HYDROLYTIC ENZYMES OF BLOOD IN HEALTH AND DISEASE

目的：探讨止血、释放激肽、 补体系统在血浆中，我们将检查某些功能：1） 活化补体第一成分血清抑制剂 （Cl-抑制剂），2）高分子量激肽原（HMW-K），3）Hageman 因子和从Hageman因子（HFf）裂解的片段， 方法：1）分离的畸形C1-抑制蛋白的能力 从某些遗传性血管神经性水肿患者的血浆中， 抑制活化Hageman因子、血浆激肽释放酶和纤溶酶， 测试. 正常的C1-抑制剂阻断这些止血酶，以及 Cl，但畸形C1-抑制剂不抑制C1。 碎片释放 从正常和畸形的C1-抑制剂在溴化氰裂解 将比较它们的抑制和结合特性， 每一种酶。 2)HMW-K轻链的单克隆抗体将作为 制备、荧光化并用于定位组织和细胞中的HMW-K， 血液和内皮细胞。 抗高分子量K抗体对血小板的影响 如果HMW-K存在于血小板膜上，则将检测其功能。 将检测抗HMW-K对以下细胞定向迁移率的影响： 在改良的Boyden室中培养多形核细胞。 如果发现HMW-K 内皮细胞，它可能会影响形态的可能性， 这些细胞，或在从细胞释放凝血活性中发挥作用， 将检查细胞。 3)用抗Hageman因子片段的抗体 (HFf)，我们将检查易受 血栓栓塞，以及正常人，看看是否HFf在血浆中可能是 与血栓栓塞倾向有关。 的可能性 Hageman因子激活的初始步骤可能涉及非酶促 单链Hageman因子中内部硫酯键的断裂 分子将被检测。

To explore interrelated functions of hemostatic, kinin-releasing and complement systems in plasma we will examine certain functions of: 1) the serum inhibitor of the activated first component of complement (Cl-Inhibitor), 2) high molecular weight kininogen (HMW-K), 3) Hageman factor and fragments cleaved from Hageman factor (HFf) in the following approaches: 1) The capacity of dysmorphic C1-Inhibitor proteins isolated from plasma of certain persons with hereditary angioneurotic edema to inhibit activated Hageman factor, plasma kallikrein, and plasmin will be tested. Normal C1-Inhibitor blocks these hemostatic enzymes, as well as Cl, but dysmorphic C1-Inhibitors do not inhibit C1. Fragments released from normal and dysmorphic C1-Inhibitors during cyanogen bromide cleavage will be compared as to their inhibitory and binding properties with respect to each enzyme. 2) Monoclonal antibody to light chain of HMW-K will be prepared, fluorescinated and used to locate HMW-K in tissues and cells of blood and endothelial cells. The effect of anti-HMW-K upon platelet functions will be tested, if HMW-K is present on platelet membranes. Anti-HMW-K will be tested for its effect on directed mobility of polymorphonuclear cells in a modified Boyden Chamber. If HMW-K is found on endothelial cells, the possibility that it may affect the morphology of these cells, or play a role in the release of coagulant activity from the cells will be examined. 3) With antibody to Hageman factor fragments (HFf), we will examine plasma from individuals susceptible to thromboembolism, as well as normal persons, to see if HFf in plasma may be associated with a thromboembolic tendency. The possibility that the initial step in the activation of Hageman factor may involve non-enzymatic cleavage of an internal thioester bond in the single-chain Hageman factor molecule will be examined.