HYDROLYTIC ENZYMES OF BLOOD IN HEALTH AND DISEASE

探讨止血、激肽释放和血管紧张素转换酶的相互作用 我们将研究血浆中的补体系统的某些功能：1) 血清中激活的补体第一组分抑制物 2)高分子激肽原(HMW-K)，3)Hageman 因子和从Hageman因子(HFF)切割的片段如下 方法：1)分离的变形的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到 抑制活化的Hageman因子、血浆激肽释放酶和纤溶酶 测试过。正常的C1-抑制剂可阻断这些止血酶，以及 CL，但变态的C1-抑制剂不抑制C1。释放的碎片 在溴化氰裂解过程中来自正常和变形的C1-抑制剂 会比较它们的抑制和结合特性 对每一种酶。2)抗HMW-K轻链的单抗 制备、荧光标记并用于HMW-K在小鼠组织和细胞中的定位 血液和内皮细胞。抗HMW-K抗体对血小板的影响 如果血小板膜上存在HMW-K，将进行功能测试。 将测试抗HMW-K对血管内皮细胞定向迁移的影响 改良博伊登小室中的多形核细胞。如果在以下位置发现HMW-K 内皮细胞，它可能会影响血管内皮细胞的形态 这些细胞，或在释放凝血剂活性方面发挥作用 将对细胞进行检查。3)具有Hageman因子片段的抗体 (HFF)，我们将检查易感人群的血浆 血栓栓塞症，以及正常人，看看血浆中的HFF是否可能 与血栓栓子倾向有关。有没有可能 Hageman因子激活的初始步骤可能涉及非酶 单链Hageman因子中内部硫酯键的断裂 将对分子进行检查。

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.