VITAMIN K-DEPENDENT PLASMA PROTEINS

维生素 K 依赖性血浆蛋白

• 批准号: 3335043
• 负责人: Gary L Nelsestuen
• 金额: $ 15.98万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 1976
• 资助国家: 美国
• 起止时间: 1976-05-01 至 1990-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3335043
• 关键词: calcium binding protein; coagulation factor V; coagulation factor VIII; complement; cow; fluorescence; gamma carboxyglutamate; light scattering; membrane activity; membrane proteins; nuclear magnetic resonance spectroscopy; phospholipids; protein C; protein S; prothrombin; vitamin K

This research has three general objectives that are related to understanding the physical and kinetic properties of assembling the blood coagulation cascade. The first is to understand the function of the vitamin K-dependent amino acid, Gamma-carboxyglutamic acid in calcium binding and protein-membrane association. The protein structures that are involved in, and change as a result of, calcium ion binding will be investigated by use of nmr and lanthanide substitution experiments utilizing the vitamin K-dependent, membrane-binding portion of prothrombin. The lanthanide experiments will also make extensive use of fluorescence techniques. The structure at the protein-membrane interface will be studied by measuring dynamic and equilibrium properties of protein-membrane association and by substitution of lanthanides for calcium and determining various distances and changes in the complex detected by fluorescence techniques. The second major area is the structure and function of membrane complexes containing more than one protein. The initial hypothesis is that the membrane-binding site of the vitamin K-dependent proteins functions as a portion of an overall binding interaction. That is, these proteins normally bind simultaneously to a membrane plus at least one other membrane-bound protein. Specific protein-protein complexes on the membrane surface to be studied include: protein S with protein C(a) and with complement C4-binding protein. Other examples include protein C(a) with factor Va, factor VIIIa or thrombomodulin. Equilibrium, dynamic and hydrodynamic aspects of these interactions will be measured to determine if the proteins associate with each other while they are membrane bound. Light scattering, quasielastic light scattering and various fluorescence techniques will be used. Protein binding to phospholipid monolayers will also be used to document the properties of the protein-lipid associations. The third area of study involves proteins of the contract phase of coagulation. At least some of these proteins bind to lipid membranes and undergo activation. The various properties of the protein-lipid associations describe above will be measured to help understand the physical forces involved in binding. Analysis of the kinetics of factor activation will be correlated with protein-lipid binding to try to understand how the surface influences this kinetic event. These studies should increase our knowledge of protein-membrane interactions in general and how such interactions are specifically responsible for enhancing the rates of the blood coagulation process.

本研究有三个总体目标， 了解血液聚集的物理和动力学特性 凝血级联反应 第一，要了解 维生素K依赖性氨基酸，钙中的γ-羧基谷氨酸 结合和蛋白质-膜缔合。 这些蛋白质结构 参与钙离子结合并因钙离子结合而发生变化， 通过核磁共振和镧系元素取代实验研究 利用维生素K依赖性的膜结合部分， 凝血酶原 镧系元素实验也将广泛使用 荧光技术 蛋白质-膜界面的结构 将通过测量的动态和平衡性能进行研究， 蛋白质-膜结合和镧系元素取代钙 以及确定通过以下步骤检测的复合物中的各种距离和变化： 荧光技术 第二个主要领域是结构和 膜复合物含有一个以上的蛋白质的功能。 的 最初的假设是维生素的膜结合位点 K依赖性蛋白作为整体结合的一部分发挥作用， 互动 也就是说，这些蛋白质通常同时结合到 膜加上至少一种其他膜结合蛋白。 具体 待研究的膜表面上的蛋白质-蛋白质复合物包括： 蛋白S与蛋白C（a）和补体C4结合蛋白。 其他 实例包括蛋白C（a）与因子Va、因子VIIIa或 血栓调节蛋白 平衡、动力学和流体动力学方面， 将测量相互作用以确定蛋白质是否与 当它们被膜结合时彼此结合。 光散射，准弹性 将使用光散射和各种荧光技术。 蛋白 与磷脂单层的结合也将用于记录 蛋白质-脂质结合的性质。 第三个研究领域 涉及凝血收缩期的蛋白质。 中的至少一些 这些蛋白质与脂膜结合并经历活化。 的各种 上述蛋白质-脂质缔合的性质将 测量有助于理解结合中涉及的物理力。 因子活化的动力学分析将与 蛋白质-脂质结合，试图了解表面如何影响这一点， 动力学事件 这些研究应该增加我们对 蛋白质-膜相互作用以及这种相互作用是如何 专门负责提高血液凝固的速度 过程