Miyamoto Y, Izumi M, Ishizaka S, Hayashi M
Institute of Biological Sciences, University of Tsukuba, Ibaraki, Japan.
Cell Struct Funct. 1989 Apr;14(2):151-62. doi: 10.1247/csf.14.151.
We have investigated the adsorption of cell-spreading activity in human serum onto polystyrene plates after treatment of the serum with sodium dodecyl sulfate (SDS). Vitronectin in human serum was remarkably adsorbed onto the plate after boiling the serum with 0.1% SDS for 5 min. SDS was effective over the concentration range from 0.05 to 0.25%. Increase of the vitronectin adsorption was accompanied by an increase of cell spreading on the plates. The cell-spreading activity in SDS-treated serum was impeded by anti-vitronectin antibody but not by anti-fibronectin antibody. After treatment with SDS, fibronectin-depleted serum could induce cell spreading but vitronectin-depleted serum could not. These results indicate that vitronectin alone was the cell-spreading factor in SDS-treated human serum. However, SDS-treated pure vitronectin itself did not retain the cell-spreading activity. The activity was recovered when bovine serum albumin was added to pure vitronectin before or after boiling with 0.1% SDS. Therefore, vitronectin adsorbed from SDS-treated serum might retain the cell-spreading activity with the aid of serum protein. Treatment of serum with SDS provides an easy, specific, and efficient method of coating polystyrene plates with vitronectin.
我们研究了用十二烷基硫酸钠（SDS）处理人血清后，血清中细胞铺展活性在聚苯乙烯平板上的吸附情况。用0.1% SDS将血清煮沸5分钟后，人血清中的玻连蛋白显著吸附到平板上。SDS在0.05%至0.25%的浓度范围内有效。玻连蛋白吸附的增加伴随着平板上细胞铺展的增加。SDS处理血清中的细胞铺展活性受到抗玻连蛋白抗体的抑制，但不受抗纤连蛋白抗体的抑制。用SDS处理后，去除纤连蛋白的血清仍可诱导细胞铺展，但去除玻连蛋白的血清则不能。这些结果表明，在SDS处理的人血清中，单独的玻连蛋白是细胞铺展因子。然而，SDS处理的纯玻连蛋白本身并不保留细胞铺展活性。在用0.1% SDS煮沸之前或之后向纯玻连蛋白中添加牛血清白蛋白时，活性得以恢复。因此，从SDS处理的血清中吸附的玻连蛋白可能借助血清蛋白保留细胞铺展活性。用SDS处理血清为用玻连蛋白包被聚苯乙烯平板提供了一种简便、特异且高效的方法。
