INITIATION OF CONGULATION AND FIBRINOLYSIS IN MAN

人类凝固和纤维蛋白溶解的启动

• 批准号: 3337385
• 负责人: ALLEN P KAPLAN
• 金额: $ 11.03万
• 依托单位: STATE UNIVERSITY NEW YORK STONY BROOK
• 依托单位国家: 美国
• 财政年份: 1979
• 资助国家: 美国
• 起止时间: 1979-04-01 至 1989-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3337385
• 关键词: bradykinin; chemical cleavage; chromatography; coagulation factor XI; coagulation factor XII; complement pathway; electrophoresis; enzyme linked immunosorbent assay; fibrinolysis; human tissue; kallikreins; macroglobulins; monoclonal antibody; plasmin; plasminogen; plasminogen activator; radioimmunoassay; surface property; ultracentrifugation

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.

我们的长期目标是确定启动的机制 固有的凝血 - 金蛋白系统发生在人血浆中， 开发用于评估疾病级联激活的测定法 国家。 我们希望区分酶的表达是否 在开始期间，未泄漏的酶原的活性可能很重要。这 通过对反应的动力分析，将解决问题 裂解产物的特定抑制剂和化学修饰， 特别关注精氨酸残基。 复合物的类型 在表面上形成的将使用交叉链接试剂（包括）定义 Hageman因子（HF）的可能聚集以及与已知的相互作用 含有前卡利氏蛋白或因子XI和HMW-金蛋白原的复合物。 我们将 还确定C1INH是否控制HF自动激活率和 检验以下假设，即启动表面会阻碍 绑定，由C1INH激活HF。 定量复合物的测定 血浆中活化的HF和C1INH将使用单克隆进一步完善 将确定抗血清和产生的活化HF的物种。 辅因子HMW差异的研究将集中在以下 问题：1）它是一个可活化的辅助因子，即，一个或 更多的债券增加了其内在的辅助活性或影响其能力 与其他血浆蛋白竞争以结合表面； 2）是什么 当键被不同的酶消化时，粘结裂解的顺序和 产生的凝结链的大小是多少？ 3）单克隆 将生产到最小的凝结链种类的抗体 表征该链的部分的表面相互作用或 结合底物prekallikrein和HMW-肌氨酸； 4）抗体 切割的分子可用于设计仅检测的测定法 血浆中的形式。 最后，我们将研究HF依赖性和 血浆的独立纤维蛋白水解途径，并确定血浆是否存在 prourokinase在任何一个中都起作用。 补体系统在 将确定血浆纤维蛋白溶解，新的纤溶酶原激活剂将是 寻求的，任何纤溶酶原激活剂或新的纤溶酶都将是 纯化和特征。