PHOSPHOLIPID-TARGETED FIBRINOLYTIC AGENTS

这项研究的长期目标是开发新的方法来 检测和治疗血栓形成。在这个项目中，一种新的方法将是 为检测和靶向促凝剂磷脂而开发的 曝光。这种方法是基于磷脂结合蛋白。 Annexin V，与活化的血小板高亲和力结合，并与 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剂 膜联蛋白V与单链尿激活酶、链激酶联用 纤溶酶原。杂交蛋白质将根据分子量进行评估， 酶活性，结合到促凝剂磷脂膜，结合到 静息和激活的血小板以及体外和体内的纤溶作用 活动。 2.重组杂合蛋白的制备及鉴定 目标1下描述的研究。 3.评估Annexin V作为检测、靶向和成像试剂的作用 体内血栓形成。 4.定点研究膜联蛋白V的结构/功能关系 磷脂结合位点(S)的突变定位 表面。 5.评估Annexin V作为潜在的内源性调节因子 研究动脉粥样硬化在正常和动脉粥样硬化中的表达 人类的冠状动脉。 这些研究代表了一种新的方法来解决检测和 治疗冠状动脉和其他部位的血栓形成。他们还将 有助于了解膜联蛋白家族及其相关的基础知识 可能参与血管病理和动脉粥样硬化。

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.