The Interaction of Activated Protein C and Thrombin with the Plasminogen Activator Inhibitor Released from Human Endothelial Cells
The Interaction of Activated Protein C and Thrombin with the Plasminogen Activator Inhibitor Released from Human Endothelial Cells
复制标题
活化的蛋白 C 和凝血酶与人内皮细胞释放的纤溶酶原激活剂抑制剂的相互作用
DOI：
10.1055/s-0038-1651089
复制
发表时间：
1987
影响因子：
6.7
通讯作者：
R. Bertina
中科院分区：
文献类型：
--
作者：
N. D. de Fouw;V. V. van Hinsbergh;Y. D. de Jong;F. Haverkate;R. Bertina
以两种方式研究了人活化蛋白C（APC）和凝血酶对从培养的人脐内皮细胞释放的纤溶酶原激活物抑制剂（派-1）的影响，所述培养的人脐内皮细胞在含35 S-甲硫氨酸的无血清培养基中生长：1）用酰胺分解试验测量派-1活性，和2）用抗PAI-1 IgG免疫沉淀培养基，抗蛋白C IgG或抗凝血酶IgG，然后进行SDS-PAGE和放射自显影。向内皮细胞条件培养基中加入APC或凝血酶导致派-1活性的时间和浓度依赖性损失，并导致派-1从46 kD降解成42 kD产物。在存在细胞的情况下将培养基与APC孵育后，发现95 kD的额外条带，其可以与抗PAI-1 IgG和抗蛋白C IgG免疫沉淀，表明在降解发生之前形成APC-PAI-1复合物。在内皮细胞存在下，培养基与凝血酶孵育后，未检测到复合物。阻断APC和凝血酶的活性位点可防止APC-PAI-1复合物的形成和派-1的失活和降解。去除培养液中的活性派-1后，APC和凝血酶均不能降解失活的派-1，说明APC和凝血酶均能与活性派-1发生反应，导致派-1的失活和降解。
Summary The effects of human activated protein C (APC) and thrombin on plasminogen activator inhibitor (PAI-1) released from cultured human umbilical endothelial cells, grown in serum-free 35S-methionine containing medium, were studied in two ways: 1) measurement of PAI-1 activity with an amidolytic assay, and 2) immunoprécipitation of the medium with anti-PAI-1 IgG, antiprotein C IgG or anti-thrombin IgG followed by SDS-PAGE and autoradiography. Addition of APC or thrombin to the endothelial cell conditioned medium results in a time and concentration dependent loss of PAI-1 activity and in the degradation of PAI-1 from 46 kD into a 42 kD product. After incubation of the medium with APC in the presence of cells, an additional band of 95 kD was found, which could be immunoprecipitated with both anti-PAI-1 IgG and anti-protein C IgG, indicating the formation of an APC-PAI-1 complex before degradation occurs. No complex could be detected after incubation of the medium with thrombin in the presence of endothelial cells. Blocking the active sites of APC and thrombin prevented both the formation of APC-PAI-1 complexes and the inactivation and degradation of PAI-1. After removal of the active PAI-1 from the medium, no degradation of the inactive PAI-1 by APC or thrombin could be found. It is concluded that both APC and thrombin react with the active PAI-1, resulting in inactivation and degradation of PAI-1.
登录
查看更多内容
DOI：
10.1016/s0021-9258(17)42691-3
发表时间：
1984-12
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
J. V. van Mourik;D. Lawrence;D. Loskutoff
通讯作者：
J. V. van Mourik;D. Lawrence;D. Loskutoff
内皮细胞产生纤溶酶原激活剂的潜在抑制剂，可以被变性剂激活。
DOI：
--
发表时间：
1985
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Hekman,CM;Loskutoff,DJ
通讯作者：
Loskutoff,DJ
DOI：
10.1073/pnas.80.10.2956
发表时间：
1983-01-01
期刊：
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES
影响因子：
--
作者：
LOSKUTOFF, DJ;VANMOURIK, JA;LAWRENCE, D
通讯作者：
LAWRENCE, D
影响因子：
2.9
作者：
BLAKE, MS;JOHNSTON, KH;GOTSCHLICH, EC
通讯作者：
GOTSCHLICH, EC