Johnson C M, Helgeson S C
Department of Pediatrics, Mayo Clinic and Foundation, Rochester, MN 55905.
Biochem Biophys Res Commun. 1988 May 31;153(1):46-50. doi: 10.1016/s0006-291x(88)81187-2.
We have previously reported that cultured porcine cardiac valve endothelial cells released less fibronectin into the culture supernatant when compared to other porcine endothelial cells. In this report we compared the spectrum of glycoproteins synthesized by cardiac valve endothelial cells to glycoproteins synthesized by comparison endothelial cells derived from the ascending thoracic aorta. The cells were endogenously radiolabeled and extracted with detergent. Glycoproteins in the cell extracts were then isolated on wheat germ lectin-agarose and compared using autoradiography following polyacrylamide gel electrophoresis. Fibronectin was identified by immunoblotting with specific antibody. The results showed that the outstanding difference between the endothelial cell types was the virtual absence of fibronectin in the cardiac valve endothelial cell extract.
我们之前曾报道,与其他猪内皮细胞相比,培养的猪心脏瓣膜内皮细胞向培养上清液中释放的纤连蛋白较少。在本报告中,我们将心脏瓣膜内皮细胞合成的糖蛋白谱与源自胸主动脉升部的对照内皮细胞合成的糖蛋白谱进行了比较。细胞进行内源性放射性标记并用去污剂提取。然后将细胞提取物中的糖蛋白在麦胚凝集素-琼脂糖上分离,并在聚丙烯酰胺凝胶电泳后使用放射自显影进行比较。通过用特异性抗体进行免疫印迹鉴定纤连蛋白。结果表明,内皮细胞类型之间的显著差异是心脏瓣膜内皮细胞提取物中几乎不存在纤连蛋白。