HUMAN THROMBINS

凝血酶在止血和创伤中的中枢生物调节作用 治愈。它也是心血管疾病的主要参与者。 (如冠状动脉血栓形成、中风、肺栓塞)，并已 与其他疾病过程有关(例如，动脉粥样硬化、癌症、 炎症反应)。与其他丝氨酸蛋白酶不同，促凝剂 α-凝血酶是一种多功能的兼有酶和 非酶(类激素)活性。十多年来发展的方法 之前由我们实验室用于制备人α-凝血酶已使 约700种出版物和对化学和化学的快速探索 凝血酶生物学。我们的研究主要集中在 结构-功能关系；我们已经确定了3个(可能是5个) 决定凝血酶特异性和生物学特性的结构区域 功能。我们下一个五年的目标是：i)生产 5至10克/年的人α-凝血酶和其他形式(例如，浸泡- α、伽马、β-凝血酶)，ii)进一步绘制地形图 凝血酶结构/功能区域通过特定的切割(例如， XI-凝血酶)和与此相对应的抗体合成肽 区域或蛋白质结构域，iii)使用重组水飞蓟素 标准化α-凝血酶活性(没有简单、绝对的方法来 日期)，并评估具有约束性的要求 工程类似物，iv)开发具有生物活性的标记凝血酶 (例如，荧光α-凝血酶、生物素结合物)用于生物 探测器，五)继续进行重大协作研究(例如，X射线 细胞生物学的结晶学)。我们的整体方法具有成本效益。 科学，跨越多个学科，以最大限度地提高我们的 投稿(例如，前三年期间的42份出版物)。

Thrombin has central bioregulatory functions in hemostasis and wound healing. It also is a major participant in cardiovascular diseases (e.g., coronary thrombosis, stroke, pulmonary embolism) and has been implicated in other disease processes (e.g., atherosclerosis, cancer, inflammatory responses). Unlike other serine proteinases, procoagulant alpha-thrombin is multi-functional possessing enzymic as well as nonenzymic (hormone-like) activities. Methods developed over 10 years ago by our laboratory for preparing human alpha-thrombin have enabled some 700 publications and the rapid exploration of the chemistry and biology of thrombin. Our research has focused primarily on structure-function relationships; we have identified 3 (possibly 5) structural regions accounting for thrombin specificity and biological functions. Our objectives for the next 5-year period are to: i) produce 5 to 10 g/year of human alpha-thrombin and other forms (e.g., DIP- alpha,gamma,epsilon-thrombins), ii) further topographical mapping of thrombin structural/functional regions by specific cleavages (e.g., xi-thrombin) and antibodies synthetic-peptides corresponding to such regions or protein domains, iii) use erecombinant hirudins for standardizing alpha-thrombin activities (no simple, absolute method to date) and assess binding requirements with synthetic and genetically engineered analogues, iv) develop biologically-active tagged thrombins (e.g., fluorescent alpha-thrombin, biotin conjugates) for biological probes, and V) continue major collaborative studies (e.g., x-ray crystallography of cell biology). Our overall approach is cost-effective science, working throughout many disciplines in order to maximize our contributions (e.g., 42 publications over the preceding 3-year period).