Sugahara K, Yamada S, Yoshida K, de Waard P, Vliegenthart J F
Department of Biological Chemistry, Faculty of Pharmaceutical Sciences, Kyoto University, Japan.
J Biol Chem. 1992 Jan 25;267(3):1528-33.
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.
一种猪14期肠肝素制剂用于肝素糖蛋白连接区的分离，该制剂以丝氨酸作为主要氨基酸。用细菌肝素酶和类肝素酶混合物彻底消化后，分离出两种糖丝氨酸，摩尔比为96:4。通过成分分析、类肝素酶消化、在高效液相色谱上与纯糖丝氨酸共色谱以及500兆赫一维和二维1H核磁共振光谱确定了它们的结构。主要的一种结构是δGlcAβ1-3Galβ1-3Galβ1-4Xylβ1-O-Ser，次要的一种结构是δGlcAβ1-4GlcNAc(6-O-硫酸盐)α1-4GlcAβ1-3Galβ1-3Galβ1-4Xylβ1-O-Ser。已证明新型6-O-硫酸化GlcNAc残基存在于糖蛋白连接区附近。与最近在硫酸软骨素蛋白聚糖糖蛋白连接区发现的硫酸化半乳糖结构相反，这些半乳糖残基未被硫酸化。结合肝素糖胺聚糖的生物合成机制讨论了结构特征。
