C1 INHIBITOR AND PAI-1--STRUCTURE-FUNCTION STUDIES

纤溶酶原激活物抑制剂-1（派-1）和C1-抑制剂是丝氨酸 蛋白酶抑制剂（丝氨酸蛋白酶抑制剂）存在于血浆中，其控制 激活纤维蛋白溶解和内源性凝血途径 分别 派-1过量被认为是一种危险因素， 血栓形成和C1抑制剂缺乏引起血管性水肿。 塞尔平斯法案 通过与它们的靶蛋白酶形成稳定的复合物， 其没有详细地明确定义。 该项目的重点是 研究这两种丝氨酸蛋白酶抑制剂的结构与功能关系， 目的是进一步阐明丝氨酸蛋白酶抑制剂的作用机制， 鉴定丝氨酸蛋白酶抑制剂活性调节的机制。 蛋白 肽化学和免疫学技术将用于 回答四个主要问题：a）结构上的修改是什么 当C1抑制剂在其反应位点被切割时， 这与抑制活性有关吗？B）分配比是多少 C1-抑制剂和派-1与其靶蛋白酶的反应 受肝素和其他配体（如玻连蛋白？）调节; C.哪些 C1-抑制剂和派-1上的位点稳定了与其 靶向蛋白酶，这些相互作用可以用来设计 丝氨酸蛋白酶抑制剂功能的抑制剂？以及d）玻连蛋白如何稳定 派-1的活性形式以及派-1上的结合位点是什么 玻连蛋白？这项研究将有助于更好地了解 丝氨酸蛋白酶抑制剂机制，以及调节蛋白水解反应， 过程，如血栓形成和溶解，以及指示 治疗干预的可能目标。

Plasminogen activator inhibitor-1 (PAI-1) and C1-inhibitor are serine proteinase inhibitors (serpins) present in plasma which control the activation of the fibrinolytic and intrinsic coagulation pathways respectively. PAI-1 excess is implicated as a risk factor for thrombosis, and C1-inhibitor deficiency causes angioedema. Serpins act by forming a stable complex with their target proteinase by mechanisms which are not clearly defined in detail. The focus of this project is to study the structurefunction relationships of these two serpins, with the aim of further elucidating the mechanism of serpin action, and identifying mechanisms for the regulation of serpin activity. Protein and peptide chemistry, and immunological techniques will be used to answer four main questions: a) what are the structural modifications observed on C1-inhibitor when it is cleaved at its reactive site, and how does this relate to inhibitory activity?; b) how is the partition ratio for the reaction of C1-inhibitor and PAI-1 with their target proteinases regulated by heparin and other ligands such as vitronectin?; c) which sites on C1-inhibitor and PAI-1 stabilize the interaction with their target proteinases, and can these interactions be used to design inhibitors of serpin function?;. and d) how does vitronectin stabilize the active form of PAI-1 and what are the binding sites on PAI-1 for vitronectin? This research will lead to a better understanding of the serpin mechanism, and the regulation of the proteolytic reactions of processes such as thrombus formation and lysis, as well as indicating possible targets for therapeutic intervention.