Involvement of Glutaredoxin and Thioredoxin Systems in The

Total Page:16

File Type:pdf, Size:1020Kb

Involvement of Glutaredoxin and Thioredoxin Systems in The Involvement of Glutaredoxin and Thioredoxin Systems in the Nitrogen-Fixing Symbiosis between Legumes and Rhizobia Geneviève Alloing, Karine Mandon, Eric Boncompagni, Françoise Montrichard, Pierre Frendo To cite this version: Geneviève Alloing, Karine Mandon, Eric Boncompagni, Françoise Montrichard, Pierre Frendo. In- volvement of Glutaredoxin and Thioredoxin Systems in the Nitrogen-Fixing Symbiosis between Legumes and Rhizobia. Antioxidants , MDPI, 2018, 7 (12), pp.182. 10.3390/antiox7120182. hal- 02138466 HAL Id: hal-02138466 https://hal.archives-ouvertes.fr/hal-02138466 Submitted on 26 May 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. antioxidants Review Involvement of Glutaredoxin and Thioredoxin Systems in the Nitrogen-Fixing Symbiosis between Legumes and Rhizobia Geneviève Alloing 1, Karine Mandon 1, Eric Boncompagni 1, Françoise Montrichard 2 and Pierre Frendo 1,* 1 Université Côte d’Azur, INRA, CNRS, ISA, France; [email protected] (G.A.); [email protected] (K.M.); [email protected] (E.B.) 2 IRHS, INRA, AGROCAMPUS-Ouest, Université d’Angers, SFR 4207 QUASAV, 42 rue Georges Morel, 49071 Beaucouzé CEDEX, France; [email protected] * Correspondence: [email protected] Received: 29 October 2018; Accepted: 1 December 2018; Published: 5 December 2018 Abstract: Leguminous plants can form a symbiotic relationship with Rhizobium bacteria, during which plants provide bacteria with carbohydrates and an environment appropriate to their metabolism, in return for fixed atmospheric nitrogen. The symbiotic interaction leads to the formation of a new organ, the root nodule, where a coordinated differentiation of plant cells and bacteria occurs. The establishment and functioning of nitrogen-fixing symbiosis involves a redox control important for both the plant-bacteria crosstalk and the regulation of nodule metabolism. In this review, we discuss the involvement of thioredoxin and glutaredoxin systems in the two symbiotic partners during symbiosis. The crucial role of glutathione in redox balance and S-metabolism is presented. We also highlight the specific role of some thioredoxin and glutaredoxin systems in bacterial differentiation. Transcriptomics data concerning genes encoding components and targets of thioredoxin and glutaredoxin systems in connection with the developmental step of the nodule are also considered in the model system Medicago truncatula–Sinorhizobium meliloti. Keywords: thioredoxin; glutaredoxin; legume plant; symbiosis; redox homeostasis; stress 1. Introduction Most terrestrial plants establish symbiotic relationships with fungi or bacteria that provide nutrients for their growth [1,2]. Nitrogen and phosphorous are critical determinants of plant growth and productivity. Amongst the plant families, leguminous plants can achieve a nitrogen-fixing symbiosis with soil bacteria of the family Rhizobiaceae to reduce atmospheric nitrogen (N2) to ammonia [3]. The ability to reduce N2 is restricted to bacteria and archaea which produce the enzyme nitrogenase. Legumes are an economically important plant family, for their contribution to animal and human nutrition on one hand, and for their ecosystemic services in cropping systems on the other hand, by participating to nitrogen enrichment of soils and thereby to a reduced use of nitrogen fertilizers. The study of these symbioses is therefore a major challenge to promote a more environmentally-friendly agriculture. The nitrogen-fixing symbiosis (NFS) between rhizobia bacteria and legumes leads to the formation of new root organs, called nodules [4,5]. The development of the nodule requires many crucial steps to achieve the fixation of atmospheric nitrogen. The first step is the cross recognition between bacteria and the plant partner. This recognition involves the nodulation (Nod) factors produced by the bacteria that play a major role in the symbiotic specificity between the two partners. In parallel to bacterial recognition, Nod factors promote the development of a new meristem in the plant root that leads to the Antioxidants 2018, 7, 182; doi:10.3390/antiox7120182 www.mdpi.com/journal/antioxidants Antioxidants 2018, 7, 182 2 of 17 establishment of the root nodule. Subsequently, the formation of infection threads allows the transport of bacteriaAntioxidants from 2018 the, 7 surface, x FOR PEER of theREVIEW root to the plant cells that will host bacteria, in an endosymbiotic2 of 17 way. The accommodation of numerous bacteria inside plant cells and nitrogen fixation requirements involve plant root that leads to the establishment of the root nodule. Subsequently, the formation of infection modificationsthreads allows of the the cellular transport structure of bacteria and from physiology the surface of bothof the partners root to the for plant maintaining cells that will the symbiotichost interaction.bacteria, These in an modifications endosymbiotic areway. achieved The accommodation through differentiation of numerous bacteria of the plant inside cells plant which cells and includes cell enlargement,nitrogen fixation DNA requirements endoreduplication, involve modifications and significant of the cellular reprogramming structure and of physiology cellular structure of both and metabolismpartners [6 ].for Cellularmaintaining and the biochemical symbiotic changesinteraction. are These also modifications observed during are achieved the differentiation through of differentiation of the plant cells which includes cell enlargement, DNA endoreduplication, and bacteria into N2-fixing bacteroids. Amongst them, a high aerobic metabolism provides ATP and reductantssignificant necessary reprogramming to sustain nitrogenaseof cellular structure activity, whereasand metabolism the nitrogen-fixing [6]. Cellular enzymeand biochemical is irreversibly changes are also observed during the differentiation of bacteria into N2-fixing bacteroids. Amongst inactivated by oxygen. Thus nitrogen-fixation efficiency depends on oxygen protective mechanisms them, a high aerobic metabolism provides ATP and reductants necessary to sustain nitrogenase involvingactivity, the formationwhereas the of nitrogen-fixing an oxygen barrier enzyme cell is layerirreversibly around inactivated the infected by oxygen. cells and Thus the nitrogen- production of a symbioticfixation hemoglobin, efficiency depends called on leghemoglobin. oxygen protective This mechanisms later is involved involving in the the formation protection of an of oxygen nitrogenase from denaturation,barrier cell layer and around in the the supply infected of cells ample and amountthe production of oxygen of a symbiotic to bacteria hemoglobin, for respiration. called The supplyleghemoglobin. of energy from This the later plant is involved to nitrogen-fixing in the protection bacteroids, of nitrogenase and thefrom export denaturation, of ammonia and in the from the bacteroidssupply to of the ample roots amount also requireof oxygen major to bacteria metabolic for respiration. adaptations The supply in the nodules.of energy from In conclusion, the plant the noduleto functioningnitrogen-fixing depends bacteroids, on and a strict the export regulation of ammo ofnia the from development the bacteroids and to the roots metabolism also require of plant major metabolic adaptations in the nodules. In conclusion, the nodule functioning depends on a strict and bacteria cells. regulation of the development and the metabolism of plant and bacteria cells. The nodulesThe nodules are consideredare considered as as “indeterminate” “indeterminate” or or “determinate” “determinate” according according to their to theirmode modeof of developmentdevelopment [7]. [7]. In theIn the determinate determinate nodules, nodules, such as as those those of of soybean soybean (Glycine (Glycine max), max the), nodular the nodular meristemsmeristems are transientlyare transiently active. active. This This results results inin sphericalspherical nodules, nodules, containi containingng cells cellswith witha similar a similar developmentaldevelopmental state state to each to each other. other. In In the the indeterminate indeterminate nodules, nodules, such such as those as those formed formed by pea by (Pisum pea ( Pisum sativumsativum), alfalfa), alfalfa (Medicago (Medicago sativa sativa) or) barrelor barrel medic medic (M. (M. truncatula truncatula),), thethe meristemsmeristems persist persist throughout throughout the plant’sthe life, plant’s giving life, an giving elongated an elongated nodule. nodule. Consequently, Consequently, the functionalthe functional nodule nodule presents presents three three zones: zones: (I) the meristematic zone, (II) the infection zone, and (III) the N2-fixing zone (Figure 1A,B). At (I) the meristematic zone, (II) the infection zone, and (III) the N -fixing zone (Figure1A,B). At later later stage, there is a rupture in the symbiotic interaction, which occurs2 in the senescence zone (zone stage, there is a rupture in the symbiotic interaction, which occurs in the senescence zone (zone IV). IV). FigureFigure 1. The 1. indeterminateThe
Recommended publications
  • Corynebacterium Glutamicum Mycoredoxin 3 Protects Against Multiple Oxidative Stresses
    Advance Publication J. Gen. Appl. Microbiol. doi 10.2323/jgam.2019.10.003 ©2020 Applied Microbiology, Molecular and Cellular Biosciences Research Foundation Full Paper 1 Corynebacterium glutamicum Mycoredoxin 3 protects against multiple oxidative stresses 2 and displays thioredoxin-like activity 3 (Received September 24, 2019; Accepted October 28, 2019; J-STAGE Advance publication date: October 30, 2020) 4 Tao Su#, Chengchuan Che#, Ping Sun, Xiaona Li, Zhijin Gong, Jinfeng Liu, Ge Yang * 5 6 College of Life Sciences, Qufu Normal University, Qufu, Shandong 273165, China; 7 8 Running title: Feature of C. glutamicum mycoredoxin 3 9 10 11 # These authors contributed equally to this work. 12 13 * Corresponding authors: 14 Ge Yang 15 16 E-mail [email protected] 17 Tel: 86-13953760056 18 19 20 21 22 23 24 25 26 27 Abstract 28 Glutaredoxins (Grxs) and thioredoxins (Trxs) play a critical role in resistance to oxidative 29 conditions. However, physiological and biochemical roles of Mycoredoxin 3 (Mrx3) that shared a 30 high amino acid sequence similarity to Grxs remain unknown in Corynebacterium glutamicum. 31 Here we showed that mrx3 deletion strains of C. glutamicum was involved in the protection 32 against oxidative stress. Recombinant Mrx3 not only catalytically reduced the disulfide bonds in 33 ribonucleotide reductase (RNR), insulin and 5, 5’-dithiobis-(2-nitro-benzoicacid) (DTNB), but 34 also reduced the mixed disulphides between mycothiol (MSH) and substrate, which was 35 exclusively linked to the thioredoxin reductase (TrxR) electron transfer pathway by a dithiol 36 mechanism. Site-directed mutagenesis confirmed that the conserved Cys17 and Cys20 in Mrx3 37 were necessary to maintain its activity.
    [Show full text]
  • Generate Metabolic Map Poster
    Authors: Peter D. Karp Suzanne Paley Julio Collado-Vides John L Ingraham Ingrid Keseler Markus Krummenacker Cesar Bonavides-Martinez Robert Gunsalus An online version of this diagram is available at BioCyc.org. Biosynthetic pathways are positioned in the left of the cytoplasm, degradative pathways on the right, and reactions not assigned to any pathway are in the far right of the cytoplasm. Transporters and membrane proteins are shown on the membrane. Carol Fulcher Ian Paulsen Socorro Gama-Castro Robert LaRossa Periplasmic (where appropriate) and extracellular reactions and proteins may also be shown. Pathways are colored according to their cellular function. EcoCyc: Escherichia coli K-12 substr. MG1655 Cellular Overview Connections between pathways are omitted for legibility. Anamika Kothari Amanda Mackie Alberto Santos-Zavaleta succinate phosphate succinate N-acetyl-DL-methionine + L-ornithine glutathione + L-methionine S-oxide D-fructofuranose γ Ag+ molybdate ferroheme b L,L-homocystine asp lys cys L-alanyl- -D- D-mannopyranose 6-phosphate 2+ 2+ H D-methionine 2-deoxy-D-glucose succinate formate formate succinate D-tartrate putrescine agmatine cadaverine L-tartrate D-fructofuranose 6-phosphate + nitrate nitrate Cu thiosulfate deoxycholate L,L-homocystine D-cystine D-cycloserine methyl β-D-glucoside putrescine asp spermidine (S)-2-hydroxybutanoate (S)-2-hydroxybutanoate arg L-homoserine lactone magnesium hydrogenphosphate magnesium hydrogenphosphate antimonous acid glutamyl-meso- Co2+ Cd2+ lactulose poly-β-1,6- met cob(I)inamide 2,3-dioxo-
    [Show full text]
  • The Role of Intermembrane Space Redox Factors in Glutathione Metabolism and Intracellular Redox Equilibrium Hatice Kubra Ozer University of South Carolina
    University of South Carolina Scholar Commons Theses and Dissertations 2015 The Role of Intermembrane Space Redox Factors In Glutathione Metabolism And Intracellular Redox Equilibrium Hatice Kubra Ozer University of South carolina Follow this and additional works at: https://scholarcommons.sc.edu/etd Part of the Chemistry Commons Recommended Citation Ozer, H. K.(2015). The Role of Intermembrane Space Redox Factors In Glutathione Metabolism And Intracellular Redox Equilibrium. (Doctoral dissertation). Retrieved from https://scholarcommons.sc.edu/etd/3702 This Open Access Dissertation is brought to you by Scholar Commons. It has been accepted for inclusion in Theses and Dissertations by an authorized administrator of Scholar Commons. For more information, please contact [email protected]. THE ROLE OF INTERMEMBRANE SPACE REDOX FACTORS IN GLUTATHIONE METABOLISM AND INTRACELLULAR REDOX EQUILIBRIUM by Hatice Kubra Ozer Bachelor of Science Uludag University, 2004 Master of Food Science and Nutrition Clemson University, 2010 Submitted in Partial Fulfillment of the Requirements For the Degree of Doctor of Philosophy in Chemistry College of Arts and Sciences University of South Carolina 2015 Accepted by: Caryn E. Outten, Major Professor F. Wayne Outten, Committee Chair Erin Connolly, Committee Member Andrew B. Greytak, Committee Member Lacy K. Ford, Senior Vice Provost and Dean of Graduate Studies © Copyright by Hatice Kubra Ozer, 2015 All Rights Reserved. ii ACKNOWLEDGEMENTS First, I would like to thank my advisor, Dr. Caryn E. Outten for her patience, guidance, expertise, and confidence in me to complete the work contained herein. She has been an excellent mentor during my graduate program. She is also the only person beside myself who is guaranteed to have read every word of this manuscript and to review every presentations in the past and her insight was irreplaceable.
    [Show full text]
  • Role of GSH and Iron-Sulfur Glutaredoxins in Iron Metabolism—Review
    molecules Review Role of GSH and Iron-Sulfur Glutaredoxins in Iron Metabolism—Review 1, 1, 1 1 Trnka Daniel y , Hossain Md Faruq y , Jordt Laura Magdalena , Gellert Manuela and Lillig Christopher Horst 2,* 1 Institute for Medical Biochemistry and Molecular Biology, University Medicine, University of Greifswald, 17475 Greifswald, Germany; [email protected] (T.D.); [email protected] (H.M.F.); [email protected] (J.L.M.); [email protected] (G.M.) 2 Christopher Horst Lillig, Institute for Medical Biochemistry and Molecular Biology, University Medicine Greifswald, Ferdinand-Sauerbruch-Straße, 17475 Greifswald, Germany * Correspondence: [email protected]; Tel.: +49-3834-865407; Fax: +49-3834-865402 These authors contributed equally to this work. y Academic Editor: Pál Perjési Received: 29 July 2020; Accepted: 22 August 2020; Published: 25 August 2020 Abstract: Glutathione (GSH) was initially identified and characterized for its redox properties and later for its contributions to detoxification reactions. Over the past decade, however, the essential contributions of glutathione to cellular iron metabolism have come more and more into focus. GSH is indispensable in mitochondrial iron-sulfur (FeS) cluster biosynthesis, primarily by co-ligating FeS clusters as a cofactor of the CGFS-type (class II) glutaredoxins (Grxs). GSH is required for the export of the yet to be defined FeS precursor from the mitochondria to the cytosol. In the cytosol, it is an essential cofactor, again of the multi-domain CGFS-type Grxs, master players in cellular iron and FeS trafficking. In this review, we summarize the recent advances and progress in this field. The most urgent open questions are discussed, such as the role of GSH in the export of FeS precursors from mitochondria, the physiological roles of the CGFS-type Grx interactions with BolA-like proteins and the cluster transfer between Grxs and recipient proteins.
    [Show full text]
  • Biochemical and Biophysical Characterization of a Mycoredoxin
    International Journal of Biological Macromolecules 107 (2018) 1999–2007 Contents lists available at ScienceDirect International Journal of Biological Macromolecules j ournal homepage: www.elsevier.com/locate/ijbiomac Biochemical and biophysical characterization of a mycoredoxin protein glutaredoxin A1 from Corynebacterium pseudotuberculosis a a a b a Raphael J. Eberle , Liege A. Kawai , Fabio R. de Moraes , Ljubica Tasic , Raghuvir K. Arni , a,∗ Monika A. Coronado a Multiuser Center for Biomolecular Innovation, Departament of Physics, Instituto de Biociências Letras e Ciências Exatas (Ibilce), Universidade Estadual Paulista (UNESP), São Jose do Rio Preto, SP, 15054-000, Brazil b Institute of Chemistry, University of Campinas (UNICAMP), Campinas, SP, 13083-970, Brazil a r t i c l e i n f o a b s t r a c t Article history: Glutaredoxin A1 from Corynebacterium pseudotuberculosis was shown to be a mycoredoxin protein. In Received 25 July 2017 this study, we established a process to overexpress and purify glutaredoxin A1. The aim of this study Received in revised form 6 October 2017 was the investigation of the Glutaredoxin A1 from C. pseudotuberculosis behavior under different redox Accepted 11 October 2017 environments and the identification of lead molecules, which can be used for specific inhibitor develop- Available online 16 October 2017 ment for this protein family. A quantitative assay was performed measuring the rate of insulin reduction spectrophotometrically at 640 nm through turbidity formation from the precipitation of the free insulin. Keywords: Glutaredoxin A1, at 5 ␮M concentration, accelerated the reduction process of 0.2 mM insulin and 1 mM Corynebacterium pseudotuberculosis Glutaredoxin DTT.
    [Show full text]
  • Tremblay Robinlee.Pdf
    Expression and Characterisation ofa Gene Enc oding RbpD, an RNA- Bind ing Protein in Anabaena sp. strain PeC 7120 by Rob in lee Tremblay A lhesis submitted to the Scltool of Graduale Studies in partial fulfilment of the requirements fOl" the degree of Master of Science Department of BiochemistrylFacultyof Science Memorial University of Newfoundland January 2000 SI.JOM'S Newfoun dland Abs t ra ct The RNA-binding protein RbpD, from the cyano bacterium Anaba ena sp, strain Pe C 7120 was expressed in £Sch~ ric h ia coli and successfully purified using me IMPACT I system (New England Biolabs). The rbp D gene was cloned into the pCYBt expre ssion vector by using polymerase chain reaction to introduce Ndel and SapI restriction sites at the 5' end 3' ends of the gene respect ively. The 3'.-end mutagenesis also chan ged the stop codon into a cysteine codon. The resulting gene encoded a fusion protein consisting of RbpD, the Saccharomyces cerev isiae VMA intein and a chitin binding domain.. Expressi on of the fusion protein was observed in £ coli strain MCI061 but Western blot analysis using an intein-directed ant ibody indicated that significant in vivo fmeln-direcred splicing of the fusion protein occurred. We were unable to eliminate this problem; no fusion protein expression was observed in 8 other E coli strains tested. Wild -type RbpD was purified following binding of the fusion protein 10 a chitin column and overnight cleavage in the presence of a reducing agent, dlthicthrehc l. A number of modifications to the manufacturer' s purification protocol were found to be necessary for success ful purification.
    [Show full text]
  • Purification and Characterisation of a Protease (Tamarillin) from Tamarillo Fruit
    Purification and characterisation of a protease (tamarillin) from tamarillo fruit Item Type Article Authors Li, Zhao; Scott, Ken; Hemar, Yacine; Zhang, Huoming; Otter, Don Citation Li Z, Scott K, Hemar Y, Zhang H, Otter D (2018) Purification and characterisation of a protease (tamarillin) from tamarillo fruit. Food Chemistry. Available: http://dx.doi.org/10.1016/ j.foodchem.2018.02.091. Eprint version Post-print DOI 10.1016/j.foodchem.2018.02.091 Publisher Elsevier BV Journal Food Chemistry Rights NOTICE: this is the author’s version of a work that was accepted for publication in Food Chemistry. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. Changes may have been made to this work since it was submitted for publication. A definitive version was subsequently published in Food Chemistry, [, , (2018-02-16)] DOI: 10.1016/j.foodchem.2018.02.091 . © 2018. This manuscript version is made available under the CC-BY-NC-ND 4.0 license http://creativecommons.org/licenses/by-nc-nd/4.0/ Download date 29/09/2021 23:19:14 Link to Item http://hdl.handle.net/10754/627180 Accepted Manuscript Purification and characterisation of a protease (tamarillin) from tamarillo fruit Zhao Li, Ken Scott, Yacine Hemar, Huoming Zhang, Don Otter PII: S0308-8146(18)30327-3 DOI: https://doi.org/10.1016/j.foodchem.2018.02.091 Reference: FOCH 22475 To appear in: Food Chemistry Received Date: 25 October 2017 Revised Date: 13 February 2018 Accepted Date: 16 February 2018 Please cite this article as: Li, Z., Scott, K., Hemar, Y., Zhang, H., Otter, D., Purification and characterisation of a protease (tamarillin) from tamarillo fruit, Food Chemistry (2018), doi: https://doi.org/10.1016/j.foodchem.
    [Show full text]
  • Supplementary Information
    Supplementary information (a) (b) Figure S1. Resistant (a) and sensitive (b) gene scores plotted against subsystems involved in cell regulation. The small circles represent the individual hits and the large circles represent the mean of each subsystem. Each individual score signifies the mean of 12 trials – three biological and four technical. The p-value was calculated as a two-tailed t-test and significance was determined using the Benjamini-Hochberg procedure; false discovery rate was selected to be 0.1. Plots constructed using Pathway Tools, Omics Dashboard. Figure S2. Connectivity map displaying the predicted functional associations between the silver-resistant gene hits; disconnected gene hits not shown. The thicknesses of the lines indicate the degree of confidence prediction for the given interaction, based on fusion, co-occurrence, experimental and co-expression data. Figure produced using STRING (version 10.5) and a medium confidence score (approximate probability) of 0.4. Figure S3. Connectivity map displaying the predicted functional associations between the silver-sensitive gene hits; disconnected gene hits not shown. The thicknesses of the lines indicate the degree of confidence prediction for the given interaction, based on fusion, co-occurrence, experimental and co-expression data. Figure produced using STRING (version 10.5) and a medium confidence score (approximate probability) of 0.4. Figure S4. Metabolic overview of the pathways in Escherichia coli. The pathways involved in silver-resistance are coloured according to respective normalized score. Each individual score represents the mean of 12 trials – three biological and four technical. Amino acid – upward pointing triangle, carbohydrate – square, proteins – diamond, purines – vertical ellipse, cofactor – downward pointing triangle, tRNA – tee, and other – circle.
    [Show full text]
  • TRACE: Tennessee Research and Creative Exchange
    University of Tennessee, Knoxville TRACE: Tennessee Research and Creative Exchange Doctoral Dissertations Graduate School 8-2009 Structure-Function Studies of the Large Subunit of Ribonucleotide Reductase from Homo sapiens and Saccharomyces cerevisiae James Wesley Fairman University of Tennessee - Knoxville Follow this and additional works at: https://trace.tennessee.edu/utk_graddiss Part of the Biochemistry, Biophysics, and Structural Biology Commons Recommended Citation Fairman, James Wesley, "Structure-Function Studies of the Large Subunit of Ribonucleotide Reductase from Homo sapiens and Saccharomyces cerevisiae. " PhD diss., University of Tennessee, 2009. https://trace.tennessee.edu/utk_graddiss/49 This Dissertation is brought to you for free and open access by the Graduate School at TRACE: Tennessee Research and Creative Exchange. It has been accepted for inclusion in Doctoral Dissertations by an authorized administrator of TRACE: Tennessee Research and Creative Exchange. For more information, please contact [email protected]. To the Graduate Council: I am submitting herewith a dissertation written by James Wesley Fairman entitled "Structure- Function Studies of the Large Subunit of Ribonucleotide Reductase from Homo sapiens and Saccharomyces cerevisiae." I have examined the final electronic copy of this dissertation for form and content and recommend that it be accepted in partial fulfillment of the equirr ements for the degree of Doctor of Philosophy, with a major in Biochemistry and Cellular and Molecular Biology. Chris G. Dealwis,
    [Show full text]
  • Glutaredoxin Function for the Carboxyl-Terminal Domain of the Plant-Type 5؅-Adenylylsulfate Reductase
    Proc. Natl. Acad. Sci. USA Vol. 95, pp. 8404–8409, July 1998 Plant Biology Glutaredoxin function for the carboxyl-terminal domain of the plant-type 5*-adenylylsulfate reductase JULIE-ANN BICK*, FREDRIK ÅSLUND†,YICHANG CHEN*, AND THOMAS LEUSTEK*‡ *Biotech Center and Plant Science Department, Rutgers University, New Brunswick, NJ 08901-8250; and †Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, MA 02115 Edited by Bob B. Buchanan, University of California, Berkeley, CA, and approved May 14, 1998 (received for review November 21, 1997) ABSTRACT 5*-Adenylylsulfate (APS) reductase (EC the basis of their similar catalytic requirements and molecular 1.8.99.-) catalyzes the reduction of activated sulfate to sulfite weights (4, 6). The major difference is that the product of the in plants. The evidence presented here shows that a domain of sulfotransferase is believed to be an organic thiosulfate (1), the enzyme is a glutathione (GSH)-dependent reductase that whereas the reductase may produce sulfite (4), although the functions similarly to the redox cofactor glutaredoxin. The actual reaction product has not been determined for either APR1 cDNA encoding APS reductase from Arabidopsis thali- enzyme. APS reductase is distinguished from the cysH product ana is able to complement the cysteine auxotrophy of an by its preference for APS over PAPS and by its ability to Escherichia coli cysH [3*-phosphoadenosine-5*-phosphosul- function in an E. coli thioredoxinyglutaredoxin double mutant fate (PAPS) reductase] mutant, only if the E. coli strain (4). These properties may be due to its unusual two-domain produces glutathione. The purified recombinant enzyme structure, consisting of a reductase domain (R domain) at the (APR1p) can use GSH efficiently as a hydrogen donor in vitro, amino terminus, showing homology with the cysH product; and ' showing a Km[GSH] of 0.6 mM.
    [Show full text]
  • A Strategy to Address Dissociation-Induced Compositional and Transcriptional Bias for Single-Cell Analysis of the Human Mammary Gland
    bioRxiv preprint doi: https://doi.org/10.1101/2021.02.11.430721; this version posted February 11, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. A strategy to address dissociation-induced compositional and transcriptional bias for single-cell analysis of the human mammary gland Lisa K. Engelbrecht1,9,*,‡, Alecia-Jane Twigger1,2,*, Hilary M. Ganz1, Christian J. Gabka4, Andreas R. Bausch5,8, Heiko Lickert6,7,8, Michael Sterr6, Ines Kunze6, Walid T. Khaled2,3 & Christina H. Scheel1,10,‡ Affiliations 1 Institute of Stem Cell Research, Helmholtz Center for Health and Environmental Research Munich, Neuherberg 85764, Germany 2 Department of Pharmacology, University of Cambridge, Cambridge CB2 1PD, UK 3 Cancer Research UK Cambridge Cancer Center, Cambridge CB2 0RE, UK 4 Nymphenburg Clinic for Plastic and Aesthetic Surgery, Munich 80637, Germany 5 Chair of Biophysics E27, Technical University Munich, 85747 Garching, Germany 6 Institute of Diabetes and Regeneration Research, Helmholtz Center for Health and Environmental Research Munich, Neuherberg 85764, Germany 7 German Center for Diabetes Research (DZD), 85764 Neuherberg, Germany 8 Technical University of Munich, 80333 Munich, Germany. 9 Present address: Chair of Biophysics E27, Technical University Munich, 85747 Garching, Germany 10 Present address: Department of Dermatology, St. Josef Hospital, Ruhr-University Bochum, 44791 Bochum, Germany * These authors contributed equally to this work. ‡ Corresponding author. Correspondence to: [email protected] or [email protected] 1 bioRxiv preprint doi: https://doi.org/10.1101/2021.02.11.430721; this version posted February 11, 2021.
    [Show full text]
  • ANABOLISM III: Biosynthesis Amino Acids & Nucleotides
    BI/CH 422/622 ANABOLISM OUTLINE: Photosynthesis Carbohydrate Biosynthesis in Animals Biosynthesis of Fatty Acids and Lipids Biosynthesis of Amino Acids and Nucleotides Nitrogen fixation nitrogenase Nitrogen assimilation Glutamine synthetase Glutamate synthase Amino-acid Biosynthesis non-essential essential Nucleotide Biosynthesis RNA precursors purines pyrimidines DNA precursors deoxy-nucleotides Biosynthesis of secondary products of amino acids ANABOLISM III: Biosynthesis Amino Acids & Nucleotides Dr. Kornberg: Lecture 04.26.17 (0:00-5:06) 5 min 1 Biosynthesis Amino Acids & Nucleotides How are Ribonucleic Acid Precursors So far: converted to Deoxyribonucleic Acid GMPàGDPàGTP Precursors? ….....and how is dTTP made? AMPàADPàATP 2’C-OH bond is directly reduced to 2’-H UMPàUDPàUTPà bond …without activating the carbon for CDPßCTP dehydration, etc.! catalyzed by ribonucleotide reductase Specific kinases, Non-specific kinase, e.g., UMP kinase, nucleoside GMP kinase, diphosphate kinase Very unique enzyme in all of biochemistry – use of free Adenylate kinase (works on both oxy- and radicals etc. deoxy-ribose GDPàdGDP nucleosides) Mechanism: Two H atoms are donated ADPàdADP by NADPH and carried by thioredoxin or glutaredoxin to the active site. UDPàdUDP –Substrates are the NDPs and the products CDPàdCDP are dNDP. Biosynthesis Amino Acids & Nucleotides Source of Reducing Structure of Ribonucleotide Reductase a2 are regulatory Electrons for and half the Ribonucleotide catalytic site; need to be reduced. Reductase b 2 are the other half (a b ) of the active site, 2 2 and the free- radical generators • NADPH serves as the electron donor. • Funneled through glutathione or JoAnne Stubbe thioredoxin pathways (1946– ) 2 •Most forms of enzyme have two catalytic/ regulatory subunits and two radical- generating subunits.
    [Show full text]