INTERACTIONS CONTROLLING VITAMIN K-DEPENDENT COAGULATION

本研究的长期目标是了解凝血系统中关键酶之间的相互作用。希望这些研究的信息将有助于改善治疗，不仅对出血性疾病的个人，而且对那些受更广泛的健康问题，血栓形成。 为了实现这些目标，研究将包括以下具体研究目标：1）鉴定与维生素K环氧化物还原酶协作产生维生素K氢醌的任何辅因子或其他辅助蛋白; 2）研究控制因子IX与胶原IV相互作用的结构元件及其在凝血中的作用; 3）表征药理剂量的因子Vila恢复止血的机制。 这些目标将通过使用多方面的方法来实现，从基础生物化学， 分子生物学到动物模型。研究将采用分子生物学，组织培养和生物化学方法来研究维生素K环氧化物还原酶与其他细胞蛋白质的相互作用。类似的分子生物学和组织培养方法将用于产生凝血因子IX和因子VII变体，其将在各种模型系统中进行评价，包括基于细胞的凝血模型和动物模型，包括因子IX缺陷小鼠和具有不结合胶原IV的因子IX的小鼠。将通过活体显微镜评价这些分子的体内作用。

The long-term goal of this research is to understand the controlling interactions among key enzymes in the blood coagulation system. It is hoped that information derived from these studies will help improve treatments for, not only individuals with bleeding disorders, but also for those affected by the broader health problem, thrombosis. To accomplish these goals the studies will include the following specific research aims: 1) identify any cofactors or other accessory proteins that collaborate with vitamin K epoxide reductase in producing vitamin K hydroquinone; 2) investigate the structural elements controlling factor IX interactions with collagen IV and its role in coagulation; and 3) characterize the mechanism by which pharmacologic doses of factor Vila restore hemostasis. These aims will be achieved by using multifaceted approaches ranging from basic biochemistry and molecular biology to animal models. Studies will employ molecular biology, tissue culture, and biochemical methods to investigate the interaction of vitamin K epoxide reductase with other cellular proteins. Similar molecular biology and tissue culture methods will be used to create coagulation factor IX and factor VII variants which will be evaluated in a variety of model systems including a cell-based coagulation model and animal models including factor IX deficient mice and mice with factor IX that does not bind collagen IV. In vivo effects of these molecules will be evaluated by intravital microscopy.