Demonstration of a keratan sulfate-containing proteoglycan in atherosclerotic aorta.
Demonstration of a keratan sulfate-containing proteoglycan in atherosclerotic aorta.
复制标题
动脉粥样硬化主动脉中含有硫酸角质素的蛋白多糖的演示。
DOI：
10.1161/01.atv.12.1.83
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发表时间：
1992
期刊：
影响因子：
--
通讯作者：
Caterson,B
中科院分区：
文献类型：
--
作者：
Robbins,RA;Wagner,WD;Register,TC;Caterson,B
蛋白聚糖分离从任何严重正常或动脉粥样硬化鸽腹后提取4 M盐酸胍和离子交换和尺寸排阻色谱纯化。来自正常和动脉粥样硬化组织的小尺寸蛋白聚糖（Kav 0.4，在Sepharose CL-4 B上）主要包含具有220-330 kd的完整分子大小的硫酸皮肤素蛋白聚糖和45 kd的核心蛋白。除了硫酸皮肤素蛋白聚糖外，该制剂还含有可被单克隆抗体（MAb）5-D-4识别的蛋白聚糖，表明存在角膜和软骨硫酸角质素共有的硫酸化聚-N-乙酰乳糖胺序列。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳显示60-150 kd的多分散蛋白多糖，被单克隆抗体5-D-4识别。与正常动脉相比，动脉粥样硬化的MAb 5-D-4的免疫反应性显著增加。在内皮-β-D-半乳糖苷酶处理来自动脉粥样硬化主动脉的蛋白多糖后，通过Western印迹分析和酶联免疫吸附试验观察到MAb 5-D-4反应性降低，表明该材料是硫酸角质素。完整蛋白聚糖的内切-β-D-半乳糖苷酶处理产生28和38 kd的核心蛋白。这些研究表明，存在一个或多个硫酸角质素蛋白聚糖在大体正常和动脉粥样硬化动脉。免疫化学数据表明，硫酸角质素蛋白聚糖的硫酸化可能是动脉粥样硬化的主动脉。
Proteoglycans were isolated from either grossly normal or atherosclerotic pigeon aortas after extraction with 4 M guanidine hydrochloride and purification by ion-exchange and size-exclusion chromatography. The small-size proteoglycans (Kav 0.4, on Sepharose CL-4B) from both normal and atherosclerotic tissue contained primarily a dermatan sulfate proteoglycan with an intact molecular size of 220-330 kd and a 45-kd core protein. In addition to the dermatan sulfate proteoglycan, the preparation contained a proteoglycan recognized by monoclonal antibody (MAb) 5-D-4, indicating the presence of sulfated poly-N-acetyllactosamine sequences common to corneal and cartilage keratan sulfate. Electrophoresis on sodium dodecyl sulfate-polyacrylamide gel revealed a polydisperse proteoglycan of 60-150 kd that was recognized by MAb 5-D-4. Significantly greater immunoreactivity with MAb 5-D-4 was observed for atherosclerotic compared with normal artery. After endo-beta-D-galactosidase treatment of the proteoglycan from atherosclerotic aorta, diminished MAb 5-D-4 reactivity observed by both Western blot analysis and enzyme-linked immunosorbent assay demonstrated that the material was keratan sulfate. Endo-beta-D-galactosidase treatment of the intact proteoglycan generated core proteins of 28 and 38 kd. These studies suggest the presence of one or more keratan sulfate proteoglycans in grossly normal and atherosclerotic arteries. Immunochemical data suggest that sulfation of the keratan sulfate proteoglycan may be greater in atherosclerotic aorta.