人黑色素瘤细胞分泌的组织纤溶酶原激活剂(t-PA)的纯化

 本文报道了培养的人黑色素瘤细胞分泌的组织纤溶酶原激活剂(t-PA)的纯化方法。Bowes株人黑色素瘤细胞的分泌产物,经CM-Sephadex C--50层析,赖氨酸-Sepharose 4B,苯甲眯-sepharose 4B亲和层析后,即可得到纯化470倍的蛋白纯品。样品经聚丙烯酰胺凝胶电泳鉴定为均一单带,测得其分子量约为72kD。纯化的t-PA与尿激酶相比较,发现前者有更高亲和纤维蛋白的能力。

Purification of Tissue Plasminogen Activator Secreted by Cultured Human Melanoma Cell

Tissue-type plasminogen activator secreted by a cultured human melanoma cell line was purified. The purification procedure consisted of chromatography on CM -Sephadex C-50, Lysine-Sepharose 4B and Benzamidine-Sepharose 4B affinity chromatography in the presence of 0.01% Tween 80. The purified protein was obtained from the culture medium, with purification factor of 470. The purified t-PA was proven to be homogeneous by polyacrylamide gel electrophoresis, with molecular weight about 72kD. The purified t-PA has a higher affinity for fibrin than urokina-se. In addition, the melanoma plasminogen activator appeared to be immunologically different from urokinase.