FIBRINOGEN STRUCTURE, FUNCTION AND PHYSIOLOGY

成纤维细胞的生长发挥了内皮细胞的几个重要作用 包括增加扩散，刺激迁移，上调 几种蛋白质的合成和刺激血管生成。内皮 细胞特性也受到与纤维蛋白原相互作用的影响。和 纤维蛋白可支持粘附，刺激蛋白质合成， 分泌，改变炎症细胞相互作用和支持 血管生成我们最近证明成纤维细胞的生长 因子-2（FGF-2）特异性结合纤维蛋白原和纤维蛋白， 与纤维蛋白原的相互作用增加了其刺激 内皮细胞增殖这些发现构成了 提出的研究，其目标是确定结构基础， 纤维蛋白（原）和FGF-2之间的相互作用及其表征 功能影响。第一个具体目标是描述 FGF-2和特定实验之间的相互作用将确定 结合位点，并确定是否FGF家族的其他成员 也与纤维蛋白原结合。这种等离子体激元电阻技术将用于 测定结合和解离速率常数， 化学计量定义的结合。初步实验表明 FGF-2与纤维蛋白原的结合提供了保护， 蛋白水解降解，这可能是重要的，在延长行动 在凝血酶和纤溶酶活跃的炎症部位。实验 目的2将探讨纤维蛋白原保护FGF-2的程度 与纤维蛋白原的血浆衍生物结合， 与这些衍生品绑定。目标3中的实验将扩展 观察到FGF-2与纤维蛋白原的结合增强了其对纤维蛋白原的抑制作用。 刺激内皮细胞增殖的能力。的参与 在这种活性增强中已知的内皮细胞受体将 使用特异性阻断单克隆抗体、肽 和去整合素。需要形成FGF-2-纤维蛋白原复合物 将使用非FGF-2结合的方法来检查增强增殖的作用。 FGF-2突变体的纤维蛋白原片段。这些将用于确定 如果缺乏结合纤维蛋白（原）能力的FGF-2突变体可以支持 纤维蛋白基质中的血管生成。进一步的实验将确定 纤维蛋白原也改变FGF-2对内皮细胞迁移的影响 以及u-PA的合成。获得的结果将提供信息 关于FGF-2与 纤维蛋白（原），这将是重要的了解正常血管 发育和功能，对损伤的反应，以及 内皮细胞暴露于纤维蛋白（原）和成纤维细胞 生长因子

Fibroblast growth exert several important effects of endothelial cells including increasing proliferation, stimulating migration, up-regulating synthesis of several proteins and stimulation angiogenesis. Endothelial cell properties are also influenced by interaction with fibrinogen. and fibrin which can support adhesion, stimulate protein synthesis and secretion, alter inflammatory cell interactions and support angiogenesis. We have recently demonstrated that fibroblast growth factor-2 (FGF-2) binds specifically to fibrinogen and fibrin, and that the interaction with fibrinogen increases its capacity to stimulate endothelial cell proliferation. These findings form the basis for the studies proposed which have a goal of defining the structural basis of the interaction between fibrin(ogen) and FGF-2 and characterizing its functional implications. The first specific aim will be to characterize the interaction between FGF-2 and specific experiments will identify the binding sites involved and determine if other members of the FGF family also bind to fibrinogen. Such plasmon resistance techniques will be used to determine the association and dissociation rate constants and stoichiometry defining the binding. Preliminary experiments indicate that the binding of FGF-2 to fibrinogen provides protection from proteolytic degradation, and this may be important in prolonging action at inflammatory sites where thrombin and plasmin are active. Experiments in Aim 2 will explore the extent to which fibrinogen protects FGF-2 binds to plasmic derivatives of fibrinogen and retains activity when bound to such derivatives. The experiments in Aim 3 will extend the observation that the binding of FGF-2 to fibrinogen potentiates its capacity to stimulate endothelial cell proliferation. The involvement of known endothelial cell receptors in this potentiation of activity will be characterized using specific blocking monoclonal antibodies, peptides and disintegrins. The need for formation for an FGF-2-fibrinogen complex in potentiating proliferation will be examined using non-FGF-2-binding fibrinogen fragments of FGF-2 mutants. These will be used to determine if FGF-2 mutants lacking the ability to bind fibrin(ogen) can support angiogenesis in a fibrin matrix. Further experiments will determine if fibrinogen also alters effects of FGF-2 on endothelial cell migration and synthesis of u-PA. The results obtained will provide information regarding fundamental aspects of the association of FGF-2 and fibrin(ogen) which will be important for understanding normal vascular development and function, response to injury, and disease conditions in which endothelial cells are exposed to both fibrin(ogen) and fibroblast growth factors.