HYDROLYTIC ENZYMES OF BLOOD IN HEALTH AND DISEASE

止血、激肽释放和 血浆中的纤维蛋白溶解酶和血清补体系统将 来深入了解其中一些 酶由血清C1抑制剂调节， 几种这样的酶，但对C1的调节至关重要， 激活补体的第一成分。 此外，止血 涉及高分子量激肽原的事件可能影响 血管功能，以及这种激肽原和 将检查内皮。 为此，有几种方法将 使用： 1. 将检查纯化的正常C1抑制剂，以确定是否 分子的不同结构域可以参与位点特异性的 与血浆蛋白酶的相互作用已知由 这种抑制剂。 这种可能性很可能是因为我们早些时候 研究显示异常C1抑制剂的异质性行为 遗传性血管神经性水肿患者的蛋白质 相对于由正常C1-抑制剂调节的每种酶。 为了做到这一点，正常的C1-抑制剂的片段产生与 溴化氰，或通过有限的蛋白水解消化，将 检查它们的抑制活性和它们形成 复合物-与这些蛋白酶（因子XIIa，激肽释放酶，纤溶酶， C1 2. 有多种可能的解释改变表达 C1抑制基因在遗传性血管神经性水肿中的作用。 一 在这个实验室中使用的方法来深入了解这一点， 问题将是比较来自外周血单核细胞的mRNA， 从正常人到不同类型的人 遗传性血管神经性水肿 的数量和质量 将来自这些患者的mRNA与来自正常人的mRNA进行比较。 个体 3. 将对培养的人脐静脉内皮细胞进行检测， 位于人高密度脂蛋白重链或轻链上的抗原 分子量激肽原。 由于轻链参与 与某些物质结合并产生促凝作用 活性，可能在内皮细胞膜上发现， 可能会转移到这些细胞在可定义的状态下， 功能会被激发。 其他研究将审查 轻链和重链抗原的血浆浓度比 正常人血浆中HMWK的含量-以及病理性 涉及脉管系统的状态。

lnterrelated functions of hemostatic, kinin-releasing, and fibrinolytic enzymes in plasma and the serum complement system will be examined to gain insight into the ways in which some of those enzymes are regulated by the serum C1-inhibitor, which regulates several such enzymes but is critical to the regulation of C1, the activated first component of complement. In addition, hemostatic events involving high molecular weight kininogen may affect vascular function, and the interactions between this kininogen and endothelium will be examined. To do this, several approaches will be used: 1. Purified normal C1-inhibitor will be examined to determine if different domains of the molecule may participate in site-specific interactions with the plasma proteases known to be regulated by this inhibitor. This possibility is likely because our earlier studies showed heterogeneous behavior of abnormal C1-inhibitor proteins from persons with hereditary angioneurotic edema with respect to each of the enzymes regulated by normal C1-inhibitor. To do this, fragments of normal C1-inhibitor generated with cyanogen bromide, or by limited proteolytic digestion, will be examined for their inhibitory activity and their ability to form complexes-with these proteases (factor Xlla, kallikrein, plasmin, C1s. 2. There are multiple possible explanations for altered expression of the C1-inhibitor gene in hereditary angioneurotic edema. One approach to be used in this laboratory to gain insights into this problem will be to compare mRNA from peripheral blood monocytes of normal persons to that from persons with different types of hereditary angioneurotic edema. Both the quantity and quality of mRNA from these patients will be compared to that from normal individuals. 3. Cultured human umbilical endothelial cells will be tested for antigens located on the heavy or light chains of human high molecular weight kininogen. Since light chain participates in binding to certain substances and in generating clot-promoting activity, it might be identified on endothelial membranes, and might transfer to these cells under definable states in which its function would be provoked. Additional studies will examine the ratios of plasma concentrations of light and heavy chain antigens of HMWK in plasma from normal persons- and those with pathologic states involving the vasculature.