Search

Your search keyword '"Emeline Fabre"' showing total 23 results
Start Over Author "Emeline Fabre"
23 results on '"Emeline Fabre"'

1. Structural Sampling of Glycan Interaction Profiles Reveals Mucosal Receptors for Fimbrial Adhesins of Enterotoxigenic Escherichia coli

Author

Manfred Wuhrer, Julie Bouckaert, Stefan Oscarson, André M. Deelder, Yann Guérardel, Emeline Fabre, Johan D. M. Olsson, Manfred S. Weiss, Lieven Buts, Arjen R. de Boer, Emanuela Lonardi, and Kristof Moonens

Subjects
adhesin, glycan array, E. coli, enterotoxigenic, F17G, FedF, FimH, charge, arginine, sulfate, Biology (General), QH301-705.5
Abstract

Fimbriae are long, proteinaceous adhesion organelles expressed on the bacterial envelope, evolutionarily adapted by Escherichia coli strains for the colonization of epithelial linings. Using glycan arrays of the Consortium for Functional Glycomics (CFG), the lectin domains were screened of the fimbrial adhesins F17G and FedF from enterotoxigenic E. coli (ETEC) and of the FimH adhesin from uropathogenic E. coli. This has led to the discovery of a more specific receptor for F17G, GlcNAcb1,3Gal. No significant differences emerged from the glycan binding profiles of the F17G lectin domains from five different E. coli strains. However, strain-dependent amino acid variations, predominantly towards the positively charged arginine, were indicated by sulfate binding in FedF and F17G crystal structures. For FedF, no significant binders could be observed on the CFG glycan array. Hence, a shotgun array was generated from microvilli scrapings of the distal jejunum of a 3-week old piglet about to be weaned. On this array, the blood group A type 1 hexasaccharide emerged as a receptor for the FedF lectin domain and remarkably also for F18-fimbriated E. coli. F17G was found to selectively recognize glycan species with a terminal GlcNAc, typifying intestinal mucins. In conclusion, F17G and FedF recognize long glycan sequences that could only be identified using the shotgun approach. Interestingly, ETEC strains display a large capacity to adapt their fimbrial adhesins to ecological niches via charge-driven interactions, congruent with binding to thick mucosal surfaces displaying an acidic gradient along the intestinal tract.

Published
2013
Full Text
View/download PDF

2. The endogenous galactofuranosidase GlfH1 hydrolyzes mycobacterial arabinogalactan

Author

Albertus Viljoen, Yann Guérardel, Alexandre Mery, Maju Joe, Laurent Kremer, Sydney A. Villaume, Emeline Fabre, Christophe Mariller, Kaoru Takegawa, Stéphane P. Vincent, Lin Shen, Todd L. Lowary, Iman Halloum, Loïc P. Chêne, Université de Lille, CNRS, Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 [UGSF], Unité de Glycobiologie Structurale et Fonctionnelle (UGSF) - UMR 8576, Institut de Recherche en Infectiologie de Montpellier [IRIM], Université de Namur [Namur] [UNamur], University of Alberta, Unité de Glycobiologie Structurale et Fonctionnelle UMR 8576 [UGSF], Kyushu University, Institut National de la Santé et de la Recherche Médicale [INSERM], Unité de Glycobiologie Structurale et Fonctionnelle (UGSF), Université de Lille-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Institut de Recherche en Infectiologie de Montpellier (IRIM), Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Chimie Bio-Organic [Namur, Belgium], Université de Namur [Namur] (UNamur), Dynamique des interactions membranaires normales et pathologiques (DIMNP), Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM)-Université Montpellier 2 - Sciences et Techniques (UM2)-Université Montpellier 1 (UM1), Kyushu University [Fukuoka], Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 (UGSF), Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM), Université de Namur [Namur], Université Montpellier 1 (UM1)-Université Montpellier 2 - Sciences et Techniques (UM2)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Université de Lille-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Recherche Agronomique (INRA), Université de Lille-Centre National de la Recherche Scientifique (CNRS), and Kremer, Laurent

Subjects
0301 basic medicine, cell envelope, mycobacteria, [SDV]Life Sciences [q-bio], Sequence Homology, Glycobiology and Extracellular Matrices, Biochemistry, Galactans, Cell wall, Mycobacterium tuberculosis, 03 medical and health sciences, chemistry.chemical_compound, Arabinogalactan, galactofuranose, Glycoside hydrolase, Amino Acid Sequence, Amoeba, Molecular Biology, [SDV.MP] Life Sciences [q-bio]/Microbiology and Parasitology, Phylogeny, chemistry.chemical_classification, polysaccharide, glycosidas, arabinogalactan, catabolism, galactofuranosidase, Rv3096, 030102 biochemistry & molecular biology, biology, Chemistry, Hydrolysis, Cell Biology, Galactan, biology.organism_classification, Galactosyltransferases, [SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, 3. Good health, [SDV] Life Sciences [q-bio], Kinetics, 030104 developmental biology, Enzyme, [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology, glycosidase, Cell envelope, [SDV.MP.BAC] Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, Biogenesis
Abstract

International audience; Despite the impressive progress made over the past 20 years in our understanding of mycolylarabinogalactan-peptidoglycan (mAGP) biogenesis, the mechanisms by which the tubercle bacillus Mycobacterium tuberculosis adapts its cell wall structure and /composition in response to various environmental conditions, especially during infection, remain poorly understood. Being the central portion of the mAGP complex, arabinogalactan (AG) is believed to be the constituent of the mycobacterial cell envelope that undergoes the least structural changes in its structure, but no reports exist supportings this assumption. Herein, using [MS2] recombinantly expressed mycobacterial protein, bioinformatics analyses, and kinetic and biochemical assays, we demonstrate that the AG can be remodeled by a mycobacterial endogenous enzyme. In particular, we identified found that the mycobacterial protein GlfH1 (Rv3096), which protein exhibits an exo-β-D-galactofuranose hydrolase activity and is capable of hydrolyzing the galactan chain of AG by recurrent cleavage of the terminal β-(1,5) and β-(1,6)-Galf linkages. The characterization of this galactosidase represents the a first step towards understanding the remodeling of mycobacterial AG.

Published
2020

3. Mannosylation of fungal glycoconjugates in the Golgi apparatus

Author

Emeline Fabre, Chantal Fradin, and Thomas Hurtaux

Subjects
Microbiology (medical), Glycosylation, Glycoconjugate, Saccharomyces cerevisiae, Golgi Apparatus, Mannosyltransferases, Microbiology, Fungal Proteins, symbols.namesake, chemistry.chemical_compound, Candida albicans, chemistry.chemical_classification, Sphingolipids, biology, Golgi apparatus, biology.organism_classification, Sphingolipid, Cell biology, carbohydrates (lipids), Infectious Diseases, Biochemistry, chemistry, Mannosylation, symbols, lipids (amino acids, peptides, and proteins), Glycoconjugates, Mannose
Abstract

Glycosylation is a crucial step in the modification of proteins or sphingolipids that then play a prominent role in fungal biology. Glycosylation controls the structure and plasticity of the fungal cell wall and fungi-host interactions. Non-pathogenic and pathogenic yeasts, such as Saccharomyces cerevisiae and Candida albicans, respectively, have been useful models for analyzing the mannosylation of proteins and sphingolipids, which mainly takes place in the Golgi apparatus. Studies of these yeasts have identified different mannosyltransferases that belong to separate families of glycosyltransferases. The characterization of mannosyltransferases and their activities is essential for deciphering cell wall biogenesis, for identifying mannosides involved in virulence and for designing inhibitors that target specific mannosylation processes.

Published
2014

4. Characterization of the recombinant Candida albicans β-1,2-mannosyltransferase that initiates the β-mannosylation of cell wall phosphopeptidomannan

Author

Ghenima Sfihi-Loualia, Bernadette Coddeville, Julie Bouckaert, Thomas Hurtaux, Jean-Maurice Mallet, Frédéric Krzewinski, Yann Guérardel, Emeline Fabre, Florence Delplace, Daniel Poulain, Chantal Fradin, Romaric Dubois, Emmanuel Maes, Marilyne Pourcelot, Unité de Glycobiologie Structurale et Fonctionnelle UMR 8576 (UGSF), Université de Lille-Centre National de la Recherche Scientifique (CNRS), Université Pierre et Marie Curie - Paris 6 (UPMC), Physiopathologie des Candidoses, Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille, Droit et Santé, Inflammation: mécanismes et régulation et interactions avec la nutrition et les candidoses, Institut National de la Recherche Agronomique (INRA)-Université de Lille-Centre National de la Recherche Scientifique (CNRS), Université de Lille, Droit et Santé-Institut National de la Santé et de la Recherche Médicale (INSERM), Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 (UGSF), and Université de Lille-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Recherche Agronomique (INRA)

Subjects
Phosphopeptides, Mannosyltransferase, Mannose, Biology, Mannosyltransferases, Biochemistry, Pichia pastoris, Microbiology, Mannans, 03 medical and health sciences, chemistry.chemical_compound, Cell Wall, Candida albicans, Molecular Biology, 030304 developmental biology, 0303 health sciences, [CHIM.ORGA]Chemical Sciences/Organic chemistry, 030302 biochemistry & molecular biology, Cell Biology, biology.organism_classification, Recombinant Proteins, Corpus albicans, Yeast, chemistry, Mannosylation
Abstract

The presence of β-mannosides in their cell walls confers specific features on the pathogenic yeasts Candida albicans and Candida glabrata compared with non-pathogenic yeasts. In the present study, we investigated the enzymatic properties of Bmt1 (β-mannosyltransferase 1), a member of the recently identified β-mannosyltransferase family, from C. albicans. A recombinant soluble enzyme lacking the N-terminal region was expressed as a secreted protein from the methylotrophic yeast Pichia pastoris. In parallel, functionalized natural oligosaccharides isolated from Saccharomyces cerevisiae and a C. albicans mutant strain, as well as synthetic α-oligomannosides, were prepared and used as potential acceptor substrates. Bmt1p preferentially utilizes substrates containing linear chains of α-1,2-linked mannotriose or mannotetraose. The recombinant enzyme consecuti-vely transfers two mannosyl units on to these acceptors, leading to the production of α-mannosidase-resistant oligomannosides. NMR experiments further confirmed the presence of a terminal βMan (β-1,2-linked mannose) unit in the first enzyme product. In the future, a better understanding of specific β-1,2-mannosyltransferase molecular requirements will help the design of new potential antifungal drugs.

Published
2013

5. Identification and characterisation of citrullinated antigen-specific B cells in peripheral blood of patients with rheumatoid arthritis

Author

Theo Rispens, Yoann Rombouts, Arnaud Zaldumbide, Emeline Fabre, Ellen I. H. van der Voort, Tom W J Huizinga, Gertjan Wolbink, Rob C. Hoeben, Dominique Baeten, Priscilla F Kerkman, Hergen Spits, Hans Scherer, René E. M. Toes, Landsteiner Laboratory, Cell Biology and Histology, Experimental Immunology, and Clinical Immunology and Rheumatology

Subjects
0301 basic medicine, Immunology, B-Lymphocyte Subsets, medicine.disease_cause, Autoantigens, Peptides, Cyclic, General Biochemistry, Genetics and Molecular Biology, Immunophenotyping, Autoimmunity, Flow cytometry, Arthritis, Rheumatoid, 03 medical and health sciences, 0302 clinical medicine, Rheumatology, Antigen, Immunity, medicine, Humans, Immunology and Allergy, Cells, Cultured, Autoantibodies, 030203 arthritis & rheumatology, biology, medicine.diagnostic_test, business.industry, medicine.disease, In vitro, 030104 developmental biology, Immunoglobulin G, Rheumatoid arthritis, Biotinylation, biology.protein, Antibody, business
Abstract

ObjectivesImmunity to citrullinated antigens is a hallmark of rheumatoid arthritis (RA). We set out to elucidate its biology by identifying and characterising citrullinated antigen-specific B cells in peripheral blood of patients with RA.MethodsDifferentially labelled streptavidin and extravidin tetramers were conjugated to biotinylated CCP2 or control antigens and used in flow cytometry to identify citrullinated antigen-specific B cells in peripheral blood. Tetramer-positive and tetramer-negative B cells were isolated by fluorescence activated cell sorting (FACS) followed by in vitro culture and analysis of culture supernatants for the presence of antibodies against citrullinated protein antigens (ACPA) by ELISA. Cells were phenotypically characterised by flow cytometry.ResultsBy combining differentially labelled CCP2 tetramers, we successfully separated citrullinated antigen-specific B cells from non-specific background signals. Isolated tetramer-positive B cells, but not tetramer-negative cells, produced large amounts of ACPA upon in vitro stimulation. Phenotypic analyses revealed that citrullinated antigen-specific B cells displayed markers of class-switched memory B cells and plasmablasts, whereas only few cells displayed a naïve phenotype. The frequency of tetramer-positive cells was high (up to 1/500 memory B cells with a median of 1/12 500 total B cells) and correlated with ACPA serum titres and spontaneous ACPA production in culture.ConclusionsWe developed a technology to identify and isolate citrullinated antigen-specific B cells from peripheral blood of patients with RA. Most cells have a memory phenotype, express IgA or IgG and are present in relatively high frequencies. These data pave the path for a direct and detailed molecular characterisation of ACPA-expressing B cells and could lead to the identification of novel therapeutic targets.

Published
2016

6. Candida albicans β-1,2 mannosyl transferase Bmt3: Preparation and evaluation of a β (1,2), α (1,2)-tetramannosyl fluorescent substrate

Author

Emmanuel Maes, Emeline Fabre, Daniel Poulain, Chantal Fradin, Laurent Cattiaux, Ghenima Sfihi-Loualia, Marilyne Pourcelot, Jean-Maurice Mallet, Florence Delplace, Boualem Sendid, Thomas Hurtaux, Yann Guérardel, Anaïs Mée, Peptides, glycoconjugués et métaux en biologie (LBM-E1), Laboratoire des biomolécules (LBM UMR 7203), Université Pierre et Marie Curie - Paris 6 (UPMC)-Département de Chimie - ENS Paris, École normale supérieure - Paris (ENS Paris)-École normale supérieure - Paris (ENS Paris)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Département de Chimie - ENS Paris, École normale supérieure - Paris (ENS Paris)-École normale supérieure - Paris (ENS Paris)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 (UGSF), Université de Lille-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Recherche Agronomique (INRA), Université Pierre et Marie Curie - Paris 6 (UPMC), Université Lille Nord de France (COMUE), Physiopathologie des Candidoses, Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille, Droit et Santé, ENSA Phytotechnie, INRA Agronomie, Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés (LISBP), Centre National de la Recherche Scientifique (CNRS)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut National de la Recherche Agronomique (INRA), École normale supérieure - Paris (ENS-PSL), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-École normale supérieure - Paris (ENS-PSL), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Département de Chimie - ENS Paris, Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Unité de Glycobiologie Structurale et Fonctionnelle UMR 8576 (UGSF), Université de Lille-Centre National de la Recherche Scientifique (CNRS), Institut National de la Recherche Agronomique (INRA)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS), Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Département de Chimie - ENS Paris, École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Département de Chimie - ENS Paris, Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université Pierre et Marie Curie - Paris 6 (UPMC), Institut National de la Recherche Agronomique (INRA)-Université de Lille-Centre National de la Recherche Scientifique (CNRS), and Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)

Subjects
0301 basic medicine, Glycosylation, Stereochemistry, Clinical Biochemistry, Molecular Conformation, Pharmaceutical Science, Biochemistry, Chemical synthesis, Mannosyltransferases, Substrate Specificity, 03 medical and health sciences, chemistry.chemical_compound, Drug Discovery, Candida albicans, Transferase, Molecular Biology, Fluorescent Dyes, chemistry.chemical_classification, biology, [CHIM.ORGA]Chemical Sciences/Organic chemistry, Organic Chemistry, Glycoside, Substrate (chemistry), biology.organism_classification, Fluorescence, 3. Good health, 030104 developmental biology, chemistry, Mannosides, Thiol, Molecular Medicine
Abstract

International audience; We describe for the first time the chemical synthesis of a tetramannoside, containing both α (1 → 2) and β (1 → 2) linkages. Dodecylthio (lauryl) glycosides were prepared from odorless dodecyl thiol and used as donors for the glycosylation steps. This tetramannoside, was coupled to a mantyl group, and revealed to be a perfect substrate of β-mannosyltransferase Bmt3, confirming the proposed specificity and allowing the preparation of a pentamannoside sequence (β Man (1,2) β Man (1,2) α Man (1,2) α Man (1,2) α Man) usable as a novel substrate for further elongation studies.

Published
2016

7. Evaluation of monovalent and multivalent iminosugars to modulate Candida albicans β-1,2-mannosyltransferase activities

Author

Sébastien G. Gouin, Boualem Sendid, Florence Delplace, Yoan Brissonnet, Ghenima Sfihi-Loualia, Jean-Maurice Mallet, Yann Guérardel, Emeline Fabre, Julie Bouckaert, Thomas Hurtaux, Chimie Et Interdisciplinarité : Synthèse, Analyse, Modélisation (CEISAM), Université de Nantes - UFR des Sciences et des Techniques (UN UFR ST), Université de Nantes (UN)-Université de Nantes (UN)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Unité de Glycobiologie Structurale et Fonctionnelle UMR 8576 (UGSF), Université de Lille-Centre National de la Recherche Scientifique (CNRS), Université Lille Nord de France (COMUE), Université Pierre et Marie Curie - Paris 6 (UPMC), Physiopathologie des Candidoses, Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille, Droit et Santé, Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés (LISBP), Institut National de la Recherche Agronomique (INRA)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS), Université de Nantes - Faculté des Sciences et des Techniques, Université de Nantes (UN)-Université de Nantes (UN)-Centre National de la Recherche Scientifique (CNRS), Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 (UGSF), Université de Lille-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Recherche Agronomique (INRA), Centre National de la Recherche Scientifique (CNRS)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut National de la Recherche Agronomique (INRA), Université de Nantes (UN)-Université de Nantes (UN)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC), and Institut National de la Recherche Agronomique (INRA)-Université de Lille-Centre National de la Recherche Scientifique (CNRS)

Subjects
0301 basic medicine, Mannosides, Mannosyltransferase, 1-Deoxynojirimycin, Iminosugar, Gene Expression, 010402 general chemistry, 01 natural sciences, Biochemistry, Mannosyltransferases, Pichia, Analytical Chemistry, Substrate Specificity, Fungal Proteins, 03 medical and health sciences, chemistry.chemical_compound, Biosynthesis, Cell Wall, Candida albicans, Cloning, Molecular, Enzyme Inhibitors, Mannan, Enzyme Assays, chemistry.chemical_classification, Glucosamine, biology, Chemistry, [CHIM.ORGA]Chemical Sciences/Organic chemistry, Organic Chemistry, General Medicine, biology.organism_classification, Yeast, Recombinant Proteins, 0104 chemical sciences, 3. Good health, Imino Sugars, Kinetics, 030104 developmental biology, Enzyme
Abstract

International audience; β-1,2-Linked oligomannosides substitute the cell wall of numerous yeast species. Several of those including Candida albicans may cause severe infections associated with high rates of morbidity and mortality, especially in immunocompromised patients. β-1,2-Mannosides are known to be involved in the pathogenic process and to elicit an immune response from the host. In C. albicans, the synthesis of β-mannosides is under the control of a family of nine genes coding for putative β-mannosyltransferases. Two of them, CaBmt1 and CaBmt3, have been shown to initiate and prime the elongation of the β-mannosides on the cell-wall mannan core. In the present study, we have assessed the modulating activities of monovalent and multivalent iminosugar analogs on these enzymes in order to control the enzymatic bio-synthesis of β-mannosides. We have identified a monovalent deoxynojirimycin (DNJ) derivative that inhibits the CaBmt1-catalyzed initiating activity, and mono-, tetra- and polyvalent deoxymannojirimycin (DMJ) that modulate the CaBmt1 activity toward the formation of a single major product. Analysis of the aggregating properties of the multivalent iminosugars showed their ability to elicit clusterization of both CaBmt1 and CaBmt3, without affecting their activity. These results suggest promising roles for multivalent iminosugars as controlling agents for the biosynthesis of β-1,2 mannosides and for monovalent DNJ derivative as a first target for the design of future β-mannosyltransferase inhibitors.

Published
2016

8. Identification of the Mycobacterium marinum Apa antigen O-mannosylation sites reveals important glycosylation variability with the M. tuberculosis Apa homologue

Author

Yoann Rombouts, Emeline Fabre, Kay-Hooi Khoo, Elisabeth Elass-Rochard, Colette Brassart, Yann Guérardel, Adeline Burguière, Sz-Wei Wu, Bernadette Coddeville, and Laurent Kremer

Subjects
Antigens, Bacterial, Glycosylation, biology, Biophysics, Mannose, Mycobacterium tuberculosis, biology.organism_classification, Biochemistry, Microbiology, chemistry.chemical_compound, Bacterial Proteins, chemistry, Antigen, Concanavalin A, Mycobacterium marinum, biology.protein, Amino Acid Sequence, Pathogen, Peptide sequence, Glycoproteins
Abstract

The 45/47 kDa Apa, an immuno-dominant antigen secreted by Mycobacterium tuberculosis is O-mannosylated at multiple sites. Glycosylation of Apa plays a key role in colonization and invasion of the host cells by M. tuberculosis through interactions of Apa with the host immune system C-type lectins. Mycobacterium marinum (M.ma) a fish pathogen, phylogenetically close to M. tuberculosis, induces a granulomatous response with features similar to those described for M. tuberculosis in human. Although M.ma possesses an Apa homologue, its glycosylation status is unknown, and whether this represents a crucial element in the pathophysiology induced by M.ma remains to be addressed. To this aim, we have identified two concanavalin A-reactive 45/47 kDa proteins from M.ma, which have been further purified by a two-step anion exchange chromatography process. Advanced liquid chromatography-nanoESI mass spectrometry-based proteomic analyses of peptides, derived from either tryptic digestion alone or in combination with the Asp-N endoproteinase, established that M.ma Apa possesses up to seven distinct O-mannosylated sites with mainly single mannose substitutions, which can be further extended at the Ser/Thr/Pro rich region near the N-terminus. This opens the way to further studies focussing on the involvement and biological functions of Apa O-mannosylation using the M.ma/zebrafish model.

Published
2012

9. Candida albicans β-1,2-mannosyltransferase Bmt3 prompts the elongation of the cell-wall phosphopeptidomannan

Author

Florence Delplace, Julie Bouckaert, Chantal Fradin, Yann Guérardel, Ghenima Sfihi-Loualia, Emeline Fabre, Jean-Maurice Mallet, Marilyne Pourcelot, Daniel Poulain, Emmanuel Maes, Anaïs Mée, Thomas Hurtaux, Unité de Glycobiologie Structurale et Fonctionnelle UMR 8576 (UGSF), Université de Lille-Centre National de la Recherche Scientifique (CNRS), Lille Inflammation Research International Center - U 995 (LIRIC), Institut Pasteur de Lille, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille-Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille), Laboratoire des biomolécules (LBM UMR 7203), Chimie Moléculaire de Paris Centre (FR 2769), École normale supérieure - Paris (ENS-PSL), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Ecole Superieure de Physique et de Chimie Industrielles de la Ville de Paris (ESPCI Paris), Université Paris sciences et lettres (PSL)-Institut de Chimie du CNRS (INC)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Paris (ENS-PSL), Université Paris sciences et lettres (PSL)-Institut de Chimie du CNRS (INC)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Département de Chimie - ENS Paris, Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Ministere de l'Enseignement Superieur et de la Recherche, Centre Commun de Mesure RMN de l'Universite de Lille 1, European Community (FEDER), Region Nord-Pas de Calais (France), Universite Lille1, Agence Nationale de la Recherche (ANR-MIEN-BMT), Region Nord-Pas de Calais, European Union (FEDER), Ministere Francais de la Recherche, Universite Lille1-Sciences et Technologies, CNRS, TGE RMN THC [Fr3050], Institut National de la Recherche Agronomique (INRA)-Université de Lille-Centre National de la Recherche Scientifique (CNRS), Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille-Institut Pasteur de Lille, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP), Centre National de la Recherche Scientifique (CNRS)-Sorbonne Université (SU)-Département de Chimie - ENS Paris, École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Chimie Moléculaire de Paris Centre (FR 2769), Institut de Chimie du CNRS (INC)-École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM), Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 (UGSF), Université de Lille-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Recherche Agronomique (INRA), Lille Inflammation Research International Center (LIRIC), École normale supérieure - Paris (ENS Paris)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Ecole Nationale Supérieure de Chimie de Paris- Chimie ParisTech-PSL (ENSCP)-ESPCI ParisTech-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Paris (ENS Paris)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Ecole Nationale Supérieure de Chimie de Paris- Chimie ParisTech-PSL (ENSCP)-ESPCI ParisTech-Centre National de la Recherche Scientifique (CNRS)-Département de Chimie - ENS Paris, École normale supérieure - Paris (ENS Paris)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés (LISBP), Institut National de la Recherche Agronomique (INRA)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS), Physiopathologie des Candidoses, Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille, Droit et Santé, Université Pierre et Marie Curie - Paris 6 (UPMC), Institut National de la Recherche Agronomique (INRA)-Centre National de la Recherche Scientifique (CNRS)-Université de Lille, Université Lille Nord de France (COMUE), Peptides, glycoconjugués et métaux en biologie (LBM-E1), Université Pierre et Marie Curie - Paris 6 (UPMC)-Département de Chimie - ENS Paris, École normale supérieure - Paris (ENS Paris)-École normale supérieure - Paris (ENS Paris)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Département de Chimie - ENS Paris, École normale supérieure - Paris (ENS Paris)-École normale supérieure - Paris (ENS Paris)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), and ENSA Phytotechnie, INRA Agronomie

Subjects
0301 basic medicine, Phosphopeptides, Mannosides, Mannosyltransferase, Glycoconjugate, [SDV]Life Sciences [q-bio], Biology, yeast, Biochemistry, Mannosyltransferases, Pichia pastoris, Substrate Specificity, Fungal Proteins, Mannans, 03 medical and health sciences, mannosyltransferase, Cell Wall, Candida albicans, [CHIM]Chemical Sciences, Candida, chemistry.chemical_classification, Fungal protein, biology.organism_classification, Yeast, 3. Good health, 030104 developmental biology, Enzyme, chemistry, beta-mannose
Abstract

International audience; beta-1,2-Linked mannosides are expressed on numerous cell-wall glycoconjugates of the opportunistic pathogen yeast Candida albicans. Several studies evidenced their implication in the host-pathogen interaction and virulence mechanisms. In the present study, we characterized the in vitro activity of CaBmt3, a beta-1,2-mannosyltransferase involved in the elongation of beta-1,2-oligomannosides oligomers onto the cell-wall polymannosylated N-glycans. A recombinant soluble enzyme Bmt3p was produced in Pichia pastoris and its enzyme activity was investigated using natural and synthetic oligomannosides as potential acceptor substrates. Bmt3p was shown to exhibit an exquisite enzymatic specificity by adding a single terminal beta-mannosyl residue to alpha-1,2-linked oligomannosides capped by a Man beta 1-2Man motif. Furthermore, we demonstrated that the previously identified CaBmt1 and CaBmt3 efficiently act together to generate Man beta 1-2Man beta 1-2[Man alpha 1-2](n) sequence from alpha-1,2-linked oligomannosides onto exogenous and endogenous substrates.

Published
2015

10. Polymeric iminosugars improve the activity of carbohydrate-processing enzymes

Author

Franck Daligault, Emeline Fabre, Gabrielle Potocki-Veronese, Sébastien G. Gouin, David Teze, Simon Ladevèze, David Deniaud, Sergej Šesták, Charles Tellier, Yoan Brissonnet, Magali Remaud-Simeon, LUNAM Université [Nantes Angers Le Mans], Chimie Et Interdisciplinarité : Synthèse, Analyse, Modélisation (CEISAM), Université de Nantes - UFR des Sciences et des Techniques (UN UFR ST), Université de Nantes (UN)-Université de Nantes (UN)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC), Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés (LISBP), Centre National de la Recherche Scientifique (CNRS)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut National de la Recherche Agronomique (INRA), UMR 5504, Centre National de la Recherche Scientifique (CNRS), Unité de Glycobiologie Structurale et Fonctionnelle UMR 8576 (UGSF), Institut National de la Recherche Agronomique (INRA)-Université de Lille-Centre National de la Recherche Scientifique (CNRS), Université des Sciences et Technologies (Lille 1) (USTL), Unité de fonctionnalité et ingénierie de protéines (UFIP), Université de Nantes (UN)-Université de Nantes (UN)-Centre National de la Recherche Scientifique (CNRS), Slovak Academy of Sciences (SAS), Centre National de la Recherche Scientifique, Ministere Delegue a l'Enseignement Superieur et a la Recherche, Université de Nantes (UN)-Université de Nantes (UN)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Institut National de la Recherche Agronomique (INRA)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS), Toulouse Biotechnology Institute (TBI), Université de Lille-Centre National de la Recherche Scientifique (CNRS), and Université de Lille, Sciences et Technologies

Subjects
Glycoside Hydrolases, Phosphorylases, Biomedical Engineering, Pharmaceutical Science, Bioengineering, Ligands, Polymerization, Enzyme activator, Coated Materials, Biocompatible, Hydrolase, Glycoside hydrolase, Fucosidase, Pharmacology, chemistry.chemical_classification, alpha-L-Fucosidase, Galactosidases, biology, Chemistry, Artificial enzyme, Phosphoric Diester Hydrolases, [CHIM.ORGA]Chemical Sciences/Organic chemistry, Organic Chemistry, Dextrans, Imino Sugars, Enzyme Activation, Enzyme, Biochemistry, Biocatalysis, biology.protein, Biotechnology
Abstract

International audience; Multivalent iminosugars have recently emerged as powerful tools to inhibit the activities of specific glycosidases. In this work, biocompatible dextrans were coated with iminosugars to form linear and ramified polymers with unprecedently high valencies (from 20 to 900) to probe the evolution of the multivalent inhibition as a function of ligand valency. This study led to the discovery that polyvalent iminosugars can also significantly enhance, not only inhibit, the enzymatic activity of specific glycoside-hydrolase, as observed on two galactosidases, a fucosidase, and a bacterial mannoside phosphorylase for which an impressive 70-fold activation was even reached. The concept of glycosidase activation is largely unexplored, with a unique recent example of small-molecules activators of a bacterial O-GlcNAc hydrolase. The possibility of using these polymers as "artificial enzyme effectors" may therefore open up new perspectives in therapeutics and biocatalysis.

Published
2015

11. Understanding the Polymerization Mechanism of Glycoside-Hydrolase Family 70 Glucansucrases

Author

Gilles Joucla, Magali Remaud-Simeon, David Harrison, Gabrielle Potocki-Véronèse, Pierre Monsan, Emeline Fabre, and Claire Moulis

Subjects
Sucrose, Stereochemistry, Molecular Sequence Data, Oligosaccharides, Biochemistry, Dextransucrase, chemistry.chemical_compound, Glucansucrase, Glycoside hydrolase, Amino Acid Sequence, Molecular Biology, DNA Primers, Sequence Homology, Amino Acid, biology, Isomaltooligosaccharide, Glycosyltransferases, Active site, Cell Biology, biology.organism_classification, Alternansucrase, Protein Structure, Tertiary, Kinetics, Dextran, Carbohydrate Sequence, Models, Chemical, chemistry, Glucosyltransferases, Leuconostoc mesenteroides, Multigene Family, Mutation, biology.protein, Leuconostoc
Abstract

Glucan formation catalyzed by two GH-family 70 enzymes, Leuconostoc mesenteroides NRRL B-512F dextransucrase and L. mesenteroides NRRL B-1355 alternansucrase, was investigated by combining biochemical and kinetic characterization of the recombinant enzymes and their respective products. Using HPAEC analysis, we showed that two molecules act as initiator of polymerization: sucrose itself and glucose produced by hydrolysis, the latter being preferred when produced in sufficient amounts. Then, elongation occurs by transfer of the glucosyl residue coming from sucrose to the non-reducing end of initially formed products. Dextransucrase preferentially produces an isomaltooligosaccharide series, whose concentration is always low because of the high ability of these products to be elongated and form high molecular weight dextran. Compared with dextransucrase, alternansucrase has a broader specificity. It produces a myriad of oligosaccharides with various alpha-1,3 and/or alpha-1,6 links in early reaction stages. Only some of them are further elongated. Overall alternan polymer is smaller in size than dextran. In dextransucrase, the A repeats often found in C-terminal domain of GH family 70 were found to play a major role in efficient dextran elongation. Their truncation result in an enzyme much less efficient to catalyze high molecular weight polymer formation. It is thus proposed that, in dextransucrase, the A repeats define anchoring zones for the growing chains, favoring their elongation. Based on these results, a semi-processive mechanism involving only one active site and an elongation by the non-reducing end is proposed for the GH-family 70 glucansucrases.

Published
2006

12. Glucansucrases of GH family 70: What are the determinants of their specifities?

Author

Gabrielle Potocki-Véronèse, Stephane Emond, Pierre Monsan, Gilles Joucla, Emeline Fabre, Claire Moulis, Magali Remaud-Simeon, and Gaëtan Richard

Subjects
chemistry.chemical_classification, Sequence analysis, Stereochemistry, Biochemistry, Catalysis, Alternansucrase, Amino acid, Dextransucrase, Enzyme, chemistry, Glycoside hydrolase, Site-directed mutagenesis, Biotechnology, Glucan
Abstract

Glucansucrases from family 70 of glycoside-hydrolases catalyse the synthesis of α-glucans with various types of osidic linkages from sucrose. Among these enzymes, alternansucrase (ASR) and dextransucrase E (DSR-E) catalyse the formation of unusual α-glucans. ASR catalyses the synthesis of linear glucan with α-1,3 and α-1,6 alternating linkages and DSR-E synthesizes a glucan containing α-1,6 linkages in the linear chain and α-1,2 branches. The sequence analysis of these enzymes enabled the identification of structural elements suspected to be involved in the enzyme specificities. Biochemical characterization of ASR and DSR-E variants obtained from gene truncations or site-directed mutagenesis experiments showed that the specificity of these enzymes to form different types of osidic linkage is controlled by two different approaches. For ASR, the double specificity is controlled by only one catalytic domain where important amino acids involved in the enzyme specificity have been identified. In the case of DS...

Published
2006

13. Molecular Characterization of DSR-E, an α-1,2 Linkage-Synthesizing Dextransucrase with Two Catalytic Domains

Author

Pierre Monsan, Magali Remaud-Simeon, René-Marc Willemot, Emeline Fabre, Sophie Bozonnet, Marguerite Dols-Laffargue, Sandra Pizzut, Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés (LISBP), Institut National de la Recherche Agronomique (INRA)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), and Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Molecular Sequence Data, Oligosaccharides, Sequence alignment, Microbiology, Dextransucrase, Glucosyltransferases, Catalytic Domain, Glucansucrase, Amino Acid Sequence, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], Cloning, Molecular, Molecular Biology, Peptide sequence, biology, Nucleic acid sequence, Dextrans, Sequence Analysis, DNA, biology.organism_classification, Enzymes and Proteins, Open reading frame, Biochemistry, Leuconostoc mesenteroides, biology.protein, Sequence Alignment, Leuconostoc
Abstract

A novel Leuconostoc mesenteroides NRRL B-1299 dextransucrase gene, dsrE , was isolated, sequenced, and cloned in Escherichia coli , and the recombinant enzyme was shown to be an original glucansucrase which catalyses the synthesis of α-1,6 and α-1,2 linkages. The nucleotide sequence of the dsrE gene consists of an open reading frame of 8,508 bp coding for a 2,835-amino-acid protein with a molecular mass of 313,267 Da. This is twice the average mass of the glucosyltransferases (GTFs) known so far, which is consistent with the presence of an additional catalytic domain located at the carboxy terminus of the protein and of a central glucan-binding domain, which is also significantly longer than in other glucansucrases. From sequence comparison with family 70 and α-amylase enzymes, crucial amino acids involved in the catalytic mechanism were identified, and several original sequences located at some highly conserved regions in GTFs were observed in the second catalytic domain.

Published
2002

14. Structural Sampling of Glycan Interaction Profiles Reveals Mucosal Receptors for Fimbrial Adhesins of Enterotoxigenic Escherichia coli

Author

Julie Bouckaert, Stefan Oscarson, Emeline Fabre, Lieven Buts, Arjen R. de Boer, Manfred S. Weiss, André M. Deelder, Manfred Wuhrer, J.D.M. Olsson, Yann Guérardel, Emanuela Lonardi, and Kristof Moonens

Subjects
Glycan, glycan array, Fimbria, arginine, sulfate, medicine.disease_cause, E. coli, Article, General Biochemistry, Genetics and Molecular Biology, Microbiology, Sulfate binding, Glycomics, 03 medical and health sciences, adhesin, charge, FimH, Enterotoxigenic Escherichia coli, enterotoxigenic, F17G, medicine, Escherichia coli, lcsh:QH301-705.5, 030304 developmental biology, 0303 health sciences, General Immunology and Microbiology, biology, 030306 microbiology, Lectin, Bacterial adhesin, lcsh:Biology (General), Others, biology.protein, FedF, General Agricultural and Biological Sciences
Abstract

Fimbriae are long, proteinaceous adhesion organelles expressed on the bacterial envelope, evolutionarily adapted by Escherichia coli strains for the colonization of epithelial linings. Using glycan arrays of the Consortium for Functional Glycomics (CFG), the lectin domains were screened of the fimbrial adhesins F17G and FedF from enterotoxigenic E. coli (ETEC) and of the FimH adhesin from uropathogenic E. coli. This has led to the discovery of a more specific receptor for F17G, GlcNAcb1,3Gal. No significant differences emerged from the glycan binding profiles of the F17G lectin domains from five different E. coli strains. However, strain-dependent amino acid variations, predominantly towards the positively charged arginine, were indicated by sulfate binding in FedF and F17G crystal structures. For FedF, no significant binders could be observed on the CFG glycan array. Hence, a shotgun array was generated from microvilli scrapings of the distal jejunum of a 3-week old piglet about to be weaned. On this array, the blood group A type 1 hexasaccharide emerged as a receptor for the FedF lectin domain and remarkably also for F18-fimbriated E. coli. F17G was found to selectively recognize glycan species with a terminal GlcNAc, typifying intestinal mucins. In conclusion, F17G and FedF recognize long glycan sequences that could only be identified using the shotgun approach. Interestingly, ETEC strains display a large capacity to adapt their fimbrial adhesins to ecological niches via charge-driven interactions, congruent with binding to thick mucosal surfaces displaying an acidic gradient along the intestinal tract.

Published
2013
Full Text
View/download PDF

15. Mantyl tagged oligo alpha (1 -> 2) mannosides as Candida albicans beta-mannosyl transferases substrates: a comparison between synthetic strategies

Author

Florence Delplace, Jean-Maurice Mallet, Emeline Fabre, Marilyne Pourcelot, Ghenima Sfihi-Loualia, Chantal Fradin, Frédéric Krzewinski, Yann Guérardel, Laurent Cattiaux, Daniel Poulain, Université Pierre et Marie Curie - Paris 6 (UPMC), Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 (UGSF), Université de Lille-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Recherche Agronomique (INRA), Physiopathologie des Candidoses, Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille, Droit et Santé, Inflammation: mécanismes et régulation et interactions avec la nutrition et les candidoses, Université de Lille, Droit et Santé-Institut National de la Santé et de la Recherche Médicale (INSERM), Agence Nationale de la Recherche CaBMT, Ministere de l'Enseignement Superieur, Unité de Glycobiologie Structurale et Fonctionnelle UMR 8576 (UGSF), Institut National de la Recherche Agronomique (INRA)-Université de Lille-Centre National de la Recherche Scientifique (CNRS), and Université de Lille-Centre National de la Recherche Scientifique (CNRS)

Subjects
chemistry.chemical_classification, 0303 health sciences, Mannosides, biology, 010405 organic chemistry, Chemistry, Stereochemistry, General Chemical Engineering, [SDV]Life Sciences [q-bio], Substrate (chemistry), General Chemistry, biology.organism_classification, 01 natural sciences, 0104 chemical sciences, carbohydrates (lipids), 03 medical and health sciences, Enzyme, Biochemistry, Click chemistry, Transferase, [CHIM]Chemical Sciences, Reactivity (chemistry), Candida albicans, 030304 developmental biology
Abstract

International audience; Fluorescent oligomannosides are important tools for the evaluation of mannosyl transferase activities and selectivities. In a project dealing with Candida albicans beta-mannosyl transferases, three mantyl tagged alpha (1 -> 2) oligomannosides were prepared by different ways: using all ester strategy compatible with the presence of an azido group suitable for direct click chemistry; and alternatively using the more classic benzyl protecting groups. Although more elegant, the all ester strategy has shown important limitations: reduced reactivity of mannosyl donors, and 3 -> 2 ester migration. Preliminary enzymatic studies have shown that the synthetic oligomannosides are efficient substrate of b-mannosyl transferases.

Published
2013

16. Mapping the Expressed Glycome and Glycosyltransferases of Zebrafish Liver Cells as a Relevant Model System for Glycosylation Studies

Author

Christian Slomianny, Jorick Vanbeselaere, Christophe Biot, Yann Guérardel, Anne Harduin-Lepers, Kay-Hooi Khoo, Emeline Fabre, Lan-Yi Chang, Nao Yamakawa, Unité de Glycobiologie Structurale et Fonctionnelle UMR 8576 (UGSF), Institut National de la Recherche Agronomique (INRA)-Université de Lille-Centre National de la Recherche Scientifique (CNRS), Institute of Biological Chemistry (IBC Sinica), Academia Sinica, Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés (LISBP), Institut National de la Recherche Agronomique (INRA)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Physiologie Cellulaire : Canaux ioniques, inflammation et cancer - U 1003 (PHYCELL), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille, Unité de Catalyse et Chimie du Solide - UMR 8181 (UCCS), Centrale Lille Institut (CLIL)-Université d'Artois (UA)-Centrale Lille-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Université de Lille, Université Lille Nord de France (COMUE), Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 (UGSF), Université de Lille-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Recherche Agronomique (INRA), Université de Lille-Centre National de la Recherche Scientifique (CNRS), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS), Université d'Artois (UA)-Centrale Lille-Institut de Chimie du CNRS (INC)-Université de Lille-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut National de la Recherche Agronomique (INRA), Physiologie Cellulaire (PHYCEL) - U1003 (PHYCEL), Université Lille Nord de France (COMUE)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille, Unité de Catalyse et de Chimie du Solide - UMR 8181 (UCCS), Université d'Artois (UA)-Ecole Centrale de Lille-Ecole Nationale Supérieure de Chimie de Lille (ENSCL)-Université de Lille-Centre National de la Recherche Scientifique (CNRS), and Université de Lille-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Lille Nord de France (COMUE)

Subjects
MESH: Glycomics, Glycosylation, [SDV.BIO]Life Sciences [q-bio]/Biotechnology, ved/biology.organism_classification_rank.species, [SDV.BID.SPT]Life Sciences [q-bio]/Biodiversity/Systematics, Phylogenetics and taxonomy, Biochemistry, Epitope, chemistry.chemical_compound, MESH: Glycolipids, MESH: Animals, Glycomics, Zebrafish, Cells, Cultured, ComputingMilieux_MISCELLANEOUS, chemistry.chemical_classification, 0303 health sciences, biology, [SDV.BID.EVO]Life Sciences [q-bio]/Biodiversity/Populations and Evolution [q-bio.PE], 030302 biochemistry & molecular biology, [SDV.BBM.MN]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular Networks [q-bio.MN], MESH: Glycosylation, [SDV.BIBS]Life Sciences [q-bio]/Quantitative Methods [q-bio.QM], [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM], Cell biology, Liver, MESH: Models, Animal, Models, Animal, MESH: Glycosyltransferases, Cellular model, MESH: Cells, Cultured, [SDV.CAN]Life Sciences [q-bio]/Cancer, 03 medical and health sciences, Polysaccharides, [SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], Animals, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], MESH: Zebrafish, Model organism, 030304 developmental biology, ved/biology, Glycosyltransferases, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, General Chemistry, biology.organism_classification, Glycome, carbohydrates (lipids), MESH: Polysaccharides, chemistry, Glycolipids, Glycoprotein, MESH: Liver
Abstract

International audience; The emergence of zebrafish as a model organism for human diseases was accompanied by the development of cellular model systems that extended the possibilities for in vitro manipulation and in vivo studies after cell implantation. The exploitation of zebrafish cell systems is, however, still hampered by the lack of genomic and biochemical data. Here, we lay a path toward the efficient use of ZFL, a zebrafish liver-derived cell system, as a platform for studying glycosylation. To achieve this, we established the glycomic profile of ZFL by a combination of mass spectrometry and NMR. We demonstrated that glycoproteins were substituted by highly sialylated multiantennary N-glycans, some of them comprising the unusual zebrafish epitope Galβ1-4[Neu5Ac(α2,3)]Galβ1-4[Fuc(α1,3)]GlcNAc, and core 1 multisialylated O-glycans. Similarly, these analyses established that glycolipids were dominated by sialylated gangliosides. In parallel, analyzing the expression patterns of all putative sialyl- and fucosyltransferases, we directly correlated the identified structures to the set of enzymes involved in ZFL glycome. Finally, we demonstrated that this cell system was amenable to metabolic labeling using functionalized monosaccharides that permit in vivo imaging of glycosylation processes. Altogether, glycomics, genomics, and functional studies established ZFL as a relevant cellular model for the study of glycosylation.

Published
2012

17. Functional and Structural Characterization of α-(1→2) Branching Sucrase Derived from DSR-E Glucansucrase

Author

Yannick Malbert, Lionel Mourey, Tjaard Pijning, Pierre Monsan, Sandrine Morel, Yoann Brison, Magali Remaud-Simeon, Emeline Fabre, Bauke W. Dijkstra, Samuel Tranier, Gabrielle Potocki-Véronèse, Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés (LISBP), Centre National de la Recherche Scientifique (CNRS)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut National de la Recherche Agronomique (INRA), University of Groningen [Groningen], Institut de pharmacologie et de biologie structurale (IPBS), Centre National de la Recherche Scientifique (CNRS)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées, Biophys Chem Lab, Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS), Institut Universitaire de France, region Midi-Pyrenees (France), European Molecular Biology Organization, Institut National de la Recherche Agronomique (INRA)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS), Université de Toulouse (UT)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS), and X-ray Crystallography

Subjects
Protein Folding, IN-VITRO FERMENTATION, ALPHA-AMYLASE FAMILY, 2 CATALYTIC DOMAINS, LEUCONOSTOC-MESENTEROIDES, GLYCOSYL HYDROLASES, CRYSTAL-STRUCTURE, GNOTOBIOTIC-RATS, OLIGOSACCHARIDES, DEXTRANSUCRASE, SPECIFICITY, Stereochemistry, [SDV]Life Sciences [q-bio], Mutation, Missense, Crystallography, X-Ray, Biochemistry, Dextransucrase, Sucrase, Structure-Activity Relationship, 03 medical and health sciences, Bacterial Proteins, parasitic diseases, Glucansucrase, Leuconostoc, Glycoside hydrolase, Molecular Biology, 030304 developmental biology, 0303 health sciences, biology, Chemistry, 030302 biochemistry & molecular biology, Dextrans, Cell Biology, biology.organism_classification, Enzyme structure, Protein Structure, Tertiary, Amino Acid Substitution, Leuconostoc mesenteroides, Protein Structure and Folding, biology.protein, Protein folding
Abstract

Delta N-123-glucan-binding domain-catalytic domain 2 (Delta N-123-GBD-CD2) is a truncated form of the bifunctional glucansucrase DSR-E from Leuconostoc mesenteroides NRRL B-1299. It was constructed by rational truncation of GBD-CD2, which harbors the second catalytic domain of DSR-E. Like GBD-CD2, this variant displays alpha-(1-->2) branching activity when incubated with sucrose as glucosyl donor and (oligo-)dextran as acceptor, transferring glucosyl residues to the acceptor via a ping-pong bi-bi mechanism. This allows the formation of prebiotic molecules containing controlled amounts of alpha-(1-->2) linkages. The crystal structure of the apo alpha-(1-->2) branching sucrase Delta N-123-GBD-CD2 was solved at 1.90 angstrom resolution. The protein adopts the unusual U-shape fold organized in five distinct domains, also found in GTF180-Delta N and GTF-SI glucansucrases of glycoside hydrolase family 70. Residues forming subsite -1, involved in binding the glucosyl residue of sucrose and catalysis, are strictly conserved in both GTF180-Delta N and Delta N-123-GBD-CD2. Subsite +1 analysis revealed three residues (Ala-2249, Gly-2250, and Phe-2214) that are specific to Delta N-123-GBD-CD2. Mutation of these residues to the corresponding residues found in GTF180-Delta N showed that Ala-2249 and Gly-2250 are not directly involved in substrate binding and regiospecificity. In contrast, mutant F2214N had lost its ability to branch dextran, although it was still active on sucrose alone. Furthermore, three loops belonging to domains A and B at the upper part of the catalytic gorge are also specific to Delta N-123-GBD-CD2. These distinguishing features are also proposed to be involved in the correct positioning of dextran acceptor molecules allowing the formation of alpha-(-->2) branches.

Published
2012

18. Synthesis of dextrans with controlled amounts of α-1,2 linkages using the transglucosidase GBD-CD2

Author

Emeline Fabre, Pierre Monsan, Magali Remaud-Simeon, Claire Moulis, Yoann Brison, Jean-Charles Portais, Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés (LISBP), Centre National de la Recherche Scientifique (CNRS)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut National de la Recherche Agronomique (INRA), Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 (UGSF), Université de Lille-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Recherche Agronomique (INRA), Institut National de la Recherche Agronomique (INRA)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS), Unité de Glycobiologie Structurale et Fonctionnelle UMR 8576 (UGSF), Université de Lille-Centre National de la Recherche Scientifique (CNRS), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS), and Institut National de la Recherche Agronomique (INRA)-Université de Lille-Centre National de la Recherche Scientifique (CNRS)

Subjects
Sucrose, Stereochemistry, Kinetics, Applied Microbiology and Biotechnology, 03 medical and health sciences, chemistry.chemical_compound, Hydrolysis, Bacterial Proteins, Glucansucrase, Side chain, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], 030304 developmental biology, chemistry.chemical_classification, 0303 health sciences, biology, 030302 biochemistry & molecular biology, Dextrans, General Medicine, biology.organism_classification, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM], Dextran, Enzyme, Glucose, chemistry, Leuconostoc mesenteroides, biology.protein, Glucosidases, Leuconostoc, Biotechnology
Abstract

GBD-CD2 is an alpha-1,2 transglucosidase engineered from DSR-E, a glucansucrase naturally produced by Leuconostoc mesenteroides NRRL B-1299. This enzyme catalyses from sucrose, the alpha-1,2 transglucosylation of glucosyl moieties onto alpha-1,6 dextran chains. Steady-state kinetic studies showed that hydrolysis and transglucosylation reactions occurred at the early stage of the reaction in the presence of 70 kDa dextran as acceptor and sucrose. The transglucosylation reaction catalysed by GBD-CD2 follows a Ping Pong Bi Bi mechanism with a high kcat value of 970 s(-1). The amount of the synthesised alpha-1,2 side chains was found to be directly dependent on the initial molar ratio [Sucrose]/[Dextran]. Dextrans with controlled alpha-1,2 linkage contents ranging from 13% to 40% were synthesised. The procedure resulted in the production of dextrans with the highest content of alpha-1,2 linkages ever reported.

Published
2010

19. New chromogenic substrates for feruloyl esterases

Author

David Navarro, Laurence Lesage-Meessen, Michèle Asther, Emeline Fabre, Hugues Driguez, Michael J. O’Donohue, Marcel Asther, Laurence Marmuse, Sébastien Fort, Centre de Recherches sur les Macromolecules Vegetales, Unité mixte de recherche de biotechnologie des champignons filamenteux, Université de la Méditerranée - Aix-Marseille 2-Institut National de la Recherche Agronomique (INRA)-Université de Provence - Aix-Marseille 1, Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés (LISBP), Institut National de la Recherche Agronomique (INRA)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS), ADEME: 0401C0042, ANR PNRB2005- 11, and Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Chromatography, Molecular Structure, 010405 organic chemistry, Chromogenic, Chemistry, [CHIM.ORGA]Chemical Sciences/Organic chemistry, Organic Chemistry, Chromogenic Substrates, Substrate (chemistry), 010402 general chemistry, 01 natural sciences, Biochemistry, Feruloyl esterase activity, Solid medium, 0104 chemical sciences, Nitrophenols, Chromogenic Compounds, Molecular Probes, Physical and Theoretical Chemistry, Carboxylic Ester Hydrolases
Abstract

International audience; Indolyl and nitrophenyl 5-O-hydroxycinnamoyl-α-L-arabinofuranosides were prepared by chemo-enzymatic syntheses. These probes were designed as substrates to be used in assays of feruloyl esterase activity (EC 3.1.1.77). Color development in the assays only occurs when feruloyl esterase activity releases an intermediate chromogenic arabinoside that is a suitable substrate for α-L-arabinofuranosidase (EC 3.2.1.55), which in turn releases the free chromogenic group. The usefulness of these compounds was evaluated in both qualitative solid media-based assays and quantitative liquid assays that can be performed in microtiter plates using feruloyl esterases and arabinofuranosidases from various origins.

Published
2008

20. A1.25 Visualisation and characterisation of citrullinated antigen-specific B cells from peripheral blood of patients with rheumatoid arthritis

Author

Dominique Baeten, Rem Toes, EH van der Voort, Twj Huizinga, Hans Scherer, Emeline Fabre, Priscilla F Kerkman, and Kristine Germar

Subjects
education.field_of_study, Cluster of differentiation, biology, business.industry, Immunology, Population, Context (language use), medicine.disease, General Biochemistry, Genetics and Molecular Biology, medicine.anatomical_structure, Immune system, Rheumatology, Antigen, Rheumatoid arthritis, biology.protein, Immunology and Allergy, Medicine, Antibody, business, education, B cell
Abstract

Background and objectives In Rheumatoid arthritis (RA), anti-citrullinated protein antibodies (ACPA) represent highly disease-specific biomarkers found in the majority of patients. As ACPA have been implicated in disease pathogenesis, it is of crucial relevance to understand the underlying B cell response. In this context, visualisation of citrullinated antigen-specific B cells would allow for a detailed characterisation of the immune response to better understand its development and maintenance. Unfortunately, visualisation of autoreactive B cells in humans has proven extremely difficult. Materials and methods Using the CCP2-antigen and its arginine control variant, we developed a multicolour tetramer-based staining method to visualise citrullinated antigen-specific B cells in peripheral blood of RA patients by flow-cytometry. Specificity of the staining was verified by culturing tetramer-positive and -negative B cells isolated by FACS. Citrullinated antigen-reactive B cells were further phenotyped using cell surface markers associated with developmental and functional B cell characteristics. Finally, the frequency of citrullinated antigen-reactive B cells was correlated to ACPA serum levels and in vitro ACPA production. Results The staining procedure successfully separated citrullinated antigen-reactive B cells from non-specific background signals. Already fourteen FACS-sorted tetramer-positive B cells produced detectable amounts of ACPA, whereas no ACPA production was observed in cultures of up to 5000 tetramer-negative B cells. The majority of citrullinated antigen-reactive B cells had a post-germinal centre memory or plasmablast phenotype. Up to 1 in 200 memory B cells were directed against citrullinated antigens, and their frequency correlated with spontaneous ACPA production in culture and ACPA serum titres in vivo . Conclusions We show, for the first time, the specific and reliable identification of citrullinated antigen-specific B cells in high frequencies in peripheral blood of RA patients. The majority of this population has a memory phenotype and closely reflects the dynamics of the in vivo ACPA response. These data provide the basis for a detailed characterisation of this disease-specific immune response on a single cell level and could lead to the identification of novel therapeutic targets.

Published
2015

21. Role of the two catalytic domains of DSR-E dextransucrase and their involvement in the formation of highly alpha-1,2 branched dextran

Author

Pierre Monsan, Sophie Bozonnet, René-Marc Willemot, Michel Vignon, Emeline Fabre, Magali Remaud-Simeon, Audrey Arcache, Centre de Recherches sur les Macromolécules Végétales (CERMAV), and Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)

Subjects
Sucrose, Biology, Microbiology, Dextransucrase, 03 medical and health sciences, chemistry.chemical_compound, Structure-Activity Relationship, Catalytic Domain, Hydrolase, Glycosyltransferase, Glucansucrase, Molecular Biology, Glucans, ComputingMilieux_MISCELLANEOUS, 030304 developmental biology, chemistry.chemical_classification, 0303 health sciences, Binding Sites, 030306 microbiology, Dextrans, Oligosaccharide, biology.organism_classification, Enzymes and Proteins, Dextran, chemistry, Biochemistry, Leuconostoc mesenteroides, Glucosyltransferases, biology.protein, Binding domain
Abstract

The dsrE gene from Leuconostoc mesenteroides NRRL B-1299 was shown to encode a very large protein with two potentially active catalytic domains (CD1 and CD2) separated by a glucan binding domain (GBD). From sequence analysis, DSR-E was classified in glucoside hydrolase family 70, where it is the only enzyme to have two catalytic domains. The recombinant protein DSR-E synthesizes both α-1,6 and α-1,2 glucosidic linkages in transglucosylation reactions using sucrose as the donor and maltose as the acceptor. To investigate the specific roles of CD1 and CD2 in the catalytic mechanism, truncated forms of dsrE were cloned and expressed in Escherichia coli . Gene products were then small-scale purified to isolate the various corresponding enzymes. Dextran and oligosaccharide syntheses were performed. Structural characterization by 13 C nuclear magnetic resonance and/or high-performance liquid chromatography showed that enzymes devoid of CD2 synthesized products containing only α-1,6 linkages. On the other hand, enzymes devoid of CD1 modified α-1,6 linear oligosaccharides and dextran acceptors through the formation of α-1,2 linkages. Therefore, each domain is highly regiospecific, CD1 being specific for the synthesis of α-1,6 glucosidic bonds and CD2 only catalyzing the formation of α-1,2 linkages. This finding permitted us to elucidate the mechanism of α-1,2 branching formation and to engineer a novel transglucosidase specific for the formation of α-1,2 linkages. This enzyme will be very useful to control the rate of α-1,2 linkage synthesis in dextran or oligosaccharide production.

Published
2004

22. Glucansucrases: Structural Basis, Mechanistic Aspects, and New Perspectives for Engineering

Author

Sandra Pizzut, Magali Remaud-Simeon, Sophie Bozonnet, Gilles Joucia, Cécile Albenne, Gabrielle Potocki-Véronèse, Pierre Monsan, Pierre Escalier, and Emeline Fabre

Subjects
0106 biological sciences, 0303 health sciences, 03 medical and health sciences, Chemistry, 010608 biotechnology, General Medicine, Computational biology, 01 natural sciences, Medicinal chemistry, 030304 developmental biology
Published
2004

23. Oligosaccharides in Food and Agriculture

Author

Gillian Eggleston, Gregory L. Côté, Jeffery S. Rohrer, Casey C. Grimm, Deborah A. Grimm, Christine J. Bergman, Robert A. Rastall, Arland T. Hotchkiss, Estibaliz Olano-Martin, William E. Grace, Glenn R. Gibson, Kiaus Buchholz, Juergen Seibel, Scott M. Holt, Candace Miller-Fosmore, Magali Remaud-Simeon, Cècile Albenne, Gilles Joucia, Emeline Fabre, Sophie Bozonnet, Sandra Pizzut, Pierre Escalier, Gabrielle Potocki-Vèronèse, Pierre Monsan, Tetsuya Nakada, Tomayaki Nishimoto, Hiroto Chaen, Shigeharu Fukuda, Bryan W. Wolf, JoMay Chow, Maureen K. Snowden, Keith A. Garleb, Bryan C. Tungland, James A. Douglas, Merilyn Manley-Harris, John J.C. Scheffer, Natalie A. Wong, John F. Robyt, Kwan Hwa Park, Soo-Bok Lee, Seung-Heon Yoon, Frank Barresi, Angela Eads, Melanie Keuyon, P.G. Morel du Boil, Michael Grisham, Gillian Eggleston, Gregory L. Côté, Jeffery S. Rohrer, Casey C. Grimm, Deborah A. Grimm, Christine J. Bergman, Robert A. Rastall, Arland T. Hotchkiss, Estibaliz Olano-Martin, William E. Grace, Glenn R. Gibson, Kiaus Buchholz, Juergen Seibel, Scott M. Holt, Candace Miller-Fosmore, Magali Remaud-Simeon, Cècile Albenne, Gilles Joucia, Emeline Fabre, Sophie Bozonnet, Sandra Pizzut, Pierre Escalier, Gabrielle Potocki-Vèronèse, Pierre Monsan, Tetsuya Nakada, Tomayaki Nishimoto, Hiroto Chaen, Shigeharu Fukuda, Bryan W. Wolf, JoMay Chow, Maureen K. Snowden, Keith A. Garleb, Bryan C. Tungland, James A. Douglas, Merilyn Manley-Harris, John J.C. Scheffer, Natalie A. Wong, John F. Robyt, Kwan Hwa Park, Soo-Bok Lee, Seung-Heon Yoon, Frank Barresi, Angela Eads, Melanie Keuyon, P.G. Morel du Boil, and Michael Grisham

Published
2003

Catalog

Books, media, physical & digital resources
See catalog results