Epitope mapping of monoclonal antibodies directed to PAI-1 using PAI-1/PAI-2 chimera and PAI-1-derived synthetic peptides
Epitope mapping of monoclonal antibodies directed to PAI-1 using PAI-1/PAI-2 chimera and PAI-1-derived synthetic peptides
复制标题
DOI：
10.1016/s0049-3848(99)00238-8
复制
发表时间：
2000-04-01
影响因子：
7.5
通讯作者：
Magdolen, V
中科院分区：
文献类型：
--
作者：
Muehlenweg, B;Guthaus, E;Magdolen, V
纤溶酶原激活物抑制剂-1（Plasminogen activator inhibitor type-1，PAI-1）是纤溶系统的关键调节蛋白，参与多种生理和病理生理过程。由纤溶酶原激活物抑制剂1型和另一种纤溶酶原激活物抑制剂纤溶酶原激活物抑制剂2型组成的嵌合体。与相应野生型蛋白质的不同部分，并合成纤溶酶原激活物抑制剂1型衍生的20-mer和10-mer线性肽。14株单克隆抗体中有9株识别纤溶酶原激活物抑制剂-1的76-188位氨基酸之间的抗原决定簇，该抗原决定簇包括与玻连蛋白、肝素的结合位点和部分纤维蛋白结合区。其中9株单克隆抗体与纤溶酶原激活物抑制剂-1的四联体突变体反应（N152 H，K156 T，Q321 L，M356 I），7例检测到纤溶酶原激活物抑制剂1型缺失突变（Delta F111-H114）。其余五种单克隆抗体中的两种分别识别位于氨基酸209-227和氨基酸352-371之间的表位，而其他三种抗体仅与野生型纤溶酶原激活物抑制剂1型反应。另外的实验显示，14种mAb中的两种中和了纤溶酶原激活物抑制剂1型，一种单克隆抗体增加了纤溶酶原激活物抑制剂1型对尿激酶型纤溶酶原激活物的活性，尿激酶型纤溶酶原激活物是其靶蛋白酶之一，（C）2000 Elsevier Science Ltd.版权所有。
Plasminogen activator inhibitor type-1 is a key regulatory protein of the fibrinolytic system that is involved ill a variety of physiological and pathophysiological processes, A panel of 14 monoclonal antibodies directed against plasminogen activator inhibitor type-1 was analyzed regarding epitope specificity on plasminogen activator inhibitor type-1, For this purpose, chimera consisting of plasminogen activator inhibitor type-1 and another plasminogen activator inhibitor, plasminogen activator inhibitor type-2. with different portions of the respective wild-type proteins, were generated and plasminogen activator inhibitor type-1-derived 20-mer and 10-mer linear peptides were synthesized. Nine of the 14 monoclonal antibodies recognized an epitope located in the region between amino acid 76-188 of plasminogen activator inhibitor type-1, which encompasses the binding sites for vitronectin, heparin, and part of the fibrin binding region, Of these nine monoclonal antibodies, sis reacted with a quadruple plasminogen activator inhibitor type-1 mutant (N152H, K156T, Q321L, M356I), and seven detected a plasminogen activator inhibitor type-1 deletion mutant (Delta F111-H114). Two of the remaining five monoclonal antibodies recognized epitopes located between amino acid 209-227 and amino acid 352-371, respectively, while the other three antibodies reacted with wildtype plasminogen activator inhibitor type-1, only. Additional experiments revealed that two of the 14 mAbs neutralized and one monoclonal antibodies increased plasminogen activator inhibitor type-1 activity toward urokinase-type plasminogen activator, one of its target proteases, (C) 2000 Elsevier Science Ltd. All rights reserved.