Detailed Investigation of the Immunodominant Role of O-Antigen Stoichiometric O-Acetylation as Revealed by Chemical Synthesis, Immunochemistry, Solution Conformation and STD-NMR Spectroscopy forShigella flexneri 3a

Autor: Laurence A. Mulard, Ana Ardá, F. Javier Cañada, Armelle Phalipon, Pilar Blasco, Françoise Thouron, Catherine Guerreiro, Jesús Jiménez-Barbero, Farida Nato, Sylvie Dartevelle, Julien Boutet
Přispěvatelé: Chimie des Biomolécules - Chemistry of Biomolecules, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Centro de Investigaciones Biológicas (CSIC), Consejo Superior de Investigaciones Científicas [Madrid] (CSIC), Pathogénie microbienne moléculaire, Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM), Ingénierie des Anticorps (plate-forme) - Antibody Engineering (Platform), Cell Biology and Stem Cells Unit (CICbioGUNE), Technologic Park of Bizkaia, This work was supported by the Agence Nationale de la Recherche (ANR, Grant NT05‐1 42479), the MENRT (fellowship to J.B.), the European Commission Seventh Framework Program (FP7/2007‐2013) under Grant agreement No. 261472‐STOPENTERICS. We also thank MINECO for financial support (Grants CTQ2012‐32025 and CTQ2015‐64597‐C2‐1‐P, and CTQ2015‐64597‐C2‐2‐P)., The authors thank F. Bonhomme (UMR CNRS 3523) for obtaining the NMR and ESI spectra, M. Tanguy (PMM) for the mAb characterization, and F. Traincard (PFIA, Institut Pasteur) for useful technical comments., European Project: 261472,EC:FP7:HEALTH,FP7-HEALTH-2010-single-stage,STOPENTERICS(2010), Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris], Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Pasteur [Paris], Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM)
Rok vydání: 2016
Předmět:
Magnetic Resonance Spectroscopy
Glycosylation
glycosylation
MESH: Shigella flexneri
Stereochemistry
carbohydrates
MESH: Mice
Inbred BALB C
010402 general chemistry
01 natural sciences
Chemical synthesis
Catalysis
Shigella flexneri
Mice
chemistry.chemical_compound
NMR spectroscopy
Antigen
Immunochemistry
medicine
Animals
antibodies
MESH: Animals
MESH: Mice
MESH: O Antigens
Mice
Inbred BALB C
[SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry
Molecular Biology/Structural Biology [q-bio.BM]
biology
MESH: Immunochemistry
MESH: Magnetic Resonance Spectroscopy
[CHIM.ORGA]Chemical Sciences/Organic chemistry
010405 organic chemistry
Chemistry
Organic Chemistry
epitope specificity
Bacillary dysentery
O Antigens
General Chemistry
Nuclear magnetic resonance spectroscopy
biology.organism_classification
medicine.disease
0104 chemical sciences
[SDV.IMM.IA]Life Sciences [q-bio]/Immunology/Adaptive immunology
Biochemistry
Acetylation
[SDV.IMM.VAC]Life Sciences [q-bio]/Immunology/Vaccinology
Zdroj: Chemistry-A European Journal
Chemistry-A European Journal, 2016, 22 (31), pp.10892-10911. ⟨10.1002/chem.201600567⟩
Chemistry-A European Journal, Wiley-VCH Verlag, 2016, 22 (31), pp.10892-10911. ⟨10.1002/chem.201600567⟩
ISSN: 0947-6539
1521-3765
DOI: 10.1002/chem.201600567
Popis: International audience; Shigella flexneri 3a causes bacillary dysentery. Its O-antigen has the {2)-[α-d-Glcp-(1→3)]-α-l-Rhap-(1→2)-α-l-Rhap-(1→3)-[Ac→2]-α-l-Rhap-(1→3)-[Ac→6]≈40 % -β-d-GlcpNAc-(1→} ([(E)ABAc CAc D]) repeating unit, and the non-O-acetylated equivalent defines S. flexneri X. Propyl hepta-, octa-, and decasaccharides sharing the (E')A'BAc CD(E)A sequence, and their non-O-acetylated analogues were synthesized from a fully protected BAc CD(E)A allyl glycoside. The stepwise introduction of orthogonally protected mono- and disaccharide imidate donors was followed by a two-step deprotection process. Monoclonal antibody binding to twenty-six S. flexneri types 3a and X di- to decasaccharides was studied by an inhibition enzyme-linked immunosorbent assay (ELISA) and STD-NMR spectroscopy. Epitope mapping revealed that the 2C -acetate dominated the recognition by monoclonal IgG and IgM antibodies and that the BAc CD segment was essential for binding. The glucosyl side chain contributed to a lesser extent, albeit increasingly with the chain length. Moreover, tr-NOESY analysis also showed interaction but did not reveal any meaningful conformational change upon antibody binding.
Databáze: OpenAIRE
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