Isolation of tissue-type plasminogen activator-inhibitor complexes from human plasma. Evidence for a rapid plasminogen activator inhibitor.
Isolation of tissue-type plasminogen activator-inhibitor complexes from human plasma. Evidence for a rapid plasminogen activator inhibitor.
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从人血浆中分离组织型纤溶酶原激活剂-抑制剂复合物。
DOI：
10.1016/0304-4165(84)90040-0
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发表时间：
1984
期刊：
影响因子：
--
通讯作者：
M. Philips
中科院分区：
文献类型：
--
作者：
S. Thorsen;M. Philips
通过 SDS-聚丙烯酰胺凝胶电泳和酶谱分析纤溶酶原激活剂-抑制剂复合物。该复合物在纤维蛋白指示剂凝胶中表现为纤维蛋白溶解活性带。通过抗 t-PA-Sepharose 上的免疫吸附，然后在 Sephadex G-150 上进行凝胶过滤，从血浆中纯化出与来自培养的人内皮细胞的 t-PA 抑制剂复合物 (Mr95 000–135 000) 共迁移的高分子量 t-PA 形式。通过纤维蛋白平板法测定时，高分子量 t-PA 形式没有纤溶活性。当用 1.5 M NH4OH/39 mM SDS 处理时，它转化为与 t-PA (Mr72 000) 具有相同电泳迁移率的形式。这些观察结果表明，血浆高分子量 t-PA 形式是一种酶抑制剂复合物。该复合物与许多已知的血浆丝氨酸蛋白酶抑制剂没有表现出免疫交叉反应性。 t-PA 和 u-PA 均与血浆中 p 摩尔浓度的抑制剂快速形成复合物。对氨基苯甲脒阻断了反应，表明激活剂的活性中心确实参与了复合物的形成。血浆抑制剂与t-PA的复合物和高分子量t-PA具有相同的电泳迁移率。血浆中的快速纤溶酶原激活剂抑制剂与培养的人内皮细胞中的纤溶酶原激活剂抑制剂表现出显着的相似性。除了上述高分子量 t-PA 形式外，还从血浆中分离出了其他三种 t-PA 形式。我们的结果表明，它们代表游离的 t-PA 和分别与 C1-酯酶抑制剂和 α2-抗纤溶酶复合的 t-PA。
Plasminogen activator-inhibitor complexes were analyzed by SDS-polyacrylamide gel electrophoresis and enzymography. The complexes appeared as fibrinolytically active bands in the fibrin-indicator gel. A high-molecular-weight t-PA form comigrating with a t-PA-inhibitor complex (Mr95 000–135 000) from cultured human endothelial cells was purified from plasma by immunoadsorption on anti-t-PA-Sepharose followed by gel filtration on Sephadex G-150. The high-molecular-weight t-PA form was fibrinolytically inactive when assayed by the fibrin-plate method. It was converted to a form with the same electrophoretic mobility as t-PA (Mr72 000) when treated with 1.5 M NH4OH/39 mM SDS. These observations suggested that the plasma high-molecular-weight t-PA form was an enzyme-inhibitor complex. The complex did not show immunological cross-reactivity with a number of known plasma serine proteinase inhibitors. Both t-PA and u-PA rapidly formed complexes with an inhibitor which was present in plasma in pmolar concentrations.p-Aminobenzamidine blocked the reaction, indicating that the active center of the activator was indeed implicated in complex formation. The complex between the plasma inhibitor and t-PA and the high-molecular-weight t-PA had the same electrophoretic mobilities. The rapid plasminogen activator inhibitor in plasma showed remarkable similarity to a plasminogen activator inhibitor from cultured human endothelial cells. In addition to the high-molecular-weight t-PA form described above, three other t-PA forms were isolated from plasma. Our results indicated that they represented free t-PA and t-PA in complex with respectively C1-esterase inhibitor and α2-antiplasmin.