Ototani N, Yosizawa Z
Carbohydr Res. 1978 Jul;64:143-53. doi: 10.1016/s0008-6215(00)83696-4.
Alkaline extraction of whale intestine, followed by pronase digestion and precipitation of heparin (omega-heparin) with dodecyltrimethylammonium chloride gave a supernatant fraction containing dermatan sulfate. Ethanol at 20% concentration precipitated dermatan sulfate from the supernatant fraction. The crude dermatan sulfate was further fractionated by ion-exchange column chromatography on Dowex-1 (C1- form), eluting stepwise with aqueous sodium chloride. The fractions eluted with 1.5M and 1.75M sodium chloride contained a typical dermatan sulfate. Chemical and enzymic studies of these preparations revealed that the sulfate groups were located solely at O-4 of the 2-acetamido-2-deoxy-D-galactose resides. L-Iduronic acid was assumed to be distributed uniformly in the backbone of the polysaccharide chain, with D-glucuronic acid being located in the linkage region to the protein core. A new method for determining the ratio of D-glucuronic to L-iduronic acid is also described.
用碱性溶液提取鲸肠，随后用链霉蛋白酶消化，并用十二烷基三甲基氯化铵沉淀肝素（ω-肝素），得到含有硫酸皮肤素的上清液部分。20%浓度的乙醇可从该上清液部分中沉淀出硫酸皮肤素。粗硫酸皮肤素通过在Dowex-1（Cl-型）上进行离子交换柱色谱进一步分级，用氯化钠水溶液逐步洗脱。用1.5M和1.75M氯化钠洗脱的部分含有典型的硫酸皮肤素。对这些制剂的化学和酶学研究表明，硫酸基团仅位于2-乙酰氨基-2-脱氧-D-半乳糖残基的O-4位。假定L-艾杜糖醛酸均匀分布在多糖链的主链中，而D-葡萄糖醛酸位于与蛋白质核心的连接区域。还描述了一种测定D-葡萄糖醛酸与L-艾杜糖醛酸比例的新方法。
