Analysis of Functional Domains of Tissue Factor Pathway Inhibitor

组织因子途径抑制剂的功能域分析

• 批准号: 09680606
• 负责人: KATO Hisao
• 金额: $ 1.73万
• 依托单位: National Cardiovascular Center Research Institute
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09680606/
• 关键词: protease inhibitor; blood coagulation; thrombosis; arteriosclerosis; heparin; 抗血栓; protease inhibitor; blood coagulation; thrombosis; arteriosclerosis; heparin; 抗血栓

(1) Degradation of TFPI by thrombinIt has been shown that TFPI exists in plasma as a free-form and liporptoein-associated forms and also as truncated forms which were caused by proteolysis. We investigated the actions of various proteases on recombinant TFPI and found that thrombin inactivated the functions of TFPI by secific cleavage of three peptide bonds of TFPI(2) Interaction of TFPI with heparinWe analyzed the interaction of TEPI with heparin by using modified heparin in which N-sulafate, 2-0-sulfate or 6-0-sulfate residues were specifically removed.and by using shorter chains of polysaccharide. The results indicate that all sulfate residues are essential for the interaction and that sugar chain with 14 units has almost the same ability to interact with TFPI as heparin. We also investigated the interaction of two heparin binding sites of TFPI with heparin. We found that Arg257 and Arg259 were essential for the interaction with heparin by using synthetic peptides with C-terminal basic part. We isolated recombinant K3 domain by Baculo virus system and examined the interaction with heparin.

（1）凝血酶对TFPI的降解已表明，TFPI在血浆中存在作为自由形式和脂肪蛋白相关的形式，也是由蛋白水解引起的截断形式。我们研究了各种蛋白酶对重组TFPI的作用，发现凝血酶通过将TFPI的三种肽键（2）与肝素相互作用的TFPI（2）相互作用的三种肽键（2）与肝素与肝素的相互作用分析N-Sulate-Sulate-Sulate-Sulate-Sulate-Sulate-Sulate-S-0-0-0-0-0-0-0-0-0-0-0--0--专门删除。使用较短的多糖链。结果表明，所有硫酸盐残基对于这种相互作用都是必不可少的，并且具有14个单位的糖链与TFPI相互作用的能力与肝素相同。我们还研究了TFPI的两个肝素结合位点与肝素的相互作用。我们发现ARG257和ARG259通过将合成肽与C末端碱性部分使用，对于与肝素相互作用至关重要。我们通过Baculo病毒系统分离了重组K3结构域，并检查了与肝素的相互作用。