HUMAN FACTOR VII & TISSUE FACTOR

人为因素七

• 批准号: 3348953
• 负责人: Walter Kisiel
• 金额: $ 16.3万
• 依托单位: UNIVERSITY OF NEW MEXICO
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-12-01 至 1993-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3348953
• 关键词: activation product; biological products; blood coagulation tests; calcium binding protein; chemical binding; coagulation factor IX; coagulation factor VII; coagulation factor X; coagulation factor XII; enzyme complex; enzyme mechanism; enzyme substrate; genetic manipulation; hemostasis; human tissue; laboratory rabbit; membrane activity; mutant; peptidases; protein purification; protein structure function; proteolysis; thrombin; thromboplastin; tissue /cell culture; vascular endothelium; zymogens

Human blood coagulation factor VII circulates in blood as a precursor of a serine-protease. The expression of the proteolytic activity of factor VII, or its activated form, factor VIIa, occurs as a result of complex formation with a specific cell surface glycoprotein designated as tissue factor. While many of the steps involved in the coagulation cascade have been studied on a molecular basis, little is known as to how the activity of factor VII towards protein substrates is augmented several order of magnitude by complex formation with tissue factor in the presence of calcium ions, and how this activity, once formed, is regulated in vivo. The specific aims of the proposed research address the issues of: (a) the interaction, procoagulant activity and substrate specificity of human factor VIIa on cultured endothelial cells in both the quiescent and perturbed states, (b) the intrinsic proteolytic activity of zymogen factor VII and mechanisms of its proteolytic activation in vivo, and (c) the biochemical properties of plasma-derived, recombinant wild-type and site-specific mutants of human factor VII. One of the long-range goals of this project is to obtain a better understanding of the relative rates of the factor VIIa-tissue factor mediated activation of factor IX and factor X as it occurs during hemostasis. Another, equally important goal of this project is to identify the protease(s) and any cofactor(s) necessary for optimal proteolytic activation of zymogen factor VII in vivo and elucidate the mechanism of this reaction. The proposed research will utilize standard protein isolation and characterization procedures, established coagulation assays, currently employed tissue culture techniques and protocols, affinity chromatography and amino acid sequence methodology. Hopefully, information gathered in this study will provide insight into the precise role of factor VII in both normal and pathological states, and inferences as to the role of extrinsic coagulation in the pathogenesis of various thromboembolic disorders including transient cerebral ischemic attacks, vein and artery occlusion, atherosclerosis, and deep venous thromboembolism. It is further hoped that, based on the results of these studies, future therapies will evolve that restrict the deleterious effects of tissue factor and be effective in the prophylaxis of thromboembolic disorders.

人体血液凝血因子VII在血液中循环作为一个 丝氨酸保护的前体。 蛋白水解的表达 因子VII的活性或其活化形式的因素VIIA发生 由于与特定细胞表面的复杂形成 糖蛋白被指定为组织因子。 而许多步骤 参与凝血级联 分子基础，几乎不知道因子的活性 VII朝向蛋白质底物的vii被增强 在存在下与组织因子形成复合物的大小 钙离子以及一旦形成的这种活性如何受到调节 体内。 拟议研究的具体目的是针对 问题：（a）互动，proc凝活动和 人类因子VIIA对培养内皮的底物特异性 静态和扰动状态的细胞，（b）固有 酶原因子VII及其机制的蛋白水解活性 体内蛋白水解活化，（c）生化特性 血浆衍生的重组野生型和位点特异性突变体的 人类因素VII。 该项目的远程目标之一 是为了更好地了解 VIIA组织因子因子介导的因子IX和 因子X在止血期间发生。 另一个，同样 该项目的重要目标是确定蛋白酶和 最佳蛋白水解激活所必需的任何辅助因子 Zymogen因子VII体内并阐明了这种机制 反应。 拟议的研究将利用标准蛋白 隔离和表征程序，已建立的凝结 测定，目前采用的组织培养技术和方案， 亲和色谱和氨基酸序列方法论。 希望这项研究收集的信息将提供洞察力 进入正常和病理中因子VII的精确作用 国家，以及关于外部凝血作用的推论 包括 瞬时脑缺血性发作，静脉和动脉阻塞， 动脉粥样硬化和深层静脉血栓栓塞。进一步 希望根据这些研究的结果，未来的疗法 将进化限制组织因子的有害影响 并有效地预防血栓栓塞性疾病。