Conformational Preferences at the Glycosidic Linkage of Saccharides in Solution as Deduced from NMR Experiments and MD Simulations: Comparison to Crystal Structures

Abstract Glycans are central to information content and regulation in biological systems. These carbohydrate molecules are active either as oligo‐ or polysaccharides, often in the form of glycoconjugates. The monosaccharide entities are joined by glycosidic linkages and stereochemical arrangements are of utmost importance in determining conformation and flexibility of saccharides. The conformational preferences and population distributions at the glycosidic torsion angles φ and ψ have been investigated for O ‐methyl glycosides of three disaccharides where the substitution takes place at a secondary alcohol, viz., in α‐ l ‐Fuc p ‐(1→3)‐β‐ d ‐Glc p ‐OMe, α‐ l ‐Fuc p ‐(1→3)‐α‐ d ‐Gal p ‐OMe and α‐ d ‐Glc p ‐(1→4)‐α‐ d ‐Gal p ‐OMe, corresponding to disaccharide structural elements present in bacterial polysaccharides. Stereochemical differences at or adjacent to the glycosidic linkage were explored by solution state NMR spectroscopy using one‐dimensional 1 H, 1 H‐NOESY NMR experiments to obtain transglycosidic proton‐proton distances and one‐ and two‐dimensional heteronuclear NMR experiments to obtain 3 J CH transglycosidic coupling constants related to torsion angles φ and ψ . Computed effective proton‐proton distances from molecular dynamics (MD) simulations showed excellent agreement to experimentally derived distances for the α‐(1→3)‐linked disaccharides and revealed that for the bimodal distribution at the ψ torsion angle for the α‐(1→4)‐linked disaccharide experiment and simulation were at variance with each other, calling for further force field developments. The MD simulations disclosed a highly intricate inter‐residue hydrogen bonding pattern for the α‐(1→4)‐linked disaccharide, including a nonconventional hydrogen bond between H5′ in the glucosyl residue and O3 in the galactosyl residue, supported by a large downfield 1 H NMR chemical shift displacement compared to α‐ d ‐Glc p ‐OMe. Comparison of population distributions of the glycosidic torsion angles φ and ψ in the disaccharide entities to those of corresponding crystal structures highlighted the potential importance of solvation on the preferred conformation.