title: Rapid identification of common hexapyranose monosaccharide units by a simple TOCSY matching approach.
abstract: Solution NMR spectroscopy is a well established technique for non-destructive characterization of the structures and conformations of complex oligo- and polysaccharides. One of the key experiments involves the use of 2D TOCSY to collect the 1H spins into groups that can be associated with the individual saccharide units that are present in the molecule under study. It is well known that the magnetization transfer rate through the 1H spin system during the TOCSY spin lock period is sensitive to the intervening 3J(H,H) scalar couplings, and therefore also to the saccharide stereochemistry. Here, we have investigated the potential to extract information on the stereochemistry of hexapyranose monosaccharide units directly from TOCSY spectra. Through a systematic experimental investigation of the magnetization transfer initiated from the anomeric 1H resonance in D-glucose, D-galactose and D-mannose it is shown that a 100 ms spin lock time provides optimal spectroscopic discrimination between these three commonly occurring building blocks. A simple matching scheme is proposed as a new tool for rapid attribution of the TOCSY traces originating from the anomeric 1H resonances towards the underlying monosaccharide type. The scheme appears robust with regard to structural variations and fairly tolerant to incidental overlap. Its application provides useful guidance during the subsequent NMR assignment process, as demonstrated with the PS7F polysaccharide from Streptococcus pneumonia. In addition, we show that our scheme affords a clear-cut distinction between the alpha- and beta-epimers of D-mannose-type units, which can be difficult to discriminate by NMR analysis. Application to the N-glycan 100.2 demonstrates the potential and wide applicability of this new discrimination approach.
authors: NMR and Structure Analysis Unit, Department of Organic Chemistry, Universiteit Gent, Krijgslaan 281, S4, 9000 Gent (Belgium), Fax: (+32) 9-264-4972.
authors: Katelijne
authors: K
authors: Gheysen
authors: NMR and Structure Analysis Unit, Department of Organic Chemistry, Universiteit Gent, Krijgslaan 281, S4, 9000 Gent (Belgium), Fax: (+32) 9-264-4972.|High Resolution NMR Centre (HNMR), Department of Polymer Science and Structural Chemistry (POSC), Vrije Universiteit Brussel, Pleinlaan 2, 1050 Brussel (Belgium).
authors: Camelia
authors: C
authors: Mihai
authors: GlaxoSmithKline, Biologicals, Rue de l'Institut 89, 1330 Rixensart (Belgium).
authors: Karel
authors: K
authors: Conrath
authors: NMR and Structure Analysis Unit, Department of Organic Chemistry, Universiteit Gent, Krijgslaan 281, S4, 9000 Gent (Belgium), Fax: (+32) 9-264-4972.
authors: José C
authors: JC
authors: Martins
mesh_terms: D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009005:Monosaccharides / Q000032:analysis* / Q000737:chemistry*; D013997:Time Factors
chemical_list: D009005:Monosaccharides