Quantifying Siglec-sialylated ligand interactions: a versatile <sup>19</sup> F-T <sub>2</sub> CPMG filtered competitive NMR displacement assay
Unai Atxabal, Andrea Fernández, María Jesús Moure, Klaudia Sobczak, Corwin M. Nycholat, Verónica Almeida‐Marrero, Iker Oyenarte, James C. Paulson, Andrés de la Escosura, Tomás Torres⊗, Niels C. Reichardt, Jesús Jiménez‐Barbero, June Ereño‐Orbea
Abstract
-relaxation NMR and on the design, synthesis, and use of a strategic spy molecule, to assess and quantify sialoside ligand binding to Siglecs. We show that the use of this specific approach allows the quantification of Siglec binding for natural and modified sialosides, multivalent sialosides, and sialylated glycoproteins in solution, which differ in binding affinities in more than two orders of magnitude, thus providing invaluable insights into sialoglycan-mediated interactions.
Topics & Concepts
SIGLECSialic acidChemistryLigand (biochemistry)CD22Immune systemBiophysicsCellBiochemistryBiologyReceptorImmunologyCD19Glycosylation and Glycoproteins ResearchCarbohydrate Chemistry and SynthesisGalectins and Cancer Biology