Moffat C F, McLean M W, Long W F, Williamson F B
Department of Molecular and Cell Biology, University of Aberdeen, Scotland.
Eur J Biochem. 1991 Dec 5;202(2):531-41. doi: 10.1111/j.1432-1033.1991.tb16405.x.
Saccharides produced by the action of heparinase II on native pig mucosal heparin (heparin IS), de-N-sulphated heparin (heparin IH), N-acetylheparin (heparin IA), de-N/O-sulphated heparin (heparin IVH), de-O-sulphated heparin (heparin IVS) and de-O-sulphated N-acetylheparin (heparin IVA) were analysed by reversed-phase HPLC using Spherisorb ODS2. Fractions obtained by gel filtration with Bio-Gel P-4 were similarly examined. Heparin IS gave delta UA-2S----GlcNS-6S (IS) as the major unsaturated disaccharide and lesser amounts of delta UA----GlcNS-6S (IIS), delta UA-2S----GlcNS (IIIS), delta UA----GlcNS (IVS), delta UA-2S----GlcNAc-6S (IA), delta UA----GlcNAc-6S (IIA), delta UA-2S----GlcNAc (IIIA) and delta UA----GlcNAc (IVA). Heparins IA, IVA and IVS gave as the predominant unsaturated disaccharide that corresponding to the major repeat structure of the polymer. These were respectively delta UA-2S----GlcNAc-6S (IA), delta UA-GlcNAc (IVA) and delta UA----GlcNS (IVS). Minor disaccharides from the heterogeneous structure in native pig heparin and from residual O-sulphates after the de-O-sulphating process were detected. Heparin IH was degraded more slowly than any of the N-substituted heparins. The predominant unsaturated disaccharide was IH, which was derived from the major repeating unit. In addition, disaccharides IIH, IIIH, IA, IIA and IVA were detected. Heparin IVH showed little degradation, the unsaturated disaccharide IVH not being detected after 24 h. Disaccharide IVA was obtained from the heterogeneous sequence in heparin IVH. Several higher oligosaccharides were identified in the gel-filtration fractions including saccharides from the linkage region (for heparin IS and IVA) and the anti-thrombin binding site (for heparin IS only). A tetrasaccharide and hexasaccharide, with the structures delta UA----GlcNAc----UA----GlcNAc and delta UA----GlcNAc----UA----GlcNAc----UA----GlcNAc, were present in the HPLC profiles of heparins IA and IVA.
使用Spherisorb ODS2通过反相高效液相色谱法分析了肝素酶II作用于天然猪黏膜肝素（肝素IS）、去N - 硫酸化肝素（肝素IH）、N - 乙酰肝素（肝素IA）、去N/O - 硫酸化肝素（肝素IVH）、去O - 硫酸化肝素（肝素IVS）和去O - 硫酸化N - 乙酰肝素（肝素IVA）所产生的糖类。对用Bio - Gel P - 4进行凝胶过滤得到的级分进行了类似的检测。肝素IS产生δUA - 2S----GlcNS - 6S（IS）作为主要的不饱和二糖，以及少量的δUA----GlcNS - 6S（IIS）、δUA - 2S----GlcNS（IIIS）、δUA----GlcNS（IVS）、δUA - 2S----GlcNAc - 6S（IA）、δUA----GlcNAc - 6S（IIA）、δUA - 2S----GlcNAc（IIIA）和δUA----GlcNAc（IVA）。肝素IA、IVA和IVS产生的主要不饱和二糖与聚合物的主要重复结构相对应。它们分别是δUA - 2S----GlcNAc - 6S（IA）、δUA - GlcNAc（IVA）和δUA----GlcNS（IVS）。检测到了天然猪肝素中异质结构产生的次要二糖以及去O - 硫酸化过程后残留的O - 硫酸盐产生的次要二糖。肝素IH的降解比任何一种N - 取代肝素都要慢。主要的不饱和二糖是IH，它源自主要的重复单元。此外，还检测到了二糖IIH、IIIH以及IA、IIA和IVA。肝素IVH几乎没有降解，24小时后未检测到不饱和二糖IVH。二糖IVA是从肝素IVH的异质序列中获得的。在凝胶过滤级分中鉴定出了几种更高的寡糖，包括来自连接区域（针对肝素IS和IVA）和抗凝血酶结合位点（仅针对肝素IS）的糖类。在肝素IA和IVA的高效液相色谱图谱中存在结构为δUA----GlcNAc----UA----GlcNAc的四糖和结构为δUA----GlcNAc----UA----GlcNAc----UA----GlcNAc的六糖。
