INITIATION OF CONGULATION AND FIBRINOLYSIS IN MAN

我们的长期目标是确定发起 内源性凝血-激肽系统发生在人的血浆中， 开发用于评估疾病中这种级联反应激活的分析方法 各州。我们希望区分酶的表达是否 未裂解的发酵原的活性在萌发过程中可能很重要；这 这一问题将通过对反应的动力学分析来解决 裂解产物的特定抑制剂和通过化学修饰， 特别关注精氨酸残留物。复合体的类型 在表面上形成的将使用交联剂进行定义，包括 Hageman因子(HF)的可能聚集及其与已知分子的相互作用 含有前激肽释放酶或凝血因子XI和高分子激肽原的复合体。我们会 还要确定C1INH是否控制HF自动激活的速度和 验证启动表面阻碍抑制的假说 被C1INH结合、激活的HF。络合物的定量测定方法 血浆中活化的HF和C1INH将用单抗进一步提纯 将确定产生的抗血清和活性HF的种类。 辅因子HMW激肽原的研究将集中在以下几个方面 问题：1)它是一种可激活的辅因子，即切割一个或 更多的键增加其固有的辅因子活性或影响其能力 与其他血浆蛋白竞争结合到表面；2)什么是 当它被不同的酶消化时，键的断裂顺序 产生的凝血剂链的大小是多少；3)克隆 针对凝血链最小种类的抗体将被产生以 表征表面相互作用所需链的部分或 结合底物前激肽释放酶和高分子激肽原；4)抗体 裂解的分子可以用来设计一种只检测裂解的检测方法 在血浆中形成。最后，我们将研究依赖于HF的 独立的血浆纤溶途径并确定一种血浆 尿激酶原在这两种情况下都起到了作用。补体系统在英语学习中的作用 将测定血浆纤溶活性，新的纤溶酶原激活剂将被 任何纤溶酶原激活剂或新的纤溶酶都将 提纯和鉴定。

Our long term objectives are to determine the mechanism by which initiation of the intrinsic coagulation-kinin system occurs in human plasma and to develop assays by which to assess activation of this cascade in disease states. We hope to distinguish whether the expression of enzymatic activity in uncleaved zymogens might be significant during initiation; this issue will be approached by kinetic analysis of the reactions using specific inhibitors of the cleaved products and by chemical modifications, particularly focusing upon arginine residues. The types of complexes formed upon surfaces will be defined using cross linking reagents including possible aggregation of Hageman factor (HF) and interaction with known complexes containing prekallikrein or factor XI and HMW-kininogen. We will also determine whether C1INH controls the rate of HF autoactivation and test the hypothesis that initiating surfaces impede the inhibition of bound, activated HF by C1INH. The assay for quantitating complexes of activated HF and C1INH in plasma will be further refined using monoclonal antisera and the species of activated HF produced will be determined. Studies of the cofactor HMW kininogen will focus upon the following issues: 1) Is it an activatable cofactor, i.e., does cleavage of one or more bonds increase its intrinsic cofactor activity or affect its ability to compete with other plasma proteins for binding to surfaces; 2) What is the order of bond cleavages when it is digested by different enzymes and what is the size of the coagulant chain that is produced; 3) Monoclonal antibodies to the smallest species of coagulant chain will be produced to characterize portions of that chain requisite for surface interaction or binding the substrates prekallikrein and HMW-kininogen; 4) Antibodies to the cleaved molecule can be used to devise an assay detecting only cleaved forms in plasma. Finally, we will investigate the HF-dependent and independent fibrinolytic pathways of plasma and determine whether a plasma prourokinase plays a role in either. The role of the complement system on plasma fibrinolysis will be determined, new plasminogen activators will be sought, and any plasminogen activators or new fibrinolytic enzymes will be purified and characterized.