DOCSLIB.ORG

Cytotoxic and Metabolic Effects of Adenosine and Adenine on Human Lymphoblasts1

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Cytotoxic and Metabolic Effects of Adenosine and Adenine on Human Lymphoblasts1 [CANCER RESEARCH 38, 2357-2362, August 1978] 0008-5472/78/0038-OOOOS02.00 Cytotoxic and Metabolic Effects of Adenosine and Adenine on Human Lymphoblasts1 Floyd F. Snyder,2 Michael S. Hershfield,3 and J. Edwin Seegmiller Department of Medicine, University of California, San Diego, La Jolla, California 92093 ABSTRACT and further studies have shown that cytotoxic concentra tions of adenosine reduce pyrimidine ribonucleotides (12, The metabolic and growth inhibitory effects of adeno- 17, 21, 31) and NAD+ (17, 31) and cause an accumulation of sine toward the human lymphoblast line WI-L2 were po orotic acid (12, 31). Adenosine also increases cellular ade tentiated by the adenosine deaminase inhibitors erythro- nine nucleotides (12, 17, 31), including a transient increase 9-(2-hydroxy-3-nonyl) adenine (EHNA) and coformycin. in cAMP4 concentration (34, 35). In addition to studies in EHNA, 5 nu, or coformycin, 3.5 ¿iM,atconcentrations that cultured cells, increased adenine ribonucleotide pools have inhibited adenosine deaminase activity more than 90%, been found in erythrocytes (1, 25) and lymphocytes (28) of had little effect on cell growth or the metabolic parameters adenosine deaminase-deficient, immune-defective patients. studied. Adenosine, 50 /¿M,plusEHNA, 5 JUM,arrested cell The selective toxicity of adenosine to dividing lymphoid growth in both parent and adenosine kinase-deficient cells, including inhibition of both the response of human lymphoblasts, implicating the nucleoside as the mediator peripheral blood lymphocytes to mitogen and the growth of of the cytostatic effect. Adenosine, 50 /KM,in combination lymphoblastoid cell lines, is considered a possible basis for with the adenosine deaminase inhibitors reduced 14C02 the severe combined immunodeficiency disease associated generation from [1-14C]glucose by 38%, depleted 5-phos- with a hereditary absence of adenosine deaminase activity phoribosy 1-1-pyrophosphate by more than 90%, and re (11,24). duced pyrimidine ribonucleotide concentrations. Uridine, It has been suggested that the toxic effects of adenosine 10 or 100 ftM, reversed adenosine plus EHNA growth are caused by an increased cellular adenine nucleotide inhibition in WI-L2 but not in adenosine kinase mutants. pool, but some evidence has suggested a mechanism(s) of Adenine, 500 //M, which may be converted to the same toxicity that does not require conversion of adenosine or intracellular nucleotides as adenosine, reduced the adenine to nucleotides. Certain analogs of adenosine that growth rate by 50% in both parent and adenine phospho- cannot be phosphorylated are cytotoxic (19, 32), and we ribosyltransferase-deficient lymphoblasts. Although ade have found the toxicity of adenosine to persist in adenosine nine also depleted cells of 5-phosphoribosyl-l-pyrophos- kinase-less mutants of the WI-L2 human splenic lympho phate and reduced pyrimidine ribonucleotide by 50%, the blast line; mutants lacking adenine phosphoribosyltransfer- mechanisms of adenine and adenosine toxicity differ. In ase were as sensitive as was their parent line to growth contrast to the ability of uridine to reverse adenosine inhibition by adenine (16). We now report more detailed cytostasis, growth inhibition by adenine was not reversed studies on the biochemical effects of adenosine and ade by uridine, indicating that pyrimidine ribonucleotide de nine on these lymphoblast lines. In these studies we have pletion is not the primary mechanism of adenine toxicity. used 2 potent inhibitors of adenosine deaminase, coformy cin (26) and EHNA (27). We have compared the effects of INTRODUCTION adenosine and adenine since both purines can be con verted to the same intracellular nucleotides (Chart 1) and The mechanism(s) of cytoxicity for the naturally occurring might therefore be expected to have similar mechanisms of nuceoside, adenosine, and its related base adenine is not toxicity if their effects are related only to expanded adenine understood but remains important, with growing interest in nucleotide pools. the use of adenosine deaminase inhibitors in combination chemotherapy. Both adenosine (9, 10, 18, 30) and adenine (20) block mitogen-induced transformation of human lym MATERIALS AND METHODS phocytes. Adenine toxicity toward mouse L-cells was par Biochemicals. Radiochemicals were purchased from tially overcome by certain pyrimidines (2), and adenosine Amersham/Searle Corp., (Arlington Heights, III.): [8- toxicity was reversed by uridine in normal (12) but not in 14C]adenine, 59 mCi/mmol; [8-14C] adenosine, 59 mCi/ adenosine deaminase-deficient fibroblasts (4). Hilz and mmol; [8-14C]hypoxanthine, 59 mCi/mmol; [1-14C]glucose, Kaukel (17) first documented a decrease in intracellular 9.37 mCi/mmol; [6-14C]glucose, 5.0 mCi/mmol; and sodium DTP concentration in adenosine-inhibited HeLa cells (21), [14C]formate, 59 mCi/mmol. Adenine, adenosine, hypoxan- thine, uridine, and PP-ribose-P (sodium salt) were pur 1This work was supported by Grants AM-1362, AM-5646, and GM-17702 chased from P-L Biochemicals (Milwaukee, Wis.). EHNA from NIH and grants from the National Foundation and the Kroc Foundation. 2 Present address: Division of Pediatrics and Medical Biochemistry, Fac was provided by Wellcome Research Labs (Research Tri- ulty of Medicine, The University of Calgary, Calgary, Alberta T2N 1N4, Canada. To whom requests for reprints should be addressed. 3 Present address: Department of Medicine, Duke University Medical 4The abbreviations used are: cAMP, cyclic adenosine 3':5'-monophos- Center, Durham, N. C. 27710. phate; EHNA, eryr/ii-o-9-(2-hydroxy-3-nonyl) adenine; PP-ribose-P, 5-phos- Received December 13, 1977; accepted May 11,1978. phoribosyl-1 -pyrophosphate. AUGUST 1978 2357 Downloaded from cancerres.aacrjournals.org on September 24, 2021. © 1978 American Association for Cancer Research. F. F. Snyder et al. ATP conditioned to more than 3 months of growth in medium II supplemented with 10% horse serum. This serum was ADP previously shown to deaminate less than 5 /¿molof 50 /XM II adenosine per 25 hr (30). The growth rate of WI-L2 lympho- blasts in this medium was inhibited 50% by approximately EHNA, Cofonnycln 200 /¿Madenosine, and 5000 /UM adenosine completely /If(ADENOSINE « INOSINE ' (AD arrested growth (Chart 2). The adenosine deaminase inhib DAP \ t ? TG A itors EHNA, 5 /iM, and coformycin, 3.5 /¿M(1 pig/ml), ADENINE HYPOXANTHINE inhibited lymphoblast adenosine deaminase activity more Chart 1. Purine interconversion showing sites of selected lymphoblast than 95% in either extracts or whole cells but had little mutation and enzyme inhibition. DAP, 2,6-diaminopurine resistant and effect on growth. WI-L2 grew at approximately 90% of the deficient in adenine phosphoribosyltransferase; MTV, 6-methylthioinosine resistant and deficient in adenosine kinase; TG . 6-thioguanme resistant and normal growth rate for 8 weeks in the presence of 5 /¿M deficient in hypoxanthine-guanine phosphoribosyltransferase; EHNA and EHNA with weekly subculturing and addition of EHNA. coformycin, inhibitors of adenosine deaminase; AMPS, adenylosuccinate. EHNA increased the sensitivity of WI-L2 lymphoblasts to growth inhibition by adenosine greater than 10-fold, such angle Park, N. C.), and coformycin was provided by Dr. H. that 100 ^M adenosine completely arrested growth in the Umezawa, Institute for Microbial Chemistry, Tokyo, Japan. presence of 5 /¿MEHNA (Chart 2). Complete growth inhibi Lymphoblasts. The human splenic lymphoblast line Wl- tion by 50 t¿Madenosine and 5 /¿MEHNA occurred after 24 L2 (23) was grown in suspension culture supplemented with hr of exposure or approximately 1 cell doubling (Chart 3/4) 2 nriM glutamine and 10% horse or fetal calf serum (Flow and was reversible for at least 72 hr of culture (Chart 30), Laboratories, Rockville, Md.) as previously described (16, demonstrating retention of cell viability despite growth 31). The isolation and characterization of clonal lympho arrest. The combination of 50 /J.M adenosine and 3.5 /¿M blast lines derived from WI-L2 deficient in adenosine kinase coformycin also arrested growth of WI-L2 lymphoblasts (EC 3.7.1.20) (MTI), both adenosine kinase and hypoxan after 24 hr. thine-guanine phosphoribosyltransferase (EC 2.4.2.8) (MTI- In studies with lymphoblast extracts, we found the appar TG), and adenine phosphoribosyltransferase (EC 2.4.2.7) ent Km's of adenosine kinase and adenosine deaminase for (DAP) were previously described (16). Cell counts were adenosine to be approximately 2 to 4 and 40 to 50 /¿M, measured by a Model ZB, Coulter counter. respectively. The maximal velocity of the deaminase was Metabolic Studies. Adenosine kinase, adenine phospho approximately 10-fold greater than that of the kinase. The ribosyltransferase, and hypoxanthine-guanine phosphori metabolism of extracellular adenosine by lymphoblasts ap bosyltransferase were assayed in lymphoblast extracts as peared to be governed by the characterisitics of these 2 previously described (16, 30). Adenosine metabolism was enzymes. Thus deamination was the principal route of studied in the intact lymphoblast as previously described adenosine metabolism for intact WI-L2 exposed to 80 /¿M (30). Intracellular PP-ribose-P concentrations (15, 16, 31) adenosine; the rates of deamination and phosphorylation and de novo purine synthesis (15) were measured as before. were 1035 and 74 pmol/106 cells/min, respectively. At a cAMP concentrations were measured according to the lower concentration of adenosine, 4 ¿¿Morapproximately method of Wastili
Load more

Recommended publications
  • REDUCTION of PURINE CONTENT in COMMONLY CONSUMED MEAT PRODUCTS THROUGH RINSING and COOKING by Anna Ellington (Under the Directio
    REDUCTION OF PURINE CONTENT IN COMMONLY CONSUMED MEAT PRODUCTS THROUGH RINSING AND COOKING by Anna Ellington (Under the direction of Yen-Con Hung) Abstract The commonly consumed meat products ground beef, ground turkey, and bacon were analyzed for purine content before and after a rinsing treatment. The rinsing treatment involved rinsing the meat samples using a wrist shaker in 5:1 ratio water: sample for 2 or 5 minutes then draining or centrifuging to remove water. The total purine content of 25% fat ground beef significantly decreased (p<0.05) from 8.58 mg/g protein to a range of 5.17-7.26 mg/g protein after rinsing treatments. After rinsing and cooking an even greater decrease was seen ranging from 4.59-6.32 mg/g protein. The total purine content of 7% fat ground beef significantly decreased from 7.80 mg/g protein to a range of 5.07-5.59 mg/g protein after rinsing treatments. A greater reduction was seen after rinsing and cooking in the range of 4.38-5.52 mg/g protein. Ground turkey samples showed no significant changes after rinsing, but significant decreases were seen after rinsing and cooking. Bacon samples showed significant decreases from 6.06 mg/g protein to 4.72 and 4.49 after 2 and 5 minute rinsing and to 4.53 and 4.68 mg/g protein after 2 and 5 minute rinsing and cooking. Overall, this study showed that rinsing foods in water effectively reduces total purine content and subsequent cooking after rinsing results in an even greater reduction of total purine content.
    [Show full text]
  • 35 Disorders of Purine and Pyrimidine Metabolism
    35 Disorders of Purine and Pyrimidine Metabolism Georges van den Berghe, M.- Françoise Vincent, Sandrine Marie 35.1 Inborn Errors of Purine Metabolism – 435 35.1.1 Phosphoribosyl Pyrophosphate Synthetase Superactivity – 435 35.1.2 Adenylosuccinase Deficiency – 436 35.1.3 AICA-Ribosiduria – 437 35.1.4 Muscle AMP Deaminase Deficiency – 437 35.1.5 Adenosine Deaminase Deficiency – 438 35.1.6 Adenosine Deaminase Superactivity – 439 35.1.7 Purine Nucleoside Phosphorylase Deficiency – 440 35.1.8 Xanthine Oxidase Deficiency – 440 35.1.9 Hypoxanthine-Guanine Phosphoribosyltransferase Deficiency – 441 35.1.10 Adenine Phosphoribosyltransferase Deficiency – 442 35.1.11 Deoxyguanosine Kinase Deficiency – 442 35.2 Inborn Errors of Pyrimidine Metabolism – 445 35.2.1 UMP Synthase Deficiency (Hereditary Orotic Aciduria) – 445 35.2.2 Dihydropyrimidine Dehydrogenase Deficiency – 445 35.2.3 Dihydropyrimidinase Deficiency – 446 35.2.4 Ureidopropionase Deficiency – 446 35.2.5 Pyrimidine 5’-Nucleotidase Deficiency – 446 35.2.6 Cytosolic 5’-Nucleotidase Superactivity – 447 35.2.7 Thymidine Phosphorylase Deficiency – 447 35.2.8 Thymidine Kinase Deficiency – 447 References – 447 434 Chapter 35 · Disorders of Purine and Pyrimidine Metabolism Purine Metabolism Purine nucleotides are essential cellular constituents 4 The catabolic pathway starts from GMP, IMP and which intervene in energy transfer, metabolic regula- AMP, and produces uric acid, a poorly soluble tion, and synthesis of DNA and RNA. Purine metabo- compound, which tends to crystallize once its lism can be divided into three pathways: plasma concentration surpasses 6.5–7 mg/dl (0.38– 4 The biosynthetic pathway, often termed de novo, 0.47 mmol/l). starts with the formation of phosphoribosyl pyro- 4 The salvage pathway utilizes the purine bases, gua- phosphate (PRPP) and leads to the synthesis of nine, hypoxanthine and adenine, which are pro- inosine monophosphate (IMP).
    [Show full text]
  • Purine Metabolism in Cultured Endothelial Cells
    PURINE METABOLISM IN MAN-III Biochemical, Immunological, and Cancer Research Edited by Aurelio Rapado Fundacion Jimenez Diaz Madrid, Spain R.W.E. Watts M.R.C. Clinical Research Centre Harrow, England and Chris H.M.M. De Bruyn Department of Human Genetics University of Nijmegen Faculty of Medicine Nijmegen, The Netherlands PLENUM PRESS · NEW YORK AND LONDON Contents of Part Β I. PURINE METABOLISM PATHWAYS AND REGULATION A. De Novo Synthesis; Precursors and Regulation De Novo Purine Synthesis in Cultured Human Fibroblasts 1 R.B. Gordon, L. Thompson, L.A. Johnson, and B.T. Emmerson Comparative Metabolism of a New Antileishmanial Agent, Allopurinol Riboside, in the Parasite and the Host Cell 7 D. J. Nelson, S.W. LaFon, G.B. Elion, J.J. Marr, and R.L. Berens Purine Metabolism in Rat Skeletal Muscle 13 E. R. Tully and T.G. Sheehan Alterations in Purine Metabolism in Cultured Fibroblasts with HGPRT Deficiency and with PRPPP Synthetase Superactivity 19 E. Zoref-Shani and 0. Sperling Purine Metabolism in Cultured Endothelial Cells 25 S. Nees, A.L. Gerbes, B. Willershausen-Zönnchen, and E. Gerlach Determinants of 5-Phosphoribosyl-l-Pyrophosphate (PRPP) Synthesis in Human Fibroblasts 31 K.0, Raivio, Ch. Lazar, H. Krumholz, and M.A. Becker Xanthine Oxidoreductase Inhibition by NADH as a Regulatory Factor of Purine Metabolism 35 M.M. Jezewska and Z.W. Kaminski vii viii CONTENTS OF PART Β Β. Nucleotide Metabolism Human Placental Adenosine Kinase: Purification and Characterization 41 CM. Andres, T.D. Palella, and I.H. Fox Long-Term Effects of Ribose on Adenine Nucleotide Metabolism in Isoproterenol-Stimulated Hearts .
    [Show full text]
  • Monophosphate- Binding Protein Complex with Subsequent Poly(A) RNA Synthesis in Embryonic Chick Cartilage
    Specific nuclear binding of adenosine 3',5'-monophosphate- binding protein complex with subsequent poly(A) RNA synthesis in embryonic chick cartilage. W M Burch Jr, H E Lebovitz J Clin Invest. 1980;66(3):532-542. https://doi.org/10.1172/JCI109885. Research Article We used embryonic chick pelvic cartilage as a model to study the mechanism by which cyclic AMP increases RNA synthesis. Isolated nuclei were incubated with [32P]-8-azidoadenosine 3,5'-monophosphate ([32P]N3cAMP) with no resultant specific nuclear binding. However, in the presence of cytosol proteins, nuclear binding of [32P]N3cAMP was demonstrable that was specific, time dependent, and dependent on a heat-labile cytosol factor. The possible biological significance of the nuclear binding of the cyclic AMP-protein complex was identified by incubating isolating nuclei with either cyclic AMP or cytosol cyclic AMP-binding proteins prepared by batch elution DEAE cellulose chromatography (DEAE peak cytosol protein), or both, in the presence of cold nucleotides and [3H]uridine 5'-triphosphate. Poly(A) RNA production occurred only in nuclei incubated with cyclic AMP and the DEAE peak cytosol protein preparation. Actinomycin D inhibited the incorporation of [3H]uridine 5'-monophosphate into poly(A) RNA. The newly synthesized poly(A) RNA had a sedimentation constant of 23S. Characterization of the cytosol cyclic AMP binding proteins using [32P]N3-cAMP with photoaffinity labeling three major cAMP-binding complexes (41,000, 51,000, and 55,000 daltons). The 51,000 and 55,000 dalton cyclic AMP binding proteins were further purified by DNA-cellulose chromatography. In the presence of cyclic AMP they stimulated poly(A) RNA synthesis in isolated nuclei.
    [Show full text]
  • Inosine Binds to A3 Adenosine Receptors and Stimulates Mast Cell Degranulation
    Inosine binds to A3 adenosine receptors and stimulates mast cell degranulation. X Jin, … , B R Duling, J Linden J Clin Invest. 1997;100(11):2849-2857. https://doi.org/10.1172/JCI119833. Research Article We investigated the mechanism by which inosine, a metabolite of adenosine that accumulates to > 1 mM levels in ischemic tissues, triggers mast cell degranulation. Inosine was found to do the following: (a) compete for [125I]N6- aminobenzyladenosine binding to recombinant rat A3 adenosine receptors (A3AR) with an IC50 of 25+/-6 microM; (b) not bind to A1 or A2A ARs; (c) bind to newly identified A3ARs in guinea pig lung (IC50 = 15+/-4 microM); (d) lower cyclic AMP in HEK-293 cells expressing rat A3ARs (ED50 = 12+/-5 microM); (e) stimulate RBL-2H3 rat mast-like cell degranulation (ED50 = 2.3+/-0.9 microM); and (f) cause mast cell-dependent constriction of hamster cheek pouch arterioles that is attenuated by A3AR blockade. Inosine differs from adenosine in not activating A2AARs that dilate vascular smooth muscle and inhibit mast cell degranulation. The A3 selectivity of inosine may explain why it elicits a monophasic arteriolar constrictor response distinct from the multiphasic dilator/constrictor response to adenosine. Nucleoside accumulation and an increase in the ratio of inosine to adenosine may provide a physiologic stimulus for mast cell degranulation in ischemic or inflamed tissues. Find the latest version: https://jci.me/119833/pdf Inosine Binds to A3 Adenosine Receptors and Stimulates Mast Cell Degranulation Xiaowei Jin,* Rebecca K. Shepherd,‡ Brian R. Duling,‡ and Joel Linden‡§ *Department of Biochemistry, ‡Department of Molecular Physiology and Biological Physics, and §Department of Medicine, University of Virginia Health Sciences Center, Charlottesville, Virginia 22908 Abstract Mast cells are found in the lung where they release media- tors that constrict bronchiolar smooth muscle.
    [Show full text]
  • Adenosine Challenge Information for Patients Your Doctor Has Recommended That You Have an Adenosine Challenge
    Adenosine challenge Information for patients Your doctor has recommended that you have an adenosine challenge. The purpose of this test is to see if you have an accessory pathway called ‘Wolff-Parkinson-White (WPW) syndrome’. What is an accessory pathway? This is an extra electrical connection between the top chambers (atria) and bottom chambers (ventricles) of the heart. This extra electrical connection may allow electrical signals to bypass the normal route in your heart and form a short circuit. This can result in your heart beating abnormally fast for periods of time, which is called supra-ventricular tachycardia (SVT). This is not usually dangerous, but can cause unpleasant symptoms, such as a racing heart (palpitations), dizziness, chest pain, shortness of breath or, rarely, may cause you to collapse. Although the extra connection is present from birth (congenital), symptoms may not develop until later in life. In some cases, WPW syndrome may be life-threatening, particularly if it occurs alongside a type of irregular heartbeat called atrial fibrillation. However, this is rare and treatment can completely remove this risk. page 2 How is an accessory pathway diagnosed? Adenosine is the drug used in this test. It belongs to a group of medicines called anti-arrhythmics. Adenosine blocks electrical signals through the atrio-ventricular (AV) node. This means signals cannot travel from the top to the bottom chambers of the heart for a few seconds, until the drug effects wear off. If an accessory pathway (extra connection) is present, the electrical signals can still travel down to the ventricles, and this will show up on the ECG.
    [Show full text]
  • Adenine-Based Purines and Related Metabolizing Enzymes: Evidence for Their Impact on Tumor Extracellular Vesicle Activities
    cells Review Adenine-Based Purines and Related Metabolizing Enzymes: Evidence for Their Impact on Tumor Extracellular Vesicle Activities Patrizia Di Iorio 1,2 and Renata Ciccarelli 1,2,* 1 Department of Medical, Oral and Biotechnological Sciences, ‘G. D’Annunzio’ University of Chieti-Pescara, 66100 Chieti, Italy; [email protected] 2 Center for Advanced Studies and Technology (CAST), ‘G. D’Annunzio’ University of Chieti-Pescara, 66100 Chieti, Italy * Correspondence: [email protected] Abstract: Extracellular vesicles (EVs), mainly classified as small and large EVs according to their size/origin, contribute as multi-signal messengers to intercellular communications in normal/pathological conditions. EVs are now recognized as critical players in cancer processes by promoting transformation, growth, invasion, and drug-resistance of tumor cells thanks to the release of molecules contained inside them (i.e., nucleic acids, lipids and proteins) into the tumor microenvironment (TME). Interestingly, secre- tion from donor cells and/or uptake of EVs/their content by recipient cells are regulated by extracellular signals present in TME. Among those able to modulate the EV-tumor crosstalk, purines, mainly the adenine-based ones, could be included. Indeed, TME is characterized by high levels of ATP/adenosine and by the presence of enzymes deputed to their turnover. Moreover, ATP/adenosine, interacting with their own receptors, can affect both host and tumor responses. However, studies on whether/how the purinergic system behaves as a modulator of EV biogenesis, release and functions in cancer are still poor. Thus, this review is aimed at collecting data so far obtained to stimulate further research in this regard.
    [Show full text]
  • Biological Activity of Pyrimidine Derivativies: a Review
    Organic and Medicinal Chemistry International Journal ISSN 2474-7610 Review Article Organic & Medicinal Chem IJ Volume 2 Issue 2 - April 2017 Copyright © All rights are reserved by Ajmal R Bhat DOI: 10.19080/OMCIJ.2017.02.555581 Biological Activity of Pyrimidine Derivativies: A Review Ajmal R. Bhat* Department of Chemistry, S. B. B.S. University, India Submission: March 20, 2017; Published: April 03, 2017 *Corresponding author: Ajmal R Bhat, Department of Chemistry, S. B. B.S. University, Jalandhar Punjab-144030, India, Tel: Email: Abstract The Pyrimidine derivativies in the chemistry of biological systems has attracted much attention due to availability in the substructures of therapeutic natural products. As a result of their prominent and remarkable pharmacological activity, pyrimidine derivatives has been found the most prominent structures in nucleic acid. The present review gives brief information about biological activity of annulated pyrimidine derivatives. Keywords: Pyrimidine derivativies; Anti-inflammatory drugs; anticancer activity; Anti-HIV agents; Antihypertensive drugs Introduction moieties which also impart pharmacological properties (Figures 1-6). The wide applicability associated with these heterocycles pharmaceutical chemistry is having most important focus for Progressive and prospective research in the field of and its novel compounds encouraged the chemists to contribute the design and formulation of new and effective drugs and their and synthesis large number of biologically active novel drugs every research work is to develop and prepare pharmaceutical successful application in applied field. The main concern to substances and preparation, which are new, effective and and introduce some efficient methods. original and to overcome with more accuracy over a drug already known.
    [Show full text]
  • Cyclic Nucleotide Phosphodiesterases in Heart and Vessels
    Cyclic nucleotide phosphodiesterases in heart and vessels: A therapeutic perspective Pierre Bobin, Milia Belacel-Ouari, Ibrahim Bedioune, Liang Zhang, Jérôme Leroy, Véronique Leblais, Rodolphe Fischmeister, Grégoire Vandecasteele To cite this version: Pierre Bobin, Milia Belacel-Ouari, Ibrahim Bedioune, Liang Zhang, Jérôme Leroy, et al.. Cyclic nucleotide phosphodiesterases in heart and vessels: A therapeutic perspective. Archives of cardiovascular diseases, Elsevier/French Society of Cardiology, 2016, 109 (6-7), pp.431-443. 10.1016/j.acvd.2016.02.004. hal-02482730 HAL Id: hal-02482730 https://hal.archives-ouvertes.fr/hal-02482730 Submitted on 23 Mar 2020 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Cyclic nucleotide phosphodiesterases in heart and vessels: A therapeutic perspective Abbreviated title: Cyclic nucleotide phosphodiesterases in heart and vessels French title: Phosphodiestérases des nucléotides cycliques dans le cœur et les vaisseaux : une perspective thérapeutique. Pierre Bobin, Milia Belacel-Ouari, Ibrahim Bedioune, Liang Zhang, Jérôme Leroy, Véronique Leblais, Rodolphe Fischmeister*, Grégoire Vandecasteele* UMR-S 1180, INSERM, Université Paris-Sud, Université Paris-Saclay, Châtenay-Malabry, France * Corresponding authors. UMR-S1180, Faculté de Pharmacie, Université Paris-Sud, 5 rue J.-B. Clément, F-92296 Châtenay-Malabry Cedex, France.
    [Show full text]
  • Lethality of Adenosine for Cultured Mammalian Cells by Interference with Pyrimidine Biosynthesis
    J. Cell Set. 13, 429-439 (i973) 429 Printed in Great Britain LETHALITY OF ADENOSINE FOR CULTURED MAMMALIAN CELLS BY INTERFERENCE WITH PYRIMIDINE BIOSYNTHESIS K. ISHII* AND H. GREEN Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts, 02139, U.S.A. SUMMARY Adenosine at low concentration is toxic to mammalian cells in culture. This may escape notice because some sera (such as calf or human) commonly used in culture media, contain adenosine deaminase. In the absence of serum deaminase, adenosine produced inhibition of growth of a number of established cell lines at concentrations as low as 5 x io~* M, and killed at 2 x io~5 M. This effect required the presence of cellular adenosine kinase, since a mutant line deficient in this enzyme was 70-fold less sensitive to adenosine. The toxic substance is therefore derived from adenosine by phosphorylation, and is probably one of the adenosine nucleotides. The toxic effect of adenosine in concentrations up to 2 x io~* M was completely prevented by the addition of uridine or of pyrimidines potentially convertible to uridine, suggesting that the adenosine was interfering with endogenous synthesis of uridylate. In the presence of adenosine, the conversion of labelled aspartate to uridine nucleotides was reduced by 80-85 %> and labelled orotate accumulated in both the cells and in the culture medium. The lethality of adenosine results from inhibition by one of its nucleotide products of the synthesis of uridylate at the stage of phosphoribosylation of orotate. INTRODUCTION Though adenosine is not an intermediate on the endogenous pathway of purine biosynthesis, it can be efficiently utilized through the purine salvage pathways as the sole purine source in cultured mammalian cells whose endogenous purine synthesis is blocked by aminopterin (Green & Ishii, 1972).
    [Show full text]
  • A Previously Undescribed Pathway for Pyrimidine Catabolism
    A previously undescribed pathway for pyrimidine catabolism Kevin D. Loh*†, Prasad Gyaneshwar*‡, Eirene Markenscoff Papadimitriou*§, Rebecca Fong*, Kwang-Seo Kim*, Rebecca Parales¶, Zhongrui Zhouʈ, William Inwood*, and Sydney Kustu*,** *Department of Plant and Microbial Biology, 111 Koshland Hall, University of California, Berkeley, CA 94720-3102; ¶Section of Microbiology, 1 Shields Avenue, University of California, Davis, CA 95616; and ʈCollege of Chemistry, 8 Lewis Hall, University of California, Berkeley, CA 94720-1460 Contributed by Sydney Kustu, January 19, 2006 The b1012 operon of Escherichia coli K-12, which is composed of tive N sources. Here we present evidence that the b1012 operon seven unidentified ORFs, is one of the most highly expressed codes for proteins that constitute a previously undescribed operons under control of nitrogen regulatory protein C. Examina- pathway for pyrimidine degradation and thereby confirm the tion of strains with lesions in this operon on Biolog Phenotype view of Simaga and Kos (8, 9) that E. coli K-12 does not use either MicroArray (PM3) plates and subsequent growth tests indicated of the known pathways. that they failed to use uridine or uracil as the sole nitrogen source and that the parental strain could use them at room temperature Results but not at 37°C. A strain carrying an ntrB(Con) mutation, which Behavior on Biolog Phenotype MicroArray Plates. We tested our elevates transcription of genes under nitrogen regulatory protein parental strain NCM3722 and strains with mini Tn5 insertions in C control, could also grow on thymidine as the sole nitrogen several genes of the b1012 operon on Biolog (Hayward, CA) source, whereas strains with lesions in the b1012 operon could not.
    [Show full text]
  • Inosine in Biology and Disease
    G C A T T A C G G C A T genes Review Inosine in Biology and Disease Sundaramoorthy Srinivasan 1, Adrian Gabriel Torres 1 and Lluís Ribas de Pouplana 1,2,* 1 Institute for Research in Biomedicine, Barcelona Institute of Science and Technology, 08028 Barcelona, Catalonia, Spain; [email protected] (S.S.); [email protected] (A.G.T.) 2 Catalan Institution for Research and Advanced Studies, 08010 Barcelona, Catalonia, Spain * Correspondence: [email protected]; Tel.: +34-934034868; Fax: +34-934034870 Abstract: The nucleoside inosine plays an important role in purine biosynthesis, gene translation, and modulation of the fate of RNAs. The editing of adenosine to inosine is a widespread post- transcriptional modification in transfer RNAs (tRNAs) and messenger RNAs (mRNAs). At the wobble position of tRNA anticodons, inosine profoundly modifies codon recognition, while in mRNA, inosines can modify the sequence of the translated polypeptide or modulate the stability, localization, and splicing of transcripts. Inosine is also found in non-coding and exogenous RNAs, where it plays key structural and functional roles. In addition, molecular inosine is an important secondary metabolite in purine metabolism that also acts as a molecular messenger in cell signaling pathways. Here, we review the functional roles of inosine in biology and their connections to human health. Keywords: inosine; deamination; adenosine deaminase acting on RNAs; RNA modification; translation Citation: Srinivasan, S.; Torres, A.G.; Ribas de Pouplana, L. Inosine in 1. Introduction Biology and Disease. Genes 2021, 12, 600. https://doi.org/10.3390/ Inosine was one of the first nucleobase modifications discovered in nucleic acids, genes12040600 having been identified in 1965 as a component of the first sequenced transfer RNA (tRNA), tRNAAla [1].
    [Show full text]