Linhardt R J, Rice K G, Kim Y S, Lohse D L, Wang H M, Loganathan D
Division of Medicinal and Natural Products Chemistry, College of Pharmacy, University of Iowa, Iowa City 52242.
Biochem J. 1988 Sep 15;254(3):781-7. doi: 10.1042/bj2540781.
A new method of determining the oligosaccharide composition of commercial glycosaminoglycan heparin is described in which heparin was first depolymerized using heparin lyase (EC 4.2.2.7), and then analysed by a single h.p.l.c. step. All 20 of the porcine and bovine heparins examined were found to contain a small number of major oligosaccharide components, which on average comprised 86% of their mass. The five most abundant oligosaccharides have defined chemical structures. Although the relative abundance of oligosaccharides varied, the heparins examined were surprisingly similar. Porcine, bovine, low-Mr, and high and low antithrombin III (ATIII)-affinity heparins, however, each had distinctly different proportions of these major oligosaccharide components. The concentrations of one of these five oligosaccharides, containing a portion of the ATIII binding site, correlated with the anticoagulant activity of the ATIII-affinity-fractionated porcine-mucosal heparins from which it was derived. An additional oligosaccharide of undetermined structure was found in significant quantities in both bovine heparin and high ATIII-affinity porcine-mucosal heparin. The correlation between oligosaccharide concentration and anticoagulant activity suggests that the oligosaccharide is derived from a structural variant of the ATIII-binding site. Finally, for the heparins examined chondroitin/dermatan sulphate formed 0.6-7.4% of their mass.
本文描述了一种测定商业糖胺聚糖肝素寡糖组成的新方法，该方法首先使用肝素裂解酶（EC 4.2.2.7）使肝素解聚，然后通过单一的高效液相色谱步骤进行分析。在所检测的所有20种猪源和牛源肝素中，均发现含有少量主要寡糖成分，这些成分平均占其质量的86%。五种最丰富的寡糖具有明确的化学结构。尽管寡糖的相对丰度有所不同，但所检测的肝素出人意料地相似。然而，猪源、牛源、低分子量以及高和低抗凝血酶III（ATIII）亲和力的肝素，各自这些主要寡糖成分的比例明显不同。这五种寡糖中的一种含有部分ATIII结合位点，其浓度与从中衍生出的ATIII亲和力分级猪黏膜肝素的抗凝血活性相关。在牛源肝素和高ATIII亲和力猪黏膜肝素中均大量发现了一种结构未确定的额外寡糖。寡糖浓度与抗凝血活性之间的相关性表明，该寡糖源自ATIII结合位点的结构变体。最后，在所检测的肝素中，硫酸软骨素/硫酸皮肤素占其质量的0.6 - 7.4%。
