CYSTEINYL-RICH DOMAIN IN BLOOD CLOTTING FACTORS

组织损伤需要血液的快速和局部反应 凝血因子和细胞增殖用于修复。这个 许多血液中的富含半胱氨基和生长因子样结构域 凝血因子可能在这一过程中发挥核心作用。 然而，这些域的功能角色目前是 未知的，没有特征的。这项提案的目标是 了解这些富含半胱氨基的结构域的作用 在凝血途径的机制中，刺激 用于血管系统的纤溶酶原激活剂和增殖物 在伤害中的反应。该提案包括化学合成， 核磁共振构象研究及生物活性评价 凝血因子IX和凝血因子X富含半胱氨酸基的结构域 路径。 这些凝血因子的人工合成富含半胱氨基的结构域 它们的变种将用固相法制备。这个 这些合成分子的溶液构象将被研究。 通过2D-核磁共振对构效关系的研究进行关联。 为了确定这些富含半胱氨基的结构域的功能作用 在凝血途径中，我们建议研究钙离子- 结合活性，组织纤溶酶原激活剂的激活 和尿激酶，以及抑制因子IX与其结合 内皮细胞受体对内皮细胞的依赖 凝血途径。合成的类似物被测试为它们的 对组织有丝分裂、刺激作用的选择性活性 纤溶酶原激活剂，或对结合的抑制活性 因子IX与其在内皮细胞上的受体的结合。 我们的长期目标是化学、物理和生物 这些富含半胱氨酸基的生长因子结构域及其结构域的研究 类比将有助于在分子水平上理解 各种凝血因子的相互作用和激活。 这最终可能导致设计和合成合适的 作为体内调节的抑制物的类似物 一种特定的凝血因子和组织纤溶酶原激活剂。 这些抑制剂可能会提供一种非常不同的方法来 肿瘤进展或血栓形成的血管中断。

Tissue injury requires rapid and focal response of blood coagulation factors and cellular proliferation for repair. The cysteinyl-rich and growth factor-like domains in many blood coagulation factors may play a central role in such a process. However, the functional roles of these domains are presently unknown and uncharacterized. The goals of this proposal are the attainment of knowledge of the role of these cysteinyl-rich domains in the mechanism of blood coagulation pathway, stimulation of plasminogen activator for vasculature, and the proliferative responses in injury. The proposal includes chemical synthesis, conformational study by NMR and biological evaluation of the cysteinyl-rich domains in Factor IX and Factor X of the coagulation pathway. Synthetic cysteinyl-rich domains of these blood coagulation factors and their variants will be prepared by the solid-phase method. The solution conformation of these synthetic molecules will be studied by 2D-NMR to correlate the structure-activity studies. To determine the functional role of these cysteinyl-rich domains in the blood coagulation pathway, we propose to study the Ca2+- binding activity, the activation of tissue plasminogen activator and urokinase, and the inhibition of binding of Factor IX to its endothelial cell receptor on the endothelial cell-dependent coagulation pathway. Synthetic analogs are tested for their selective activity on mitogenicity, stimulation of tissue plasminogen activator, or inhibitory activity against the binding of Factor IX to its receptor on the endothelial cell. Our long term goal is that the chemical, physical and biological studies of these cysteinyl-rich, growth factor domains and their analogs will aid the understanding, on a molecular level, of the interaction and activation of various blood coagulation factors. This may ultimately lead to the design and synthesis of suitable analogs that function as inhibitors for the in vivo regulation of a specific coagulation factor and tissue plasminogen activator. Such inhibitors may provide a very different approach to the interruption of vasculative of tumor progression or thrombosis.