MOLECULAR MECHANISMS OF PLATELET ACTIVATION

我们研究的总体目标是描绘生化途径。 它们在由激动剂结合到 表面受体，并导致血小板分泌，激活 纤维蛋白原结合部位和聚集。作为我正在进行的研究的一部分 在这条调查路线上，我们准备并隔离了一个刺激 单抗(M.Ab)，命名为F-11。M.A.b F-11作为激动剂 可诱导人血小板的分泌和聚集。临床部 激活血小板的抗体的重要性已经得到了很好的证明，但 作为受体的表面抗原的详细特征 在这个过程中，由这种抗体触发的生化途径 还没有被划定。我们希望对M.Ab的详细研究。F-11 将开始填补这些空白。M.AbF-11的使用提供了两种截然不同的 优点：(1)抗体可识别的蛋白质。血小板上的F-11 表面已在我们实验室进行了鉴定和部分纯化。 因此，我们的研究现在集中在一个激活过程，由一个 分子实体已知的受体。(2)激活 用纯化的抗结核分枝杆菌抗体抗体制备人血小板。F-11的滞后时间为6至 20分钟。此等待时间段允许详细测量 导致血小板分泌和聚集的分子事件序列。 因此，在此提交的研究计划旨在实现 具体目标如下：(A)提纯非变性形式的 F-11受体，(B)F-11抗原的特性和研究 至于它是否是唯一的受体，对于一个人来说，是受体复合体的一部分 天然产生的血小板激动剂，或已知的 信号转导系统，(C)生化途径的测定(S) 在F-11激活血小板的过程中工作，(D)发展 用于纯化的F-11受体的多克隆和单克隆抗体 在血小板F-11抗原的功能区，(E)克隆、测序 以及F-11受体基因的染色体定位。我们预计 这些研究目标的实现将提供新的和 关于血小板作用的基本机制的重要信息 激活。这些研究对健康的影响是 尤其与涉及相互作用的病理生理状态有关 抗体与循环中的血小板有关。

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.