Rice K G, Rottink M K, Linhardt R J
Division of Medicinal and Natural Products Chemistry, College of Pharmacy, University of Iowa, Iowa City 52242.
Biochem J. 1987 Jun 15;244(3):515-22. doi: 10.1042/bj2440515.
Heparin-derived oligosaccharides, prepared by using flavobacterial heparinase, having a high degree of heterogeneity (sequence variability) were resolved into sharp well-defined bands by using polyacrylamide gel electrophoresis (PAGE). The use of a stacking gel and a high-density-pore-gradient resolving gel was primarily responsible for the success of this separation. Low-Mr standards of known structure and having a degree of polymerization (dp) 2-6 were used to establish that the separation on gradient PAGE was primarily dependent on molecular size. High-Mr oligosaccharides (dp 8-20) were prepared using strong-anion-exchange h.p.l.c. and were used to help characterize the gradient PAGE separation. Kinetic profiles were obtained for the depolymerization of heparin and heparan sulphate with heparinase and heparitinase respectively. The utility of this approach in sequencing oligosaccharides derived from glycosaminoglycans is discussed.
利用黄杆菌属肝素酶制备的具有高度异质性（序列变异性）的肝素衍生寡糖，通过聚丙烯酰胺凝胶电泳（PAGE）被分离成清晰明确的条带。使用堆积凝胶和高密度孔梯度分辨凝胶是这种分离成功的主要原因。使用已知结构且聚合度（dp）为2 - 6的低分子量标准品来确定梯度PAGE上的分离主要取决于分子大小。使用强阴离子交换高效液相色谱法制备了高分子量寡糖（dp 8 - 20），并用于帮助表征梯度PAGE分离。分别获得了肝素酶和硫酸乙酰肝素酶使肝素和硫酸乙酰肝素解聚的动力学曲线。讨论了这种方法在对糖胺聚糖衍生的寡糖进行测序中的实用性。
