High-speed gel-permeation chromatography of glycosaminoglycans: its application to the analysis of heparan sulfate of embryonic carcinoma and its degradation products by tumor cell-derived heparanase.
High-speed gel-permeation chromatography of glycosaminoglycans: its application to the analysis of heparan sulfate of embryonic carcinoma and its degradation products by tumor cell-derived heparanase.
复制标题
糖胺聚糖高速凝胶渗透色谱：其在分析胚胎癌硫酸乙酰肝素及其肿瘤细胞源性乙酰肝素酶降解产物中的应用。
DOI：
10.1016/0003-2697(83)90616-4
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发表时间：
1983
影响因子：
2.9
通讯作者：
Nicolson,GL
中科院分区：
文献类型：
--
作者：
Irimura,T;Nakajima,M;DiFerrante,N;Nicolson,GL
建立了一种快速凝胶渗透色谱系统，该系统采用两个0.7 × 75 cm不锈钢柱，柱内装有Fractogel（Toyopolitan）TSK HW-55（S）。应用糖胺聚糖，用0.2 M氯化钠溶液洗脱，并通过210 nm处的紫外吸收或放射性监测。在55°C和1.0 ml/min的流速下获得聚糖的最佳分离度。分子量（Mr）范围为600- 60，000的酸性和中性聚糖在45分钟内洗脱。使用标准糖胺聚糖、甲壳素低聚糖和猪甲状腺球蛋白糖肽发现保留时间和分子量之间呈线性关系。该系统用于分析由PYS-2胚胎癌细胞合成的硫酸乙酰肝素和通过将其与从转移性B16黑色素瘤细胞提取的糖苷酶孵育产生的降解产物。结果表明，B16黑色素瘤细胞含有至少两种不同的硫酸乙酰肝素降解活性，其中之一似乎是糖苷内切酶。
A high-speed gel-permeation chromatographic system for analyzing glycosaminoglycans which uses two 0.7 × 75-cm stainless-steel columns containing Fractogel (Toyopearl) TSK HW-55(S), was developed. Glycosaminoglycans were applied and eluted with a 0.2 m sodium chloride solution and monitored by ultraviolet absorption at 210 nm or radioactivity. The best resolution of glycans was obtained at 55°C at a flow rate of 1.0 ml/min. Acidic and neutral glycans in the molecular weight (Mr) range 600–60,000 eluted within 45 min. A linear relationship was found between retention time and molecular weight using standard glycosaminoglycans, chitin oligosaccharides, and a porcine thyroglobulin glycoprotide. This system was used to analyze the heparan sulfate synthesized by PYS-2 embryonic carcinoma cells and the degradation products produced by incubating it with extracted glycosidases from metastatic B16 melanoma cells. The results indicated that B16 melanoma cells contain at least two different heparan sulfate degradative activities, one of which appears to be an endoglycosidase.