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。 的 许多血液中富含半胱氨酰和生长因子样结构域 凝血因子可能在这一过程中起中心作用。 然而，这些结构域的功能作用目前还不清楚。 未知的和不确定的。 本提案的目标是 了解这些富含半胱氨酰的结构域的作用 在凝血途径的机制中， 用于血管系统的纤溶酶原激活剂， 受伤的反应。 该提案包括化学合成， 核磁共振构象研究和生物学评价 凝血因子IX和凝血因子X中富含半胱氨酸的结构域 通路 这些凝血因子的合成富半胱氨酰结构域 并且它们的变体将通过固相法制备。 的 研究了这些合成分子的溶液构象 通过2D-NMR来关联结构-活性研究。 为了确定这些富含半胱氨酰的结构域的功能作用， 在凝血途径中，我们建议研究Ca 2 +- 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 内皮细胞依赖性内皮细胞受体 凝血途径 测试合成类似物的 对促有丝分裂、组织刺激的选择性活性 纤溶酶原激活剂，或对结合的抑制活性 与内皮细胞上的受体结合。 我们的长期目标是， 这些富含半胱氨酰的生长因子结构域及其 类似物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。 这可能最终导致设计和合成合适的 作为抑制剂用于体内调节 特异性凝血因子和组织纤溶酶原激活剂。 这样的抑制剂可以提供一种非常不同的方法， 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.