title: Lectins as probes for assessing the accessibility of N-linked glycans in relation to the conformational changes of fibronectin.
abstract: Fibronectin, a ≈ 450-kDa protein with 4-9% (w/w) glycosylation, is a key component of extracellular matrices and has a high conformational lability regarding its functions. However, the accessibility and the role of glycosylated moieties associated with the conformational changes of fibronectin are poorly understood. Using lectins as probes, we developed an approach comprising dynamic light scattering, turbidimetry measurements, and isothermal titration calorimetry to assess the accessibility of glycosylated moieties of fibronectin undergoing thermal-induced conformational changes. Among a set of 14 lectins, fibronectin mainly reacted with mannose-binding lectins, specifically concanavalin A. When temperature was raised from 25 to 50 °C, fibronectin underwent progressive unfolding, but the conformation of concanavalin A was unaffected. Dynamic light scattering, turbidimetry measurements, and isothermal titration calorimetry showed increased concanavalin A binding to fibronectin during progressive thermal-induced unfolding of the protein core. Such data suggest that mannosylated residues are progressively exposed as fibronectin unfolds. Because oligosaccharide moieties can be differently exposed to cells, and the cell's responses could be modified physiologically or pathologically, modulation of fibronectin sugar chains could be relevant to its biological functions. Thus, lectins might be useful tools to probe the glycosylation accessibility accompanying changes in protein core folding, for which a better understanding would be of value for biological and biomedical research.
authors.affiliation: Equipe de Recherche sur les Relations Matrice Extracellulaire-Cellules, ERRMECe (EA1391), Institut des matériaux, I-MAT (FD4122), Université de Cergy-Pontoise, F-95000 Cergy-Pontoise, 2 Avenue Adolphe Chauvin, Cergy-Pontoise Cedex, 95302, France.
authors.forename: Rémy
authors.identifier: http://orcid.org/0000-0002-2956-7532
authors.initials: R
authors.lastname: Agniel
authors.affiliation: Equipe de Recherche sur les Relations Matrice Extracellulaire-Cellules, ERRMECe (EA1391), Institut des matériaux, I-MAT (FD4122), Université de Cergy-Pontoise, F-95000 Cergy-Pontoise, 2 Avenue Adolphe Chauvin, Cergy-Pontoise Cedex, 95302, France.
authors.forename: Charlotte
authors.initials: C
authors.lastname: Vendrely
authors.affiliation: Equipe de Recherche sur les Relations Matrice Extracellulaire-Cellules, ERRMECe (EA1391), Institut des matériaux, I-MAT (FD4122), Université de Cergy-Pontoise, F-95000 Cergy-Pontoise, 2 Avenue Adolphe Chauvin, Cergy-Pontoise Cedex, 95302, France.|Orange, Direction Nord de France, 2 rue Trémière, Villeneuve d'Ascq, F-59650, France.
authors.forename: Laurent
authors.initials: L
authors.lastname: Poulouin
authors.affiliation: Equipe de Recherche sur les Relations Matrice Extracellulaire-Cellules, ERRMECe (EA1391), Institut des matériaux, I-MAT (FD4122), Université de Cergy-Pontoise, F-95000 Cergy-Pontoise, 2 Avenue Adolphe Chauvin, Cergy-Pontoise Cedex, 95302, France.
authors.forename: Rümeyza
authors.initials: R
authors.lastname: Bascetin
authors.affiliation: Equipe de Recherche sur les Relations Matrice Extracellulaire-Cellules, ERRMECe (EA1391), Institut des matériaux, I-MAT (FD4122), Université de Cergy-Pontoise, F-95000 Cergy-Pontoise, 2 Avenue Adolphe Chauvin, Cergy-Pontoise Cedex, 95302, France.
authors.forename: Hamanou
authors.initials: H
authors.lastname: Benachour
authors.affiliation: Equipe de Recherche sur les Relations Matrice Extracellulaire-Cellules, ERRMECe (EA1391), Institut des matériaux, I-MAT (FD4122), Université de Cergy-Pontoise, F-95000 Cergy-Pontoise, 2 Avenue Adolphe Chauvin, Cergy-Pontoise Cedex, 95302, France.
authors.forename: Olivier
authors.initials: O
authors.lastname: Gallet
authors.affiliation: Equipe de Recherche sur les Relations Matrice Extracellulaire-Cellules, ERRMECe (EA1391), Institut des matériaux, I-MAT (FD4122), Université de Cergy-Pontoise, F-95000 Cergy-Pontoise, 2 Avenue Adolphe Chauvin, Cergy-Pontoise Cedex, 95302, France.
authors.forename: Johanne
authors.initials: J
authors.lastname: Leroy-Dudal
mesh_terms: D002151:Calorimetry; D000067493:Dynamic Light Scattering; D005353:Fibronectins / Q000378:metabolism*; D006031:Glycosylation; D006801:Humans; D037241:Mannose-Binding Lectins / Q000378:metabolism*; D009391:Nephelometry and Turbidimetry; D011485:Protein Binding; D011487:Protein Conformation; D017510:Protein Folding
chemical_list: C067625:FN1 protein, human; D005353:Fibronectins; D037241:Mannose-Binding Lectins
keywords: N-linked glycosylation; concanavalin A; fibronectin; lectins