Microanalysis of glycosaminoglycan-derived oligosaccharides labeled with a fluorophore 2-aminobenzamide by high-performance liquid chromatography: application to disaccharide composition analysis and exosequencing of oligosaccharides.
A series of disaccharides derived from chondroitin sulfate and heparin/heparan sulfate were derivatized at their reducing ends with a fluorophore 2-aminobenzamide to develop a sensitive microanalytical method for glycosaminoglycans. The resulting labeled compounds derived from chondroitin sulfate or heparin/heparan sulfate were well-separated and quantified by HPLC equipped with a fluorescence detector. The detection limit was a low picomole level. This method was applied to the analysis of the disaccharide composition of tetra- and hexasaccharides derived from chondroitin sulfate and heparin/heparan sulfate as well as these glycosaminoglycan polysaccharides. The method was also successfully applied to the exosequencing of chondrohexasaccharides, where the fluorophore-labeled oligosaccharides were degraded exolytically from the nonreducing ends using bacterial eliminases. The resultant labeled fragments were identified by HPLC.
Kinoshita
A
A
Department of Biochemistry, Kobe Pharmaceutical University, Kobe, Higashinada-ku, 658-8558, Japan.
Sugahara
K
K
D002240:Carbohydrate Sequence; D002809:Chondroitin Sulfates / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods* / Q000706:statistics & numerical data; D004187:Disaccharides / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification; D005456:Fluorescent Dyes; D006025:Glycosaminoglycans / Q000737:chemistry*; D006493:Heparin / Q000737:chemistry; D006497:Heparan Sulfate / Q000737:chemistry; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D062367:ortho-Aminobenzoates
D002809:Chondroitin Sulfates; D004187:Disaccharides; D005456:Fluorescent Dyes; D006025:Glycosaminoglycans; D006493:Heparin; D006497:Heparan Sulfate; D009844:Oligosaccharides; D062367:ortho-Aminobenzoates; C000219:anthranilamide