Børsum T
Institute for Surgical Research, University of Oslo, Norway.
Thromb Haemost. 1990 Apr 12;63(2):303-11.
Human endothelial cells isolated from umbilical cords were solubilized in Triton X-100 and examined by crossed immunoelectrophoresis using rabbit antiserum against endothelial cells. Endogenous labelling of the endothelial cell proteins with 14C-mannose followed by crossed immunoelectrophoresis and autoradiography revealed about 10 immunoprecipitates. Four of these endothelial cell glycoproteins were labelled by lactoperoxidase catalyzed iodination and thus were surface located. Three of the surface located glycoproteins showed reduced electrophoretic mobility after incubation of the endothelial cells with neuraminidase and were therefore sialoglycoproteins. Amphiphilicity of endothelial cell glycoproteins was studied by crossed hydrophobic interaction immunoelectrophoresis with phenyl-Sepharose in the intermediate gel. Amphiphilic proteins also show increasing electrophoretic migration velocity with decreasing concentration of Triton X-100 in the first dimension gels. Five of the endothelial cell glycoproteins were shown to be amphiphilic using these two techniques. Two monoclonal antibodies against the platelet glycoprotein complex IIb-IIIa and glycoprotein IIIa, respectively, reacted with the same precipitate of endothelial cells. When a polyclonal antibody against the platelet glycoprotein complex IIb-IIIa was incorporated into the intermediate gel the position of two endothelial cell precipitates were lowered. One of these was a sialoglycoprotein.
从脐带分离出的人内皮细胞用曲拉通X-100溶解，并使用抗内皮细胞兔抗血清通过交叉免疫电泳进行检测。用14C-甘露糖对内皮细胞蛋白质进行内源性标记，随后进行交叉免疫电泳和放射自显影，结果显示约有10种免疫沉淀物。其中4种内皮细胞糖蛋白通过乳过氧化物酶催化碘化进行标记，因此位于细胞表面。在用神经氨酸酶孵育内皮细胞后，3种位于细胞表面的糖蛋白显示出电泳迁移率降低，因此是唾液酸糖蛋白。通过在中间凝胶中用苯基琼脂糖进行交叉疏水相互作用免疫电泳研究内皮细胞糖蛋白的两亲性。两亲性蛋白质在第一维凝胶中也随着曲拉通X-100浓度降低而显示出电泳迁移速度增加。使用这两种技术显示5种内皮细胞糖蛋白具有两亲性。两种分别针对血小板糖蛋白复合物IIb-IIIa和糖蛋白IIIa的单克隆抗体与内皮细胞的同一沉淀物发生反应。当将抗血小板糖蛋白复合物IIb-IIIa的多克隆抗体加入中间凝胶时，两种内皮细胞沉淀物的位置降低。其中一种是唾液酸糖蛋白。
