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.

该研究计划旨在了解凝血酶的产生方式以及如何调节凝血酶的生成。我们对这些问题的方法是通过与1）凝结酶复合物的物理特性，它们的成分以及这些复合物如何聚集成有效的酶催化剂的四个独立方向的融合。 2）混合了多种凝血催化剂/抑制剂的研究，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。 3）在最小修饰的生物系统（全血）中研究此过程，以评估纯化系统得出的假设的正确性。 4）创建数学模型，可用于根据定量定义血液凝结的过程及其调节，以帮助实验设计1,2,3，并有助于评估药理学药物，以及对止血和血栓疾病的诊断和治疗。本研究的目的是了解促凝剂和抗凝维生素-K依赖性复合物及其在凝血酶产生过程中的调节的性质。研究将采用物理化学技术，包括水动力学和荧光光谱，（后者在溶液和表面上）在封闭的系统和流量下以研究合成膜和细胞的复合物。将使用合成和天然底物进行反应，以监测质状状态和稳态动力学事件。天然和重组抑制剂将用于研究与凝血酶产生相关的促凝剂和抗凝剂过程的调节。我们将通过对单个反应的研究与从多反应中心系统获得的详细信息集成可用的详细信息。相反，在整个血液系统中提到的过程将在纯化的系统分析中引起适当的关注。我们预计会对与复杂反应相关的生物学相关化学反应进行定量评估，这些化学在血液凝结事件中同时发生。这些数据在解释正常生理和开发方法以纠正与血栓形成和血友病相关的凝血病理学方面具有重要意义。我们开发的技术将为评估止血和血栓性疾病中潜在的药理干预措施提供工具。