Isolation and characterization of a heparin with high anticoagulant activity from the clam Tapes phylippinarum:: evidence for the presence of a high content of antithrombin III binding site
Isolation and characterization of a heparin with high anticoagulant activity from the clam Tapes phylippinarum:: evidence for the presence of a high content of antithrombin III binding site
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DOI：
10.1093/glycob/cwh128
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发表时间：
2004-12-01
期刊：
影响因子：
4.3
通讯作者：
Volpi, N
中科院分区：
文献类型：
--
作者：
Cesaretti, M;Luppi, E;Volpi, N
具有高抗凝活性的肝素（活化部分凝血活酶时间为347 +/- 56.4，抗Xa活性为317 +/- 48.3）是从海洋蛤种Tapes phylippinarum中分离出来的，其含量与2.1 mg/g干动物相似。琼脂糖凝胶电泳显示慢移动肝素组分（22 +/- 6.8%）和78 +/- 5.4%的快速移动物质含量高。通过PAGE分析计算平均分子量为13，600，而通过高效尺寸排阻色谱法获得数均分子量Mn值为10，700，重均分子量Mw为14，900，分散指数Mn/Mw为1.386。通过用肝素酶（EC www.example.com）解聚肝素样品4.2.2.7，然后通过强阴离子交换-HPLC分离所得的不饱和寡糖，进行蛤肝素的结构分析，结果显示存在大量的（比从牛肠获得的标准药物肝素高130%）的具有部分ATIII结合区的寡糖序列，在T中的Δ UA2S（1->4）-α-D-GlcN2S6S（1->4）-α-L-IdoA（1->4）-α-D-GlcNAc6S（1->4）-β-D-GlcA（1->4）-α-D-GlcN2S3S6S。phylippinarum肝素，与牛粘膜肝素和猪粘膜肝素样品相比，以前发表。此外，正如寡糖组成分析所预期的，由于存在大mol %（80.6%）的三硫酸化二糖Δ UA 2S（1->4）-α-D-GlcN 2S 6S，软体动物肝素是一种比先前报道的牛粘膜肝素（73.5%）和猪粘膜肝素样品（72.8%）硫酸化程度更高的多糖。据我们所知，这是第一篇描述蛤肝素的ATIII结合位点与人和猪肠粘膜肝素和牛肠粘膜肝素的ATIII结合位点基本相同，但与牛肺肝素中发现的ATIII结合位点不同的文章。
Heparin with high anticoagulant activity (activated partial thromboplastin time of 347 +/- 56.4 and anti-Xa activity of 317 +/- 48.3) was isolated from the marine clam species Tapes phylippinarum in an amount of similar to2.1 mg/g dry animals. Agarose-gel electrophoresis showed a high content of the slow-moving heparin component (22 +/- 6.8%) and 78 +/- 5.4% of the fast-moving species. An average molecular mass of 13,600 was calculated by PAGE analysis, whereas a number average molecular weight Mn value of 10,700, a weight average molecular weight Mw of 14,900, and a dispersity index Mn/Mw of 1.386 were obtained by high-performance size-exclusion chromatography. Structural analysis of clam heparin, performed by depolymerizing heparin samples with heparinase (EC 4.2.2.7) and then separating the resulting unsaturated oligosaccharides by strong anion exchange-HPLC revealed the presence of large amounts (more than 130% than standard pharmaceutical heparin obtained from bovine intestine) of the oligosaccharide sequence bearing part of the ATIII-binding region, DeltaUA2S (1-->4)-alpha-D-GlcN2S6S (1-->4)-alpha-L-IdoA (1-->4)-alpha-D-GlcNAc6S (1-->4)-beta-D-GlcA (1-->4)-alpha-D-GlcN2S3S6S in the T. phylippinarum heparin, in comparison with bovine mucosal heparin and a sample of porcine mucosal heparin previously published. Furthermore, as expected from the oligosaccharide compositional analysis, due to the presence of a great mol % (80.6%) of the trisulfated disaccharide DeltaUA2S(1-->4)-alpha-D-GlcN2S6S, mollusc heparin is a more sulfated polysaccharide than bovine mucosal heparin (73.5%) and a sample of porcine mucosal (72.8%) heparin previously reported. To our knowledge, this is the first article describing a clam heparin having the ATIII binding site mainly identical to that of human and porcine intestinal mucosal heparins and bovine intestinal mucosal heparin but different from that found in beef lung heparin.