Distribution of proteoglycans antigenically related to corneal keratan sulfate proteoglycan.
Distribution of proteoglycans antigenically related to corneal keratan sulfate proteoglycan.
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DOI：
10.1016/s0021-9258(18)60856-7
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发表时间：
1987-08
期刊：
影响因子：
--
通讯作者：
J. Funderburgh;B. Caterson;G. Conrad
中科院分区：
文献类型：
--
作者：
J. Funderburgh;B. Caterson;G. Conrad
用三种与硫酸角膜角质素蛋白多糖反应的抗体检测了11例牛和13例鸡胚组织中的抗原相关分子。两种单克隆抗体识别硫酸角质素链上的硫酸化表位，一种多克隆抗体结合角膜硫酸角质素蛋白聚糖核心蛋白上的抗原位点。竞争性免疫分析检测到核心蛋白和硫酸角质素抗原盐酸胍提取物的大部分组织。大多数牛组织的硫酸角质素抗原仅部分用盐酸胍提取，但其余的可通过溴化氰处理的胍提取残留物溶解。硫酸角质素和核心蛋白抗原与纯化的硫酸角质素蛋白聚糖在离子交换高效液相色谱（HPLC）上共洗脱。内切-β-半乳糖苷酶消化HPLC纯化的硫酸角质素抗原消除了酶联免疫吸附测定中单克隆抗硫酸角质素抗体的结合。除脑和软骨外，所有11种牛组织提取物均能同时与抗硫酸角质素单克隆抗体和抗核心蛋白多克隆抗体结合。结合对硫酸角质素竞争和内切β-半乳糖苷酶消化敏感。这些结果表明，广泛存在的蛋白聚糖或硫酸化糖蛋白轴承硫酸角质素样碳水化合物和核心蛋白类似的角膜硫酸角质素蛋白聚糖。
Three antibodies reacting with corneal keratan sulfate proteoglycan were used to detect antigenically related molecules in 11 bovine and 13 embryonic chick tissues. Two monoclonal antibodies recognized sulfated epitopes on the keratan sulfate chain and a polyclonal antibody bound antigenic sites on the core protein of corneal keratan sulfate proteoglycan. Competitive immunoassay detected core protein and keratan sulfate antigens in guanidine HCl extracts of most tissues. Keratan sulfate antigens of most bovine tissues were only partially extracted with guanidine HCl, but the remainder could be solubilized by CNBr treatment of the guanidine-extracted residue. Keratan sulfate and core protein antigens co-eluted with purified corneal keratan sulfate proteoglycan on ion exchange high-performance liquid chromatography (HPLC). Endo-beta-galactosidase digestion of the HPLC-purified keratan sulfate antigens eliminated the binding of monoclonal anti-keratan sulfate antibodies in enzyme-linked immunosorbent assay. Extracts of all 11 bovine tissues, except those from brain and cartilage, could bind both anti-keratan sulfate monoclonal antibodies and anti-core protein polyclonal antibody simultaneously. Binding was sensitive to competition with keratan sulfate and to digestion with endo-beta-galactosidase. These results suggest widespread occurrence of a proteoglycan or sulfated glycoprotein bearing keratan sulfate-like carbohydrate and a core protein resembling that of corneal keratan sulfate proteoglycan.