DOCSLIB.ORG

Identification and Characterization of the Arabidopsis Gaut1:Gaut7

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Identification and Characterization of the Arabidopsis Gaut1:Gaut7 IDENTIFICATION AND CHARACTERIZATION OF THE ARABIDOPSIS GAUT1:GAUT7 PECTIN HOMOGALACTURONAN:GALACTURONOSYLTRANSFERASE COMPLEX by MELANI AGUSTINA ATMODJO (Under the Direction of Debra Mohnen) ABSTRACT Pectin is a family of structurally complex polysaccharides defined by the presence of α- 1,4-linked galacturonic acid (GalA) residues. It is present in all plant cell walls and contributes substantially to normal plant growth and development. Prior research has elucidated the intricate structure of pectin, yet a detailed understanding of pectin biosynthesis remains elusive. The membrane-bound Arabidopsis GALACTURONOSYLTRANSFERASE 1 (GAUT1) catalyzes the biosynthesis of homogalacturonan (HG), a linear homopolymer of α-1,4-linked GalA that makes up ~65% of total pectin. GAUT1 is an α-1,4-galacturonosyltransferase (GalAT) that transfers GalA from UDP-GalA onto HG acceptors to synthesize HG oligomers and polymers. This study presents the identification of an Arabidopsis HG:GalAT complex composed of GAUT1 and its homolog GAUT7, both of which are Golgi-localized, type II transmembrane proteins. Co-immunoprecipitation using anti-GAUT1 and anti-GAUT7 polyclonal antibodies and bimolecular fluorescent complementation demonstrated protein-protein interaction between GAUT1 and GAUT7. Transcript analyses indicate GAUT1 and GAUT7 co-expression in most tissues and developmental stages of Arabidopsis. Upon non-reducing SDS-PAGE, both GAUT1 and GAUT7 resolve at ~185 kDa, a size larger than their monomeric forms (~60 kDa and ~75 kDa, respectively) observed by reducing SDS-PAGE, thus indicating disulfide bond involvement in GAUT1:GAUT7 complex formation. Proteomics analyses identified GAUT1 and GAUT7 as the only components of the non-reducing SDS-PAGE-stable GAUT1:GAUT7 complex. The observed size of monomeric Arabidopsis GAUT1 (~60 kDa) is smaller than its predicted mass (77.4 kDa). N-terminal sequencing of GAUT1, mass spectrometry protein sequence coverage, and primary structure-dependent GAUT1 reactivity against a series of anti-GAUT1 antibodies, all suggest post-translational cleavage of Arabidopsis GAUT1 between Met167 and Arg168, resulting in a mature GAUT1 with no transmembrane domain. Transient expression of several GFP fusion constructs of GAUT1 in N. benthamiana leaves, in the presence or absence of GAUT7, showed a GAUT7-dependent Golgi accumulation of GAUT1-GFP. It is concluded that Golgi retention of GAUT1 is due to the formation of a complex with GAUT7. Models of the GAUT1:GAUT7 HG:GalAT complex, with GAUT1 as the catalytic subunit and GAUT7 as the membrane anchor, are proposed. Twelve additional proteins were also identified by proteomic analyses and proposed as GAUT1:GAUT7 complex putative interacting proteins. INDEX WORDS: plant cell wall, pectin, homogalacturonan, biosynthesis, glycosyltransferase, galacturonosyltransferase, Golgi apparatus, Arabidopsis thaliana, GAUT, protein complex, membrane anchor IDENTIFICATION AND CHARACTERIZATION OF THE ARABIDOPSIS GAUT1:GAUT7 PECTIN HOMOGALACTURONAN:GALACTURONOSYLTRANSFERASE COMPLEX by MELANI AGUSTINA ATMODJO B.S., Bogor Agricultural University, Indonesia, 1997 M.S., University of Adelaide, Australia, 2001 A Dissertation Submitted to the Graduate Faculty of The University of Georgia in Partial Fulfillment of the Requirements for the Degree DOCTOR OF PHILOSOPHY ATHENS, GEORGIA 2010 © 2010 Melani Agustina Atmodjo All Rights Reserved IDENTIFICATION AND CHARACTERIZATION OF THE ARABIDOPSIS GAUT1:GAUT7 PECTIN HOMOGALACTURONAN:GALACTURONOSYLTRANSFERASE COMPLEX by MELANI AGUSTINA ATMODJO Major Professor: Debra Mohnen Committee: Maor Bar-Peled Michael Pierce Lance Wells Electronic Version Approved: Maureen Grasso Dean of the Graduate School The University of Georgia December 2010 DEDICATION This doctoral work is dedicated to my husband Frank V. Gagliano, my family back home in Bandung, Indonesia, and my new family here in Athens and Crawford, Georgia, U.S. Their continuous support, love, and prayers are what kept me going and made all this possible. iv ACKNOWLEDGEMENTS I would like to thank my major professor, Dr. Debra Mohnen, for giving me the opportunity to learn and work on this project. I appreciate her guidance, friendship, and encouragement throughout my time in her laboratory, as well as the knowledge, experience, and inspiration that she shared me. I would also like to thank my committee members Dr. Maor Bar- Peled, Dr. Michael Pierce, and Dr. Lance Wells, for their support, advice, and constructive criticism that helped me get to where I am now. It was a great honor for me to have the chance to collaborate with these excellent researchers: Dr. Yumiko Sakuragi (University of Copenhagen, Denmark), Dr. Henrik Scheller (Joint BioEnergy Institute), Xiang Zhu (CCRC), Dr James Atwood III (Bioinquire), and Dr. Ron Orlando (CCRC). I am indebted as well to many people who generously provided me with assistance and materials to do my research: Drs. Carl Bergmann and Gerardo Gutierrez-Sanchez (CCRC–for exo- and endopolygalacturonase enzymes), Stefan Eberhard (CCRC–for Arabidopsis suspension culture, RG-I and RG-II polysaccharides), Sarah Inwood (CCRC–for technical assistance and enzyme), Drs. Jae-Min Lim and Lance Wells (CCRC–for LC-MS/MS identification of GAUT1 and GAUT7), Dr. Mary Tierney (The University of Vermont-for pBI101 vector containing AtPRP3 promoter), Dr. Malcolm O’Neill (CCRC–for RG-II monomer and dimer), Dr. Henk Schols (Wageningen University–for RG-I hydrolase enzyme), Dr. Simon Turner (University of Manchester, UK-for KOR1 antibody), and Sangita Mohanty (CCRC–for technical assistance). v My gratitude goes to all past and present members of the Mohnen lab, whom I have been privileged to know, work with, and learn from over the years, and to all the undergraduate students who contributed to my research, especially Amy Burrell and Ioana Petrascu, for their excellent work. I would also like to acknowledge people at CCRC for their administrative (especially Sheilah Dixon-Huckabee, Karen Howard, and Carol Connelly) and technical assistance, scientific advice, and friendships. vi TABLE OF CONTENTS Page ACKNOWLEDGEMENTS.............................................................................................................v INTRODUCTION ...........................................................................................................................1 CHAPTER 1 LITERATURE REVIEW: BIOSYNTHESIS OF PECTIN HOMOGALACTURONAN .................................................................................................4 Introduction ...............................................................................................................4 Overview of Pectin Structure and Functions.............................................................5 Biosynthesis of Homogalacturonan.........................................................................11 Synthesis of Homogalacturonan Backbone.............................................................13 Arabidopsis Galacturonosyltransferase 1 (GAUT1) and the GAUT Gene Family.18 Modifications of Homogalacturonan During Synthesis .........................................29 Conclusions and Relevance ....................................................................................41 2 LITERATURE REVIEW: EUKARYOTIC POLYSACCHARIDE BIOSYNTHESIS – MECHANISM AND THE ROLE OF PROTEIN COMPLEXES...................................43 Introduction .............................................................................................................43 Cellulose Biosynthesis in Higher Plants .................................................................45 Starch Biosynthesis in Higher Plants ......................................................................64 Hyaluronan Biosynthesis in Vertebrates .................................................................81 Heparan Sulfate Biosynthesis in Vertebrates ..........................................................93 vii Protein N-Glycosylation: Role of the Oligosaccharyltransferase Complex..........109 Conclusions and Relevance...................................................................................120 3 GAUT1 AND GAUT7 ARE THE CORE OF A PLANT CELL WALL PECTIN BIOSYNTHETIC HOMOGALACTURONAN:GALACTURONOSYLTRANFERASE COMPLEX........................................................................................................................124 Abstract .................................................................................................................125 Introduction ...........................................................................................................126 Results and Discussion..........................................................................................128 Methods.................................................................................................................146 Supporting Information .........................................................................................149 DISCUSSIONS AND CONCLUSIONS .....................................................................................179 REFERENCES ............................................................................................................................199 APPENDICES .............................................................................................................................238 A Characterization of Anti-GAUT1, Anti-GAUT7, and Anti-GAUT4 Polyclonal Antibodies..................................................................................................................238
Load more

Recommended publications
  • Bacteria Belonging to Pseudomonas Typographi Sp. Nov. from the Bark Beetle Ips Typographus Have Genomic Potential to Aid in the Host Ecology
    insects Article Bacteria Belonging to Pseudomonas typographi sp. nov. from the Bark Beetle Ips typographus Have Genomic Potential to Aid in the Host Ecology Ezequiel Peral-Aranega 1,2 , Zaki Saati-Santamaría 1,2 , Miroslav Kolaˇrik 3,4, Raúl Rivas 1,2,5 and Paula García-Fraile 1,2,4,5,* 1 Microbiology and Genetics Department, University of Salamanca, 37007 Salamanca, Spain; [email protected] (E.P.-A.); [email protected] (Z.S.-S.); [email protected] (R.R.) 2 Spanish-Portuguese Institute for Agricultural Research (CIALE), 37185 Salamanca, Spain 3 Department of Botany, Faculty of Science, Charles University, Benátská 2, 128 01 Prague, Czech Republic; [email protected] 4 Laboratory of Fungal Genetics and Metabolism, Institute of Microbiology of the Academy of Sciences of the Czech Republic, 142 20 Prague, Czech Republic 5 Associated Research Unit of Plant-Microorganism Interaction, University of Salamanca-IRNASA-CSIC, 37008 Salamanca, Spain * Correspondence: [email protected] Received: 4 July 2020; Accepted: 1 September 2020; Published: 3 September 2020 Simple Summary: European Bark Beetle (Ips typographus) is a pest that affects dead and weakened spruce trees. Under certain environmental conditions, it has massive outbreaks, resulting in attacks of healthy trees, becoming a forest pest. It has been proposed that the bark beetle’s microbiome plays a key role in the insect’s ecology, providing nutrients, inhibiting pathogens, and degrading tree defense compounds, among other probable traits. During a study of bacterial associates from I. typographus, we isolated three strains identified as Pseudomonas from different beetle life stages. In this work, we aimed to reveal the taxonomic status of these bacterial strains and to sequence and annotate their genomes to mine possible traits related to a role within the bark beetle holobiont.
    [Show full text]
  • Glucan Phosphorylase-Catalyzed Enzymatic Reactions Using Analog Substrates to Synthesize Non-Natural Oligo- and Polysaccharides
    catalysts Review α-Glucan Phosphorylase-Catalyzed Enzymatic Reactions Using Analog Substrates to Synthesize Non-Natural Oligo- and Polysaccharides Jun-ichi Kadokawa Department of Chemistry, Biotechnology, and Chemical Engineering, Graduate School of Science and Engineering, Kagoshima University, 1-21-40 Korimoto, Kagoshima 860-0065, Japan; [email protected]; Tel.: +81-99-285-7743 Received: 9 October 2018; Accepted: 16 October 2018; Published: 19 October 2018 Abstract: As natural oligo- and polysaccharides are important biomass resources and exhibit vital biological functions, non-natural oligo- and polysaccharides with a well-defined structure can be expected to act as new functional materials with specific natures and properties. α-Glucan phosphorylase (GP) is one of the enzymes that have been used as catalysts for practical synthesis of oligo- and polysaccharides. By means of weak specificity for the recognition of substrates by GP, non-natural oligo- and polysaccharides has precisely been synthesized. GP-catalyzed enzymatic glycosylations using several analog substrates as glycosyl donors have been carried out to produce oligosaccharides having different monosaccharide residues at the non-reducing end. Glycogen, a highly branched natural polysaccharide, has been used as the polymeric glycosyl acceptor and primer for the GP-catalyzed glycosylation and polymerization to obtain glycogen-based non-natural polysaccharide materials. Under the conditions of removal of inorganic phosphate, thermostable GP-catalyzed enzymatic polymerization of analog monomers occurred to give amylose analog polysaccharides. Keywords: analog substrate; α-glucan phosphorylase; non-natural oligo- and polysaccharides 1. Introduction Oligo- and polysaccharides are widely distributed in nature and enact specific important biological functions in accordance with their chemical structures [1].
    [Show full text]
  • An Evolving Hierarchical Family Classification for Glycosyltransferases
    doi:10.1016/S0022-2836(03)00307-3 J. Mol. Biol. (2003) 328, 307–317 COMMUNICATION An Evolving Hierarchical Family Classification for Glycosyltransferases Pedro M. Coutinho1, Emeline Deleury1, Gideon J. Davies2 and Bernard Henrissat1* 1Architecture et Fonction des Glycosyltransferases are a ubiquitous group of enzymes that catalyse the Macromole´cules Biologiques transfer of a sugar moiety from an activated sugar donor onto saccharide UMR6098, CNRS and or non-saccharide acceptors. Although many glycosyltransferases catalyse Universite´s d’Aix-Marseille I chemically similar reactions, presumably through transition states with and II, 31 Chemin Joseph substantial oxocarbenium ion character, they display remarkable diversity Aiguier, 13402 Marseille in their donor, acceptor and product specificity and thereby generate a Cedex 20, France potentially infinite number of glycoconjugates, oligo- and polysacchar- ides. We have performed a comprehensive survey of glycosyltransferase- 2Structural Biology Laboratory related sequences (over 7200 to date) and present here a classification of Department of Chemistry, The these enzymes akin to that proposed previously for glycoside hydrolases, University of York, Heslington into a hierarchical system of families, clans, and folds. This evolving York YO10 5YW, UK classification rationalises structural and mechanistic investigation, harnesses information from a wide variety of related enzymes to inform cell biology and overcomes recurrent problems in the functional prediction of glycosyltransferase-related open-reading frames. q 2003 Elsevier Science Ltd. All rights reserved Keywords: glycosyltransferases; protein families; classification; modular *Corresponding author structure; genomic annotations The biosynthesis of complex carbohydrates and are involved in glycosyl transfer and thus conquer polysaccharides is of remarkable biological import- what Sharon has provocatively described as “the ance.
    [Show full text]
  • A Genomic Study of Soybean Iron Deficiency Chlorosis Jamie Aileen O'rourke Iowa State University
    Iowa State University Capstones, Theses and Retrospective Theses and Dissertations Dissertations 2008 A genomic study of soybean iron deficiency chlorosis Jamie Aileen O'Rourke Iowa State University Follow this and additional works at: https://lib.dr.iastate.edu/rtd Part of the Agricultural Science Commons, Agronomy and Crop Sciences Commons, and the Horticulture Commons Recommended Citation O'Rourke, Jamie Aileen, "A genomic study of soybean iron deficiency chlorosis" (2008). Retrospective Theses and Dissertations. 15824. https://lib.dr.iastate.edu/rtd/15824 This Dissertation is brought to you for free and open access by the Iowa State University Capstones, Theses and Dissertations at Iowa State University Digital Repository. It has been accepted for inclusion in Retrospective Theses and Dissertations by an authorized administrator of Iowa State University Digital Repository. For more information, please contact [email protected]. A genomic study of soybean iron deficiency chlorosis by Jamie Aileen O’Rourke A dissertation submitted to the graduate faculty in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY Major: Genetics Program of Study Committee: Randy C. Shoemaker, Co-Major Professor Steve Whitham, Co-Major Professor Silvia Cianzio Thomas Baum David Hannapel Iowa State University Ames, Iowa 2008 Copyright © Jamie Aileen O’Rourke, 2008. All rights reserved. 3316180 3316180 2008 ii Dedicated to: To my parents, Tamia and Dennis, and my sisters, Kelly and Darcy. For always believing in and supporting my
    [Show full text]
  • Plant Nucleotide Sugar Formation, Interconversion, and Salvage by Sugar Recycling*
    Plant Nucleotide Sugar Formation, Interconversion, and Salvage by Sugar Recycling∗ Maor Bar-Peled1,2 and Malcolm A. O’Neill2 1Department of Plant Biology and 2Complex Carbohydrate Research Center, University of Georgia, Athens, Georgia 30602; email: [email protected], [email protected] Annu. Rev. Plant Biol. 2011. 62:127–55 Keywords First published online as a Review in Advance on nucleotide sugar biosynthesis, nucleotide sugar interconversion, March 1, 2011 nucleotide sugar salvage, UDP-glucose, UDP-xylose, The Annual Review of Plant Biology is online at UDP-arabinopyranose mutase plant.annualreviews.org This article’s doi: Abstract 10.1146/annurev-arplant-042110-103918 Nucleotide sugars are the universal sugar donors for the formation Copyright c 2011 by Annual Reviews. by Oak Ridge National Lab on 05/20/11. For personal use only. of polysaccharides, glycoproteins, proteoglycans, glycolipids, and All rights reserved glycosylated secondary metabolites. At least 100 genes encode proteins 1543-5008/11/0602-0127$20.00 involved in the formation of nucleotide sugars. These nucleotide ∗ Annu. Rev. Plant Biol. 2011.62:127-155. Downloaded from www.annualreviews.org Dedicated to Peter Albersheim for his inspiration sugars are formed using the carbohydrate derived from photosynthesis, and his pioneering studies in determining the the sugar generated by hydrolyzing translocated sucrose, the sugars structure and biological functions of complex carbohydrates. released from storage carbohydrates, the salvage of sugars from glycoproteins and glycolipids, the recycling of sugars released during primary and secondary cell wall restructuring, and the sugar generated during plant-microbe interactions. Here we emphasize the importance of the salvage of sugars released from glycans for the formation of nucleotide sugars.
    [Show full text]
  • Starch Phosphorylase: an Overview of Biochemical Characterization, Immobilization and Peptide Mapping
    British Biotechnology Journal 5(3): 103-122, 2015, Article no.BBJ.2015.011 ISSN: 2231–2927 SCIENCEDOMAIN international www.sciencedomain.org Starch Phosphorylase: An Overview of Biochemical Characterization, Immobilization and Peptide Mapping Ritu Jain1, Sarika Garg2 and Anil Kumar1* 1School of Biotechnology, Devi Ahilya University, Khandwa Road, Indore-452001, India. 2Department of Neuroscience, The University of Montreal Hospital Research Centre (CRCHUM), Bureau/Office: CRCHUM R09.476, Tour Viger, 900, rue St-Denis, Montréal, Québec H2X 0A9, Canada. Authors’ contributions This work was carried out in collaboration between all authors. Authors RJ and SG have equal contribution in this manuscript. Author RJ prepared the first draft of the manuscript. Authors RJ and SG both searched literature. The manuscript has been finalized by authors SG and AK. Author AK also designed the study and managed the analyses of the study. All authors read and approved the final manuscript. Article Information DOI: 10.9734/ BBJ/2015/14522 Editor(s): (1) Samuel Peña-Llopis, Developmental Biology and Internal Medicine, Oncology Division, University of Texas Southwestern Medical Center, USA. Reviewers: (1) Anonymous, University of Genoa, Italy. (2) Anonymous, Kagoshima University, Japan. (3) Muhammad Yasin Naz, Dept. of Fundamental & Applied Sciences, Universiti Teknologi Petronas, 31750 Tronoh, Malaysia. (4) Anonymous, Center for Research for Food and Development, México. (5) Ana Lucia Ferrarezi, Department of Biology, Unesp, Brazil. Complete Peer review History: http://www.sciencedomain.org/review-history.php?iid=803&id=11&aid=7562 Received 4th October 2014 th Review Article Accepted 9 December 2014 Published 31st December 2014 ABSTRACT Starch phosphorylase (EC 2.4.1.1) is the key enzyme which catalyzes the reversible conversion of starch and inorganic phosphate into glucose-1-phosphate (G-1-P) and has been reported in many higher plants.
    [Show full text]
  • The Decomposition of Starch Through Phosphorolysis by Streptoccus
    The decomposition of starch through phosphorolysis by Streptoccus bovis by Richard N Ushijima A THESIS Submitted to the Graduate Faculty in partial fulfillment of the requirements for the degree of Master of Science in Bacteriology at Montana State College Montana State University © Copyright by Richard N Ushijima (1957) Abstract: The possibility of starch utilization by Streptococcus bovis through a starch phosphorylase was investigated. large quantities of cells were grown on a starch medium, and the cells were harvested by centrifugation. The cell-free enzyme extract was prepared by grinding the cells with glass beads in a micro-Waring blendor. Starch phosphorylase was detected in the reaction mixture containing soluble starch, inorganic phosphate, enzyme preparation, and sodium hydrosulfite. Since a maltose phosphorylase was also found, the formation of maltose was prevented by adjusting the mixture to pH 8.6, which inhibited alpha amylase activity. The experiments also revealed the presence of a phosphoglueomutase, which converts glucose-l-phosphate to glucose-6-phosphate, a product uti-lizable through the glycolytic pathways. I THE''DECOMPOSITION' OF' STARCH THROUGH ' ^ PHOSPHOROLYSIS BI STREPTOCOCCUS'BOVIS by RICHARD N„ USHIJHja A THESIS Submitted to the Graduate Faculty in partial fulfillment of the requirements for the degree of Master of Science in Bacteriology at Montana State College Approved^ Headj Major Department Chairman, Examining Committee Hi Bozeman, Montana <ii ‘‘il.j ' ') I' V r L u 5 A- CCg J- " 2 ' TABLE OF CONTENTS ACKNOWLEDGMENTS 3 ABSTRACT U INTRODUCTION 5 METHODS AND MATERIALS 0\ CN Os Culture Preparation of large cultures Preparation of cell-free bacterial extracts IO Phosphorus determination 11 Chromatography 12 Glucose determination 12 EXPERIMENTAL RESULTS lh Phosphorylase ill Amylase 15> Starch phosphorylase 20 Phosphoglucomutase 2h DISCUSSION 2? SUMMARY 31 LITERATURE CITED 32 123688 ACKmimEDGMEHTS The author takes this opportunity to express his gratitude to Dr, R» H.
    [Show full text]
  • Immobilization of Starch Phosphorylase from Cabbage Leaves: Production of Glucose-1-Phosphate
    Brazilian Journal of Chemical ISSN 0104-6632 Printed in Brazil Engineering www.abeq.org.br/bjche Vol. 25, No. 02, pp. 229 - 235, April - June, 2008 IMMOBILIZATION OF STARCH PHOSPHORYLASE FROM CABBAGE LEAVES: PRODUCTION OF GLUCOSE-1-PHOSPHATE N. Garga and A. Kumar* School of Biotechnology, Devi Ahilya University, Phone: +(91) 731-2470372, 2470373, Fax: +(91) -731-2470372, Khandwa Road, Indore-452001, India. E mail: [email protected] aPresent address: Division of Plant Physiology, Institute of Biochemistry and Biology, University of Potsdam, Karl-Liebknecht-Str. 24-25, Haus 20 D-14476, Golm, Germany. (Received: October 10, 2006 ; Accepted: January 8, 2008) Abstract - Starch phosphorylase has been isolated from cabbage (Elephantopus scabar) leaves and partially purified using ammonium sulfate fractionation. The partially purified enzyme was desalted using Sephadex- G-25 chromatography. In the direction of polysaccharide synthesis, the enzyme showed optimum activity at pH 6.0 with two half pH optima at pH 5.3 and pH 7.1 whereas in the direction of glucose-1-phosphate formation, it showed optimum pH at pH 7.0 with half pH-optima at pH 6.4 and 7.6. The optimum temperature for the enzyme activity has been found to be 37oC with two half temperature optima at 34oC and 40oC. The partially purified enzyme has been immobilized using egg shell as solid support. The percentage retention of the enzyme on egg shell was nearly 56%. After immobilization, specific activity of the enzyme increased from 0. 0225 to 0.0452. Upon immobilization, there was a slight alkaline shift in the optimum pH when assayed in both the directions.
    [Show full text]
  • Characterization of the Starch Branching Enzymes in Arabidopsis Thaliana Qinglei Gan Iowa State University
    Iowa State University Capstones, Theses and Retrospective Theses and Dissertations Dissertations 2007 Characterization of the starch branching enzymes in Arabidopsis thaliana Qinglei Gan Iowa State University Follow this and additional works at: https://lib.dr.iastate.edu/rtd Part of the Biochemistry Commons, and the Molecular Biology Commons Recommended Citation Gan, Qinglei, "Characterization of the starch branching enzymes in Arabidopsis thaliana" (2007). Retrospective Theses and Dissertations. 15042. https://lib.dr.iastate.edu/rtd/15042 This Thesis is brought to you for free and open access by the Iowa State University Capstones, Theses and Dissertations at Iowa State University Digital Repository. It has been accepted for inclusion in Retrospective Theses and Dissertations by an authorized administrator of Iowa State University Digital Repository. For more information, please contact [email protected]. Characterization of the starch branching enzymes in Arabidopsis thaliana by Qinglei Gan A thesis submitted to the graduate faculty in partial fulfillment of the requirements for the degree of MASTER OF SCIENCE Major: Biochemistry Program of Study Committee: Alan M. Myers, Co-major Professor Martha G. James, Co-major Professor Eve S. Wurtele Paul M. Scott Iowa State University Ames, Iowa 2007 Copyright © Qinglei Gan, 2007. All rights reserved. UMI Number: 1446041 UMI Microform 1446041 Copyright 2007 by ProQuest Information and Learning Company. All rights reserved. This microform edition is protected against unauthorized copying under Title 17, United States Code. ProQuest Information and Learning Company 300 North Zeeb Road P.O. Box 1346 Ann Arbor, MI 48106-1346 ii TABLE OF CONTENTS LIST OF FIGURES iii LIST OF TABLES v CHAPTER 1.
    [Show full text]
  • Functional and Structural Insights Into Glycoside Hydrolase Family 130 Enzymes: Implications in Carbohydrate Foraging by Human Gut Bacteria
    TTHHÈÈSSEE En vue de l'obtention du DOCTORAT DE L’UNIVERSITÉ DE TOULOUSE Délivré par Discipline ou spécialité : Ingénieries Moléculaires et Enzymatiques Présentée et soutenue par Le Titre : JURY Ecole doctorale : Sciences Ecologiques, Vétérinaires, Agronomiques et Bioingénieries (SEVAB) Unité de recherche : LISBP, UMR CNRS 5504, UMR INRA 792, INSA Toulouse Directeur(s) de Thèse : Gabrielle POTOCKI-VÉRONÈSE, Directrice de Recherche INRA Elisabeth LAVILLE, Chargée de Recherche INRA THÈSE En vue de l’obtention du DOCTORAT de L’INSTITUT NATIONAL DES SCIENCES APPLIQUÉES DE TOULOUSE Présentée et soutenue publiquement le mardi 28 avril 2015 Spécialité : Sciences Ecologiques, Vétérinaires, Agronomiques et Bioingénieries Filière : Ingénieries Microbienne et Enzymatique Par Simon LADEVÈZE Functional and Structural insights into glycoside hydrolase family 130 enzymes: implications in carbohydrate foraging by human gut bacteria Directrices de thèse: Mme Gabrielle POTOCKI-VÉRONÈSE, Directrice de Recherche INRA Mme Elisabeth LAVILLE, Chargée de Recherche INRA 1 2 NOM: LADEVÈZE PRÉNOM: Simon TITRE: Apports Fonctionnels et Structuraux à la famille des glycoside hydrolase 130: implications dans la dégradation des glycanes par les bactéries de l'intestin humain. SPÉCIALITÉ: Sciences Ecologiques, Vétérinaires, Agronomiques et Bioingénieries FILIÈRE: Ingénieries Moléculaires et Enzymatiques ANNÉE: 2015 LIEU: INSA, TOULOUSE DIRECTRICES DE THÈSE: Mmes Gabrielle POTOCKI-VÉRONÈSE et Elisabeth LAVILLE RÉSUMÉ: Les relations entre bactéries intestinales, aliments et hôte jouent un rôle crucial dans le maintien de la santé humaine. La caractérisation fonctionnelle d’Uhgb_MP, une enzyme de la famille 130 des glycoside hydrolases découverte par métagénomique fonctionnelle, a révélé une nouvelle fonction de dégradation par phosphorolyse des polysaccharides de la paroi végétale et des glycanes de l'hôte tapissant l'épithélium intestinal.
    [Show full text]
  • Enabling Tools for Toxoplasma Glycobiology 02Sept2018
    bioRxiv preprint doi: https://doi.org/10.1101/401828; this version posted September 8, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. Enabling tools for Toxoplasma glycobiology Elisabet Gas-Pascual1,2,*, H. Travis Ichikawa1,5,*, M. Osman Sheikh3, M. Isabella Serji1,6, Bowen Deng1,2, Msano Mandalasi1,2, Giulia Bandini4, John Samuelson4, Lance Wells1,3, and Christopher M. West1,2,7 1Department of Biochemistry & Molecular Biology, 2Center for Tropical and Emerging Global Diseases, 3Complex Carbohydrate Research Center, University of Georgia, Athens, GA 30602 USA 4Department of Molecular and Cell Biology, Henry M. Goldman School of Dental Medicine, Boston University, 72 East Concord Street, Boston, MA 02118 USA 5 current address: New Materials Institute, University of Georgia, Athens, GA 30602 USA 6 current address: George Washington University School of Public Health, Washington, D.C. 20009 USA 7 to whom requests for information or materials should be sent at Dept. of Biochemistry & Molecular Biology, 120 E. Green St., Davison Life Sciences – B129, University of Georgia, Athens GA 30602 USA, telephone +1-706-542-4259, email [email protected] * These authors contributed equally to this project Keywords: CRISPR/Cas, glycomics, Toxoplasma gondii, glycosyltransferase, glycobiology, mass spectrometry, parasitology Abbreviations: AAL, Aleuria aurantia
    [Show full text]
  • A Fungal Pathogen Causing Phomopsis Seed Decay in Soybean Shuxian Li1*† , Omar Darwish2†, Nadim W
    Li et al. BMC Genomics (2017) 18:688 DOI 10.1186/s12864-017-4075-x RESEARCHARTICLE Open Access Analysis of the genome sequence of Phomopsis longicolla: a fungal pathogen causing Phomopsis seed decay in soybean Shuxian Li1*† , Omar Darwish2†, Nadim W. Alkharouf2, Bryan Musungu3,4 and Benjamin F. Matthews5 Abstract Background: Phomopsis longicolla T. W. Hobbs (syn. Diaporthe longicolla) is a seed-borne fungus causing Phomopsis seed decay in soybean. This disease is one of the most devastating diseases reducing soybean seed quality worldwide. To facilitate investigation of the genomic basis of pathogenicity and to understand the mechanism of the disease development, the genome of an isolate, MSPL10–6, from Mississippi, USA was sequenced, de novo assembled, and analyzed. Results: ThegenomeofMSPL10–6wasestimatedtobeapproximately62MbinsizewithanoverallG+C content of 48.6%. Of 16,597 predicted genes, 9866 genes (59.45%) had significant matches to genes in the NCBI nr database, while 18.01% of them did not link to any gene ontology classification, and 9.64% of genes did not significantly match any known genes. Analysis of the 1221 putative genes that encoded carbohydrate-activated enzymes (CAZys) indicated that 715 genes belong to three classes of CAZy that have a direct role in degrading plant cell walls. A novel fungal ulvan lyase (PL24; EC 4.2.2.-) was identified. Approximately 12.7% of the P. longicolla genome consists of repetitive elements. A total of 510 potentially horizontally transferred genes were identified. They appeared to originate from 22 other fungi, 26 eubacteria and 5 archaebacteria. Conclusions: The genome of the P. longicolla isolate MSPL10–6 represented the first reported genome sequence in the fungal Diaporthe-Phomopsis complex causing soybean diseases.
    [Show full text]