ACTIVATION OF PROTHROMBIN

凝血酶原的激活

• 批准号: 6657100
• 负责人: KENNETH G MANN
• 金额: $ 18.67万
• 依托单位: UNIVERSITY OF VERMONT & ST AGRIC COLLEGE
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-01 至 2003-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6657100
• 关键词: anticoagulants; blood coagulation; electron microscopy; enzyme complex; fibrinolysis; hemostatics; human tissue; immunologic assay /test; laboratory mouse; mathematical model; plasminogen activator; prothrombin; thrombin; thrombosis; vitamin K

This research program is aimed at understanding how thrombin is generated and how thrombin generation is regulated. Our approach to these questions comes via the convergence of four separate directions associated with 1) the physical properties of coagulation enzyme complexes, their constituents and how these complexes can assemble into efficient enzyme catalysts. 2) Studies in which multiple coagulation catalysts/inhibitors are mixed to attempt to duplicate the performance of the combined catalyst system associated with the tissue factor pathway of thrombin expression. 3) To study this process in minimally modified biological systems (whole blood) to evaluate the correctness of hypothesis derived from purified systems. 4) To create mathematical models which can be used to define, on a quantitative basis, the process of blood clotting and its regulation both to aid in experimental designs 1,2,3, and also to aid in the evaluation of the pharmacologic agents and the diagnosis and treatment of hemostatic and thrombotic diseases. The aim of the present investigation is to understand the nature of procoagulant and anticoagulant vitamin-K dependent complexes and their regulation during the process of thrombin generation. Studies will employ physical chemistry techniques including hydrodynamics and fluorescence spectroscopy, (the latter both in solution and on surfaces) in closed systems and under flow to study complexes on synthetic membranes and cells. Reactions will be followed using both synthetic and natural substrates to monitor both presteady state and steady state kinetic events. Natural and recombinant inhibitors will be used to study the regulation of procoagulant and anticoagulant processes associated with thrombin generation. We will integrate the detailed information available through studies of individual reactions with that obtained from multi-reaction center systems. Conversely, the processes noted to occur in the whole blood system will direct appropriate attention in the purified system analyses. We anticipate developing a quantitative evaluation of the biologically relevant chemistry associated with the complex reactions which occur simultaneously during a blood clotting event. These data have significance in interpreting normal physiology and in developing approaches to correct the coagulation pathology associated with thrombosis and hemophilia. The techniques we develop will provide tools for the evaluation of potential pharmacological intervention in hemostatic and thrombotic disease.

该研究项目旨在了解凝血酶是如何产生的以及凝血酶的产生是如何调节的。我们解决这些问题的方法是通过四个不同方向的融合来实现的：1）凝固酶复合物的物理特性、它们的成分以及这些复合物如何组装成有效的酶催化剂。 2)混合多种凝血催化剂/抑制剂以尝试复制与凝血酶表达的组织因子途径相关的组合催化剂系统的性能的研究。 3) 在经过最低限度改造的生物系统（全血）中研究这一过程，以评估源自纯化系统的假设的正确性。 4) 创建数学模型，可用于定量定义血液凝固过程及其调节，以帮助实验设计 1,2,3，并帮助评估药物和药物止血及血栓性疾病的诊断和治疗。本研究的目的是了解促凝血和抗凝血维生素 K 依赖性复合物的性质及其在凝血酶生成过程中的调节。研究将采用物理化学技术，包括流体动力学和荧光光谱（后者在溶液中和表面上）在封闭系统和流动下研究合成膜和细胞上的复合物。将使用合成和天然底物跟踪反应，以监测稳态前和稳态动力学事件。天然和重组抑制剂将用于研究与凝血酶生成相关的促凝血和抗凝血过程的调节。我们将把通过个体反应研究获得的详细信息与从多反应中心系统获得的信息整合起来。相反，在全血系统中发生的过程将在纯化系统分析中引起适当的关注。我们期望对与血液凝固过程中同时发生的复杂反应相关的生物相关化学进行定量评估。这些数据对于解释正常生理学和开发纠正与血栓形成和血友病相关的凝血病理学的方法具有重要意义。我们开发的技术将为评估止血和血栓性疾病的潜在药物干预提供工具。