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.

这项研究计划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独立方向的融合来实现的：1）凝固酶复合物的物理性质，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)在最小修饰的生物系统（全血）中研究该过程，以评价纯化系统假设的正确性。4)建立数学模型，可用于定量定义凝血过程及其调节，以帮助实验设计1，2，3，并帮助评价药理学药物以及诊断和治疗止血和血栓性疾病。本研究的目的是了解凝血酶生成过程中促凝血和抗凝维生素K依赖性复合物的性质及其调节。研究将采用物理化学技术，包括流体力学和荧光光谱学，（后者在溶液和表面上）在封闭系统和流动下研究合成膜和细胞上的复合物。反应将使用合成和天然底物来监测稳定前状态和稳定状态动力学事件。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过程的调节。我们将把通过研究单个反应获得的详细信息与从多反应中心系统获得的信息结合起来。相反，在全血系统中观察到的过程将在纯化系统分析中引起适当的注意。我们期望开发一种定量评价的生物学相关的化学与复杂的反应，同时发生在血液凝固事件。这些数据在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方面具有重要意义。我们开发的技术将为止血和血栓性疾病的潜在药物干预的评价提供工具。