title: Surface-based and mass spectrometric approaches to deciphering sugar-protein interactions in a galactose-specific agglutinin.
abstract: Interest in powerful, nanosized tools to analyze in detail glycan-protein interactions has increased significantly over recent years. Here, we report two complementary approaches to characterize such interactions with high sensitivity, low sample consumption, and without the need for sample labeling, namely, surface plasmon resonance (SPR) and an approach that combines limited proteolysis and mass spectrometry. Combination of these two approaches to investigate glycan-protein interactions allows (1) to characterize interactions through kinetic and thermodynamic parameters, (2) to capture efficiently the carbohydrate-binding protein, and (3) to identify the interacted protein and its carbohydrate binding site by mass spectrometry. As a proof of principle, the interaction of the galactose-specific legume lectin Erythrina cristagalli agglutinin with several sugars has been characterized in-depth by means of these two approaches.
authors: Department of Experimental and Health Sciences, Pompeu Fabra University, Barcelona Biomedical Research Park, 08003 Barcelona, Spain.
authors: Carmen
authors: C
authors: Jiménez-Castells
authors: Sira
authors: S
authors: Defaus
authors: Adrian
authors: A
authors: Moise
authors: Michael
authors: M
authors: Przbylski
authors: David
authors: D
authors: Andreu
authors: Ricardo
authors: R
authors: Gutiérrez-Gallego
mesh_terms: D001665:Binding Sites; D005690:Galactose / Q000737:chemistry / Q000378:metabolism*; D007700:Kinetics; D037121:Plant Lectins / Q000378:metabolism; D011485:Protein Binding; D011956:Receptors, Cell Surface / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D020349:Surface Plasmon Resonance; D013816:Thermodynamics
chemical_list: D037121:Plant Lectins; D011956:Receptors, Cell Surface; C045629:erythrina lectin; C055213:saccharide-binding proteins; D005690:Galactose