Your browser doesn't support javascript.
loading
Minimal epitope for Mannitou IgM on paucimannose-carrying glycoproteins.
Robakiewicz, Stefania; Bridot, Clarisse; Serna, Sonia; Gimeno, Ana; Echeverria, Begoña; Delgado, Sandra; de Ruyck, Jérôme; Semwal, Shubham; Charro, Diego; Dansercoer, Ann; Verstraete, Kenneth; Azkargorta, Mikel; van Noort, Kim; Wilbers, Ruud H P; Savvides, Savvas N; Abrescia, Nicola G A; Arda, Ana; Reichardt, Niels C; Jiménez-Barbero, Jesús; Bouckaert, Julie.
Afiliação
  • Robakiewicz S; Unité de Glycobiologie Structurale et Fonctionnelle, UMR 8576 du CNRS et Université de Lille, 50 Avenue Halley, 59650 Villeneuve d'Ascq, France.
  • Bridot C; Unité de Glycobiologie Structurale et Fonctionnelle, UMR 8576 du CNRS et Université de Lille, 50 Avenue Halley, 59650 Villeneuve d'Ascq, France.
  • Serna S; Center for Cooperative Research in Biomaterials (CIC biomaGUNE), Basque Research and Technology Alliance (BRTA), Paseo Miramón 182, 20014 San Sebastian, Spain.
  • Gimeno A; Chemical Glycobiology lab, CIC bioGUNE, ProteoRed-ISCIII, CIBERehd, Basque Research & Technology Alliance (BRTA), Bizkaia Science and Technology park, 48160 Derio, Spain.
  • Echeverria B; Center for Cooperative Research in Biomaterials (CIC biomaGUNE), Basque Research and Technology Alliance (BRTA), Paseo Miramón 182, 20014 San Sebastian, Spain.
  • Delgado S; Chemical Glycobiology lab, CIC bioGUNE, ProteoRed-ISCIII, CIBERehd, Basque Research & Technology Alliance (BRTA), Bizkaia Science and Technology park, 48160 Derio, Spain.
  • de Ruyck J; Unité de Glycobiologie Structurale et Fonctionnelle, UMR 8576 du CNRS et Université de Lille, 50 Avenue Halley, 59650 Villeneuve d'Ascq, France.
  • Semwal S; Unité de Glycobiologie Structurale et Fonctionnelle, UMR 8576 du CNRS et Université de Lille, 50 Avenue Halley, 59650 Villeneuve d'Ascq, France.
  • Charro D; Chemical Glycobiology lab, CIC bioGUNE, ProteoRed-ISCIII, CIBERehd, Basque Research & Technology Alliance (BRTA), Bizkaia Science and Technology park, 48160 Derio, Spain.
  • Dansercoer A; Unit for Structural Biology, VIB - UGent Center for Inflammation Research, Department of Biochemistry and Microbiology, Ghent University, Technologiepark 71, 9052 Ghent, Belgium.
  • Verstraete K; Unit for Structural Biology, VIB - UGent Center for Inflammation Research, Department of Biochemistry and Microbiology, Ghent University, Technologiepark 71, 9052 Ghent, Belgium.
  • Azkargorta M; Chemical Glycobiology lab, CIC bioGUNE, ProteoRed-ISCIII, CIBERehd, Basque Research & Technology Alliance (BRTA), Bizkaia Science and Technology park, 48160 Derio, Spain.
  • van Noort K; Laboratory of Nematology, Plant Science Group, Wageningen University and Research, Droevendaalsesteeg 1, 6708 Wageningen, The Netherlands.
  • Wilbers RHP; Laboratory of Nematology, Plant Science Group, Wageningen University and Research, Droevendaalsesteeg 1, 6708 Wageningen, The Netherlands.
  • Savvides SN; Unit for Structural Biology, VIB - UGent Center for Inflammation Research, Department of Biochemistry and Microbiology, Ghent University, Technologiepark 71, 9052 Ghent, Belgium.
  • Abrescia NGA; Chemical Glycobiology lab, CIC bioGUNE, ProteoRed-ISCIII, CIBERehd, Basque Research & Technology Alliance (BRTA), Bizkaia Science and Technology park, 48160 Derio, Spain.
  • Arda A; IKERBASQUE, Basque Foundation for Science, 48009 Bilbao, Spain.
  • Reichardt NC; Chemical Glycobiology lab, CIC bioGUNE, ProteoRed-ISCIII, CIBERehd, Basque Research & Technology Alliance (BRTA), Bizkaia Science and Technology park, 48160 Derio, Spain.
  • Jiménez-Barbero J; IKERBASQUE, Basque Foundation for Science, 48009 Bilbao, Spain.
  • Bouckaert J; Unité de Glycobiologie Structurale et Fonctionnelle, UMR 8576 du CNRS et Université de Lille, 50 Avenue Halley, 59650 Villeneuve d'Ascq, France.
Glycobiology ; 31(8): 1005-1017, 2021 09 09.
Article em En | MEDLINE | ID: mdl-33909073
Paucimannosidic glycans are restricted to the core structure [Man1-3GlcNAc2Fuc0-1] of N-glycans and are rarely found in mammalian tissues. Yet, especially [Man2-3GlcNAc2Fuc1] have been found significantly upregulated in tumors, including in colorectal and liver cancer. Mannitou IgM is a murine monoclonal antibody that was previously shown to recognize Man3GlcNAc2 with an almost exclusive selectivity. Here, we have sought the definition of the minimal glycan epitope of Mannitou IgM, initiated by screening on a newly designed paucimannosidic glycan microarray; among the best binders were Man3GlcNAc2 and its α1,6 core-fucosylated variant, Man3GlcNAc2Fuc1. Unexpectedly and in contrast to earlier findings, Man5GlcNAc2-type structures bind equally well and a large tolerance was observed for substitutions on the α1,6 arm. It was confirmed that any substitution on the single α1,3-linked mannose completely abolishes binding. Surface plasmon resonance for kinetic measurements of Mannitou IgM binding, either directly on the glycans or as presented on omega-1 and kappa-5 soluble egg antigens from the helminth parasite Schistosoma mansoni, showed submicromolar affinities. To characterize the epitope in greater and atomic detail, saturation transfer difference nuclear magnetic resonance spectroscopy was performed with the Mannitou antigen-binding fragment. The STD-NMR data demonstrated the strongest interactions with the aliphatic protons H1 and H2 of the α1-3-linked mannose and weaker imprints on its H3, H4 and H5 protons. In conclusion, Mannitou IgM binding requires a nonsubstituted α1,3-linked mannose branch of paucimannose also on proteins, making it a highly specific tool for the distinction of concurrent human tumor-associated carbohydrate antigens.
Assuntos
Palavras-chave

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Schistosoma mansoni / Glicoproteínas Limite: Animals / Humans Idioma: En Revista: Glycobiology Assunto da revista: BIOQUIMICA Ano de publicação: 2021 Tipo de documento: Article País de afiliação: França

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Schistosoma mansoni / Glicoproteínas Limite: Animals / Humans Idioma: En Revista: Glycobiology Assunto da revista: BIOQUIMICA Ano de publicação: 2021 Tipo de documento: Article País de afiliação: França