Conformation And Binding Aspects Of Sulfated Carbohydrates By Liquid-State Nmr

Liquid-state NMR has long been an important tool for understanding the conformations and interactions of biomacromolecules. To date these conclusions have primarily been focused upon nucleotides and peptides with little information regarding the conformational structures of carbohydrates. Through the methods described in this manuscript the accurate conformational analysis of a sulfated fucan from Lytechinus variegatus has been assessed. This was achieved through a combination of NOESY spectroscopy and assessment of chemical exchange in hydroxyl protons undergoing fast chemical exchange with excess solvent. This is accomplished through usage of super- cooled H2O: acetone solvent and T1 inversion recovery. This methodology was optimized in L-fucose and applied to the sulfated fucan for normalization of NOE information for derivatization of distance restraints. Molecular dynamics modeling further confirmed these structures as supported. Furthermore, the structure-activity relationships between glycosaminoglycan oligosaccharides and their binding partners have been heavily studied. Usage of saturation transfer difference NMR has allowed robust conclusions towards understanding the impact of chain length, position of sulfates, and carbohydrate composition on the complex formation between the sulfated polysaccharides and their binding partners