MECHANISMS OF TISSUE FACTOR/FACTOR VII ASSEMBLY

组织因子/因子 VII 组装机制

• 批准号: 2223328
• 负责人: James H. Morrissey
• 金额: $ 16.96万
• 依托单位: OKLAHOMA MEDICAL RESEARCH FOUNDATION
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-08-01 至 1996-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2223328
• 关键词: active sites; blood coagulation; calcium binding protein; coagulation factor IX; coagulation factor VII; coagulation factor X; enzyme activity; enzyme substrate complex; epidermal growth factor; epitope mapping; fluorescent dye /probe; genetic manipulation; human subject; monoclonal antibody; phospholipids; polymerization; protein purification; protein structure function; site directed mutagenesis; synthetic peptide; thromboplastin; zymogens

Tissue factor (TF), the primary triggering agent of the blood clotting cascade in normal hemostasis, is also implicated as a key initiator in the pathogenesis of a variety of life-threatening thrombohemorrhagic disorders. TF has two known functions: (1) TF greatly accelerates the activation of zymogen factor VII (fVII to the catalytically activate form, fVIIa; and (2) TF is the protein cofactor for fVIIa enzymatic activity. Neither function of TF is well understood in molecular terms. The goal of this study is to understand how TF interacts with fVII and fVIIa to assemble the functional TF:fVIIa complex, which is responsible for initiating the clotting cascade. The interactive sites on TF and fVIIa will be identified using mutagenesis, peptides, and antibodies. A battery of assays will be used that will permit measurement of specific aspects of TF/fVII function. Importantly, results from the activity assays will be compared to direct biophysical measures of protein-protein and protein-phospholipid interactions. The demonstration that the two major functions of TF can be separated by mutagenesis underscores the feasibility of this approach. This new insight into the structure/function relationship of TF has led to the formulation of several hypotheses, the testing of which will now enable the direct investigation of the poorly understood mechanism by which TF accelerates fVII activation. An important development from these studies will be the generation of novel anti-thrombotic agents with extremely high specificity (since they will inhibit only one specific aspect of TF or fVII function).

组织因子（TF），血液凝结的主要触发因子 正常止血中的级联反应也被视为关键引发剂 各种威胁生命的血栓形象的发病机理 疾病。 TF具有两个已知功能：（1）TF极大地加速了 酶原因子VII的激活（FVII至催化激活 形式，fviia; （2）TF是FVIIA酶促的蛋白质辅因子 活动。 TF的功能都以分子术语很好地理解。 这项研究的目的是了解TF如何与FVII相互作用和 FVIIA组装功能性TF：FVIIA复合物，这是负责的 用于启动凝结的级联反应。 TF和 FVIIA将使用诱变，肽和抗体确定。 一个 将使用一系列测定，以允许特定的测量 TF/FVII功能的各个方面。 重要的是，活动的结果 将测定与蛋白质蛋白的直接生物物理测量进行比较 和蛋白质磷脂相互作用。 两者的演示 TF的主要功能可以通过诱变强调 这种方法的可行性。 对 TF的结构/功能关系导致了 几个假设，其测试现在将实现直接 研究TF加速的知识较低的机制 FVII激活。 这些研究的重要发展将是 产生非常高的新型抗血栓形成剂 特异性（因为它们将仅抑制TF的一个特定方面或 FVII函数）。