N-Acetylated amino sugars: the dependence of NMR 3 J (HNH2)-couplings on conformation, dynamics and solvent† †Electronic supplementary information (ESI) available: Calculated energies, relative populations and calculated coupling constants of GlcNAc. See DOI: 10.1039/b705761j Click here for additional data file.

N-Acetylated amino sugars are essential components of living organisms, but their dynamic conformational properties are poorly understood due to a lack of suitable experimental methodologies. Nuclear magnetic resonance (NMR) is ideally suited to these conformational studies, but accurate equations relating the conformation of key substituents (e.g., the acetamido group) to NMR observables are unavailable. To address this, density functional theory (DFT) methods have been used to calculate vicinal coupling constants in N-acetylated amino sugars and derive empirical Karplus equations for (3)J(H(N)H(2)) of N-acetyl-D-glucosamine (GlcNAc) and N-acetyl-D-galactosamine (GalNAc). The fitted Karplus parameters were found to be similar to those previously derived for peptide amide groups, but are consistently larger in magnitude. Local intramolecular interactions had a small effect on the calculated J-couplings and comparison with experimental data suggested that DFT slightly overestimated them. An implicit solvation model consistently lowered the magnitude of the calculated values, improving the agreement with the experimental data. However, an explicit solvent model, while having a small effect, worsened the agreement with experimental data. The largest contributor to experimentally-determined (3)J(H(N)H(2))-couplings is proposed to be librations of the amide group, which are well approximated by a Gaussian distribution about a mean dihedral angle. Exemplifying the usefulness of our derived Karplus equations, the libration of the amide group could be estimated in amino sugars from experimental data. The dynamical spread of the acetamido group in free alpha-GlcNAc, beta-GlcNAc and alpha-GalNAc was estimated to be 32 degrees , 42 degrees and 20 degrees , with corresponding mean dihedral angles of 160 degrees , 180 degrees and 146 degrees , respectively.