A novel sulfated structure in the carbohydrate-protein linkage region isolated from porcine intestinal heparin.
A preparation of porcine stage 14 intestinal heparin, which contains Ser as a predominant amino acid, was used for isolation of the carbohydrate-protein linkage region of heparin. Two glycoserines were isolated in a molar ratio of 96:4 after an exhaustive digestion with a mixture of bacterial heparinase and heparitinases. Their structures were determined by composition analysis, heparitinase digestion, co-chromatography with an authentic glycoserine on high performance liquid chromatography, and by 500-MHz one- and two-dimensional 1H NMR spectroscopy. The structure of the major one is delta GlcA beta 1-3Gal beta 1-3Gal beta 1-4Xyl beta 1-O-Ser and that of the minor is delta GlcA beta 1-4GlcNAc(6-O-sulfate) alpha 1-4GlcA beta 1-3Gal beta 1-3Gal beta 1-4Xyl beta 1-O-Ser. The novel 6-O-sulfated GlcNAc residue was demonstrated to occur in the vicinity of the carbohydrate-protein linkage region. The Gal residues were nonsulfated, in contrast to the sulfated Gal structures recently discovered in the carbohydrate-protein linkage region of chondroitin sulfate proteoglycans. The structural features are discussed in relation to biosynthetic mechanisms of the heparin glycosaminoglycans.
Sugahara
K
K
Department of Biological Chemistry, Faculty of Pharmaceutical Sciences, Kyoto University, Japan.
Yamada
S
S
Yoshida
K
K
de Waard
P
P
Vliegenthart
J F
JF
D000818:Animals; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002851:Chromatography, High Pressure Liquid; D006493:Heparin / Q000737:chemistry* / Q000302:isolation & purification; D019763:Heparin Lyase; D006497:Heparan Sulfate / Q000737:chemistry* / Q000302:isolation & purification; D007422:Intestines / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000737:chemistry / Q000302:isolation & purification; D011133:Polysaccharide-Lyases; D012694:Serine / Q000032:analysis; D013552:Swine
D006493:Heparin; D019763:Heparin Lyase; D006497:Heparan Sulfate; D009844:Oligosaccharides; D011133:Polysaccharide-Lyases; D012694:Serine; C021349:heparitinsulfate lyase