DOCSLIB.ORG

Effect of Carbon Monoxide Donor CORM-2 on Vitamin D Metabolism

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Effect of Carbon Monoxide Donor CORM-2 on Vitamin D Metabolism Kidney Blood Press Res 2013;37:496-505 DOI: 10.1159/000355730 © 2013 S. Karger AG, Basel Published online: November 11, 2013 www.karger.com/kbr 496 Accepted:Feger et al.: September CORM-2-induced 17, 2013 inhibition of vitamin D31423-0143/13/0375-0496$38.00/0 activation This is an Open Access article licensed under the terms of the Creative Commons Attribution- NonCommercial 3.0 Unported license (CC BY-NC) (www.karger.com/OA-license), applicable to the online version of the article only. Distribution permitted for non-commercial purposes only. Original Paper Effect of Carbon Monoxide Donor CORM-2 on Vitamin D3 Metabolism Martina Fegera,f Abul Fajola,f Aleksandra Lebedevaa,b Adrian Meissnera Diana Michaela Jakob Voelkla Ioana Alesutana Erwin Schleicherc Christoph Reichetzederd Berthold Hocherd Syed M. Qadria,e,g Florian Langa,g aDepartment of Physiology, University of Tübingen, Tübingen, Germany; bDepartment of Immunology, Institute of Experimental Medicine, St. Petersburg, Russia; cDepartment of Internal Medicine, University of Tübingen, Tübingen, Germany; dInstitute of Nutritional Science, University of Potsdam, Potsdam, Germany, eDepartment of Pharmacology, University of Saskatchewan, Saskatoon, Canada; fcontributed equally and thus share first authorship; gcontributed equally and thus share last authorship Key Words CORM-2 • 1,25-Dihydroxyvitamin D3 • Klotho • FGF23 • Phosphate • Calcium Abstract Background/Aims: Carbon monoxide (CO) interferes with cytochrome-dependent cellular functions and acts as gaseous transmitter. CO is released from CO-releasing molecules (CORM) including tricarbonyl-dichlororuthenium (II) dimer (CORM-2), molecules considered for the treatment of several disorders including vascular dysfunction, inflammation, tissue ischemia and organ rejection. Cytochrome P450-sensitive function include formation of 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) by renal 25-hydroxyvitamin D3 1-alpha-hydroxylase 2+ (Cyp27b1). The enzyme is regulated by PTH, FGF23 and klotho. 1,25(OH)2D3 regulates Ca and phosphate transport as well as klotho expression. The present study explored, whether CORM- 2 influences 1,25(OH)2D3 formation and klotho expression. Methods: Mice were treated with intravenous CORM-2 (20 mg/kg body weight). Plasma 1,25(OH)2D3 and FGF23 concentrations were determined by ELISA, phosphate, calcium and creatinine concentrations by colorimetric methods, transcript levels by quantitative RT-PCR and protein expression by western blotting. Fgf23 mRNA transcript levels were further determined in rat osteosarcoma UMR106 cells without or with prior treatment for 24 hours with 20 µM CORM-2. Results: CORM-2 injection within 24 hours significantly increased FGF23 plasma levels and decreased 1,25(OH)2D3 plasma levels, renal Cyp27b1 gene expression as well as renal klotho protein abundance and transcript levels. Moreover, treatment of UMR106 cells with CORM-2 significantly increased Fgf23 transcript levels. Conclusion: CO-releasing molecule CORM-2 enhances FGF23 expression and release and decreases klotho expression and 1,25(OH)2D3 synthesis. Copyright © 2013 S. Karger AG, Basel Prof. Dr. Florian Lang Physiologisches Institut der Universität Tübingen, Gmelinstr. 5, D-72076 Tuebingen (Germany), Tel. +49 7071 29 72194, Fax +49 7071 29 5618 E-Mail [email protected] Kidney Blood Press Res 2013;37:496-505 DOI: 10.1159/000355730 © 2013 S. Karger AG, Basel Published online: November 11, 2013 www.karger.com/kbr 497 Feger et al.: CORM-2-induced inhibition of vitamin D3 activation Introduction Carbon monoxide (CO), a gas, which is highly toxic to humans and animals when encountered in higher concentrations, binds rapidly to hemoglobin (Hb), leading to the stable formation of carboxyhemoglobin (COHb) [1]. As a result of CO binding to Hb both O2 binding at high and O2 release at low O2 partial pressure are impaired [1] leading to a reduced oxygen carrying capacity of blood after CO inhalation [2]. Besides its toxic effects at high concentrations, CO is a gaseous transmitter molecule [3]. In mammalian cells CO is generated by the enzyme heme-oxygenase [4]. CO participates in the regulation of a wide variety of cellular functions [5-9] and contributes to the regulation of cell death [10-15]. CO may elicit both vasodilation and vasoconstriction [16, 17]. In addition, CO in vitro and in vivo [18, 19]. CO can be released in an organism by the tricarbonyl-dichlororuthenium (II) dimer (CORM-2) [6]. CO-releasing moleculesfosters coagulation have been and suggested attenuates as therapeuticfibrinolysis option in various pathologies, but the risks of CO interferes with the function of cytochromes [21-23]. The cytochrome P450 enzyme 25-hydroxyvitamintreatment with those D molecules3 1-alpha-hydroxylase have been insufficiently accomplishes addressed formation [6, 20]. of 1,25-dihydroxy- vitamin D3 (1,25(OH)2D3) from its inactive precursor (25-hydroxyvitamin D3) [24]. The enzyme 25-hydroxyvitamin D3 1-alpha-hydroxylase is encoded by the Cyp27b1 gene [24]. 1,25(OH)2D3 is inactivated by 1,25-dihydroxyvitamin D3 24-hydroxylase, an enzyme encoded by the Cyp24a1 gene [25]. 1,25(OH)2D3 is a powerful regulator of CaHPO4 metabolism and lack of 1,25(OH)2D3 leads to rickets and osteomalacia [26]. Reduced levels of 1,25(OH)2D3 are further associated with the onset and progression of various diseases such as respiratory infections, cardiovascular disease and cancer [27-30]. 1,25(OH)2D3 formation is inhibited by joint action of klotho and FGF23, proteins regulating phosphate homeostasis and implicated in various disorders including arteriosclerosis, cancer metastasis, and chronic kidney disease [31-33]. The present study explored, whether the CO donor tricarbonyl-dichlororuthenium (II) Furthermore, heme-oxygenase-1 is involved3 metabolism in bone [6, metabolism 14]. To this andend, inflammation vitamin D metabolism [34, 35]. was investigated after CORM-2 injection into mice. dimer (CORM-2) influences vitamin D Materials and Methods All animal experiments were conducted according to the German law for the welfare of animals and were approved by local authorities. Male and female BALB/c mice had free access to food (ssniff Spezialdiäten, Soest, Germany) containing 7000 mg/kg phosphorus and to tap drinking water ad libitum. Where indicated, Germany), solubilized in dimethyl sulfoxide (DMSO, Sigma Aldrich), was injected intravenously (20 mg/kg bodythe CO-releasing weight). Control molecule mice tricarbonyldichlororutheniumwere injected with vehicle (10% (II) DMSO dimer in (CORM-2, saline solution). Sigma Aldrich, Taufkirchen, To determine excretion of phosphate, the mice were placed individually in metabolic cages (Techniplast, Hohenpeissenberg, Germany) for 24-hour urine collection [36, 37]. They were allowed a 2-day habituation period. Subsequently, 24-hour collection of urine was performed. To assure quantitative urine collection, metabolic cages were siliconized, and urine was collected under water-saturated oil [38]. The urinary phosphate concentration was determined colorimetrically utilizing a commercial diagnostic kit (Roche Diagnostics, Mannheim, Germany) [39]. Wiesbaden- Delkenheim, Germany) and about 50 - 200 µl of blood was withdrawn into heparinized capillaries by puncturingTo obtain the retro-orbital blood specimens, plexus mice[40]. Thewere plasma lightly concentrations anaesthetized of with phosphate isoflurane and Ca(Abott,2+ were measured by a photometric method (FUJI FDC 3500i, Sysmex, Norsted, Germany). The creatinine concentrations in plasma were measured using a commercial enzymatic kit (creatinine PAP, Labor & Technik, Berlin, Germany, based on the creatininase method). An EIA kit was employed to determine plasma concentrations of 1,25(OH)2D3 (IDS, Boldon, UK). Plasma FGF23 (C-Term) concentrations were determined utilizing a commercial ELISA Kit (Immutopics, San Clemente, USA) [41]. Kidney Blood Press Res 2013;37:496-505 DOI: 10.1159/000355730 © 2013 S. Karger AG, Basel Published online: November 11, 2013 www.karger.com/kbr 498 Feger et al.: CORM-2-induced inhibition of vitamin D3 activation cervical dislocation. The kidneys were removed and immediately snap frozen in liquid nitrogen [42]. To determineAfter protein24 hours expression of CORM-2 by treatment,western blotting, mice were kidneys anesthetized were lysed with with isoflurane ice-cold andlysis sacrificedbuffer (54.6 by mM HEPES; 2.69 mM Na4P2O7; 360 mM NaCl; 10% [vol/vol] Glycerol; 1% [vol/vol] NP40) containing phosphatase and protease inhibitors (Complete mini, Roche, Mannheim, Germany). Homogenates were Tris-Glycine), transferred to nitrocellulose membranes (Schleicher and Schuell, Dassel, Germany), blocked clarifiedfor 1 h in by blocking centrifugation buffer (5%at 12000 fat-free rpm milk for in20 Tris-bufferedmin [43]. Total saline protein (TBS) was containing separated 0.1% by SDS-PAGE Tween (TBS- (8% T)),and incubated overnight with an anti-klotho antibody (1:1000 in 5% fat-free milk in TBS-T, KM2076 (Rat), kindly provided by Kyowa Hakko Kirin Co. Ltd, Japan). After incubation with an anti-rat secondary antibody (1:2000 in 5% fat-free milk in TBS-T, Cell Signaling), the bands were visualized with enhanced chemiluminescence according to the manufacturer's instructions. Homogenates were also probed with a primary Gapdh antibody (1:2000 in 5% BSA in TBS-T, Cell Signaling) as a loading control. Densitometric analysis of klotho and Gapdh was performed using Quantity One software (Bio-Rad Laboratories, München, Germany). containing 4,5 g/l glucose (PAA Laboratories,
Load more

Recommended publications
  • Regulation of Ribulose-1, 5-Bisphosphate Carboxylase
    Plant Physiol. (1993) 102: 21-28 Regulation of Ribulose- 1,5-Bisphosphate Carboxylase/Oxygenase Activity in Response to Reduced Light lntensity in C4 Plants' Rowan F. Sage* and Jeffrey R. Seemann Department of Botany, University of Georgia, Athens, Ceorgia 30602 (R.F.S.); and Department of Biochemistry, University of Nevada, Reno, Nevada 89557 (J.R.S.) tion of Rubisco activity by reversible carbamylation occurs in lhe light-dependent regulation of ribulose-1,5-bisphosphate response to changes in light intensity as well as the concen- carboxylase/oxygenase (Rubisco) activity was studied in 16 species tration of C02and 02,whereas inhibition of Rubisco activity of C, plants representing all three biochemical subtypes and a by CAlP occurs only in response to varying PPFD (Sharkey variety of taxonomic groups. Rubisco regulation was assessed by et al., 1986; Sage et al., 1990; Seemann et al., 1990). At measuring (a) the ratio of initial to total Rubisco activity, which physiological levels of COz in C3 plants (5-10 PM), full reflects primarily the carbamylation state of the enzyme, and (b) carbamylation of Rubisco requires Rubisco activase (Salvucci, total Rubisco activity per mo1 of Rubisco catalytic sites, which 1989; Portis, 1990). In the absence of Rubisco activase, only declines when 2-carboxyarabinitol I-phosphate (CAlP) binds to carbamylated Rubisco. In all species examined, the activity ratio of 20 to 40% of Rubisco catalytic sites are carbamylated under Rubisco declined with a reduction in light intensity, although sub- physiological conditions, leading to a significant inhibition of stantial variation was apparent between species in the degree of photosynthesis (Salvucci, 1989; Portis, 1990).
    [Show full text]
  • Somatic Aging Pathways Regulate Reproductive Plasticity in Caenorhabditis Elegans Maria C Ow, Alexandra M Nichitean, Sarah E Hall*
    RESEARCH ARTICLE Somatic aging pathways regulate reproductive plasticity in Caenorhabditis elegans Maria C Ow, Alexandra M Nichitean, Sarah E Hall* Department of Biology, Syracuse University, Syracuse, United States Abstract In animals, early-life stress can result in programmed changes in gene expression that can affect their adult phenotype. In C. elegans nematodes, starvation during the first larval stage promotes entry into a stress-resistant dauer stage until environmental conditions improve. Adults that have experienced dauer (postdauers) retain a memory of early-life starvation that results in gene expression changes and reduced fecundity. Here, we show that the endocrine pathways attributed to the regulation of somatic aging in C. elegans adults lacking a functional germline also regulate the reproductive phenotypes of postdauer adults that experienced early-life starvation. We demonstrate that postdauer adults reallocate fat to benefit progeny at the expense of the parental somatic fat reservoir and exhibit increased longevity compared to controls. Our results also show that the modification of somatic fat stores due to parental starvation memory is inherited in the F1 generation and may be the result of crosstalk between somatic and reproductive tissues mediated by the germline nuclear RNAi pathway. Introduction Evidence indicating that experiences during early development affect behavior and physiology in a stress-specific manner later in life is abundant throughout the animal kingdom (Telang and Wells, *For correspondence: [email protected] 2004; Weaver et al., 2004; Binder et al., 2008; Pellegroms et al., 2009; van Abeelen et al., 2012; Zhao and Zhu, 2014; Canario et al., 2017; Dantzer et al., 2019; Vitikainen et al., 2019).
    [Show full text]
  • A Three-Component Dicamba O-Demethylase from Pseudomonas Maltophilia, Strain DI-6 GENE ISOLATION, CHARACTERIZATION, and HETEROLOGOUS EXPRESSION*
    THE JOURNAL OF BIOLOGICAL CHEMISTRY Vol. 280, No. 26, Issue of July 1, pp. 24759–24767, 2005 © 2005 by The American Society for Biochemistry and Molecular Biology, Inc. Printed in U.S.A. A Three-component Dicamba O-Demethylase from Pseudomonas maltophilia, Strain DI-6 GENE ISOLATION, CHARACTERIZATION, AND HETEROLOGOUS EXPRESSION* Received for publication, January 18, 2005, and in revised form, March 16, 2005 Published, JBC Papers in Press, April 26, 2005, DOI 10.1074/jbc.M500597200 Patricia L. Herman‡, Mark Behrens, Sarbani Chakraborty, Brenda M. Chrastil, Joseph Barycki, and Donald P. Weeks§ From the Department of Biochemistry, University of Nebraska-Lincoln, Lincoln, Nebraska 65888-0664 Dicamba O-demethylase is a multicomponent enzyme The herbicide dicamba (2-methoxy-3,6-dichlorobenzoic acid) from Pseudomonas maltophilia, strain DI-6, that cata- has been used to effectively control broadleaf weeds in crops lyzes the conversion of the widely used herbicide di- such as corn and wheat for almost 40 years. Like a number of camba (2-methoxy-3,6-dichlorobenzoic acid) to DCSA other chlorinated organic compounds, dicamba does not persist (3,6-dichlorosalicylic acid). We recently described the in the soil because it is efficiently metabolized by a consortium biochemical characteristics of the three components of of soil bacteria under both aerobic and anaerobic conditions this enzyme (i.e. reductaseDIC, ferredoxinDIC, and oxy- (1–4). Studies with different soil types treated with dicamba genaseDIC) and classified the oxygenase component of have demonstrated that 3,6-dichlorosalicylic acid (DCSA),1 a dicamba O-demethylase as a member of the Rieske non- compound without herbicidal activity, is a major product of the heme iron family of oxygenases.
    [Show full text]
  • Flavin-Containing Monooxygenases: Mutations, Disease and Drug Response Phillips, IR; Shephard, EA
    Flavin-containing monooxygenases: mutations, disease and drug response Phillips, IR; Shephard, EA For additional information about this publication click this link. http://qmro.qmul.ac.uk/jspui/handle/123456789/1015 Information about this research object was correct at the time of download; we occasionally make corrections to records, please therefore check the published record when citing. For more information contact [email protected] Flavin-containing monooxygenases: mutations, disease and drug response Ian R. Phillips1 and Elizabeth A. Shephard2 1School of Biological and Chemical Sciences, Queen Mary, University of London, Mile End Road, London E1 4NS, UK 2Department of Biochemistry and Molecular Biology, University College London, Gower Street, London WC1E 6BT, UK Corresponding author: Shephard, E.A. ([email protected]). and, thus, contribute to drug development. This review Flavin-containing monooxygenases (FMOs) metabolize considers the role of FMOs and their genetic variants in numerous foreign chemicals, including drugs, pesticides disease and drug response. and dietary components and, thus, mediate interactions between humans and their chemical environment. We Mechanism and structure describe the mechanism of action of FMOs and insights For catalysis FMOs require flavin adenine dinucleotide gained from the structure of yeast FMO. We then (FAD) as a prosthetic group, NADPH as a cofactor and concentrate on the three FMOs (FMOs 1, 2 and 3) that are molecular oxygen as a cosubstrate [5,6]. In contrast to most important for metabolism of foreign chemicals in CYPs FMOs accept reducing equivalents directly from humans, focusing on the role of the FMOs and their genetic NADPH and, thus, do not require accessory proteins.
    [Show full text]
  • Identification of Novel CYP2E1 Inhibitor to Investigate Cellular and Exosomal CYP2E1-Mediated Toxicity
    University of Tennessee Health Science Center UTHSC Digital Commons Theses and Dissertations (ETD) College of Graduate Health Sciences 6-2019 Identification of Novel CYP2E1 Inhibitor to Investigate Cellular and Exosomal CYP2E1-Mediated Toxicity Mohammad Arifur Rahman University of Tennessee Health Science Center Follow this and additional works at: https://dc.uthsc.edu/dissertations Part of the Pharmacy and Pharmaceutical Sciences Commons Recommended Citation Rahman, Mohammad Arifur (0000-0002-5589-0114), "Identification of Novel CYP2E1 Inhibitor to Investigate Cellular and Exosomal CYP2E1-Mediated Toxicity" (2019). Theses and Dissertations (ETD). Paper 482. http://dx.doi.org/10.21007/etd.cghs.2019.0474. This Dissertation is brought to you for free and open access by the College of Graduate Health Sciences at UTHSC Digital Commons. It has been accepted for inclusion in Theses and Dissertations (ETD) by an authorized administrator of UTHSC Digital Commons. For more information, please contact [email protected]. Identification of Novel CYP2E1 Inhibitor to Investigate Cellular and Exosomal CYP2E1-Mediated Toxicity Abstract Cytochrome P450 2E1 (CYP2E1)-mediated hepatic and extra-hepatic toxicity is of significant clinical importance. Diallyl sulfide (DAS) has been shown to prevent xenobiotics such as alcohol- (ALC/ETH), acetaminophen- (APAP) induced toxicity and disease (e.g. HIV-1) pathogenesis. DAS imparts its beneficial effect by inhibiting CYP2E1-mediated metabolism of xenobiotics, especially at high concentration. However, DAS also causes toxicity at relatively high dosages and with long exposure times. The objective of the first project was to find potent ASD analogs which can replace DAS as a research tool or as potential adjuvant therapy in CYP2E1-mediated pathologies.
    [Show full text]
  • The Stress of Phenylalanine on Rats to Study the Phenylketonuria at Biochemical and Molecular Level
    Journal of Applied Pharmaceutical Science Vol. 4 (04), pp. 024-029, April, 2014 Available online at http://www.japsonline.com DOI: 10.7324/JAPS.2014.40405 ISSN 2231-3354 The stress of phenylalanine on rats to study the phenylketonuria at biochemical and molecular level Mohga Shafik¹, Hayat Ibrahime¹, Ibrahim Abo elyazeid², Osama Abass³, Heba M Saad¹ ¹Faculty of science, Helwan University, Egypt, ²Center Middle East Radioactive Isotopes, Egypt, ³ Egyptian Atomic Energy Authority, Egypt. ABSTRACT ARTICLE INFO Article history: The present study was aimed to study the stress of phenylalanine on rats to study the effect of Phenylketonuria at Received on: 06/02/2014 molecular and biochemical level. In our study, the rats’ weight ranged from 132 to 190 gm. They were housed 25 Revised on: 24/02/2014 day and the diet was prepared 5% phenylalanine and the weight is recorded every week. The rats divided into 2 Accepted on: 19/04/2014 groups, control group and phenylalanine group. After feeding with 5% phenylalanine diet, we take blood samples Available online: 28/04/2014 to measure biochemical markers as (ALT, AST, creatinine, Lipid profile and S100B) and tissues for PCR. Our biochemical results showed significant increase in S100B in phenylalanine group and reduction in total Key words: cholesterol, HDL, LDL and triglyceride in phenylalanine group. The molecular study which based on comparing Phenylketonuria, the DNA obtained by RAPD-PCR showed a specific DNA bands which may be responsible for Phenylketonuria phenylalanine, RAPD, PCR and may be used for identification of disease at earlier time of injury. The excess of phenylalanine in diet lead to neural tissue damage and may cause mutation combined with the induced PKU (Phenylketonuria).
    [Show full text]
  • Relating Metatranscriptomic Profiles to the Micropollutant
    1 Relating Metatranscriptomic Profiles to the 2 Micropollutant Biotransformation Potential of 3 Complex Microbial Communities 4 5 Supporting Information 6 7 Stefan Achermann,1,2 Cresten B. Mansfeldt,1 Marcel Müller,1,3 David R. Johnson,1 Kathrin 8 Fenner*,1,2,4 9 1Eawag, Swiss Federal Institute of Aquatic Science and Technology, 8600 Dübendorf, 10 Switzerland. 2Institute of Biogeochemistry and Pollutant Dynamics, ETH Zürich, 8092 11 Zürich, Switzerland. 3Institute of Atmospheric and Climate Science, ETH Zürich, 8092 12 Zürich, Switzerland. 4Department of Chemistry, University of Zürich, 8057 Zürich, 13 Switzerland. 14 *Corresponding author (email: [email protected] ) 15 S.A and C.B.M contributed equally to this work. 16 17 18 19 20 21 This supporting information (SI) is organized in 4 sections (S1-S4) with a total of 10 pages and 22 comprises 7 figures (Figure S1-S7) and 4 tables (Table S1-S4). 23 24 25 S1 26 S1 Data normalization 27 28 29 30 Figure S1. Relative fractions of gene transcripts originating from eukaryotes and bacteria. 31 32 33 Table S1. Relative standard deviation (RSD) for commonly used reference genes across all 34 samples (n=12). EC number mean fraction bacteria (%) RSD (%) RSD bacteria (%) RSD eukaryotes (%) 2.7.7.6 (RNAP) 80 16 6 nda 5.99.1.2 (DNA topoisomerase) 90 11 9 nda 5.99.1.3 (DNA gyrase) 92 16 10 nda 1.2.1.12 (GAPDH) 37 39 6 32 35 and indicates not determined. 36 37 38 39 S2 40 S2 Nitrile hydration 41 42 43 44 Figure S2: Pearson correlation coefficients r for rate constants of bromoxynil and acetamiprid with 45 gene transcripts of ECs describing nucleophilic reactions of water with nitriles.
    [Show full text]
  • The X-Ray Crystal Structure of the Human Mono-Oxygenase Cytochrome P450 3A5-Ritonavir Complex Reveals Active Site Differences Between P450s 3A4 and 3A5 S
    Supplemental material to this article can be found at: http://molpharm.aspetjournals.org/content/suppl/2017/11/01/mol.117.109744.DC1 1521-0111/93/1/14–24$25.00 https://doi.org/10.1124/mol.117.109744 MOLECULAR PHARMACOLOGY Mol Pharmacol 93:14–24, January 2018 Copyright ª 2017 by The American Society for Pharmacology and Experimental Therapeutics The X-Ray Crystal Structure of the Human Mono-Oxygenase Cytochrome P450 3A5-Ritonavir Complex Reveals Active Site Differences between P450s 3A4 and 3A5 s Mei-Hui Hsu, Uzen Savas, and Eric F. Johnson Department of Molecular Medicine, The Scripps Research Institute, La Jolla, California Received June 26, 2017; accepted October 25, 2017 Downloaded from ABSTRACT The contributions of cytochrome P450 3A5 to the metabolic secondary and tertiary structures of 3A5 and 3A4 are similar, but clearance of marketed drugs is unclear, but its probable role is to the architectures of their active sites differ. The 3A5 active site is augment the metabolism of several drugs that are largely cleared taller and narrower than that of 3A4. As a result, ritonavir adopts a by P450 3A4. Selective metabolism by 3A4 is often a concern in distinctly different conformation to fit into the cavity of 3A5 than drug development owing to potential drug-drug interactions and seen for 3A4. These structural changes reflect amino acid differ- molpharm.aspetjournals.org the variability of 3A4 and 3A5 expression. The contribution of P450 ences that alter the conformation of the helix F through helix G 3A5 to these clearance pathways varies between individuals region in the upper portion of the cavity and ionic interactions owing to genetic differences and similarities and differences in between residues in the beta-sheet domain that reduce the width the metabolic properties of 3A5 compared with 3A4.
    [Show full text]
  • Structural Basis for Regiospecific Midazolam Oxidation by Human Cytochrome P450 3A4
    Structural basis for regiospecific midazolam oxidation by human cytochrome P450 3A4 Irina F. Sevrioukovaa,1 and Thomas L. Poulosa,b,c aDepartment of Molecular Biology and Biochemistry, University of California, Irvine, CA 92697-3900; bDepartment of Chemistry, University of California, Irvine, CA 92697-3900; and cDepartment of Pharmaceutical Sciences, University of California, Irvine, CA 92697-3900 Edited by Michael A. Marletta, University of California, Berkeley, CA, and approved November 30, 2016 (received for review September 28, 2016) Human cytochrome P450 3A4 (CYP3A4) is a major hepatic and Here we describe the cocrystal structure of CYP3A4 with the intestinal enzyme that oxidizes more than 60% of administered drug midazolam (MDZ) bound in a productive mode. MDZ therapeutics. Knowledge of how CYP3A4 adjusts and reshapes the (Fig. 1) is the benzodiazepine most frequently used for pro- active site to regioselectively oxidize chemically diverse com- cedural sedation (9) and is a marker substrate for the CYP3A pounds is critical for better understanding structure–function rela- family of enzymes that includes CYP3A4 (10). MDZ is hydrox- tions in this important enzyme, improving the outcomes for drug ylated by CYP3A4 predominantly at the C1 position, whereas metabolism predictions, and developing pharmaceuticals that the 4-OH product accumulates at high substrate concentrations have a decreased ability to undergo metabolism and cause detri- (up to 50% of total product) and inhibits the 1-OH metabolic mental drug–drug interactions. However, there is very limited pathway (11–14). The reaction rate and product distribution structural information on CYP3A4–substrate interactions available strongly depend on the assay conditions and can be modulated by to date.
    [Show full text]
  • Structural Basis for the Electron Transfer from an Open Form of NADPH-Cytochrome P450 Oxidoreductase to Heme Oxygenase
    Structural basis for the electron transfer from an open form of NADPH-cytochrome P450 oxidoreductase to heme oxygenase Masakazu Sugishimaa,1, Hideaki Satoa, Yuichiro Higashimotob, Jiro Haradaa, Kei Wadac, Keiichi Fukuyamad, and Masato Noguchia,1 aDepartment of Medical Biochemistry and bDepartment of Chemistry, Kurume University School of Medicine, Kurume, Fukuoka 830-0011, Japan; cOrganization for Promotion of Tenure Track, University of Miyazaki, Kiyotake, Miyazaki 889-1692, Japan; and dDepartment of Applied Chemistry, Graduate School of Engineering, Osaka University, Suita, Osaka 565-0871, Japan Edited by Thomas C. Pochapsky, Brandeis University, Waltham, MA, and accepted by the Editorial Board January 14, 2014 (received for review November 26, 2013) NADPH-cytochrome P450 oxidoreductase (CPR) supplies electrons hardly conceivable that the redox partners such as cytochrome to various heme proteins including heme oxygenase (HO), which is P450 could approach closely enough to FMN for intermolecular a key enzyme for heme degradation. Electrons from NADPH flow electron transfer to occur. Site-directed mutagenesis analysis first to flavin adenine dinucleotide, then to flavin mononucleotide demonstrated that the acidic clusters (Asp207-Asp208-Asp209 (FMN), and finally to heme in the redox partner. For electron trans- and Glu213-Glu214-Asp215 in rat CPR) near FMN are involved fer from CPR to its redox partner, the ‘‘closed-open transition’’ of in the association with cytochrome P450s (4), but these clusters CPR is indispensable. Here, we demonstrate that a hinge-short- were covered by the FAD domain in the closed conformation ened CPR variant, which favors an open conformation, makes a sta- (3). Therefore, a large conformational change in CPR is expec- ble complex with heme–HO-1 and can support the HO reaction, ted to take place for smooth interaction between CPR and its although its efficiency is extremely limited.
    [Show full text]
  • WO 2013/158938 Al 24 October 2013 (24.10.2013) P O P CT
    (12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property Organization I International Bureau (10) International Publication Number (43) International Publication Date WO 2013/158938 Al 24 October 2013 (24.10.2013) P O P CT (51) International Patent Classification: 94080 (US). BHAT, Riyaz; 225 Gateway Boulevard, CUP 7/64 (2006.01) South San Francisco, California 94080 (US). ZHAO, Xin¬ hua; 225 Gateway Boulevard, South San Francisco, Cali (21) International Application Number: fornia 94080 (US). BOND, Risha; 225 Gateway PCT/US20 13/037261 Boulevard, South San Francisco, California 94080 (US). (22) International Filing Date: RAKITSKY, Walter; 225 Gateway Boulevard, South San 18 April 2013 (18.04.2013) Francisco, California 94080 (US). MARANGONI, Ale¬ jandro; 225 Gateway Boulevard, South San Francisco, (25) Filing Language: English California 94080 (US). BRAKSMAYER, Diza; 225 Gate (26) Publication Language: English way Boulevard, South San Francisco, California 94080 (US). (30) Priority Data: 61/635,285 18 April 2012 (18.04.2012) US (74) Agents: TERMES, Lance et al; Alston & Bird LLP, 100 61/639,838 27 April 2012 (27.04.2012) us South Tyron Street, Bank of America Plaza, Suite 4000, 61/655,469 4 June 2012 (04.06.2012) us Charlotte, North Carolina 28280-4000 (US). 61/672,196 2012 (16.07.2012) 16 July us (81) Designated States (unless otherwise indicated, for every 61/679,026 2 August 2012 (02.08.2012) us kind of national protection available): AE, AG, AL, AM, 61/715,998 19 October 2012 (19. 10.2012) us AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, 61/769,678 26 February 2013 (26.02.2013) us BZ, CA, CH, CL, CN, CO, CR, CU, CZ, DE, DK, DM, 61/778,963 13 March 2013 (13.03.2013) us DO, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, 61/809,213 5 April 2013 (05.04.2013) us HN, HR, HU, ID, IL, IN, IS, JP, KE, KG, KM, KN, KP, (71) Applicant: SOLAZYME, INC.
    [Show full text]
  • Working Party of National Coordinators of the Test Guidelines Programme
    Organisation for Economic Co-operation and Development ENV/CBC/TG(2021)31 For Official Use English - Or. English 7 May 2021 ENVIRONMENT DIRECTORATE CHEMICALS AND BIOTECHNOLOGY COMMITTEE Working Party of National Coordinators of the Test Guidelines Programme Adverse Outcome Pathway 220 on Cyp2E1 Activation Leading to Liver Cancer Nathalie Delrue, Administrator [email protected] JT03476102 OFDE This document, as well as any data and map included herein, are without prejudice to the status of or sovereignty over any territory, to the delimitation of international frontiers and boundaries and to the name of any territory, city or area. 2 ENV/CBC/TG(2021)31 Foreword This Adverse Outcome Pathway (AOP) on Cyp2E1 Activation Leading to Liver Cancer, has been developed under the auspices of the OECD AOP Development Programme, overseen by the Extended Advisory Group on Molecular Screening and Toxicogenomics (EAGMST), which is an advisory group under the Working Party of the National Coordinators of the Test Guidelines Programme (WNT). The AOP has been reviewed internally by the EAGMST, externally by experts nominated by the WNT, and has been endorsed by the WNT and the Working Party on Hazard Assessment (WPHA) on XXX. Through endorsement of this AOP, the WNT and the WPHA express confidence in the scientific review process that the AOP has undergone and accept the recommendation of the EAGMST that the AOP be disseminated publicly. Endorsement does not necessarily indicate that the AOP is now considered a tool for direct regulatory application. The OECD's Chemicals and Biotechnology Committee agreed to declassification of this AOP on XXX.
    [Show full text]