Conformations of Flexible Oligosaccharides Molecular Simulations and NMR spectroscopy

Abstract: The conformational preferences of several oligosaccharides are investigated herein using a combination of NMR spectroscopy and molecular dynamics (MD) simulations, focusing on the torsion angles associated with the glycosidic linkages.Strategies for obtaining usable J-HMBC spectra for carbons with an adjacent 13C label are described. By employing a selective pulse or a constant time modification, spectra free from interferences are obtained for site-specifically 13C labeled oligosaccharides.Intermolecular hydrogen bonding in sucrose is investigated using MD simulations performed at different concentrations. One of the most frequent intermolecular hydrogen bonds in the simulations, O3f???HO3g, was detected using the HSQC-TOCSY NMR experiment.Based on MD simulations and NMR spectroscopy, the conformational ensemble for a trisaccharide segment of the LeaLex hexasaccharide is proposed to feature conformational exchange between conformations with positive and negative values for the ?3 torsion angle in the ?-D-GlcpNAc-(1?3)-?-D-Galp linkage.Using MD simulations, the conformation of the N-acetyl group is shown to influence the glycosidic conformation at a nearby linkage in two oligosaccharides.Short (1?6)-linked oligosaccharides are shown to exhibit conformational exchange at the ? and ? torsion angles. Notably, the former torsion angle populates states with ? ? ±90°. Conformationally sensitive homo- and heteronuclear coupling constants are determined using various NMR experiments. The experimental data, including effective distances from NOESY obtained for two of the compounds, is used to improve the representation of the ? torsion angle in the CHARMM36 force field.