COMPARATIVE ELECTROPHORETIC ANALYSIS OF HUMAN AND PORCINE PLASMINOGEN ACTIVATORS IN SDS-POLYACRYLAMIDE GELS CONTAINING PLASMINOGEN AND CASEIN
COMPARATIVE ELECTROPHORETIC ANALYSIS OF HUMAN AND PORCINE PLASMINOGEN ACTIVATORS IN SDS-POLYACRYLAMIDE GELS CONTAINING PLASMINOGEN AND CASEIN
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DOI：
10.1016/0167-4838(83)90172-3
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发表时间：
1983-01-01
期刊：
影响因子：
--
通讯作者：
SHAW, ST
中科院分区：
文献类型：
--
作者：
ROCHE, PC;CAMPEAU, JD;SHAW, ST
在含有纤溶酶原和酪蛋白的SDS[十二烷基硫酸钠]-聚丙烯酰胺梯度平板凝胶中，对猪心脏和人子宫、血浆和黑色素瘤细胞中的纤溶酶原激活物进行电泳分析。去除SDS，在缓冲液中孵育，并用考马斯亮蓝染色后，在聚丙烯酰胺凝胶中实现激活剂活性条带的直接可视化。从猪心提取的组织激活剂显示出72，000的MW，其迁移类似于人黑素瘤细胞分泌的激活剂和人子宫提取物中存在的1种激活剂组分。黑色素瘤细胞激活剂抗血清的免疫吸附实验表明，这些72 kDa的激活剂都是相关的免疫。人子宫还含有分子量为55，000的第二激活剂组分，其迁移类似于尿激酶的较高分子量组分，并与尿激酶抗血清发生交叉反应。72-kDa子宫激活剂组分显然代表组织激活剂，55-kDa组分代表尿激酶样激活剂。来自静脉闭塞血浆的优球蛋白溶液在MW范围85，000 - 96，000内显示多条纤溶酶活性带。还存在两种活化剂组分，一种MW为72，000，另一种MW为62，000。72 kDa优球蛋白激活剂被MCA抗血清吸附，该组分显然代表血管激活剂。62，000激活剂也具有弱的纤溶酶原非依赖性酪蛋白溶解活性，并且不受黑素瘤细胞激活剂或尿激酶抗血清的影响。目前还不能就其身份作出结论。
Electrophoretic analysis of plasminogen activators from pig heart and human uterus, plasma and melanoma cells was performed in SDS[sodium dodecyl sulfate]-polyacrylamide gradient slab gels containing plasminogen and casein. Direct visualization of activator activity bands in polyacrylamide gels was achieved after removal of SDS, incubation in buffer, and staining with Coomassie brilliant blue. Tissue activator extracted from pig hearts displayed a MW of 72,000 migrated similarly to activator secreted by human melanoma cells and to 1 activator component present in extracts of human uterus. Immunoadsorption experiments with melanoma cell activator antiserum indicated that these 72-kDa activators are all related immunologically. Human uterus also contained a 2nd activator component with a MW 55,000, which migrated similarly to a higher MW component of urokinase and cross-reacted with urokinase antiserum. The 72-kDa uterine activator component apparently represents a tissue activator and the 55-kDa component represents a urokinase-like activator. A euglobulin solution from venous occlusion plasma displayed multiple bands of plasmin activity in the MW range 85,000-96,000. Two activator components were also present, one of MW 72,000 and another of MW 62,000. The 72-kDa euglobulin activator was adsorbed by MCA antiserum, and this component apparently represents vascular activator. The 62,000 activator also had weak plasminogen-independent caseinolytic activity and was not affected by either melanoma cell activator or urokinase antisera. Conclusions concerning its identity cannot be made at this time.