BIOCHEMICAL PATHWAYS OF PLATELET ACTIVATION

血小板激活的生化途径

• 批准号: 3343971
• 负责人: ELIZABETH H KORNECKI
• 金额: $ 11.54万
• 依托单位: SUNY DOWNSTATE MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-08-01 至 1990-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3343971
• 关键词: G protein; adenylate cyclase; affinity chromatography; antigen antibody reaction; arachidonate; binding proteins; biological signal transduction; calcium flux; chemical binding; fibrinogen; fibrinogen receptors; fluorescent dye /probe; gel electrophoresis; human subject; immunochemistry; laboratory mouse; membrane activity; membrane proteins; monoclonal antibody; phosphatidylinositols; phosphorylation; platelet activation; platelet aggregation; prostaglandin E; protein kinase C; protein structure function; radiotracer; secretion; serotonin

The overall goal of this 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 our ongoing studies in this line of investigation, we have prepared and isolated a stimulatory monoclonal antibody (M.Ab.), named F- 11. M.Ab. F-11 acts as an agonist which induces secretion and aggregation of human platelets. The clinical significance of antibodies which activate platelets is well documented, but detailed characterization of the surface antigens that serve as receptors in this process and the 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 characterized 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 which is dependent on M.Ab. F-11 concentration. 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: 1) Purification of a non-denatured form of the platelet surface protein recognized by M.Ab. F-11; 2) 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, 3) Determination of the biochemical pathway(s) operating in the activation of platelets by F-11; and, 4) Development of polyclonal and monoclonal antibodies to the purified F-11 receptor, for use in cellular localization, tissue distribution, and for immunological mapping of functional domains of the platelet F-11 antigen. 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.

这项研究的总体目标是描述生化 通过结合开始的过程中运行的途径 表面受体的激动剂，并导致血小板分泌， 纤维蛋白原结合位点的激活和聚集。 作为 我们正在进行的调查研究中，我们已经准备了 并分离了一种刺激性单克隆抗体（M.Ab。），称为f- 11。M.AB。 F-11充当诱导分泌和的激动剂 人血小板的聚集。 临床意义的 激活血小板的抗体有充分的文献记录，但 表面抗原的详细表征 在此过程中的受体和触发的生化途径 通过这种抗体，尚未描述。 我们期望这一点 M.Ab的详细研究F-11将开始填补这些空白。 这 M.Ab的使用F-11提供了两个不同的优势：1） M.Ab识别的蛋白质血小板表面上的F-11已经 在我们的实验室中确定并部分表征。 因此，我们的 现在的研究重点是由受体启动的激活过程 其分子实体已经知道。 2）激活 M.Ab的纯化IgG的人血小板。 F-11涉及滞后 6至20分钟的周期取决于M.Ab。 F-11 专注。 该潜伏期允许详细 测量分子事件的序列，导致 血小板分泌和聚集。 因此，研究 此处提交的计划旨在实现以下特定 目标：1）纯化血小板的非定型形式 M.Ab识别的表面蛋白F-11; 2）表征 F-11抗原和有关它是否是唯一的调查 受体，一部分自然的受体复合物 发生的血小板激动剂或已知信号的成分 转导系统，3）确定生化 F-11在血小板激活中运行的途径；而且，4） 对多克隆和单克隆抗体的开发 纯化的F-11受体，用于细胞定位，组织 分布以及功能域的免疫学映射 血小板F-11抗原。 我们期望成就 这些研究目标将提供新的重要信息 关于在血小板激活过程中运行的基本机制。 这 这些研究的与健康有关的含义尤其是 与涉及相互作用的病理生理状态有关 带有循环血小板的抗体。