FIBRIN(OGEN) STRUCTURE AND INTERACTIONS

(研究摘要)：纤维蛋白原是一种多功能蛋白质 参与了许多重要的生理和病理过程 流程。在组织损伤部位形成蚕豆蛋白基质(或 其他地方)可防止失血并提供临时基质， 与不同类型的细胞相互作用，促进其沉积、迁移 和扩散。因此，它除了具有显著的止血作用外， 纤维蛋白原被认为是一种细胞黏附分子，它发挥着 在炎症、血管生成、伤口愈合、动脉粥样硬化形成中的重要作用 和肿瘤的发生。尽管进行了大量研究，但有几个基本问题 对全面了解其生物学功能具有重要意义 仍未得到解决。本申请涉及三个这样的问题：(1) 高分辨结构，(2)转化的分子机理 纤维蛋白原与纤维蛋白的相互作用机制 具有不同细胞类型的蚕豆蛋白(原)。第一个目标是建立 纤维蛋白原中心E区的高分辨结构及其意义 Alpha-C域。第二个目的是阐明其机制。 假设涉及开关的纤维蛋白原到蚕豆蛋白的转化 从α-C结构域之间的分子内到分子间结合以及 调控酶活性的D区构象变化 二次聚合位点和新的相互作用位点的表达。 第三个目标是建立相互作用的机制。 内皮细胞和白细胞与纤维蛋白原和纤维蛋白 确定所涉及的关键氨基酸残基。这些目标将 通过制备各种纤维蛋白原结构域和 通过有限的蛋白质分解和重组技术的组合， 并通过生物化学和生物化学方法研究它们的结构和相互作用 生物物理学方法。这些研究将产生基本的知识， 对不同事物的理解和控制能力有影响 纤维蛋白原依赖的过程。他们还将有一个重要的实践 健康相关应用于改良生物粘合剂的开发 (纤维蛋白胶)由纤维蛋白原的从头设计，具有选定的功能。

(Investigator's abstract): Fibrinogen is a multifunctional protein involved in a number of important physiological and pathological processes. The formation of a fabrin matrix at sites of tissue injury (or other places) prevents blood loss and provides a provisional stroma that interacts with different cell types to promote their deposition, migration and proliferation. Thus, besides its prominent function in hemostasis, fibrinogen is recognized as a cell adhesion molecule that plays an important role in inflammation, angiogenesis, wound healing, atherogenesis and tumorigenesis. Despite numerous studies, several basic problems important for a comprehensive understanding of its biological functions are still unresolved. This application deals with three such problems: (1) a high resolution structure, (2) the molecular mechanisms of conversions of fibrinogen to fibrin, and (3) the mechanisms of interactions of fabrin(ogen) with different cell types. The first aim is to establish the high resolution structure of the central E regions of fibrinogen and its alpha-C domains. The second aim is to elucidate the mechanisms of fibrinogen to fabrin conversions which is hypothesized to involve a switch from intra- to intermolecular binding between alpha-C domains as well as conformation changes in the D region that modulate activity of the secondary polymerization sites and expression of new interaction sites. The third aim focuses on establishing the mechanisms of interaction of endothelial cells and leukocytes with fibrinogen and fibrin and identification of critical amino acid residues involved. These aims will be accomplished by preparation of various fibrinogen domains and combinations there of by limited proteolysis and recombinant techniques, and studying their structures and interactions by biochemical and biophysical methods. These studies will generate basic knowledge that will have an impact on the understanding of and ability to control different fibrinogen-dependent processes. They will also have an important practical health-related application to the development of improved bioadhesives (fibrin glue) by the de novo design of fibrinogen with selected functions.