CELL/CELL INTERACTIONS IN THROMBOSIS

描述：这份续签申请提出了一系列研究，以 阐明人脐血细胞的分子基础及其调控 静脉内皮细胞(HUVEC)胞外ADPase。该项目将进行克隆， 对HUVEC ADPase进行测序和表达，并对其进行生化鉴定； 研究结构/功能关系以确定关键区域 核苷酸结合和酶活性；确定其作用机制 溶血磷脂酰胆碱、同型半胱氨酸、细胞因子 和生长因子影响酶活性；并评估转录 受ADPase基因5‘-侧翼区元件的调控。 总之，本申请中提出的研究将阐明 人体表面一种酶的分子机制 内皮细胞从激活状态立即代谢ADP 血小板。为此，我们将对分离的HUVEC进行克隆、测序和表达 ADPase及其生化特征；研究结构/功能 确定酶活性关键区域的关系；确定 溶血磷脂酰胆碱等生物物质， 同型半胱氨酸、细胞因子和生长因子影响酶活性；以及 评估5‘-侧翼区元件对转录的调控 ADPase基因。本申请中提出的实验将提供 为今后的信息化发展提供合理有效的形式 抗血栓治疗将在最早的阶段起作用 血栓形成。

DESCRIPTION: This renewal application proposes a series of studies to elucidate the molecular basis and regulation of human umbilical vein endothelial cell (HUVEC) ecto-ADPase. The project will clone, sequence and express HUVEC ADPase and characterize it biochemically; study structure/function relationships to identify regions critical for nucleotide binding and enzyme activity; determine the mechanisms by which lysophosphatidylcholine (lysoPC), homocysteine, cytokines and growth factors affect enzyme activity; and assess transcriptional regulation by elements of the 5'-flanking region of the ADPase gene. In summary, research proposed in this application will elucidate molecular mechanisms by which an enzyme on the surface of human endothelial cells immediately metabolizes ADP emerging from activated platelets. To this end we will clone, sequence and express isolated HUVEC ADPase and characterize it biochemically; study structure/function relationships to identify regions critical for enzyme activity; determine mechanisms by which biologic substances such as lysophosphatidylcholine, homocysteine, cytokines and growth factors affect enzyme activity; and assess transcriptional regulation by elements of the 5'-flanking region of the ADPase gene. Experiments proposed in this application will furnish information for future development of rational and effective forms of antithrombotic therapy which will act at the very earliest stages of thrombus formation.