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TRYPTOPHANYL TRANSF'er RNA SYNTHETASE and EXPRESSION of the TRYPTOPHAN OPERON in the Trp8 MUTANTS of ESCHERICHIA COLI I N Additi
TRYPTOPHANYL TRANSF'ER RNA SYNTHETASE AND EXPRESSION OF THE TRYPTOPHAN OPERON IN THE trp8 MUTANTS OF ESCHERICHIA COLI KOREAKI ITO, SOTA HIRAGA AND TAKASHI WRA Institute for Virus Research, Kyoto Uniuersity, Kyoto, Japan Received November 15, 1968 INaddition to their role in protein synthesis, aminoacyl tRNA synthetases in bacteria have been shown to have a regulatory role in the formation of enzymes involved in the synthesis of corresponding amino acids. This has been demon- strated at least in the case of isoleucine and valine (EIDLICand NEIDHARDT1965; YANIV,JACOB and GROS1965) and histidine (SCHLESINGERand MAGASANIK1964; ROTH, ANTONand HARTMAN1966; ROTH and AMES1966). Transfer RNA (tRNA) also appears to be involved in the regulation of these systems (FREUND- LICH 1967; SILBERT,FINK and AMES1966). The expression of the genes determining the tryptophan biosynthetic enzymes, the tryptophan operon in Escherichia coli, is subject to repression by excess L-tryptophan, the end product of the pathway. The mutants of a regulatory gene, trpR, located outside of this operon, exhibit constitutive synthesis of the biosyn- thetic enzymes (COHENand JACOB1959). In addition, we have previously reported tryptophan auxotrophic mutants in which the known structural genes of the tryptophan operon remain intact. One of these mutants, designated trpiS5, has been mapped near strA on the chromosome (HIRAGA,ITO, HAMADA and YURA 1967a). In the present paper, we report on further characterization of the trpS5 mutant and discuss the possible role of the trpS gene in the regulation of the tryptophan operon. The results suggest that trpS, located between strA and maZA, is the structural gene for tryptophanyl tRNA synthetase, and that tryptophanyl tRNA synthetase is somehow involved in the repression of the tryptophan operon. -
Appendix H EPA Hazardous Waste Law
Appendix H EPA Hazardous Waste Law This Appendix is intended to give you background information on hazardous waste laws and how they apply to you. For most U.S. Environmental Protection Agency (EPA) requirements that apply to the University, the Safety Department maintains compliance through internal inspections, record keeping and proper disposal. In Wisconsin, the Department of Natural Resources (DNR) has adopted the EPA regulations, consequently EPA and DNR regulations are nearly identical. EPA defines This Appendix only deals with "hazardous waste" as defined by the EPA. hazardous waste as Legally, EPA defines hazardous waste as certain hazardous chemical waste. This hazardous chemical Appendix does not address other types of regulated laboratory wastes, such as waste; radioactive, infectious, biological, radioactive or sharps. Chapter 8 descibes disposal procedures infectious and biohazardous waste for animals. Chapter 9 describes disposal procedures for sharps and other waste that are regulated by can puncture tissue. Chapter 11 discusses Radiation and the Radiation Safety for other agencies. Radiation Workers provides guidelines for the disposal of radioactive waste. Procedures for medical waste are written by the UW Hospital Safety Officer. The Office of Biological Safety can provide guidance for the disposal of infectious and biological waste. EPA regulations focus on industrial waste streams. As a result, many laboratory chemical wastes are not regulated by EPA as hazardous chemical waste. However, many unregulated chemical wastes do merit special handling and disposal If a waste can be procedures. Thus, Chapter 7 and Appendix A of this Guide recommend disposal defined as: procedures for many unregulated wastes as if they were EPA hazardous waste. -
APPENDIX G Acid Dissociation Constants
harxxxxx_App-G.qxd 3/8/10 1:34 PM Page AP11 APPENDIX G Acid Dissociation Constants § ϭ 0.1 M 0 ؍ (Ionic strength ( † ‡ † Name Structure* pKa Ka pKa ϫ Ϫ5 Acetic acid CH3CO2H 4.756 1.75 10 4.56 (ethanoic acid) N ϩ H3 ϫ Ϫ3 Alanine CHCH3 2.344 (CO2H) 4.53 10 2.33 ϫ Ϫ10 9.868 (NH3) 1.36 10 9.71 CO2H ϩ Ϫ5 Aminobenzene NH3 4.601 2.51 ϫ 10 4.64 (aniline) ϪO SNϩ Ϫ4 4-Aminobenzenesulfonic acid 3 H3 3.232 5.86 ϫ 10 3.01 (sulfanilic acid) ϩ NH3 ϫ Ϫ3 2-Aminobenzoic acid 2.08 (CO2H) 8.3 10 2.01 ϫ Ϫ5 (anthranilic acid) 4.96 (NH3) 1.10 10 4.78 CO2H ϩ 2-Aminoethanethiol HSCH2CH2NH3 —— 8.21 (SH) (2-mercaptoethylamine) —— 10.73 (NH3) ϩ ϫ Ϫ10 2-Aminoethanol HOCH2CH2NH3 9.498 3.18 10 9.52 (ethanolamine) O H ϫ Ϫ5 4.70 (NH3) (20°) 2.0 10 4.74 2-Aminophenol Ϫ 9.97 (OH) (20°) 1.05 ϫ 10 10 9.87 ϩ NH3 ϩ ϫ Ϫ10 Ammonia NH4 9.245 5.69 10 9.26 N ϩ H3 N ϩ H2 ϫ Ϫ2 1.823 (CO2H) 1.50 10 2.03 CHCH CH CH NHC ϫ Ϫ9 Arginine 2 2 2 8.991 (NH3) 1.02 10 9.00 NH —— (NH2) —— (12.1) CO2H 2 O Ϫ 2.24 5.8 ϫ 10 3 2.15 Ϫ Arsenic acid HO As OH 6.96 1.10 ϫ 10 7 6.65 Ϫ (hydrogen arsenate) (11.50) 3.2 ϫ 10 12 (11.18) OH ϫ Ϫ10 Arsenious acid As(OH)3 9.29 5.1 10 9.14 (hydrogen arsenite) N ϩ O H3 Asparagine CHCH2CNH2 —— —— 2.16 (CO2H) —— —— 8.73 (NH3) CO2H *Each acid is written in its protonated form. -
S42003-019-0587-Z.Pdf
Corrected: Author Correction ARTICLE https://doi.org/10.1038/s42003-019-0587-z OPEN High-resolution crystal structure of human asparagine synthetase enables analysis of inhibitor binding and selectivity Wen Zhu 1,10, Ashish Radadiya 1, Claudine Bisson2,10, Sabine Wenzel 3, Brian E. Nordin4,11, 1234567890():,; Francisco Martínez-Márquez3, Tsuyoshi Imasaki 3,5, Svetlana E. Sedelnikova2, Adriana Coricello 1,6,7, Patrick Baumann 1, Alexandria H. Berry8, Tyzoon K. Nomanbhoy4, John W. Kozarich 4, Yi Jin 1, David W. Rice 2, Yuichiro Takagi 3 & Nigel G.J. Richards 1,9 Expression of human asparagine synthetase (ASNS) promotes metastatic progression and tumor cell invasiveness in colorectal and breast cancer, presumably by altering cellular levels of L-asparagine. Human ASNS is therefore emerging as a bona fide drug target for cancer therapy. Here we show that a slow-onset, tight binding inhibitor, which exhibits nanomolar affinity for human ASNS in vitro, exhibits excellent selectivity at 10 μM concentration in HCT- 116 cell lysates with almost no off-target binding. The high-resolution (1.85 Å) crystal structure of human ASNS has enabled us to identify a cluster of negatively charged side chains in the synthetase domain that plays a key role in inhibitor binding. Comparing this structure with those of evolutionarily related AMP-forming enzymes provides insights into intermolecular interactions that give rise to the observed binding selectivity. Our findings demonstrate the feasibility of developing second generation human ASNS inhibitors as lead compounds for the discovery of drugs against metastasis. 1 School of Chemistry, Cardiff University, Cardiff, UK. 2 Department of Molecular Biology and Biotechnology, University of Sheffield, Sheffield, UK. -
Novel Derivatives of Nicotinamide Adenine Dinucleotide (NAD) and Their Biological Evaluation Against NAD- Consuming Enzymes
Novel derivatives of nicotinamide adenine dinucleotide (NAD) and their biological evaluation against NAD- Consuming Enzymes Giulia Pergolizzi University of East Anglia School of Pharmacy Thesis submitted for the degree of Doctor of Philosophy July, 2012 © This copy of the thesis has been supplied on condition that anyone who consults it is understood to recognise that its copyright rests with the author and that use of any information derived there from must be in accordance with current UK Copyright Law. In addition, any quotation or extract must include full attribution. ABSTRACT Nicotinamide adenine dinucleotide (β-NAD+) is a primary metabolite involved in fundamental biological processes. Its molecular structure with characteristic functional groups, such as the quaternary nitrogen of the nicotinamide ring, and the two high- energy pyrophosphate and nicotinamide N-glycosidic bonds, allows it to undergo different reactions depending on the reactive moiety. Well known as a redox substrate owing to the redox properties of the nicotinamide ring, β-NAD+ is also fundamental as a substrate of NAD+-consuming enzymes that cleave either high-energy bonds to catalyse their reactions. In this study, a panel of novel adenine-modified NAD+ derivatives was synthesized and biologically evaluated against different NAD+-consuming enzymes. The synthesis of NAD+ derivatives, modified in position 2, 6 or 8 of the adenine ring with aryl/heteroaryl groups, was accomplished by Suzuki-Miyaura cross-couplings. Their biological activity as inhibitors and/or non-natural substrates was assessed against a selected range of NAD+-consuming enzymes. The fluorescence of 8-aryl/heteroaryl NAD+ derivatives allowed their use as biochemical probes for the development of continuous biochemical assays to monitor NAD+-consuming enzyme activities. -
Hydrolysis and Atmospheric Oxidation Reactions of Perfluorinated Carboxylic Acid Precursors
Hydrolysis and Atmospheric Oxidation Reactions of Perfluorinated Carboxylic Acid Precursors by Derek A. Jackson A thesis submitted in conformity with the requirements for the degree of Doctor of Philosophy Graduate Department of Chemistry University of Toronto © Copyright by Derek A. Jackson 2013 Hydrolysis and Atmospheric Oxidation Reactions of Perfluorinated Carboxylic Acid Precursors Derek Andrew Jackson Doctor of Philosophy Department of Chemistry University of Toronto 2013 Abstract This dissertation explores a number of different environmentally relevant reactions that lead to the production of perfluorocarboxylic acids (PFCAs), a family of environmental pollutants that does not undergo any further degradation pathways. The compound perfluoro-2-methyl-3-pentanone (PFMP) is a new fire fighting fluid developed by 3M that is designed as a Halon replacement. The environment fate of PFMP with regards to direct photolysis, abiotic hydrolysis and hydration was determined using a combination of laboratory experiments and computational modeling. PFMP was found to undergo direct photolysis giving a lifetime of 4-14 days depending on latitude and time of year. Offline samples confirmed PFCA products and a mechanism was proposed. Polyfluorinated amides (PFAMs) are a class of chemicals produced as byproducts of polyfluorinated sulfonamide synthesis via electrochemical fluorination (ECF). Using synthesized standards of four model compounds, PFAMs were detected and quantified in a variety of legacy commercial materials synthesized by ECF. PFAMs were hypothesized to undergo biological hydrolysis reactions, suggesting their importance as historical PFOA precursors. ii The PFAMs were also investigated with regards to their environmental fate upon atmospheric oxidation. Using a smog chamber, the kinetics and degradation mechanisms of N- ethylperfluorobutyramide (EtFBA) were elucidated. -
United States Patent 19 11 4,289,699 Oba Et Al
United States Patent 19 11 4,289,699 Oba et al. 45 Sep. 15, 1981 54 PROCESS FOR THE PRODUCTION OF Primary Examiner-Donald G. Daus N-(HYDROXYPHENYL) MALEIMEDES Assistant Examiner-D. B. Springer 75 Inventors: Masayuki Oba; Motoo Kawamata; Attorney, Agent, or Firm-Fisher, Christen & Sabol Hikotada Tsuboi; Nobuhito Koga, all 57 ABSTRACT of Yokohama, Japan N-(hydroxyphenyl) maleimides of the general formula 73 Assignee: Mitsui Toatsu Chemicals, Incorporated, Tokyo, Japan 21 Appl. No.: 88,825 (22 Filed: Oct. 26, 1979 N CO-m-CH Related U.S. Application Data (HO) 62) Division of Ser. No. 956,971, Nov. 2, 1978, Pat. No. 4,231,934. where R' stands for H, CH3, C2H5, F, Cl, Br or I and in 30 Foreign Application Priority Data is an integer of 1-5 are produced by treating the corre sponding maleamic acid or by treating the ester of said Nov. 2, 1977 (JP) Japan ................................ 52-130905 N-(hydroxyphenyl) maleimide at a temperature of Nov. 4, 1977 (JP Japan ................................ 52-3504 0-150° C. in the presence of at least one dehydrating 51) Int. Cl. .......................................... C07D 207/.452 agent selected for the group consisting of oxides and (52) U.S. Cl. .......................................... 260/326.5 FM oxyacids of sulfur or phosphorus and alkali metal and 58) Field of Search .............................. 260/326.5 FM alkaline earth metal salts of the said oxyacids. The cor responding maleamic acid can be obtained by reacting 56) - References Cited an aminophenol having one or more hydroxyl groups U.S. PATENT DOCUMENTS on its phenyl nucleus with maleic anhydride. -
An Interstellar Synthesis of Phosphorus Oxoacids
ARTICLE DOI: 10.1038/s41467-018-06415-7 OPEN An interstellar synthesis of phosphorus oxoacids Andrew M. Turner1,2, Alexandre Bergantini 1,2, Matthew J. Abplanalp1,2, Cheng Zhu 1,2, Sándor Góbi 1,2, Bing-Jian Sun3, Kang-Heng Chao3, Agnes H.H. Chang3, Cornelia Meinert 4 & Ralf I. Kaiser 1,2 Phosphorus signifies an essential element in molecular biology, yet given the limited solubility of phosphates on early Earth, alternative sources like meteoritic phosphides have been proposed to incorporate phosphorus into biomolecules under prebiotic terrestrial conditions. Here, we report on a previously overlooked source of prebiotic phosphorus from interstellar 1234567890():,; phosphine (PH3) that produces key phosphorus oxoacids—phosphoric acid (H3PO4), phos- phonic acid (H3PO3), and pyrophosphoric acid (H4P2O7)—in interstellar analog ices exposed to ionizing radiation at temperatures as low as 5 K. Since the processed material of molecular clouds eventually enters circumstellar disks and is partially incorporated into planetesimals like proto Earth, an understanding of the facile synthesis of oxoacids is essential to untangle the origin of water-soluble prebiotic phosphorus compounds and how they might have been incorporated into organisms not only on Earth, but potentially in our universe as well. 1 Department of Chemistry, University of Hawaii at Manoa, Honolulu, HI 96822, USA. 2 W.M. Keck Laboratory in Astrochemistry, University of Hawaii at Manoa, Honolulu, HI 96822, USA. 3 Department of Chemistry, National Dong Hwa University, Shoufeng, 974 Hualien, Taiwan. 4 Université Côte d’Azur, CNRS, Institut de Chimie de Nice, Nice, France. Correspondence and requests for materials should be addressed to R.I.K. -
Scientific Opinion on the Safety Evaluation of the Active Substances
EFSA Journal 2013;11(6):3245 SCIENTIFIC OPINION Scientific Opinion on the safety evaluation of the active substances, iron, polyethyleneglycol, disodium pyrophosphate, monosodium phosphate and sodium chloride for use in food contact materials1 EFSA Panel on Food Contact Materials, Enzymes, Flavourings and Processing Aids (CEF)2, 3 European Food Safety Authority (EFSA), Parma, Italy ABSTRACT This scientific opinion of EFSA Panel on Food Contact Materials, Enzymes, Flavourings and Processing Aids deals with the safety evaluation of an iron based oxygen absorber, comprising polyethyleneglycol, disodium pyrophosphate, monosodium phosphate and sodium chloride. This mixture is incorporated in polyethylene (PE) or polypropylene (PP) articles intended to be in contact with foodstuffs for hot fill/pasteurisation and/or long term storage at room temperature. For dried and fatty foods, direct contact with the materials is envisaged whereas other food types will be separated from the active material by a layer that does not contain the active components. All the substances constituting the oxygen absorber system have been evaluated and authorised for use as plastic food contact materials, as food additives or as food supplements. Based on migration results, the specific migration limits for iron, polyethyleneglycol, pyrophosphoric acid salts, phosphoric acid salts and sodium chloride, and the tolerable intake of phosphorus (phosphate) are not expected to be exceeded when the oxygen absorber system is used under the intended conditions of use, notably -
Isolation of a Pyrophosphoryl Form of Pyruvate, Phosphate Dikinase from Propionibacteria* (Phosphoenolpyruvate/ATP/Enzyme) YORAM MILNER and HARLAND G
Proc. Nat. Acad. Sci. USA Vol. 69, No. 9, pp. 2463-2468, September 1972 Isolation of a Pyrophosphoryl Form of Pyruvate, Phosphate Dikinase from Propionibacteria* (phosphoenolpyruvate/ATP/enzyme) YORAM MILNER AND HARLAND G. WOOD Department of Biochemistry, Case Western Reserve University School of Medicine, Cleveland, Ohio 44106 Contributed by Harland G. Wood, June 15, 1972 ABSTRACT Pyruvate, phosphate dikinase from Pro- Enzyme-P + pyruvate ;. enzyme + P-enolpyruvate [ic] pionibacterium shermanii catalyzes the formation of P- enolpyruvate, AMP, and inorganic pyrophosphate from P-enolpyruvate synthase of Escherichia coli, which catalyzes pyruvate, ATP, and orthophosphate; the mechanism reaction 2, likewise uses a mechanism that does not involve a involves three partial reactions and three forms of the enzyme: pyrophosphoryl-enzyme, phosphoryl-enzyme, stable, free diphosphoryl-enzyme (3). and free enzyme. The phosphoryl-enzyme was prepared by -- AMP Pi incubation with P-enolpyruvate and isolated by gel- ATP + pyruvate P-enolpyruvate + + [2] chromatography. The phosphoryl-enzyme was converted These observations led us to reinvestigate the validity of the to 32P31P-enzyme and [32P]Pi by incubation with [32P]PPi; 1 mol of pyrophosphoryl-enzyme was formed per mol of Evans-Wood mechanism. Steady-state and exchange kinetics enzyme of molecular weight 150,000. The labeled enzyme have shown clearly that the pyruvate, phosphate dikinase of released its radioactivity upon incubation with Pi or propionibacteria proceeds via a tri(uni,uni) ping-pong se- AMP to produce the expected [33PJPPi or [y-y3P]ATP, re- quence, as expected from the Evans-Wood mechanism (4). We spectively. Hydrolysis of the pyrophosphoryl-enzyme with here the isolation of the form of the dilute acid yielded PPi. -
Quantification of Pyrophosphate in Soil Solution by Pyrophosphatase
Soil Biology & Biochemistry 74 (2014) 95e97 Contents lists available at ScienceDirect Soil Biology & Biochemistry journal homepage: www.elsevier.com/locate/soilbio Short communication Quantification of pyrophosphate in soil solution by pyrophosphatase hydrolysis Kasper Reitzel a,*, Benjamin L. Turner b a Department of Biology, University of Southern Denmark, Campusvej 55, 5230 Odense M, Denmark b Smithsonian Tropical Research Institute, Apartado 0843-03092, Balboa, Ancon, Panama article info abstract Article history: A commercial pyrophosphatase from Saccharomyces cerevisiae selectively hydrolyzed sodium pyro- Received 20 December 2013 phosphate, but showed no significant activity towards a range of other organic and condensed inorganic Received in revised form phosphorus compounds. Pyrophosphate determined by pyrophosphatase hydrolysis accounted for 8 February 2014 38 Æ 12% (mean Æ standard error of 19 sites) of the non-reactive phosphorus in soil solution obtained by Accepted 3 March 2014 centrifugation from a series of lowland tropical rain forest soils. Pyrophosphate concentrations were up Available online 17 March 2014 À to 89 mgPl 1 and correlated positively with microbial phosphorus, soil solution pH, and native phos- phomonoesterase activity in soil solution, but not with total soil pyrophosphate determined by NaOH Keywords: e 31 Pyrophosphate EDTA extraction and solution P NMR spectroscopy. In summary, we identify pyrophosphate as a major Phosphatase constituent of soil solution phosphorus in lowland tropical rain forests, and demonstrate that a com- Soil solution mercial pyrophosphatase can be used as a selective tool to quantify trace concentrations of pyrophos- Phosphorus phate in soil solution. Tropical forests Ó 2014 Elsevier Ltd. All rights reserved. Pyrophosphate is ubiquitous in soils, where it appears to origi- Bünemann, 2008). -
Downregulation of Carnitine Acyl-Carnitine Translocase by Mirnas
Page 1 of 288 Diabetes 1 Downregulation of Carnitine acyl-carnitine translocase by miRNAs 132 and 212 amplifies glucose-stimulated insulin secretion Mufaddal S. Soni1, Mary E. Rabaglia1, Sushant Bhatnagar1, Jin Shang2, Olga Ilkayeva3, Randall Mynatt4, Yun-Ping Zhou2, Eric E. Schadt6, Nancy A.Thornberry2, Deborah M. Muoio5, Mark P. Keller1 and Alan D. Attie1 From the 1Department of Biochemistry, University of Wisconsin, Madison, Wisconsin; 2Department of Metabolic Disorders-Diabetes, Merck Research Laboratories, Rahway, New Jersey; 3Sarah W. Stedman Nutrition and Metabolism Center, Duke Institute of Molecular Physiology, 5Departments of Medicine and Pharmacology and Cancer Biology, Durham, North Carolina. 4Pennington Biomedical Research Center, Louisiana State University system, Baton Rouge, Louisiana; 6Institute for Genomics and Multiscale Biology, Mount Sinai School of Medicine, New York, New York. Corresponding author Alan D. Attie, 543A Biochemistry Addition, 433 Babcock Drive, Department of Biochemistry, University of Wisconsin-Madison, Madison, Wisconsin, (608) 262-1372 (Ph), (608) 263-9608 (fax), [email protected]. Running Title: Fatty acyl-carnitines enhance insulin secretion Abstract word count: 163 Main text Word count: 3960 Number of tables: 0 Number of figures: 5 Diabetes Publish Ahead of Print, published online June 26, 2014 Diabetes Page 2 of 288 2 ABSTRACT We previously demonstrated that micro-RNAs 132 and 212 are differentially upregulated in response to obesity in two mouse strains that differ in their susceptibility to obesity-induced diabetes. Here we show the overexpression of micro-RNAs 132 and 212 enhances insulin secretion (IS) in response to glucose and other secretagogues including non-fuel stimuli. We determined that carnitine acyl-carnitine translocase (CACT, Slc25a20) is a direct target of these miRNAs.