TYPE-2 TISSUE FACTOR PATHWAY INHIBITOR

2 型组织因子途径抑制剂

• 批准号: 6616225
• 负责人: Walter Kisiel
• 金额: $ 29.4万
• 依托单位: UNIVERSITY OF NEW MEXICO
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-05 至 2004-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6616225
• 关键词: SCID mouse; X ray crystallography; biological models; blood coagulation; enzyme complex; enzyme inhibitors; extracellular matrix; extracellular matrix proteins; gene expression; genetic regulatory element; homeostasis; laboratory mouse; laboratory rabbit; model design /development; monoclonal antibody; mutant; neoplastic cell culture for noncancer research; protein protein interaction; protein structure function; recombinant proteins; serine proteinases; thromboplastin; tissue /cell culture; vascular endothelium

DESCRIPTION: (Investigator's abstract) Extracellular matrix (ECM) is a complex mixture of proteoglycans that function in cell adhesion, migration, growth and differentiation. In addition, the ECM serves as a reservoir for a number of growth factors, proteinases and proteinase inhibitors essential for ECM homeostasis. Recent new information from the applicant's laboratory has shown that one of the serine proteinase inhibitors found in the ECM is a 32kDa glycoprotein consisting of three tandemly-arranged Kunitz-type proteinase inhibitor domains homologous to tissue factor pathway inhibitor, an important negative regulator of the extrinsic pathway of blood coagulation. This novel inhibitor, designated as type-2 tissue factor pathway inhibitor, or TFPI-2, is a potent inhibitor of trypsin, plasmin, kallikrein and factor XIa amidolytic activity in the test tube. A variety of human endothelial cells synthesize TFPI-2, and 60-90 percent of the synthesized TFPI-2 is secreted by the endothelial cells into their ECM. Inflammatory mediators upregulate endothelial cell TFPI-2 synthesis 2-14 fold. In addition, evidence was obtained that recombinant TFPI-2 inhibited the plasmin-mediated invasion and degradation of Matrigel by a high invasive fibrosarcoma cell line in a dose-dependent manner. Furthermore, treatment of confluent endothelial cell monolayers with anti-TFPI-2 IgG detached the cells from the substrata in a time and IgG concentration dependent manner, suggesting an important role for TFPI-2 in maintaining the integrity of the ECM essential for cell attachment. In spite of these findings several questions concerning the physiological role of TFPI-2 remain unanswered, such as (1) which Kurutz domain is responsible for its inhibitory activity, (2) what proteinase(s) are regulated by ECM-bound TFPI-2, and (3) what are the regulatory mechanisms of TFPI-2 expression. The objectives of this proposal are to accomplish a comprehensive characterization of TFPI-2 in order to gain further insight into its biological role in normal and pathological ECM turnover.

描述：（研究者的摘要）细胞外基质（ECM）是一个复杂的 在细胞粘附，迁移，生长和 分化。此外，ECM充当许多 生长因子，蛋白酶和蛋白酶抑制剂对ECM必不可少的 稳态。申请人实验室的最新新信息已显示 在ECM中发现的丝氨酸蛋白酶抑制剂之一是32KDA 糖蛋白由三个串联的kunitz型蛋白酶组成 与组织因子途径抑制剂同源的抑制剂结构域，这是一个重要的 血液凝结外部途径的负调节剂。这本小说 被指定为2型组织因子途径抑制剂（TFPI-2）的抑制剂为 胰蛋白酶，纤溶酶，kallikrein和因子xia的有效抑制剂 测试管中的活性。各种人类内皮细胞合成 TFPI-2和60-90％的合成TFPI-2由 内皮细胞进入其ECM。炎症介体上调 细胞TFPI-2合成2-14倍。此外，还获得了证据表明 重组TFPI-2抑制了纤溶酶介导的浸润和降解 以剂量依赖性方式通过高浸润性纤维肉瘤细胞系的基质凝胶。 此外，用汇合的内皮细胞单层处理 抗TFPI-2 IgG一段时间从基质中分离出细胞，而IgG 浓度依赖性方式，表明TFPI-2在 维持ECM对细胞附着至关重要的完整性。尽管 这些发现有关TFPI-2的生理作用的几个问题 保持没有答案，例如（1）Kurutz域名负责 抑制活性，（2）哪种蛋白酶受ECM结合的TFPI-2调节， （3）TFPI-2表达的调节机制是什么？目标 该提议的旨在完成TFPI-2的全面表征 为了进一步了解其在正常和 病理ECM周转。