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

(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的降解研究表明，TFPI在血浆中以游离形式和脂蛋白结合形式存在，也以蛋白水解引起的截短形式存在。我们研究了各种蛋白酶对重组TFPI的作用，发现凝血酶通过特异性地切割TFPI的三个肽键而使TFPI失活。（2）TFPI与肝素的相互作用我们用特异性地去除N-硫酸酯、2-O-硫酸酯和6-O-硫酸酯残基的修饰肝素和用较短的多糖链分析了TEPI与肝素的相互作用。结果表明，所有的硫酸酯残基是必需的相互作用和糖链的14个单位几乎有相同的能力与肝素TFPI的相互作用。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，Arg 257和Arg 259是必需的与肝素的相互作用，通过使用合成肽与C-末端碱性部分。我们利用杆状病毒系统分离重组K3结构域，并检测其与肝素的相互作用。

项目成果

N Ohkura: "A novel degradation pathway of tissue factor pathway inhibitor : Incorporation into fibrin clot and degradation by thrombin." BLOOD. 90. 1883-1892 (1997)

N Ohkura：“组织因子途径抑制剂的一种新的降解途径：掺入纤维蛋白凝块并被凝血酶降解。”

N Ohkuru: "A novel degradation pathway of tissue factor pathway inhibitor : Incorporation into fibrin clot and degradation by thrombin." BLOOD. 90. 1883-1892 (1997)

N Ohkuru：“组织因子途径抑制剂的一种新的降解途径：掺入纤维蛋白凝块并被凝血酶降解。”

Z Ye: "Structural requirements of human tissue factor pathway inhibitor(TFPI)and heparin for TFPI-heparin interaction." Thromb Res. 89. 263-270 (1998)

Z Ye：“人组织因子途径抑制剂（TFPI）和肝素对 TFPI-肝素相互作用的结构要求。”

Naoki Ohkura: "A novel degradation pathway of tissue factor pathway inhibitor : Incorporation into fibrin clot and clegradation by thrombin" Blood. 90(5). 1883-1892 (1997)

Naoki Ohkura：“组织因子途径抑制剂的新型降解途径：掺入纤维蛋白凝块并通过凝血酶降解”血液。

神窪勇一: "血栓と循環" メディカルレビュー社, 8 (1998)

Yuichi Kamikubo：“血栓形成和循环”医学评论出版，8（1998）