Structure and anticoagulant activity of a fucosylated chondroitin sulfate from echinoderm - Sulfated fucose branches on the polysaccharide account for its high anticoagulant action
Structure and anticoagulant activity of a fucosylated chondroitin sulfate from echinoderm - Sulfated fucose branches on the polysaccharide account for its high anticoagulant action
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DOI：
10.1074/jbc.271.39.23973
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发表时间：
1996-09-27
影响因子：
4.8
通讯作者：
Abildgaard, U
中科院分区：
文献类型：
--
作者：
Mourao, PAS;Pereira, MS;Abildgaard, U
从海参体壁分离得到的一种多糖具有哺乳动物硫酸软骨素的骨架：[4-β-D-Glca-1->；3-β-D-GalNAc-1](N)，但在β-D-葡萄糖醛酸残基的3位被硫酸化的α-L-岩藻糖基取代(Vieira，R.P.，Mulloy，B.和Mourao，P.A.S.(1991)J.Biol)。化学。13530-13536)。温和的酸水解去除了α-L岩藻糖的硫酸化分支，裂解残基通过H-1NMR谱进行了表征，最丰富的物种是岩藻糖4-O-单硫酸盐，但也存在2，4-和3，4-二O-O-硫酸盐残基。用软骨素ABC裂解酶对剩下的多糖进行降解，结果表明，温和酸解释放的硫酸化的α-L岩藻糖残基集中在多糖链的非还原末端；耐酶多糖材料包括还原末端，携带耐酸的L岩藻糖取代物。硫酸化的α-L岩藻糖枝对多糖具有抗凝血活性。在活化的部分凝血活酶时间测定中，岩藻糖化的硫酸软骨素的比活性大于相同硫化程度的线型均聚体α-L-岩藻聚糖的比活性；这种活性在脱糖或脱硫时丧失，但不在聚合物的羧基还原时丧失。纯化试剂检测表明，岩藻糖化硫酸软骨素能增强抗凝血酶和肝素辅因子II的凝血酶抑制活性。
A polysaccharide isolated from the body wall of the sea cucumber Ludwigothurea grisea has a backbone like that of mammalian chondroitin sulfate: [4-beta-D-GlcA-1-->3-beta-D-GalNAc-1](n) but substituted at the 3-position of the beta-D-glucuronic acid residues with sulfated alpha-L-fucopyranosyl branches (Vieira, R. P., Mulloy, B., and Mourao, P. A. S. (1991) J. Biol. Chem. 266, 13530-13536). Mild acid hydrolysis removes the sulfated alpha-L-fucose branches, and cleaved residues have been characterized by H-1 NMR spectroscopy; the most abundant species is fucose 4-O-monosulfate, but 2,4- and 3,4-di-O-sulfated residues are also present. Degradation of the remaining polysaccharide with chondroitin ABC lyase shows that the sulfated alpha-L-fucose residues released by mild acid hydrolysis are concentrated toward the non-reducing end of the polysaccharide chains; enzyme-resistant polysaccharide material includes the reducing terminal and carries acid resistant L-fucose substitution. The sulfated alpha-L-fucose branches confer anticoagulant activity on the polysaccharide. The specific activity of fucosylated chondroitin sulfate in the activated partial thromboplastin time assay is greater than that of a linear homopolymeric alpha-L-fucan with about the same level of sulfation; this activity is lost on defucosylation or desulfation but not on carboxyl-reduction of the polymer. Assays with purified reagents show that the fucosylated chondroitin sulfate can potentiate the thrombin inhibition activity of both antithrombin and heparin cofactor II.