisms of this gene. Wild-type CYP2C9 and mutants were coexpressed with NADPH-cytochrome P450 reductase in Escherichia coli. The hydroxylase activities toward 7-ethoxycoumarin, flavanone and steroids were examined. Six CYP2C9 variants showed Soret peaks (450 nm) typical of P450 in reduced CO-difference spectra. CYP2C9.38 had the highest 7-ethoxycoumarin de-ethylase activity. All the CYP2C9 variants showed lower flavanone 6-hydroxylation activities than CYP2C9...
July 30, 2017: Biopharmaceutics & Drug Disposition
https://www.readbyqxmd.com/read/28741929/putidaredoxin-binds-to-the-same-site-on-cytochrome-p450cam-in-the-open-and-closed-conformation
#9
Shu-Hao Liou, William K Myers, Jason D Oswald, R David Britt, David B Goodin
Cytochrome P450 CYP101A1 (P450cam) hydroxylates camphor by receiving two distinct electrons from its unique reductase, putidaredoxin (Pdx). Upon binding ferric P450cam, Pdx is now known to trigger a conformational change in the enzyme. This Pdx-induced conversion may provide the trigger to coordinate enzyme turnover and protect the enzyme from oxidative damage, so the interactions responsible for this conversion are of significant current interest. This proposed role for Pdx requires that its interactions with P450cam are different for the open and closed conformations...
July 25, 2017: Biochemistry
https://www.readbyqxmd.com/read/28734978/insights-into-the-functional-properties-of-the-marneral-oxidase-cyp71a16-from-arabidopsis-thaliana
#10
Sarah Kranz-Finger, Osama Mahmoud, Esther Ricklefs, Nikolas Ditz, Patrick J Bakkes, Vlada B Urlacher
The Arabidopsis thaliana gene encoding CYP71A16 is part of the gene cluster for the biosynthesis and modification of the triterpenoid marneral. Previous investigations of A. thaliana have revealed that CYP71A16 catalyzes marneral oxidation, while it also can accept marnerol as substrate. The aim of the present study was to investigate functional properties of CYP71A16 in vitro. For this purpose, heterologous expression of a N-terminally modified version of CYP71A16 was established in Escherichia coli, which yielded up to 50mgL(-1) recombinant enzyme...
July 19, 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/28734977/modulation-of-the-interaction-between-human-p450-3a4-and-b-megaterium-reductase-via-engineered-loops
#11
Silvia Castrignanò, Serena D'Avino, Giovanna Di Nardo, Gianluca Catucci, Sheila J Sadeghi, Gianfranco Gilardi
Chimerogenesis involving cytochromes P450 is a successful approach to generate catalytically self-sufficient enzymes. However, the connection between the different functional modules should allow a certain degree of flexibility in order to obtain functional and catalytically efficient proteins. We previously applied the molecular Lego approach to develop a chimeric P450 3A4 enzyme linked to the reductase domain of P450 BM3 (BMR). Three constructs were designed with the connecting loop containing no glycine, 3 glycine or 5 glycine residues and showed a different catalytic activity and coupling efficiency...
July 19, 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/28730410/engineering-of-the-cytochrome-p450-monooxygenase-system-for-benzyl-maltol-hydroxylation
#12
Iori Kozono, Kousuke Mihara, Kazuyuki Minagawa, Makoto Hibi, Jun Ogawa
Maltol derivatives are utilized in a variety of fields due to their metal-chelating abilities, and modification of the 2-methyl side chain is known to effectively expand their functional diversity. In the present study, microbial enzymes were screened for hydroxylating activity towards the 2-methyl group in a maltol derivative, 3-benzyloxy-2-methyl-4-pyrone (BMAL). Novosphingobium sp. SB32149 was found to have the ability to convert BMAL into 3-benzyloxy-2-hydroxymethyl-4-pyrone (BMAL-OH). The enzymes responsible, a cytochrome P450 monooxygenase (P450nov), a ferredoxin (FDXnov), and a ferredoxin reductase (FDRnov), were identified in the SB32149 strain...
July 20, 2017: Applied Microbiology and Biotechnology
https://www.readbyqxmd.com/read/28724407/characterization-of-the-cytochrome-p450-monooxygenase-genes-p450ome-from-the-carotenogenic-yeast-xanthophyllomyces-dendrorhous
#13
Pamela Córdova, Ana-María Gonzalez, David R Nelson, María-Soledad Gutiérrez, Marcelo Baeza, Víctor Cifuentes, Jennifer Alcaíno
BACKGROUND: The cytochromes P450 (P450s) are a large superfamily of heme-containing monooxygenases involved in the oxidative metabolism of an enormous diversity of substrates. These enzymes require electrons for their activity, and the electrons are supplied by NAD(P)H through a P450 electron donor system, which is generally a cytochrome P450 reductase (CPR). The yeast Xanthophyllomyces dendrorhous has evolved an exclusive P450-CPR system that specializes in the synthesis of astaxanthin, a carotenoid with commercial potential...
July 19, 2017: BMC Genomics
https://www.readbyqxmd.com/read/28710782/identification-and-functional-analysis-of-two-p450-enzymes-of-gossypium-hirsutum-involved-in-dmnt-and-tmtt-biosynthesis
#14
Danfeng Liu, Xinzheng Huang, Weixia Jing, Xingkui An, Qiang Zhang, Hong Zhang, Jingjiang Zhou, Yongjun Zhang, Yuyuan Guo
The homoterpenes (3E)-4,8-dimethyl-1,3,7-nonatriene (DMNT) and (E,E)-4,8,12-trimethyl-1,3,7,11-tridecatetraene (TMTT) are major herbivore-induced plant volatiles (HIPV) that can attract predatory or parasitic arthropods to protect injured plants from herbivore attack. In the present study, DMNT and TMTT were confirmed to be emitted from cotton (Gossypium hirsutum) plants infested with chewing caterpillars or sucking bugs. Two CYP genes (GhCYP82L1 and GhCYP82L2) involved in homoterpene biosynthesis in G. hirsutum were newly identified and characterized...
July 15, 2017: Plant Biotechnology Journal
https://www.readbyqxmd.com/read/28706181/effects-of-an-enriched-extract-of-paeoniflorin-a-monoterpene-glycoside-used-in-chinese-herbal-medicine-on-cholesterol-metabolism-in-a-hyperlipidemic-rat-model
#15
Huiming Hu, Qiaoqiao Zhu, Jie Su, Yajun Wu, Yanchen Zhu, Yin Wang, Hui Fang, Minxia Pang, Bo Li, Suhong Chen, Guiyuan Lv
BACKGROUND Paeoniflorin is a monoterpene glycoside extracted from the roots of Paeonia lactiflora and is used in Chinese herbal medicine to treat hyperlipidemia. The aim of this study was to evaluate the effects of an enriched extract of paeoniflorin on cholesterol levels, hemodynamics, and oxidative stress in a hyperlipidemic rat model. MATERIAL AND METHODS Male Sprague-Dawley rats were fed high-cholesterol diets and treated with three different doses of paeoniflorin for 12 weeks. The effects of paeoniflorin treatment were assessed on cholesterol levels, cholesterol metabolism, red blood cell vascular flow using hemorheology, antioxidant enzymes, and expression of the rate-limiting enzyme in the mevalonate pathway, 3-hydroxy-3-methylglutharyl-coenzyme A reductase (HMG-CoAR)...
July 14, 2017: Medical Science Monitor: International Medical Journal of Experimental and Clinical Research
https://www.readbyqxmd.com/read/28700330/translocation-of-heme-oxygenase-1-contributes-to-imatinib-resistance-in-chronic-myelogenous-leukemia
#16
Bianca Schaefer, Soenke Behrends
Heme oxygenase-1 (HO-1) degrades heme to bilirubin. In addition, it is upregulated in malignant disease and has been described as an important factor for cancer prognosis and therapy. Under physiological conditions HO-1 is anchored to the endoplasmic reticulum (ER). Under stress conditions HO-1 can be cleaved and subsequently translocates to the cytosol and nucleus.In this study we systematically investigated the influence of HO-1's catabolic activity and subcellular localization on resistance against the tyrosine kinase inhibitor imatinib in leukemia cells by confocal laser scanning microscopy, hemoglobin synthesis experiments and cell viability assays...
June 27, 2017: Oncotarget
https://www.readbyqxmd.com/read/28690182/functional-characterization-of-a-soluble-nadph-cytochrome-p450-reductase-from-fusarium-graminearum
#17
Thomas Etzerodt, Karl Wetterhorn, Giuseppe Dionisio, Ivan Rayment
Fusarium head blight is a devastating disease in wheat caused by some fungal pathogens of the Fusarium genus mainly F. graminearum, due to accumulation of toxic trichothecenes. Most of the trichothecene biosynthetic pathway has been mapped, although some proteins of the pathway remain uncharacterized, including an NADPH-cytochrome P450 reductase. We subcloned a F. graminearum cytochrome P450 reductase that might be involved in the trichothecene biosynthesis. It was expressed heterologously in E. coli as N-terminal truncated form with an octahistidine tag for purification...
October 2017: Protein Expression and Purification
https://www.readbyqxmd.com/read/28677809/inhibition-of-aldose-reductase-ameliorates-alcoholic-liver-disease-by-activating-ampk-and-modulating-oxidative-stress-and-inflammatory-cytokines
#18
Changxuan Shi, Yuanfang Wang, Jing Gao, Si Chen, Xiangqian Zhao, Chengchao Cai, Chang Guo, Longxin Qiu
Aldose reductase (AR) expression is elevated in the livers of patients with alcoholic liver diseases. However, the role of AR in the development of alcoholic liver diseases remains unclear. The aim of the present study was to determine the effect of AR inhibition on ethanol‑induced hepatosteatosis in vivo and in vitro, and to identify possible underlying molecular mechanisms. Alcoholic fatty livers were induced in C57BL/6 mice by feeding the mice with Lieber‑DeCarli liquid diets. The expression of AR protein was elevated in the liver tissue of C57BL/6 mice fed with an ethanol diet and in mouse AML12 liver cells exposed to ethanol...
September 2017: Molecular Medicine Reports
https://www.readbyqxmd.com/read/28665167/altered-hepatic-drug-metabolizing-activity-in-rats-suffering-from-hypoxemia-with-experimentally-induced-acute-lung-impairment
#19
Yuki Hori, Yasumasa Shimizu, Tetsuya Aiba
1. Hepatic drug-metabolizing activity was investigated in vitro with liver microsomes prepared from rats suffering from hypoxemia with experimentally induced acute lung impairment (ALI). 2. Male Wistar rats received an intrabronchial administration of dilute hydrochloride solution for ALI induction. Pooled liver microsomes were prepared for the normal and ALI rats, and the hepatic drug metabolism mediated by cytochrome P450 (CYP) 3A was examined in an incubation study with the microsomes. 3. The NADPH-dependent metabolism of midazolam significantly increases in ALI rats as compared with that in normal rats...
June 30, 2017: Xenobiotica; the Fate of Foreign Compounds in Biological Systems
https://www.readbyqxmd.com/read/28655775/the-iron-chaperone-poly-rc-binding-protein-2-forms-a-metabolon-with-the-heme-oxygenase-1-cytochrome-p450-reductase-complex-for-heme-catabolism-and-iron-transfer
#20
Izumi Yanatori, Des R Richardson, Shinya Toyokuni, Fumio Kishi
Mammals incorporate a major proportion of absorbed iron as heme, which is catabolized by the heme oxygenase 1 (HO1)-NADPH-cytochrome P450 reductase (CPR) complex into biliverdin, carbon monoxide, and ferrous iron. Moreover, intestinal iron is incorporated as ferrous iron, which is transported via the iron importer, divalent metal transporter 1 (DMT1). Recently, we demonstrated that the iron chaperone poly(rC)-binding protein 2 (PCBP2) can directly receive ferrous iron from DMT1 or transfer iron to the iron exporter, ferroportin 1...
August 11, 2017: Journal of Biological Chemistry
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