DOCSLIB.ORG

Redox Proteomics in Selected Neurodegenerative Disorders: from Its Infancy to Future Applications Allan Butterfield University of Kentucky

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Redox Proteomics in Selected Neurodegenerative Disorders: from Its Infancy to Future Applications Allan Butterfield University of Kentucky Eastern Kentucky University Encompass Chemistry Faculty and Staff choS larship Chemistry 2012 Redox Proteomics in Selected Neurodegenerative Disorders: From Its Infancy to Future Applications Allan Butterfield University of Kentucky Marzia Perluigi Sapienza University of Rome Tanea Reed Eastern Kentucky University Tasneem Muharib University of Kentucky Christopher P. Hughes University of Kentucky See next page for additional authors Follow this and additional works at: http://encompass.eku.edu/che_fsresearch Part of the Chemistry Commons Recommended Citation Butterfield, D. A., Perluigi, M., Reed, T., Muharib, T., Hughes, C. P., Robinson, R. A., & Sultana, R. (2012). Redox Proteomics in Selected Neurodegenerative Disorders: From Its Infancy to Future Applications. Antioxidants & Redox Signaling, 17(11), 1610-1655. doi:10.1089/ars.2011.4109 This Article is brought to you for free and open access by the Chemistry at Encompass. It has been accepted for inclusion in Chemistry Faculty and Staff Scholarship by an authorized administrator of Encompass. For more information, please contact [email protected]. Authors Allan Butterfield, Marzia Perluigi, Tanea Reed, Tasneem Muharib, Christopher P. Hughes, Rena A.S. Robinson, and Rukhsana Sultana This article is available at Encompass: http://encompass.eku.edu/che_fsresearch/3 See discussions, stats, and author profiles for this publication at: https://www.researchgate.net/publication/51828850 Redox Proteomics in Selected Neurodegenerative Disorders: From Its Infancy to Future Applications Article in Antioxidants & Redox Signaling · November 2011 Impact Factor: 7.41 · DOI: 10.1089/ars.2011.4109 · Source: PubMed CITATIONS READS 72 32 7 authors, including: D. Allan Butterfield Marzia Perluigi University of Kentucky Sapienza University of Rome 628 PUBLICATIONS 39,353 CITATIONS 113 PUBLICATIONS 3,459 CITATIONS SEE PROFILE SEE PROFILE Tanea Reed Renã A S Robinson Eastern Kentucky University University of Pittsburgh 17 PUBLICATIONS 1,404 CITATIONS 30 PUBLICATIONS 355 CITATIONS SEE PROFILE SEE PROFILE All in-text references underlined in blue are linked to publications on ResearchGate, Available from: Tanea Reed letting you access and read them immediately. Retrieved on: 11 May 2016 ANTIOXIDANTS & REDOX SIGNALING Volume 17, Number 11, 2012 COMPREHENSIVE INVITED REVIEW ª Mary Ann Liebert, Inc. DOI: 10.1089/ars.2011.4109 Redox Proteomics in Selected Neurodegenerative Disorders: From Its Infancy to Future Applications D. Allan Butterfield,1 Marzia Perluigi,2 Tanea Reed,3 Tasneem Muharib,4 Christopher P. Hughes,4 Rena˜ A.S. Robinson,4 and Rukhsana Sultana1 Abstract Several studies demonstrated that oxidative damage is a characteristic feature of many neurodegenerative diseases. The accumulation of oxidatively modified proteins may disrupt cellular functions by affecting protein expression, protein turnover, cell signaling, and induction of apoptosis and necrosis, suggesting that protein oxidation could have both physiological and pathological significance. For nearly two decades, our laboratory focused particular attention on studying oxidative damage of proteins and how their chemical modifications induced by reactive oxygen species/reactive nitrogen species correlate with pathology, biochemical alterations, and clinical presentations of Alzheimer’s disease. This comprehensive article outlines basic knowledge of oxi- dative modification of proteins and lipids, followed by the principles of redox proteomics analysis, which also involve recent advances of mass spectrometry technology, and its application to selected age-related neurode- generative diseases. Redox proteomics results obtained in different diseases and animal models thereof may provide new insights into the main mechanisms involved in the pathogenesis and progression of oxidative- stress-related neurodegenerative disorders. Redox proteomics can be considered a multifaceted approach that has the potential to provide insights into the molecular mechanisms of a disease, to find disease markers, as well as to identify potential targets for drug therapy. Considering the importance of a better understanding of the cause/effect of protein dysfunction in the pathogenesis and progression of neurodegenerative disorders, this article provides an overview of the intrinsic power of the redox proteomics approach together with the most significant results obtained by our laboratory and others during almost 10 years of research on neurodegener- ative disorders since we initiated the field of redox proteomics. Antioxid. Redox Signal. 17, 1610–1655. I. Introduction 1611 II. Protein (/Lipid) Oxidation and Protein Dysfunction 1611 A. Protein carbonyls 1612 B. Protein nitration 1613 1. Peroxynitrite (ONOO - ) 1615 2. Nitrogen dioxide (NO2) 1616 C. HNE adduction to proteins 1616 D. Importance of clearance and detoxification systems 1616 1. The proteasome, parkin, ubiquitin carboxy-terminal hydrolase-L1, and HSPs 1616 2. Superoxide dismutase 1617 3. Catalase 1618 4. Peroxiredoxins 1618 5. Trx and Trx reductase 1618 6. Glutathione reductase 1618 7. Vitamins in neurodegeneration 1618 8. Involvement of iron in neurodegeneration 1618 Reviewing Editors: Michalis Aivaliotis, Valentino Bonetto, Howard Gendelman, Pietro Ghezzi, Mahin Maines, Jan Ottervald, Junji Yodoi, and Moussa Youdim 1Department of Chemistry, Center of Membrane Sciences, Sanders-Brown Center on Aging, University of Kentucky, Lexington, Kentucky. 2Department of Biochemical Sciences, Sapienza University of Rome, Rome, Italy. 3Department of Chemistry, Eastern Kentucky University of Kentucky, Richmond, Kentucky. 4Department of Chemistry, University of Pittsburgh, Pittsburgh, Pennsylvania. 1610 REDOX PROTEOMICS IN NEURODEGENERATIVE DISORDERS 1611 E. Role of iron in neurodegeneration 1620 1. Fe homeostasis in AD 1620 2. Fe homeostasis in PD 1620 3. Fe homeostasis in ALS 1620 4. Fe homeostasis in HD 1620 F. Some known consequences of protein oxidation 1621 III. Overview of Redox Proteomics 1621 A. Global, gel-based approaches 1621 B. Targeted, gel-free approach 1623 1. Enrichment of PCO modified proteins 1623 2. Enrichment of HNE modified proteins 1624 3. Enrichment of 3-NT modified proteins 1624 IV. Application of Redox Proteomics to Selected Neurodegenerative Disorders 1626 A. Alzheimer’s disease 1626 1. PCO in AD 1626 2. Identification of carbonylated proteins in brain of subjects with AD 1627 a. Sample: the brain 1627 b. Energy dysfunction 1627 c. Excitotoxicity 1628 d. Proteosomal dysfunction 1628 e. Neuritic abnormalities 1628 f. APP regulation, tau hyperphosphorylation, and cell cycle regulation 1628 g. Synaptic abnormalities and LTP 1629 h. pH maintenance 1629 i. Mitochondrial abnormalities 1629 3. Carbonylated proteins in brain of subjects with amnestic MCI 1629 4. EAD carbonylated proteins 1630 5. PCAD vs. amnestic MCI protein carbonylation in brain 1630 6. Protein-bound HNE in brain and progression of Alzheimer’s disease 1630 7. Protein-bound 3-NT in brain and progression of Alzheimer’s disease 1632 8. Nitrated brain proteins in MCI 1632 9. Nitrated proteins in EAD 1633 B. Parkinson disease 1633 1. Redox proteomics in PD 1635 C. Amyotrophic lateral sclerosis 1635 1. Redox proteomics studies in ALS transgenic mice 1636 D. Huntington disease 1637 1. Redox proteomics-transgenic mouse model of HD 1638 2. Proteomics of HD brain 1638 E. Down syndrome 1638 1. Redox proteomics in DS transgenic mice 1639 V. Conclusions and Future Directions 1639 I. Introduction neurodegenerative disorders (63, 180). Free radicals are generated in vivo from various sources, one of the major edox proteomics is the subset of proteomics in which sources being the leakage of superoxide radical from the Roxidatively or nitrosatively modified proteins are iden- mitochondria (Fig. 1). Under physiological conditions, levels .2 tified (115). Our laboratory was among the first that used of superoxide anion radicals (O2 ) are maintained in the cell redox proteomics to identify oxidatively modified brain pro- by the antioxidant enzyme, superoxide dismutase (SOD), .2 teins (91, 92, 233). Others first used redox proteomics to identify which disproportionates O2 to hydrogen peroxide (H2O2) oxidized thiols (34, 88, 157, 250). Redox proteomics has been and oxygen (Fig. 1). Further, the H2O2 formed is converted applied to numerous disorders known to be associated with to water and oxygen by the enzymes catalase, peroxidase, or oxidative stress (OS) (115). This comprehensive article focuses glutathione peroxidase (GPx). GPx uses reduced glutathione on applications and results of redox proteomics that provide (GSH) to carry out its functions, and the levels of reduced insights into selected neurodegenerative disorders. GSH are maintained by the enzyme glutathione reductase (GR), which converts oxidized glutathione (GSSG) to GSH using NADPH for reducing equivalents. In the brain, the II. Protein (/Lipid) Oxidation and Protein Dysfunction levels of catalase are greater than those for GPx. The OS induced by free radicals plays an important role in the importance of these enzymes in relation to neurodegenera- pathophysiology of a wide variety of diseases including tion will be discussed in further detail next. During 1612 BUTTERFIELD ET AL. FIG. 1. Free radicals are generated by various mechanisms. One way by which free radicals are generated is via release of superoxide anion from the mitochondria, leading to increased formation of reactive oxygen and reactive nitrogen species and, consequently, damaging the biomolecules. (To see this illustration
Load more

Recommended publications
  • Sorgodb: Superoxide Reductase Gene Ontology Curated Database
    Lucchetti-Miganeh et al. BMC Microbiology 2011, 11:105 http://www.biomedcentral.com/1471-2180/11/105 DATABASE Open Access SORGOdb: Superoxide Reductase Gene Ontology curated DataBase Céline Lucchetti-Miganeh1*, David Goudenège1, David Thybert1,2, Gilles Salbert1 and Frédérique Barloy-Hubler1 Abstract Background: Superoxide reductases (SOR) catalyse the reduction of superoxide anions to hydrogen peroxide and are involved in the oxidative stress defences of anaerobic and facultative anaerobic organisms. Genes encoding SOR were discovered recently and suffer from annotation problems. These genes, named sor, are short and the transfer of annotations from previously characterized neelaredoxin, desulfoferrodoxin, superoxide reductase and rubredoxin oxidase has been heterogeneous. Consequently, many sor remain anonymous or mis-annotated. Description: SORGOdb is an exhaustive database of SOR that proposes a new classification based on domain architecture. SORGOdb supplies a simple user-friendly web-based database for retrieving and exploring relevant information about the proposed SOR families. The database can be queried using an organism name, a locus tag or phylogenetic criteria, and also offers sequence similarity searches using BlastP. Genes encoding SOR have been re-annotated in all available genome sequences (prokaryotic and eukaryotic (complete and in draft) genomes, updated in May 2010). Conclusions: SORGOdb contains 325 non-redundant and curated SOR, from 274 organisms. It proposes a new classification of SOR into seven different classes and allows biologists to explore and analyze sor in order to establish correlations between the class of SOR and organism phenotypes. SORGOdb is freely available at http://sorgo.genouest.org/index.php. Background (called reactive oxygen species or ROS), particularly Two and a half billion years ago, the intense photosyn- hydroxyl radicals (•OH), hydrogen peroxide (H2O2)and thetic activity of cyanobacteria caused the largest envir- superoxide anion radicals (O2-).
    [Show full text]
  • ABSTRACT JI, MIKYOUNG LEE. Superoxide Reductase from The
    ABSTRACT JI, MIKYOUNG LEE. Superoxide Reductase from the Hyperthermophilic Archaeon Pyrococcus furiosus: its Function, Regulation, and Biotechnological Applications. (Under the direction of Amy M. Grunden.) The anaerobic hyperthermophilic archaeon, Pyrococcus furious, possesses a system for the detoxification of reactive oxygen species, which is different from the classical defense mechanisms present in aerobes. P. furiosus employs a novel enzyme system centered on the enzyme superoxide reductase (SOR), which reduces superoxide molecules to hydrogen peroxide without producing oxygen. Surprisingly, P. furiosus SOR, unlike many P. furiosus enzymes, was shown to function at low temperature (<25o C). A model for superoxide reduction by SOR was proposed where the electrons used by SOR to reduce superoxide are supplied by a small iron containing protein, rubredoxin (Rd), and Rd is reduced by the oxidoreductase, NAD(P)H-rubredoxin oxidoreductase (NROR). The first objective of this study was to evaluate the validity of the proposed superoxide reduction pathway by using the recombinant SOR, Rd and NROR enzymes in an in vitro assay as well as to demonstrate in vivo function via complementation studies in superoxide detoxification deficient Escherichia coli strains. The second objective was to investigate the transcriptional expression levels of genes that are involved in the SOR- centered superoxide reduction pathway in order to determine how these genes are expressed and regulated in response to various oxidative stresses. The third objective was to evaluate the efficacy of the biotechnological application of this superoxide detoxification system by expressing SOR in plant cells, which enhanced their survival at high temperature and from drought indicating that it functions successfully in vivo.
    [Show full text]
  • Role of Superoxide Reductase FA796 in Oxidative Stress Resistance in Filifactor Alocis Arunima Mishra✉, Ezinne Aja & Hansel M Fletcher
    www.nature.com/scientificreports OPEN Role of Superoxide Reductase FA796 in Oxidative Stress Resistance in Filifactor alocis Arunima Mishra✉, Ezinne Aja & Hansel M Fletcher Filifactor alocis, a Gram-positive anaerobic bacterium, is now a proposed diagnostic indicator of periodontal disease. Because the stress response of this bacterium to the oxidative environment of the periodontal pocket may impact its pathogenicity, an understanding of its oxidative stress resistance strategy is vital. Interrogation of the F. alocis genome identifed the HMPREF0389_00796 gene that encodes for a putative superoxide reductase (SOR) enzyme. SORs are non-heme, iron-containing enzymes that can catalyze the reduction of superoxide radicals to hydrogen peroxide and are important in the protection against oxidative stress. In this study, we have functionally characterized the putative SOR (FA796) from F. alocis ATCC 35896. The recombinant FA796 protein, which is predicted to be a homotetramer of the 1Fe-SOR class, can reduce superoxide radicals. F. alocis FLL141 (∆FA796::ermF) was signifcantly more sensitive to oxygen/air exposure compared to the parent strain. Sensitivity correlated with the level of intracellular superoxide radicals. Additionally, the FA796-defective mutant had increased sensitivity to hydrogen peroxide-induced stress, was inhibited in its ability to form bioflm and had reduced survival in epithelial cells. Collectively, these results suggest that the F. alocis SOR protein is a key enzymatic scavenger of superoxide radicals and protects the bacterium from oxidative stress conditions. All living cells in an oxygen-rich environment encounter oxidative stress due to the generation of reactive oxygen 1,2 species (ROS), including superoxide radicals, hydroxyl radicals and hydrogen peroxide (H2O2) .
    [Show full text]
  • Endogenous Superoxide Is a Key Effector of the Oxygen Sensitivity of A
    Endogenous superoxide is a key effector of the oxygen PNAS PLUS sensitivity of a model obligate anaerobe Zheng Lua,1, Ramakrishnan Sethua,1, and James A. Imlaya,2 aDepartment of Microbiology, University of Illinois, Urbana, IL 61801 Edited by Irwin Fridovich, Duke University Medical Center, Durham, NC, and approved March 1, 2018 (received for review January 3, 2018) It has been unclear whether superoxide and/or hydrogen peroxide fects (3, 4). Thus, these phenotypes confirmed the potential tox- play important roles in the phenomenon of obligate anaerobiosis. icity of reactive oxygen species (ROS), and they broadly supported This question was explored using Bacteroides thetaiotaomicron,a the idea that anaerobes might be poisoned by endogenous major fermentative bacterium in the human gastrointestinal tract. oxidants. Aeration inactivated two enzyme families—[4Fe-4S] dehydratases The metabolic defects of the mutant E. coli strains were sub- and nonredox mononuclear iron enzymes—whose homologs, in sequently traced to damage to two types of enzymes: dehy- contrast, remain active in aerobic Escherichia coli. Inactivation- dratases that depend upon iron-sulfur clusters and nonredox rate measurements of one such enzyme, B. thetaiotaomicron fu- enzymes that employ a single atom of ferrous iron (5–9). In both marase, showed that it is no more intrinsically sensitive to oxi- enzyme families, the metal centers are solvent exposed so that dants than is an E. coli fumarase. Indeed, when the E. coli they can directly bind and activate their substrates. Superoxide B. thetaiotaomicron enzymes were expressed in , they no longer and H2O2 are tiny molecules that cannot easily be excluded from could tolerate aeration; conversely, the B.
    [Show full text]
  • Alkyl Hydroperoxide Reductase Dependent on Thioredoxin-Like Protein from Pyrococcus Horikoshii
    Rapid Communication J. Biochem. 134, 25-29 (2003) D O I: 10.1093/j b/mvg l09 Alkyl Hydroperoxide Reductase Dependent on Thioredoxin-Like Protein from Pyrococcus horikoshii Yasuhiro Kashima and Kazuhiko Ishikawa* Special Division of Human Life Technology, National Institute of Advanced Industrial Science and Technology (AIST Kansai)1-8-31 Midorigaoka, Ikeda, Osaka 563-8577 Received February 25, 2003; accepted May 14, 2003 Pyrococcus horikoshii is an obligate anaerobic hyperthermophilic archaeon. In P. horikoshii cells, a hydroperoxide reductase homologue ORF (PH1217) was found to be induced by oxygen. The recombinant protein, which was expressed in E. coli under aerobic conditions, exhibited no activity. However, the recombinant protein prepared under semi-anaerobic conditions exhibited alkyl hydroperoxide reductase activity. Furthermore, it was clarified that it was coupled with the thioredoxin-like system in P. horikoshii. Western blot analysis revealed that the protein was induced by oxygen and hydrogen peroxide. This protein seems to be sensitive to oxygen but forms a thioredoxin-dependent system to eliminate reactive oxygen species in P. horikoshii. Key words: alkyl hydroperoxide reductase, hyperthermophilic archaea, oxidative stress, Pyrococcus horikoshii, thioredoxin system. Organisms have developed various mechanisms that AhpC-like enzyme plays a critical role in the elimination have the ability to eliminate many forms of physiological of hydrogen peroxide that is produced through reduction and chemical stress from their environments, such as of the superoxide anion by SOR. However, an enzyme for reactive oxygen species (ROS), temperature, pH, and the elimination of peroxide has not been reported in osmotic pressure. In particular, oxidative stress caused hyperthermophilic archaea.
    [Show full text]
  • Characterization of the Thioredoxin System of Methanosarcina Mazei
    Characterization of the Thioredoxin System of Methanosarcina mazei Usha Loganathan Thesis submitted to the faculty of Virginia Polytechnic Institute and State University in partial fulfillment of the requirements for the degree of Master of Science In Biological Sciences Committee: Dr. Biswarup Mukhopadhyay, Chair Dr. David L. Popham, Co-Chair Dr. Birgit Scharf December 1, 2014 Blacksburg, Virginia Key words: Methane, fuel, greenhouse gas, Methanogen, Archaea, Methanosarcina mazei, thioredoxin, NADPH thioredoxin reductase, ferredoxin thioredoxin reductase, oxidative stress, redox regulation. Copyright 2014, Usha Loganathan Characterization of the thioredoxin system of Methanosarcina mazei Usha Loganathan ABSTRACT Thioredoxin (Trx) and thioredoxin reductase (TrxR) along with an electron donor form a thioredoxin system. Such systems are widely distributed among the organisms belonging to the three domains of life. It is one of the major disulfide reducing systems, which provides electrons to several enzymes, such as ribonucleotide reductase, methionine sulfoxide reductase and glutathione peroxidase to name a few. It also plays an important role in combating oxidative stress and redox regulation of metabolism. Trx is a small redox protein, about 12 kDa in size, with an active site motif of Cys-X-X-Cys. The reduction of the disulfide in Trx is catalyzed by TrxR. Two types of thioredoxin reductases are known, namely NADPH thioredoxin reductase (NTR) with NADPH as the electron donor and ferredoxin thioredxoin reductase (FTR) which depends on reduced ferredoxin as electron donor. Although NTR is widely distributed in the three domains of life, it is absent in some archaea, whereas FTRs are mostly found in plants, photosynthetic eukaryotes, cyanobacteria, and some archaea.
    [Show full text]
  • Discovery of Industrially Relevant Oxidoreductases
    DISCOVERY OF INDUSTRIALLY RELEVANT OXIDOREDUCTASES Thesis Submitted for the Degree of Master of Science by Kezia Rajan, B.Sc. Supervised by Dr. Ciaran Fagan School of Biotechnology Dublin City University Ireland Dr. Andrew Dowd MBio Monaghan Ireland January 2020 Declaration I hereby certify that this material, which I now submit for assessment on the programme of study leading to the award of Master of Science, is entirely my own work, and that I have exercised reasonable care to ensure that the work is original, and does not to the best of my knowledge breach any law of copyright, and has not been taken from the work of others save and to the extent that such work has been cited and acknowledged within the text of my work. Signed: ID No.: 17212904 Kezia Rajan Date: 03rd January 2020 Acknowledgements I would like to thank the following: God, for sending me angels in the form of wonderful human beings over the last two years to help me with any- and everything related to my project. Dr. Ciaran Fagan and Dr. Andrew Dowd, for guiding me and always going out of their way to help me. Thank you for your patience, your advice, and thank you for constantly believing in me. I feel extremely privileged to have gotten an opportunity to work alongside both of you. Everything I’ve learnt and the passion for research that this project has sparked in me, I owe it all to you both. Although I know that words will never be enough to express my gratitude, I still want to say a huge thank you from the bottom of my heart.
    [Show full text]
  • Kinetics Studies of the Superoxide-Mediated Electron Transfer Reactions Between Rubredoxin-Type Proteins and Superoxide Reductases
    J Biol Inorg Chem (2006) 11: 433–444 DOI 10.1007/s00775-006-0090-0 ORIGINAL PAPER Franc¸oise Auche` re Æ Sofia R. Pauleta Pedro Tavares Æ Isabel Moura Æ Jose´J. G. Moura Kinetics studies of the superoxide-mediated electron transfer reactions between rubredoxin-type proteins and superoxide reductases Received: 26 August 2005 / Accepted: 3 February 2006 / Published online: 17 March 2006 Ó SBIC 2006 Abstract In this work we present a kinetic study of the Keywords Superoxide reductase Æ Rubredoxin Æ superoxide-mediated electron transfer reactions between Oxidative stress Æ Iron–sulfur protein Æ Electron transfer rubredoxin-type proteins and members of the three dif- ferent classes of superoxide reductases (SORs). SORs Abbreviations Dg: Desulfovibrio gigas Æ from the sulfate-reducing bacteria Desulfovibrio vulgaris Dv: Desulfovibrio vulgaris Æ SOD: Superoxide (Dv)andD. gigas (Dg) were chosen as prototypes of dismutase Æ SOR: Superoxide reductase Æ Tp: Treponema classes I and II, respectively, while SOR from the pallidum Æ Tris: Tris(hydroxymethyl)aminomethane syphilis spyrochete Treponema pallidum (Tp) was repre- sentative of class III. Our results show evidence for different behaviors of SORs toward electron acceptance, with a trend to specificity for the electron donor and Introduction acceptor from the same organism. Comparison of the À1 different kapp values, 176.9±25.0 min in the case of Superoxide reductases (SORs), non-heme iron enzymes the Tp/Tp electron transfer, 31.8±3.6 minÀ1 for the Dg/ initially found in sulfate-reducing bacteria, play a fun- Dg electron transfer, and 6.9±1.3 minÀ1 for Dv/Dv, damental role in the defense of microorganisms against could suggest an adaptation of the superoxide-mediated oxidative stress, by catalyzing the monovalent reduction electron transfer efficiency to various environmental of the superoxide anion, rather than the dismutation, conditions.
    [Show full text]
  • Nitrogen Fixation in a Chemoautotrophic Lucinid Symbiosis
    ARTICLES PUBLISHED: 24 OCTOBER 2016 | VOLUME: 2 | ARTICLE NUMBER: 16193 OPEN Nitrogen fixation in a chemoautotrophic lucinid symbiosis Sten König1,2†, Olivier Gros2†, Stefan E. Heiden1, Tjorven Hinzke1,3, Andrea Thürmer4, Anja Poehlein4, Susann Meyer5, Magalie Vatin2, Didier Mbéguié-A-Mbéguié6, Jennifer Tocny2, Ruby Ponnudurai1, Rolf Daniel4,DörteBecher5, Thomas Schweder1,3 and Stephanie Markert1,3* The shallow water bivalve Codakia orbicularis lives in symbiotic association with a sulfur-oxidizing bacterium in its gills. fi ’ The endosymbiont xes CO2 and thus generates organic carbon compounds, which support the host s growth. To investigate the uncultured symbiont’s metabolism and symbiont–host interactions in detail we conducted a proteogenomic analysis of purified bacteria. Unexpectedly, our results reveal a hitherto completely unrecognized feature of the C. orbicularis symbiont’s physiology: the symbiont’s genome encodes all proteins necessary for biological nitrogen fixation (diazotrophy). Expression of the respective genes under standard ambient conditions was confirmed by proteomics. Nitrogenase activity in the symbiont was also verified by enzyme activity assays. Phylogenetic analysis of the bacterial nitrogenase reductase NifH revealed the symbiont’s close relationship to free-living nitrogen-fixing Proteobacteria from the seagrass sediment. The C. orbicularis symbiont, here tentatively named ‘Candidatus Thiodiazotropha endolucinida’,may thus not only sustain the bivalve’scarbondemands.C. orbicularis may also benefit from a steady supply
    [Show full text]
  • Characterization of the Human Copper Chaperone to Superoxide Dismutase
    X-ray Absorption Spectroscopy of Copper: Characterization of the Human Copper Chaperone to Superoxide Dismutase Jay Paul Stasser B.S., Chemistry, The Evergreen State College (1993) A dissertation submitted to the faculty of the OGI School of Science & Engineering at Oregon Health & Science University in partial fulfillment of the requirements for the degree Doctor of Philosophy in Biochemistry and Molecular Biology March 2006 The dissertation “X-ray Absorption Spectroscopy of Copper: Characterization of the Human Copper Chaperone to Superoxide Dismutase” by Jay Paul Stasser has been examined and approved by the following examination committee: ________________________________________ Ninian J. Blackburn, Research Advisor Professor ________________________________________ Bradley M. Tebo Professor ________________________________________ Pierre Moënne-Loccoz Assistant Professor ________________________________________ Arthur Glasfeld Professor Reed College ii Acknowledgments I would like to thank my advisor, Dr. Ninian Blackburn, for the opportunity he gave me by accepting me as one of his students and for his guidance, advice and patience. And I would like to thank my wife and kids, Diana, Regina, and Ariel, for their love and support and the much needed distraction they provided. I would also like to acknowledge the members of the Blackburn lab, past and present, who helped me out through the years, specifically: Martina Ralle, for her help on the beamline and her advice afterwards; Amanda Barry, for the discussions about CCS and other things; Mary Mayfield-Gambill, for her plasmids; Frank Rhames and Shula Jaron, for helping to get me settled into the lab. iii Table of Contents Acknowledgments ......................................................iii List of Tables ........................................................ vii List of Figures ........................................................ viii Abstract ............................................................. xii 1 Introduction .....................................................1 1.1 Overview.
    [Show full text]
  • The Role of Protons in Superoxide Reduction by a Superoxide Reductase Analogue Roslyn M
    The Role of Protons in Superoxide Reduction by a Superoxide Reductase Analogue Roslyn M. Theisen and Julie A. Kovacs* Department of Chemistry, University of Washington, Box 351700 Seattle, WA 98105-1700 RECEIVED DATE (will be automatically inserted after manuscript is accepted) Superoxide reduction by thiolate–ligated catalytically more competent. Protons clearly play an II Me2 + [Fe (S N4(tren))] (1) involves two proton–dependent important role in the SOR mechanism. The source of steps and a single peroxide intermediate protons (in both the first and second protonation steps) has III Me2 + also been a point of discussion. Under acidic conditions, [Fe (S N4(tren))(OOH)] (2). An external proton donor is required, ruling out mechanisms involving H+ or H-atom abstraction from the ligand N–H. The initial protonation step affording 2 occurs with fairly basic + proton donors (EtOH, MeOH, NH4 ) in THF. More acidic proton donors are required to cleave the Fe–O(peroxide) bond in MeOH, and this occurs via a dissociative mechanism. Reaction rates are dependent on the pKa of the proton donor, and a III Me2 2+ common [Fe (S N4(tren))(MeOH)] (3) intermediate is involved. Acetic acid releases H2O2 from 2 under pseudo first-order conditions ([HOAc]= 138 mM, [2]= 0.49 mM) with a rate constant of 8.2 x 10–4 sec–1 at –78 ˚C in MeOH. Reduction of 3 with Cp2Co regenerates the active catalyst 1. Figure 1. Proposed mechanism of superoxide reduction by the non-heme Superoxide reductases (SORs) are non–heme iron iron enzyme superoxide reductase (SOR). – 14 enzymes that reduce superoxide (O2 ) to H2O2 in anaerobic Glu–COOH has been proposed to serve as a proton 1a-j microbes.
    [Show full text]
  • Methionine Sulfoxide Reductases of Archaea
    antioxidants Review Methionine Sulfoxide Reductases of Archaea Julie A. Maupin-Furlow Department of Microbiology and Cell Science, University of Florida, Gainesville, FL 32611-0700, USA; jmaupin@ufl.edu; Tel.: +1-352-392-4095 Received: 29 August 2018; Accepted: 11 September 2018; Published: 20 September 2018 Abstract: Methionine sulfoxide reductases are found in all domains of life and are important in reversing the oxidative damage of the free and protein forms of methionine, a sulfur containing amino acid particularly sensitive to reactive oxygen species (ROS). Archaea are microbes of a domain of life distinct from bacteria and eukaryotes. Archaea are well known for their ability to withstand harsh environmental conditions that range from habitats of high ROS, such as hypersaline lakes of intense ultraviolet (UV) radiation and desiccation, to hydrothermal vents of low concentrations of dissolved oxygen at high temperature. Recent evidence reveals the methionine sulfoxide reductases of archaea function not only in the reduction of methionine sulfoxide but also in the ubiquitin-like modification of protein targets during oxidative stress, an association that appears evolutionarily conserved in eukaryotes. Here is reviewed methionine sulfoxide reductases and their distribution and function in archaea. Keywords: archaea; methionine sulfoxide reductase; reactive oxygen species; ubiquitin-like modification; thiol relay systems 1. Introduction 1 Reactive oxygen species (ROS), such as singlet oxygen ( O2), hydrogen peroxide (H2O2), − superoxide anion (O 2) and hydroxyl radical (HO•), can cause widespread damage to cells. Proteins, amino acids, lipids, nucleic acids, and carbohydrates are generally susceptible to ROS damage [1,2]. The sulfur containing amino acids, methionine (Met) and cysteine (Cys), whether in protein or free form, are particularly sensitive to oxidation by ROS [3,4].
    [Show full text]