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ROLE OF PROTEOLYSIS BY CANP IN PLATELET ACTIVATION

CANP 蛋白水解在血小板激活中的作用

基本信息

批准号：
3448824
负责人：
JANICE R OKITA
金额：
$ 5.54万
依托单位：
VERSITI WISCONSIN, INC.
依托单位国家：
美国
项目类别：
财政年份：
1985
资助国家：
美国
起止时间：
1985-04-01 至 1988-03-31
项目状态：
已结题

项目摘要

Upon stimulation of platelets, a number of different events occur. One of these events is the activation of a calcium-activated neutral protease (CANP). This enzyme catalyzes the limited proteolysis of several proteins which are intimately involved in platelet function: glycoprotein Ib, actin-binding protein, von Willebrand factor, figrinogen and protein kinase C. In order to increase our knowledge of the mechanisms involved in platelet activation, it is essential that we understand the structure and function of these proteins and how they are modified in response to stimuli. The objectives of this proposal are to characterize CANP and to investigate mechanisms by which CANP may be involved in platelet activation. CANP has been purified from human platelets and partially characterized by other investigators. In this investigation, CANP will be purified and further characterized with respect to substrate specificity and the relationship between the two forms of CANP. One form of CANP (which requires a high concentration of calcium ions) may be converted by autoproteolysis to the other form (which requires a lower, more physiological, concentration of calcium ions). An important component of the methodology used in this project will be the development of a murine monoclonal antibody against CANP. The monoclonal antibody will be used to determine the subcellular localization of CANP by immunoelectron microscopy. This proposal will address several specific mechanisms by which CANP may be involved in platelet activation. Are the CANP-sensitive high molecular weight complexes of glycoprotein Ib and actin-binding protein degraded upon activation of platelets? Is the activation of protein kinase C by CANP involved in any of the various mechanisms of platelet activation by different stimuli? Does CANP play a role in the activation of phospholipase C? Is phospholipase A2 present in platelet membranes as an inactive proenzyme which can be activated by CANP? This investigation will lead to a better understanding of the events that take place upon platelet stimulation and aggregation and thus provide new insight into the etiology of thrombotic and hemostatic disorders.

当刺激血小板时，会发生许多不同的事件。其中之一这些事件是钙激活的中性蛋白酶的激活。 (CANP)。这种酶催化几种蛋白质的有限蛋白质分解。它们与血小板功能密切相关：糖蛋白Ib，肌动蛋白结合蛋白、血管性血友病因子、纤溶酶原和蛋白激酶 C.为了增加我们对所涉及的机制的了解血小板活化，我们必须了解其结构和功能这些蛋白质的功能以及它们是如何对刺激作出反应的。这项提案的目标是描述CANP的特征并调查 CANP可能参与血小板活化的机制。CANP有是从人的血小板中提纯出来的，并由其他调查人员。在这项研究中，CANP将被提纯并进一步具有底物专一性的特征及其与在这两种形式的CANP之间。一种形式的CANP(需要很高的钙离子浓度)可通过自身蛋白分解转化为其他形式(需要较低的、更具生理性的钙离子)。在这方面使用的方法的一个重要组成部分该项目将开发一种鼠源性抗病毒单抗。 CANP。单抗将被用来确定亚细胞 CANP的免疫电子显微镜定位。这项提案将涉及CANP可通过的几个具体机制参与了血小板的激活。是CANP敏感的大分子吗糖蛋白Ib和肌动蛋白结合蛋白的重量复合体降解血小板的激活？是CANP对蛋白激酶C的激活参与任何一种不同的血小板激活机制不同的刺激？CANP是否在激活中起作用？磷脂酶C？血小板膜上是否存在磷脂酶A2 CANP能激活哪些酶是失活的？这项调查将使我们更好地了解在血小板刺激和聚集时发生，从而提供新的深入了解血栓和止血疾病的病因学。