DOCSLIB.ORG

1 Here Are the Quiz 4 Questions You Will Answer Online Using the Quiz

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
1 Here Are the Quiz 4 Questions You Will Answer Online Using the Quiz Chemistry 6720, quiz 4 handout Here are the quiz 4 questions you will answer online using the quiz tool in canvas instructure. Some of the questions deal with citric acid cycle enzymes that use mechanistic strategies covered in the lectures. Since we did not cover all of those specific enzymes in lecture, be sure to review the citric acid cycle ahead of taking the quiz online so you can answer the appropriate questions regarding mechanistic strategies/themes for the enzymes. In doing so, practice drawing mechanisms for each enzyme of the citric acid cycle. You will probably need to look up the mechanisms for isocitrate dehydrogenase, aconitase, succinate dehydrogenase, and succinyl-CoA synthetase in a general biochemistry text. I will not be asking specific details of the mechanisms for those 4 enzymes, but you will still need to be generally familiar with how they operate to answer a few of the questions below. You do not need to know the chemistry of FAD reduction for the quiz. Be sure to note the quiz deadline in canvas to be sure you get your answers submitted by the deadline. 1. Shown at the right is a Fischer projection for D-glucose. Use the strategy described in the lecture to assign the configurations (R or S) of the chiral centers at C2, C3, C4, and C5. 2. Using the structures shown to the right, assign the methyl groups of isopropanol, and the hydrogens at C2 and C3 of succinate, as either ProR or ProS. Use the strategy covered in lecture to guide you in making the assignments. 3. Which faces of acetaldehyde, acetyl chloride, glycinate, pyruvate, and methylethylketone are shown here? 1 Chemistry 6720, quiz 4 handout 4. Match the following classes of enzymes with the product that will be formed from substrate oxidation by the enzyme. a. primary alcohol dehydrogenase = b. aldehyde dehydrogenase = c. (primary) amine dehydrogenase = d. secondary alcohol dehydrogenase = 5. What is the primary purpose of the catalytic Zn2+ ion in the liver alcohol dehydrogenase mechanism? 6. Which of the following show INCORRECT electron pushing for alcohol dehydrogenase reaction(s)? Check all the options that are INCORRECT. Correct and incorrect electron pushing diagrams will appear in the online quiz. 7. Order the following steps properly, from first to last, for the reaction catalyzed by GAPDH (steps will be provided in the online quiz; review the mechanism ahead of time to prepare). 8. Which of the following dehydrogenases catalyze the dehydrogenation of an alcohol functional group? Select all the correct answers (choices provided on online quiz). 9. Which of the following dehydrogenases catalyze reactions involving oxidative decarboxylation? a. lactate dehydrogenase b. pyruvate dehydrogenase c. malate dehydrogenase d. isocitrate dehydrogenase e. alpha-ketoglutarate dehydrogenase f. succinate dehydrogenase 10. Match the following enzymes with the unique chemistry used for the transformations. a. type I aldolase = b. Type II aldolase = c. Claisen condensation = d. carboxylation with bicarbonate as the substrate 2 Chemistry 6720, quiz 4 handout 11. Classify the following reactions based on the type of enzyme. a. ATP + glucose → glucose-6-phosphate + ADP = b. glucose-6-phosphate → fructose-6-phosphate = c. glucose-6-phosphate → glucose-1-phosphate = d. Glyceraldehyde-3-phosphate → dihydroxyacetone phosphate = e. Glyceraldehyde-3-phosphate → 1,3-diphosphoglycerate = f. 2-phosphoglycerate → phosphoenolpyruvate = 12. The following diagrams depicts the initial step for which types of reaction? 13. Which glycolytic enzymes will form a cis-enediolate during catalysis? 14. Match the indicated enzymes associated with an appropriate answer that relates to the mechanism of the transformation. Note that alcohol dehydrogenase is involved only in anaerobic glycolysis in microorganisms that produce ethanol as the fermentation product. a. alcohol dehydrogenase = b. glyceraldehyde-3-phosphate dehydrogenase = c. triose phophate isomerase = d. type I aldolase = e. phosphoglycerate kinase = 15. Which glycolytic enzymes will form a covalent adduct with the substrate or an intermediate formed during the reaction? 16. During the reaction catalyzed by aldolase, in the direction relevant to the reaction that occurs in glycolysis, what will occur in the next step for each of the intermediates shown below? Intermediate structures will be shown on the online quiz. 3 Chemistry 6720, quiz 4 handout 17. Correctly order the following sequence of events in the reaction catalyzed by triosephosphate isomerase in the direction shown below. Note some of the steps may occur in a concerted fashion, yet these steps can still be placed in the sequence based on which electron pushing step occurs first. Sequence will be shown on the online quiz. 18. Molecules of dihydroxyacetone phosphate (DHAP) were labelled with T in the positions shown below to give the molecules (shown as Fischer projections) labelled (1) and (2). Either (1) or (2) was then mixed with triose phosphate isomerase and allowed to react until equilibrium was reached. At that point, the glyceraldehy-3-phosphate (G3P) product was isolated and the amount of T present determined by liquid scintillation counting. For the reaction with (1), no T was found to be present in G3P. For the reaction with (2), one mol of T was present for each mol G3P formed. What does this tell you about the stereochemistry associated with the mechanism of the reaction? T H HO C T HO 1C H 1 C O 2C O 2 2- CH O PO 2- 3 CH2O PO3 3 2 3 (1) (2) 19. Match the following citric acid cycle enzymes with the type of reaction they catalyze. Enzymes: Aconitase, alpha-ketoglutarate dehydrogenase, citrate synthase, fumarase, isocitrate dehydrogenase, malate dehydrogenase, succinate dehydrogenase, succinyl-CoA synthetase Reaction types: Isomerization, oxidative decarboxylation requiring TPP, claisen condensation, substrate hydration, oxidative decarboxylation not requiring TPP, alcohol dehydrogenation, dehydrogenation of a C-C bond, substrate level phosphorylation 4 Chemistry 6720, quiz 4 handout 20. Match the following citric acid cycle enzymes with the unique chemistry involved in the enzymatic transformation. Enzymes can be used more than once, but there is only one correct match for each feature. Note: some of these were not covered in lecture, so you might have to do some research to answer them. Enzymes: Fumarase, aconitase, succinyl-CoA synthetase, isocitrate dehydrogenase, alpha- ketoglutarate dehydrogenase, citrate synthase Features: net substrate hydration, prochiral arm discrimination for hydroxyl group movement, substrate level phosphorylation, dehydration and rehydration, phosphorolysis = succinyl-CoA synthetase, oxidative decarboxylation not using TPP, oxidative decarboxylation using TPP, isomerization, carbon-carbon bond formation = 21. Match the following roles to the correct cofactors in the reaction catalyzed by pyruvate dehydrogenase. a. Accepts the acetyl group from reduced lipoamide b. Oxidizes FADH2 c. Initial electron acceptor in oxidation of pyruvate d. Attacks and attaches to the central carbon in pyruvate e. Oxidizes the reduced form of lipoic acid 22. During the reaction catalyzed by pyruvate dehydrogenase, what will occur in the next step for the intermediates shown below? Intermediate structures will be provided on the online quiz. 23. According to the figure provided in my lecture notes, what are the cofactor roles of biotin in biotin-dependent carboxylases? 24. and 25. The transformations of the following substrates in the citric acid cycle will be initiated how? 5 .
Load more

Recommended publications
  • Dehydrogenase Gene of Entamoeba Histolytica
    Proc. Nad. Acad. Sci. USA Vol. 89, pp. 10188-10192, November 1992 Medical Sciences Cloning and expression of an NADP+-dependent alcohol dehydrogenase gene of Entamoeba histolytica (amebiasis/protozoan parasite/fermentation/inhibitors) AJIT KUMAR*, PEI-SHEN SHENtt, STEVEN DESCOTEAUX*, JAN POHL§, GORDON BAILEYt, AND JOHN SAMUELSON*1II *Department of Tropical Public Health, Harvard School of Public Health, Boston, MA 02115; tDepartment of Biochemistry, Morehouse School of Medicine, Atlanta, GA 30310; §Microchemical Facility, Emory University, Atlanta, GA 30322; and 'Department of Pathology, New England Deaconess Hospital, Boston, MA 02215 Communicated by Elkan Blout, June 29, 1992 (receivedfor review May 1, 1992) ABSTRACT Ethanol is the major metabolic product of NAD(P)+ so that the reducing equivalents are regenerated glucose fermentation by the protozoan parasite Entmoeba (4-7). Both NADP+-dependent and NAD+-dependent alco- histolytica under the anaerobic conditions found in the lumen hol dehydrogenase (ADH) activities have been identified in of the colon. Here an internal peptide sequence determined E. histolytica trophozoites (4-7). The NADP+-dependent from a major 39-kDa amoeba protein isolated by isoeLtric ADH (EhADHi; EC 1.1.1.2) was found to be composed of focusing followed by SDS/PAGE was used to clone the gene for four -30-kDa subunits, prefer secondary alcohols to primary the E. histolytica NADP+-dependent alcohol dehydrogenase alcohols, and be inhibited by the substrate analogue pyrazole (EhADH1; EC 1.1.1.2). The EhADHi clone had an open (7). The amoeba NAD+-dependent ADH (EC 1.1.1.1) is not reading frame that was 360 amino acids long and encoded a well-characterized (4).
    [Show full text]
  • The Role of Protein Crystallography in Defining the Mechanisms of Biogenesis and Catalysis in Copper Amine Oxidase
    Int. J. Mol. Sci. 2012, 13, 5375-5405; doi:10.3390/ijms13055375 OPEN ACCESS International Journal of Molecular Sciences ISSN 1422-0067 www.mdpi.com/journal/ijms Review The Role of Protein Crystallography in Defining the Mechanisms of Biogenesis and Catalysis in Copper Amine Oxidase Valerie J. Klema and Carrie M. Wilmot * Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota, 321 Church St. SE, Minneapolis, MN 55455, USA; E-Mail: [email protected] * Author to whom correspondence should be addressed; E-Mail: [email protected]; Tel.: +1-612-624-2406; Fax: +1-612-624-5121. Received: 6 April 2012; in revised form: 22 April 2012 / Accepted: 26 April 2012 / Published: 3 May 2012 Abstract: Copper amine oxidases (CAOs) are a ubiquitous group of enzymes that catalyze the conversion of primary amines to aldehydes coupled to the reduction of O2 to H2O2. These enzymes utilize a wide range of substrates from methylamine to polypeptides. Changes in CAO activity are correlated with a variety of human diseases, including diabetes mellitus, Alzheimer’s disease, and inflammatory disorders. CAOs contain a cofactor, 2,4,5-trihydroxyphenylalanine quinone (TPQ), that is required for catalytic activity and synthesized through the post-translational modification of a tyrosine residue within the CAO polypeptide. TPQ generation is a self-processing event only requiring the addition of oxygen and Cu(II) to the apoCAO. Thus, the CAO active site supports two very different reactions: TPQ synthesis, and the two electron oxidation of primary amines. Crystal structures are available from bacterial through to human sources, and have given insight into substrate preference, stereospecificity, and structural changes during biogenesis and catalysis.
    [Show full text]
  • C1 Assimilation in Obligate Methylotrophs and Restricted Facultative Methylotrophs by JOHN COLBY* and LEONARD J
    Biochem. J. (1975) 148, 513-520 513 Printed in Great Britain Enzymological Aspects ofthe Pathways for Trimethylamine Oxidation and C1 Assimilation in Obligate Methylotrophs and Restricted Facultative Methylotrophs By JOHN COLBY* and LEONARD J. ZATMAN Department ofMicrobiology, University ofReading, Reading RG1 5AQ, U.K. (Received8 January 1975) 1. Extracts of trimethylamine-grown W6A and W3A1 (type M restricted facultative methylotrophs) contain trimethylamine dehydrogenase whereas similar extracts of Bacillus PM6 and Bacillus S2A1 (type L restricted facultative methylotrophs) contain trimethylamine mono-oxygenase and trimethylamine N-oxide demethylase but no trimethylamine dehydrogenase. 2. Extracts oftherestricted facultatives and ofthe obligate methylotroph C2A1 contain hexulose phosphate synthase-hexulose phosphate isomerase activity; hydroxypyruvate reductase was not detected. 3. Neither the restricted facultatives nor the obligates 4B6 and C2A1 contain all the enzymes ofthe hexulose phos- phatecycleofformaldehyde assimilation as originallyproposed byKemp & Quayle (1967). 4. Organisms PM6 and S2A1 lack transaldolase and use a modified cycle involving sedoheptulose 1,7-diphosphate and sedoheptulose diphosphatase. 5. The obligates 4B6 and C2A1, and the type M organisms W6A and W3A1, use a different modification of the assimilatory hexulose phosphate cycle involving the Entner-Doudoroff-pathway enzymes phosphogluconate dehydratase and phospho-2-keto-3-deoxygluconate aldolase. Thelackoffructosediphosphatealdolaseandhexosediphosphataseintheseorganismsmay
    [Show full text]
  • Novel Amine Dehydrogenase from Leucine Dehydrogenase 19
    DEVELOPMENT OF AN AMINE DEHYDROGENASE A Dissertation Presented to The Academic Faculty by Michael J. Abrahamson In Partial Fulfillment of the Requirements for the Degree Doctor of Philosophy in the School of Chemical and Biomolecular Engineering Georgia Institute of Technology December 2012 DEVELOPMENT OF AN AMINE DEHYDROGENASE Approved by: Dr. Andreas S. Bommarius, Advisor Dr. Jeffrey Skolnick School of Chemical & Biomolecular School of Biology Engineering Georgia Institute of Technology Georgia Institute of Technology Dr. Christopher W. Jones Dr. John W. Wong School of Chemical & Biomolecular Biocatalysis Center of Emphasis Engineering Chemical Research & Development Georgia Institute of Technology Pfizer Global Research & Development Dr. Yoshiaki Kawajiri School of Chemical & Biomolecular Engineering Georgia Institute of Technology Date Approved: August 13, 2012 To my parents, Joseph & Deborah ACKNOWLEDGEMENTS First and foremost, I would like to thank my parents Joseph and Deborah. Your guidance and unconditional support has been invaluable to my success throughout college. The encouragement from my entire family has been so helpful throughout graduate school. I would also like to thank my advisor, Prof. Andreas Bommarius for his direction, instruction, and patience over the last five years. Your input and optimism has kept me ‘plowing ahead’ and has been instrumental in my scientific development. I would like to thank my committee members; Prof. Christopher Jones, Prof. Yoshiaki Kawajiri, Prof. Jeffrey Skolnick, and Dr. John W. Wong. Thank you all for your time, encouragement, and support. All of the members of the Bommarius lab, past and present, have made my graduate career not only productive, but enjoyable. We have shared many great and unforgettable moments.
    [Show full text]
  • Glutamate-Secretion-In-Cancer-Cell
    Biochemical and Biophysical Research Communications 495 (2018) 761e767 Contents lists available at ScienceDirect Biochemical and Biophysical Research Communications journal homepage: www.elsevier.com/locate/ybbrc Glutamate production from ammonia via glutamate dehydrogenase 2 activity supports cancer cell proliferation under glutamine depletion * Yukiko Takeuchi a, Yasumune Nakayama b, c, Eiichiro Fukusaki b, Yasuhiro Irino a, d, a The Integrated Center for Mass Spectrometry, Kobe University Graduate School of Medicine, 7-5-1 Kusunoki-cho, Kobe 650-0017, Japan b Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamada-oka, Suita, Osaka 565-0871, Japan c Department of Applied Microbial Technology, Faculty of Biotechnology and Life Science, Sojo University, 4-22-1 Ikeda, Kumamoto 860-0082, Japan d Division of Evidence-based Laboratory Medicine, Kobe University Graduate School of Medicine, 7-5-1 Kusunoki-cho, Kobe 650-0017, Japan article info abstract Article history: Cancer cells rapidly consume glutamine as a carbon and nitrogen source to support proliferation, but the Received 9 November 2017 cell number continues to increase exponentially after glutamine is nearly depleted from the medium. In Accepted 13 November 2017 contrast, cell proliferation rates are strongly depressed when cells are cultured in glutamine-free me- Available online 14 November 2017 dium. How cancer cells survive in response to nutrient limitation and cellular stress remains poorly understood. In addition, rapid glutamine catabolism yields ammonia, which is a potentially toxic Keywords: metabolite that is secreted into the extracellular space. Here, we show that ammonia can be utilized for Ammonia glutamate production, leading to cell proliferation under glutamine-depleted conditions.
    [Show full text]
  • Is There a Role for Ethanol Assay Using Alcohol Dehydrogenase in a Basic Drugs and Alcohol Screen of Blood Following Routine Autopsies?
    Journal of Clinical Pathology and Forensic Medicine Vol. 3(2), pp. 17-19, December 2012 Available online at http://www.academicjournals.org/JCPFM DOI: 10.5897/JCPFM12.003 ISSN 2I41-2405 ©2012 Academic Journals Short Communication Is there a role for ethanol assay using alcohol dehydrogenase in a basic drugs and alcohol screen of blood following routine autopsies? William Tormey 1,2 and Tara Moore 3 1Biomedical Sciences, University of Ulster Cromore Road, Coleraine, BT52 1SA, Northern Ireland. 2Department of Chemical Pathology, Beaumont Hospital, Dublin 9, Ireland. 3Biomedical Sciences, University of Ulster, Cromore Road, Coleraine, BT52 1SA, Northern Ireland. Accepted 22 March, 2012 The external examination system of determining the cause of death as operated in Dundee, Scotland is controversial. The addition of a blood or urine specimen for drugs and alcohol processed by hospital autoanalysers will reduce error in death certification. This is even more convenient to organise with a shorter turn-around time than imaging by computed tomography (CT) or magnetic resonance imaging (MRI). These measures may reduce the autopsy rate and ameliorate the distress of families of the deceased. Key words: Autopsy, alcohol, death. INTRODUCTION For over a decade, there has been controversy over the dehydrogenase and the back reaction nicotinamide appropriateness of the high rate of coroner’s autopsies adenine dinucleotide (NAD) to NADH measured (Pounder, 2000; Rutty et al., 2001). Advocacy of the spectrophotometrically at 340 nm on the Olympus 5400 “view and grant” system of issuing the cause of death as autoanalyser for the analysis of alcohol. The data sheet operated in Dundee, Scotland has been challenged and claims that the assay can accurately quantitate alcohol imaging is suggested as a partial answer to lessen the concentrations within the range of 10 to 600 mg/dl.
    [Show full text]
  • How Is Alcohol Metabolized by the Body?
    Overview: How Is Alcohol Metabolized by the Body? Samir Zakhari, Ph.D. Alcohol is eliminated from the body by various metabolic mechanisms. The primary enzymes involved are aldehyde dehydrogenase (ALDH), alcohol dehydrogenase (ADH), cytochrome P450 (CYP2E1), and catalase. Variations in the genes for these enzymes have been found to influence alcohol consumption, alcohol-related tissue damage, and alcohol dependence. The consequences of alcohol metabolism include oxygen deficits (i.e., hypoxia) in the liver; interaction between alcohol metabolism byproducts and other cell components, resulting in the formation of harmful compounds (i.e., adducts); formation of highly reactive oxygen-containing molecules (i.e., reactive oxygen species [ROS]) that can damage other cell components; changes in the ratio of NADH to NAD+ (i.e., the cell’s redox state); tissue damage; fetal damage; impairment of other metabolic processes; cancer; and medication interactions. Several issues related to alcohol metabolism require further research. KEY WORDS: Ethanol-to­ acetaldehyde metabolism; alcohol dehydrogenase (ADH); aldehyde dehydrogenase (ALDH); acetaldehyde; acetate; cytochrome P450 2E1 (CYP2E1); catalase; reactive oxygen species (ROS); blood alcohol concentration (BAC); liver; stomach; brain; fetal alcohol effects; genetics and heredity; ethnic group; hypoxia The alcohol elimination rate varies state of liver cells. Chronic alcohol con- he effects of alcohol (i.e., ethanol) widely (i.e., three-fold) among individ- sumption and alcohol metabolism are on various tissues depend on its uals and is influenced by factors such as strongly linked to several pathological concentration in the blood T chronic alcohol consumption, diet, age, consequences and tissue damage. (blood alcohol concentration [BAC]) smoking, and time of day (Bennion and Understanding the balance of alcohol’s over time.
    [Show full text]
  • Identification, Characterization and Site-Saturation Mutagenesis of a Thermostable
    Identication, Characterization and Site-Saturation Mutagenesis of a Thermostable ω-Transaminase From Chloroexi Bacterium Chen Wang Nanjing Tech University Kexin Tang Nanjing Tech University Ya Dai Nanjing Tech University Honghua Jia ( [email protected] ) Nanjing Tech University https://orcid.org/0000-0002-3824-7768 Yan Li Nanjing Tech University Zhen Gao Nanjing Tech University Bin Wu Nanjing Tech University Research Keywords: ω-Transaminase,Chloroexibacterium, Characterization,Substrate specicity, Site- saturationmutagenesis Posted Date: March 17th, 2021 DOI: https://doi.org/10.21203/rs.3.rs-296936/v1 License: This work is licensed under a Creative Commons Attribution 4.0 International License. Read Full License Identification, characterization and site-saturation mutagenesis of a thermostable ω-transaminase from Chloroflexi bacterium Chen Wang, Kexin Tang, Ya Dai, Honghua Jia*, Yan Li*, Zhen Gao, Bin Wu College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing 211816, China Correspondent authors: [email protected]; [email protected] Abstract In present study, we have mined a ω-transaminase (ω-TA) from Chloroflexi bacterium from genome database by using an ω-TA sequence ATA117 Arrmut11 from Arthrobacter sp. KNK168 and an amine transaminase from Aspergillus terreus as templates in a BLASTP search and motif sequence alignment. The protein sequence of the ω-TA from C. bacterium shows 38% sequence identity to ATA117 Arrmut11. The gene sequence of the ω-TA was inserted into pRSF-Duet1 and functionally expressed in E. coli BL21(DE3). Results showed that the recombinant ω-TA has a specific activity of 1.19 U/mg at pH 8.5, 40 °C. The substrate acceptability test showed that ω-TA has significant reactivity to aromatic amino donors and amino receptors.
    [Show full text]
  • Original Article Compensatory Upregulation of Aldo-Keto Reductase 1B10 to Protect Hepatocytes Against Oxidative Stress During Hepatocarcinogenesis
    Am J Cancer Res 2019;9(12):2730-2748 www.ajcr.us /ISSN:2156-6976/ajcr0097527 Original Article Compensatory upregulation of aldo-keto reductase 1B10 to protect hepatocytes against oxidative stress during hepatocarcinogenesis Yongzhen Liu1, Jing Zhang1, Hui Liu1, Guiwen Guan1, Ting Zhang1, Leijie Wang1, Xuewei Qi1, Huiling Zheng1, Chia-Chen Chen1, Jia Liu1, Deliang Cao2, Fengmin Lu3, Xiangmei Chen1 1State Key Laboratory of Natural and Biomimetic Drugs, Department of Microbiology and Infectious Disease Center, School of Basic Medical Sciences, Peking University Health Science Center, Beijing 100191, P. R. China; 2Department of Medical Microbiology, Immunology and Cell Biology, Simmons Cancer Institute at Southern Illinois University School of Medicine, 913 N, Rutledge Street, Springfield, IL 62794, USA; 3Peking University People’s Hospital, Peking University Hepatology Institute, Beijing 100044, P. R. China Received May 26, 2019; Accepted November 15, 2019; Epub December 1, 2019; Published December 15, 2019 Abstract: Aldo-keto reductase 1B10 (AKR1B10), a member of aldo-keto reductase superfamily, contributes to detox- ification of xenobiotics and metabolization of physiological substrates. Although increased expression of AKR1B10 was found in hepatocellular carcinoma (HCC), the role of AKR1B10 in the development of HCC remains unclear. This study aims to illustrate the role of AKR1B10 in hepatocarcinogenesis based on its intrinsic oxidoreduction abilities. HCC cell lines with AKR1B10 overexpression or knockdown were treated with doxorubicin or hydrogen peroxide to determinate the influence of aberrant AKR1B10 expression on cells’ response to oxidative stress. Using Akr1b8 (the ortholog of human AKR1B10) knockout mice, diethylnitrosamine (DEN) induced liver injury, chronic inflammation and hepatocarcinogenesis were explored. Clinically, the pattern of serum AKR1B10 relevant to disease progres- sion was investigated in a patient cohort with chronic hepatitis B (n=30), liver cirrhosis (n=30) and HCC (n=40).
    [Show full text]
  • Kobe University Repository : Kernel
    Kobe University Repository : Kernel タイトル Glutamate production from ammonia via glutamate dehydrogenase 2 Title activity supports cancer cell proliferation under glutamine depletion 著者 Takeuchi, Yukiko / Nakayama, Yasumune / Fukusaki, Eiichiro / Irino, Author(s) Yasuhiro 掲載誌・巻号・ページ Biochemical and Biophysical Research Communications,495(1):761- Citation 767 刊行日 2018-01-01 Issue date 資源タイプ Journal Article / 学術雑誌論文 Resource Type 版区分 author Resource Version © 2017 Elsevier. This manuscript version is made available under the 権利 CC-BY-NC-ND 4.0 license http://creativecommons.org/licenses/by-nc- Rights nd/4.0/ DOI 10.1016/j.bbrc.2017.11.088 JaLCDOI URL http://www.lib.kobe-u.ac.jp/handle_kernel/90005457 PDF issue: 2021-10-11 Glutamate production from ammonia via glutamate dehydrogenase 2 activity supports cancer cell proliferation under glutamine depletion Yukiko Takeuchia, Yasumune Nakayamab, c, Eiichiro Fukusakib and Yasuhiro Irinoa, d aThe Integrated Center for Mass Spectrometry, Kobe University Graduate School of Medicine, 7- 5-1 Kusunoki-cho, Kobe 650-0017, Japan bDepartment of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamada- oka, Suita, Osaka 565-0871, Japan cDepartment of Applied Microbial Technology, Faculty of Biotechnology and Life Science, Sojo University, 4-22-1 Ikeda, Kumamoto 860-0082, Japan dDivision of Evidence-based Laboratory Medicine, Kobe University Graduate School of Medicine, 7-5-1 Kusunoki-cho, Kobe 650-0017, Japan Address for correspondence: Yasuhiro Irino, PhD. Assistant Professor, Division of Evidence-based Laboratory Medicine, Kobe University Graduate School of Medicine, 7-5-1 Kusunoki-cho, Chuo-ku, Kobe 650-0017, Japan Tel.: +81-78-382-5846; Fax: +81-78-382-5859; E-mail: [email protected] 1 Abstract Cancer cells rapidly consume glutamine as a carbon and nitrogen source to support proliferation, but the cell number continues to increase exponentially after glutamine is nearly depleted from the medium.
    [Show full text]
  • Alcohol Dehydrogenase
    Catalog Number: 100161, 151430, 159858 Alcohol Dehydrogenase Molecular Weight: 141,000 comprised of four subunits of 35,000 each.8 CAS # : 9031-72-5 Synonyms: ADH; Alcohol : NAD+ oxidoreductase Source: Yeast E.C. 1.1.1.1 Description: Alcohol dehydrogenase catalyzes the reaction: The common reaction in yeast cells is reduction of acetaldehyde to ethanol. In vitro the enzyme is generally assayed and used in a more alkaline pH region, a condition which favors a shift of equilibrium towards the oxidation of ethanol. Composition: A metalloenzyme containing four tightly bound zinc atoms per molecule.18,19 Per subunit, there are two distinct active site sulfhydryl groups which can be distinguished on the basis of differential reactivity with iodoacetate and butyl isocyanate.17 A histidine residue is considered to have an essential role.9 Optimum pH: For the oxidation of ethanol, pH 8.6-9.0 (the enzyme becomes increasingly unstable at higher pH). For the reduction of acetaldehyde a pH nearer to 7.0 is considered optimum. This reaction is kinetically complex with pH being only one factor determining optimum conditions. 1% Extinction coefficient: E 260= 12.6. Isoelectric point: 5.4.16 Solubility: Dissolves readily at 5 mg/ml in 0.01 M sodium phosphate pH 7.5 to give a clear colorless solution. Activators: Sulfyhydryl activating reagents, mercaptoethanol, dithiothreitol, cysteine, etc., and heavy metal chelating reagents. Specificity: Yeast ADH which has a more narrow specificity than that of liver enzyme, accepts ethanol, is somewhat active on the straight chain primary alcohols, and acts to a very limited extent on certain secondary and branched chain alcohols.10 NADP does not serve as coenzyme.
    [Show full text]
  • Alcohol Dehydrogenase Activity in Lactococcus Chungangensis: Application in Cream Cheese to Moderate Alcohol Uptake
    J. Dairy Sci. 98:5974–5982 http://dx.doi.org/10.3168/jds.2015-9697 © American Dairy Science Association®, 2015. Alcohol dehydrogenase activity in Lactococcus chungangensis: Application in cream cheese to moderate alcohol uptake Maytiya Konkit, Woo Jin Choi, and Wonyong Kim1 Department of Microbiology, Chung-Ang University College of Medicine, Seoul 156-756, Republic of Korea ABSTRACT teroides (Molimard and Spinnler, 1996; Bockelmann et al., 1997; Beresford et al., 2001; Bockelmann and Many human gastrointestinal facultative anaerobic Hoppe-Seyler, 2001). Members of the genus Lactococcus and aerobic bacteria possess alcohol dehydrogenase (Schleifer et al., 1985) are used extensively in the dairy (ADH) activity and are therefore capable of oxidizing and food fermentation industry, a multibillion-dollar ethanol to acetaldehyde. However, the ADH activity of biotech industry. Lactococcus lactis has been the major Lactococcus spp., except Lactococcus lactis ssp. lactis, bacteria used in cheese manufacture for over 8,000 yr, has not been widely determined, though they play an with an excess of 1,000 varieties worldwide (Sandine important role as the starter for most cheesemaking and Elliker, 1970). Currently, the genus comprises 9 technologies. Cheese is a functional food recognized species and 4 subspecies with validly published names as an aid to digestion. In the current study, the ADH T (Parte, 2014). activity of Lactococcus chungangensis CAU 28 and 11 Several metabolic properties of Lactococcus serve reference strains from the genus Lactococcus was de- special functions that directly or indirectly affect termined. Only 5 strains, 3 of dairy origin, L. lactis processes, such as flavor development and ripening of ssp.
    [Show full text]