FUNCTION OF PLATELETS AND COAGULATION FACTORS

这项资助的目的是阐明止血的基本机制。 并制定策略来改变血栓形成的自然历史 精神错乱。有两个项目。第一个问题涉及到 血小板、白细胞和血管内皮细胞的启动和控制 凝血反应。血小板上的凝血酶原激活将是 用均一因子Va及其组成多肽进行了研究。角色 Va因子在蛋白C激活中的作用将使用分离的 Va因子多肽。血栓调节蛋白与凝血因子Va、2的关系 同时参与蛋白C激活的蛋白质将被研究 利用培养的内皮细胞和抗血栓调节蛋白和抗体 因子Va。抗蛋白C、凝血因子Va及其组分的单抗 将会准备好锁链。激活因子V的血小板蛋白水解酶 会被提纯及其在启动血小板表面的作用 将使用特异性抗体研究凝血酶原激活情况 因子Va和免疫印迹技术。第二个项目涉及 与血小板摄取和释放二十烷类前体的机制 和培养的细胞。花生四烯酸辅酶A的脂肪酸专一性 合成酶将使用各种14C标记的方法来阐明 多不饱和脂肪酸及其与高亲和力脂肪酸的比较 血小板摄取能力。脂肪酸的专一性 激动剂诱导的血小板脂肪酸释放机制也将是 已澄清。花生四烯酰辅酶A合成酶将纯化至均一 并准备好抗体。自杀后的选择技巧 突变将用于选择花生四烯酸摄取和释放突变体 HSD小鼠纤维肉瘤细胞。突变株将通过克隆和分离 在涤纶布上复制电镀。突变的细胞系将在 这些脂肪酸的摄取和释放机制的阐明 细胞。

The objective of this grant is to elucidate basic mechanisms of hemostasis and to devise strategies to alter the natural history of thrombotic disorders. There are two projects. The first deals with the role of platelets, leukocytes, and endothelial cells in the initiation and control of coagulation reactions. Prothrombin activation on platelets will be studied using homogeneous factor Va and its component peptides. The role of factor Va in protein C activation will be elucidated using the isolated factor Va peptides. The relation between thrombomodulin and factor Va, two proteins that both participate in protein C activation, will be studied using cultured endothelial cells and antibodies to thrombomodulin and factor Va. Monoclonal antibodies to protein C, factor Va and its component chains will be prepared. The platelet protease that activates factor V will be purified and its role in the initiation of platelet surface prothrombin activation will be investigated using antibodies specific for factor Va and the western blotting technique. The second project deals with the mechanism of eicosanoid precursor uptake and release by platelets and cultured cells. The fatty acid specificity of arachidonoyl CoA synthetase will be elucidated using a variety of 14C-labeled polyunsaturated fatty acids and compared to the high affinity fatty acid uptake capacity of platelets. The fatty acid specificity of the agonist-induced fatty acid release mechanism of platelets will also be elucidated. Arachidonoyl CoA synthetase will be purified to homogeneity and antibodies prepared. The technique of suicide selection after mutagenesis will be used to select arachidonate uptake and release mutants of HSD mouse fibrosarcoma cells. Mutants will be isolated by cloning and replica-plating on polyester cloth. Mutant cell lines will be useful in the elucidation of mechanisms of fatty acid uptake and release in these cells.