METAL-DEPENDENT COMPLEXES IN BLOOD COAGULATION

血液凝固中的金属依赖性复合物

• 批准号: 3485593
• 负责人: BARBARA C FURIE
• 金额: $ 32.76万
• 依托单位: TUFTS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1978
• 资助国家: 美国
• 起止时间: 1978-06-01 至 1998-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3485593
• 关键词: affinity chromatography; blood coagulation; blood coagulation disorders; calcium binding protein; conformation; dialysis; fluorescence spectrometry; gamma carboxyglutamate; hemostasis; human tissue; immunochemistry; laboratory mouse; ligands; membrane proteins; metal complex; monoclonal antibody; platelets; protein sequence; prothrombin; surface antigens; thrombin; thrombosis; zymogens

The understanding of the role of metal ions in blood coagulation has been a long term goal of this laboratory. To further the understanding of the structure-function relationships of the Gamma-carboxyglutamic acid residues in prothrombin to the metal liganding processes of the protein we will study a naturally occurring variant of prothrombin which contains only three Gamma-carboxyglutamic acid residues. Normal prothrombin contains ten Gamma-carboxyglutamic acid residues. We will correlate the sequence position of the Gamma-carboxyglutamic acid residues in the variant with its metal binding and conformational properties using amino acid sequence analysis, state rate dialysis, fluorescence spectroscopy and conformation specific antibodies. The results of these analyses will be used to compare and contrast the properties of normal prothrombin and the variant prothrombin. Platelets secrete Ca(II) upon activation. There is potential for a significant increase in Ca(II) concentration at the site of platelet plug formation. In addition, Ca(II) dependent conversion of several plasma zymogens may occur on the platelet surface. We will use monoclonal conformation specific antibodies to identify platelet surface antigens whose conformations are altered depending on the presence or absence of metal ligands. We will correlate the presence of the antigen to the status of the platelet (native or activated). We will identify a possible receptor function for the platelet surface antigens recognized by these monoclonal antibodies. Finally we have identified a novel cleavage product of human prothrombin in plasma, fragment 1.2.3. We will access a potential regulatory role for this fragment in hemostasis. There is currently heightened interest in understanding the processes that regulate hemostasis and thrombosis. The studies outlined in this proposal address these biologically interesting and medically important questions.

对金属离子在血液凝血中的作用的理解一直是 这个实验室的长期目标。 进一步理解 γ-羧基谷氨酸残基的结构 - 功能关系 在将蛋白质的金属配体过程的凝血酶原中 研究一种天然存在的凝血酶原变体，仅包含 三个γ-羧基谷氨酸残基。 普通凝血酶蛋白包含十个 γ-羧基谷氨酸残基。 我们将关联序列 γ-羧基谷氨酸残基在该变体中的位置 使用氨基酸序列的金属结合和构象性能 分析，州速率透析，荧光光谱和构象 特定抗体。 这些分析的结果将用于比较 并对比正常凝血酶原和变体的特性 凝血酶原。 激活后血小板分泌Ca（II）。 有潜力 对于血小板部位的Ca（II）浓度显着升高 插头组。 此外，CA（ii）几个等离子体的依赖性转化 酶原可能发生在血小板表面。 我们将使用单克隆 构象特异性抗体以鉴定血小板表面抗原 其构象的改变，具体取决于存在或不存在 金属配体。 我们将将抗原的存在与状态相关联 血小板（天然或激活）。 我们将确定可能 这些识别的血小板表面抗原的受体功能 单克隆抗体。 最后，我们确定了一种新型的乳沟产品 血浆中的人类凝血酶原，片段1.2.3。 我们将获得潜力 该碎片在止血中的调节作用。 目前有 对了解调节止血的过程的兴趣提高了 和血栓形成。 该提案中概述的研究解决了这些 生物学上有趣且在医学上重要的问题。