MOLECULAR MECHANISMS OF PLATELET ACTIVATION

血小板激活的分子机制

• 批准号: 3074478
• 负责人: ELIZABETH H KORNECKI
• 金额: $ 6.99万
• 依托单位: SUNY DOWNSTATE MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-09-30 至 1995-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3074478
• 关键词: affinity chromatography; antibody formation; biological signal transduction; genetic mapping; human genetic material tag; human subject; laboratory mouse; laboratory rabbit; membrane reconstitution /synthesis; molecular cloning; platelet activation; platelet aggregation; protein purification; receptor binding; surface antigens

The overall goal of our research is the delineation of biochemical pathways which operate in the process initiated by the binding of an agonist to surface receptors, and leading to platelet secretion, activation of fibrinogen binding sites and aggregation. As part of my ongoing studies in this line of investigation, we have prepared and isolated a stimulatory monoclonal antibody (M.Ab.), named F-11. M.A.b F-11 acts as an agonist which induces secretion and aggregation of human platelets. The clinical significant of antibodies which activate platelets is well documented, but detailed characterization of the surface antigens that serve as receptors in this process and biochemical pathways triggered by such antibodies has not yet been delineated. We expect that detailed studies of M.Ab. F-11 will begin to fill these gaps. The use of M.Ab F-11 provides two distinct advantages: (1) The protein recognized by M.Ab. F-11 on the platelet surface has been identified and partially purified in our laboratory. Thus, our studies focus now on an activation process initiated by a receptor whose molecular entity is already known. (2) The activation of human platelets by purified IgG of M.Ab. F-11 involves a lag period of 6 to 20 minutes. This latency period permits detailed measurements of the sequence of molecular events leading to platelet secretion and aggregation. Accordingly, the Research Plan submitted here is designed to achieve the following specific goals: (a) Purification of a non-denatured form of the F-11 receptor, (b) characterization of the F-11 antigen and investigation as to whether it is a unique receptor, part of the receptor complex for one of the naturally-occurring platelet agonists, or a component of a known signal transduction system, (c) Determination of the biochemical pathway(s) operating in the activation of platelets by F-11, (d) development of polyclonal and monoclonal antibodies to the purified F-11 receptor, for use in functional domains of the platelet F-11 antigen, (e) cloning, sequencing and chromosomal localization of the gene for the F-11 receptor. We expect that accomplishment of these research goals will provide new and significant information on basic mechanisms operating during platelet activation. The health-related implications of these studies are particularly relevant to pathophysiological states involving interaction of antibodies with circulating platelets.

我们研究的总体目标是描述生化途径 在激动剂与 表面受体，并导致血小板分泌，激活 纤维蛋白原结合位点和聚集。 作为我正在进行的研究的一部分 这条调查，我们已经准备了并隔离了刺激性 单克隆抗体（M.Ab。），名为F-11。 M.A.B F-11充当激动剂 这会诱导人血小板的分泌和聚集。 临床 有意义的抗体很重要，有充分的文献记载，但 表面抗原作为受体的详细表征 在此过程和此类抗体触发的生化途径中 尚未被描述。 我们期望M.Ab的详细研究F-11 将开始填补这些空白。 M.AB F-11的使用提供了两个不同的 优点：（1）M.Ab识别的蛋白质。血小板上的F-11 在我们的实验室中已经确定并部分纯化了表面。 因此，我们的研究现在集中于A 其分子实体已经知道的受体。 （2）激活 M.Ab的纯化IgG的人血小板。 F-11涉及6至6 20分钟。 该延迟期允许详细测量 分子事件的序列导致血小板分泌和聚集。 因此，此处提交的研究计划旨在实现 以下具体目标：（a）净化非定型形式的 F-11受体，（b）F-11抗原的表征和研究 关于它是否是独特的受体，一个受体复合物的一部分 天然血小板激动剂或已知的成分的 信号转导系统，（c）测定生化途径（S） 通过F-11激活血小板，（d）开发 纯化F-11受体的多克隆和单克隆抗体，供使用 在血小板F-11抗原的功能结构域中，（E）克隆，测序 F-11受体基因的染色体定位。 我们期望 这些研究目标的实现将提供新的和 有关在血小板过程中运行的基本机制的重要信息 激活。 这些研究的与健康有关的含义是 与涉及相互作用的病理生理状态特别相关 带有循环血小板的抗体。