High-performance liquid chromatographic identification of disaccharides generated from heparan sulphate isomers using heparitinases.
Specific heparan sulphate-lyases, heparitinases I and II, were used to identify unsaturated disaccharide constituents generated from heterogeneous heparan sulphate isomers. All determinations were made using high-performance liquid chromatography with a column containing a sulphonized styrene-divinylbenzene copolymer. Unsaturated disaccharides generated from variously sulphated heparan sulphate isomers after simultaneous digestion with heparitinases I and II facilitated separation of the individual disaccharides, based on sulphate groups at the specific position of the uronic acid and glucosamine residues. The simultaneous digestion with heparitinases I and II produces unsaturated disaccharides from heparan sulphate isomers with the structure of 4-deoxy-2-O-alpha-L-threo-hex-4-enepyranosyluronic acid (1----4)-2-amino-deoxy-D-glucose, 4-deoxy-2-O-alpha-L-threo-hex-4-enepyranosyluronic acid (1----4)-2-deoxy-2-sulphamido-D-glucose, 4-deoxy-2-O-alpha-L-threo-hex-4-enepyranosyluronic acid (1----4)-2-aminodeoxy-6-O-sulpho-D-glucose, 4-deoxy-2-O-alpha-L-threo-hex-4-enepyranosyluronic acid (1----4)-2-deoxy-2-sulphamido-6-O-sulpho-D-glucose, 4-deoxy-2-O-sulpho-alpha-L-threo-hex-4-enepyranosyluronic acid (1----4)-2-amino-2-deoxy-6-O-sulpho-D-glucose and 4-deoxy-2-O-sulpho-alpha-L-threo-hex-4-enepyranosyluronic acid (1----4)-2-deoxy-2-sulphamido-6-O-sulpho-D-glucose.
Murata
K
K
Medical Center of Tokyo National University of Arts and Music, Japan.
Yokoyama
Y
Y
Yoshida
K
K
D002851:Chromatography, High Pressure Liquid; D004187:Disaccharides / Q000032:analysis* / Q000302:isolation & purification; D006025:Glycosaminoglycans / Q000032:analysis*; D006497:Heparan Sulfate / Q000032:analysis*; D006801:Humans; D007202:Indicators and Reagents; D007536:Isomerism; D007668:Kidney / Q000378:metabolism; D011133:Polysaccharide-Lyases; D013056:Spectrophotometry, Ultraviolet
D004187:Disaccharides; D006025:Glycosaminoglycans; D006497:Heparan Sulfate; D007202:Indicators and Reagents; D011133:Polysaccharide-Lyases; C021349:heparitinsulfate lyase