PHOSPHOLIPID-TARGETED FIBRINOLYTIC AGENTS

这项研究的长期目标是开发新的方法， 检测和治疗血栓形成。 在这个项目中，一种新的方法将是 开发用于检测和靶向促凝血磷脂 exposure. 这种方法是基于磷脂结合蛋白 膜联蛋白V，其以高亲和力与活化的血小板结合， 含有磷脂酰丝氨酸的膜。 具体目标是： 1.化学法制备磷脂靶向纤溶药物 膜联蛋白V与单链尿激酶、链激酶和 纤溶酶原 将评价杂合蛋白的分子量， 酶活性，与促凝血磷脂膜结合，与 静息和活化血小板，以及体外和体内纤溶 活动 2.制备和表征重组杂合蛋白，以延伸 目标1所述的研究。 3.评价膜联蛋白V作为检测、靶向和成像剂的作用 体内血栓 4.膜联蛋白V的结构与功能关系的研究 诱变以定位磷脂的结合位点 表面。 5.评价膜联蛋白V作为一种潜在的内源性调节剂， 通过研究其在正常和动脉粥样硬化中的表达， 人体冠状动脉 这些研究代表了一种新的方法来检测和 治疗冠状动脉和其他地方的血栓形成。 他们还将 有助于了解膜联蛋白家族及其 可能参与血管病理学和动脉粥样硬化。

The long-term objective of this research is to develop new approaches for detecting and treating thrombosis. In this project, a new method will be developed for detection and targeting of procoagulant phospholipid exposure. This approach is based on the phospholipid binding protein annexin V, which binds with high affinity to activated platelets and to membranes containing phosphatidylserine. The specific aims are: 1. Prepare phospholipid-targeted fibrinolytic agents by chemical crosslinking of annexin V with single-chain urokinase, streptokinase and plasminogen. The hybrid proteins will be evaluated as to molecular weight, enzyme activity, binding to procoagulant phospholipid membranes, binding to resting and activated platelets, and in vitro and in vivo fibrinolytic activity. 2. Prepare and characterize recombinant hybrid proteins in order to extend the studies described under Aim 1. 3. Evaluate annexin V as an agent for detecting, targeting and imaging thrombi in vivo. 4. Study the structure/function relationships of annexin V by site-directed mutagenesis in order to localize the binding site(s) for phospholipid surfaces. 5. Evaluate annexin V as a potential endogenous regulator in atherosclerosis by studying its expression in normal and atherosclerotic human coronary arteries. These studies represent a novel approach to the problem of detecting and treating thrombosis in the coronary arteries and elsewhere. They will also contribute to basic knowledge about the annexin protein family and its potential involvement in vascular pathology and atherosclerosis.