2-DIMENSIONAL AFFINITY RESOLUTION ELECTROPHORESIS DEMONSTRATES THAT 3 DISTINCT HEPARIN POPULATIONS INTERACT WITH ANTITHROMBIN-III
2-DIMENSIONAL AFFINITY RESOLUTION ELECTROPHORESIS DEMONSTRATES THAT 3 DISTINCT HEPARIN POPULATIONS INTERACT WITH ANTITHROMBIN-III
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DOI：
10.1021/bi00008a002
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发表时间：
1995-02-28
期刊：
影响因子：
2.9
通讯作者：
WEILER, JM
中科院分区：
文献类型：
--
作者：
EDENS, RE;FROMM, JR;WEILER, JM
肝素是一种多分散、高度硫酸化的多糖，由重复的 1 --> 4 连接的糖醛酸和葡萄糖胺糖残基组成，可与凝血蛋白、补体蛋白和生长因子结合以调节多种生物活性。肝素最广为人知的用途是抗凝剂，其活性主要源自肝素中的特定五糖序列，该序列与抗凝血酶 III 中的独特位点相互作用。对于肝素内可能与抗凝血酶 III 相互作用的其他结构或与无数其他肝素结合蛋白和肽相互作用的肝素结构知之甚少。使用一种为定量这种相互作用而开发的新技术，检查了从猪肠粘膜制备的普通糖胺聚糖肝素与抗凝血酶 III 的结合能力。二维亲和分辨率电泳是一种强大的方法，可以评估与蛋白质结合的独特肝素分子种类，从而可以确定每种蛋白质结合肝素种类的肝素分子量以及每种相互作用的解离常数。这项研究提供了第一个明确的证据，证明糖胺聚糖肝素含有至少三个与抗凝血酶 III 具有亲和力的分子群。此外，每种肝素种类对抗凝血酶III的亲和力似乎与肝素链的大小成反比变化，一些较小的寡糖比较大的寡糖对抗凝血酶III具有更大的亲和力。
Heparin is a polydisperse, highly sulfated polysaccharide consisting of repeating 1 --> 4 linked uronic acid and glucosamine sugar residues that binds to coagulation proteins, complement proteins, and growth factors to regulate a variety of biological activities. Heparin is best known as an anticoagulant, an activity that results largely from a specific pentasaccharide sequence in heparin that interacts with a unique site in antithrombin III. Little is known about additional structures within heparin that might interact with antithrombin III or the heparin structures that interact with the myriad of other heparin-binding proteins and peptides. Unfractionated glycosaminoglycan heparin that had been prepared from porcine intestinal mucosa was examined for its capacity to bind antithrombin III using a new technique developed to quantitate that interaction. Two-dimensional affinity resolution electrophoresis is a powerful method that allows assessment of unique species of heparin molecules that bind to protein, allowing determination of heparin molecular weight for each protein-binding heparin species as well as the dissociation constant of each interaction. This study provides the first definitive evidence that glycosaminoglycan heparin contains at least three populations of molecules with affinity for antithrombin III. Furthermore, the affinity of each heparin species far antithrombin III appears to vary inversely with the size of the heparin chain, with some smaller oligosaccharides having greater affinity for antithrombin III than larger oligosaccharides.