MOLECULAR BIOLOGY OF COLLAGEN RECEPTORS

描述：（研究者摘要）血小板GPVI和整合素α 2/β 1密切合作，介导有效的血小板粘附胶原蛋白， 随后血小板促凝活性。在初步调查结果中，我们有 观察到血小板GPVI的水平与α 2/β 1。α 2/β 1或GPVI本身密度的差异 可能不会损害血小板功能，否则健康的人，但他们 可能会影响风险增加的个体的临床结果， 血栓形成或出血。为了全面了解 血小板粘附胶原蛋白，我们的目标是研究的分子生物学 整合素α 2/β 1和血小板GPVI。该补助金的重点是 后者受体，血小板GPVI。为了实现我们的目标，我们开发了新的 首先，我们分离了含有人GPVI的BAC质粒， 基因，我们已经组装了完整的人类基因序列。二是 克隆并测序了小鼠血小板GPVI cDNA， 序列被用来分离含有完整鼠源性DNA的BAC质粒。 GPVI基因。有了这些资源，那么，这个应用程序的具体目标 方法：1）分析GPVI血小板含量与血小板密度的关系， 整合素α 2/β 1在正常人中的作用，并评估 受体变异对血小板促凝活性和血小板粘附的影响 2）表征GPVI上胶原蛋白的结合位点， （3）鉴定人GPVI基因，并确定GPVI基因的结构。 血小板GPVI表达变异的分子基础;和4）开发一种 用转基因技术建立GPVI缺陷小鼠模型。成功 建议的研究完成后， 了解血小板胶原蛋白受体的遗传学和生理学， GPVI。

DESCRIPTION: (Investigator's abstract) Platelet GPVI and integrin alpha 2/beta 1 cooperate closely to mediate effective platelet adhesion to collagens and subsequent platelet procoagulant activity. In preliminary findings, we have observed that the levels of platelet GPVI vary in parallel with those of alpha 2/beta 1. Differences in the density of alpha 2/beta 1 or GPVI in themselves may not compromise platelet function in otherwise healthy individuals, but they can influence clinical outcomes in individuals who are at increased risk for thrombosis or bleeding. In order to develop a comprehensive understanding of platelet adhesion to collagens, our goal is to study the molecular biology of both integrin alpha 2/beta 1 and platelet GPVI. The focus of this grant is the latter receptor, platelet GPVI. To accomplish our goal, we have developed new resources: First, we have isolated a BAC plasmid containing the human GPVI gene, and we have assembled the complete human gene sequence. Second, we have cloned and sequenced murine platelet GPVI cDNA, and the knowledge of this sequence was employed to isolate BAC plasmids containing the complete murine GPVI gene. With these resources, then, the Specific Aims of this application are: 1) To analyze the platelet content of GPVI vis-a-vis the density of integrin alpha 2/beta 1 among normal individuals and to assess the effect of receptor variation on platelet procoagulant activity and platelet adhesion to collagens; 2) To characterize the binding sites on GPVI for collagen and convulxin (CVX); 3) To characterize the human GPVI gene and define the molecular basis for variation in platelet GPVI expression; and 4) To develop a mouse model for GPVI deficiency using transgenic technology. The successful completion of the proposed studies will lead to a substantial increase in our understanding of the genetics and physiology of the platelet collagen receptor, GPVI.