Calatroni A, Cetta G, Pallavicini G
Ital J Biochem. 1977 Mar-Apr;26(2):122-32.
Glycosaminoglycan-protein complexes were extracted from bovine duodenal mucosa with distilled water, resulting in solubilization of a fraction of the total proteoglycan of the tissue. The extracted material was purified by anion exchange chromatography on DEAE-Sephadex A-25, and then characterized by chemical analysis and by fractionation on Dowex 1. By using these procedures, two major fractions were identified, which were eluted from Dowex with 1.0-1.25 M NaC1 and with 1.5-1.75 M NaC1 respectively. Analyses showed that both fractions were mainly composed of glucosamine-containing, hyaluronidase-resistant polysaccharides, which were identified by their N-sulphate: D-glucosamine and total sulphate: D-glucosamine ratios as heparan-sulphate in the less acidic fraction, and as heparin in the more acidic fraction. Dermatan sulphate molecules were also present in both preparations, with an approximate ratio 1:3 to the glucosamine-containing polysaccharides. Solubility behaviour of the complexes formed by the isolated polyanionic molecules with cetylpyridinium chloride was strongly modified by papain digestion of the duodenal material. This reduction of molecular size of papain treatment suggests that the molecules extracted with water from duodenal mucosa are complex proteoglycans, perhaps in the native state.
用蒸馏水从牛十二指肠黏膜中提取糖胺聚糖 - 蛋白质复合物,结果使组织中一部分总蛋白聚糖溶解。提取的物质通过在DEAE - Sephadex A - 25上进行阴离子交换色谱法纯化,然后通过化学分析和在Dowex 1上分级进行表征。通过使用这些程序,鉴定出两个主要级分,它们分别用1.0 - 1.25M NaCl和1.5 - 1.75M NaCl从Dowex上洗脱。分析表明,两个级分主要由含葡萄糖胺的、对透明质酸酶有抗性的多糖组成,通过它们的N - 硫酸盐:D - 葡萄糖胺和总硫酸盐:D - 葡萄糖胺比率,在酸性较弱的级分中鉴定为硫酸乙酰肝素,在酸性较强的级分中鉴定为肝素。硫酸皮肤素分子也存在于两种制剂中,与含葡萄糖胺的多糖的比例约为1:3。十二指肠物质经木瓜蛋白酶消化后,分离出的聚阴离子分子与十六烷基吡啶氯化物形成的复合物的溶解性行为发生了强烈改变。木瓜蛋白酶处理导致分子大小减小,这表明从十二指肠黏膜用水提取的分子是复合蛋白聚糖,可能处于天然状态。