HUMAN THROMBINS

凝血酶在止血和创伤过程中具有重要的生物调节作用 治愈它也是心血管疾病的主要参与者 (e.g.,冠状动脉血栓形成、中风、肺栓塞）， 与其它疾病过程有关（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， α-凝血酶是多功能的，具有酶活性以及 非酶（类酶）活性。10年来开发的方法 我们的实验室以前用于制备人α-凝血酶的方法已经使 大约700篇出版物和对化学的快速探索， 凝血酶生物学我们的研究主要集中在 结构-功能关系;我们已确定3个（可能5个） 凝血酶特异性和生物学特性的结构区域 功能协调发展的我们未来五年的目标是：i）生产 5至10克/年的人α-凝血酶和其它形式（例如，DIP- α，γ，ε-凝血酶），ii）进一步的拓扑图绘制， 凝血酶结构/功能区通过特异性切割（例如， XI-凝血酶）和抗体合成的肽 区域或蛋白质结构域，iii）使用重组水蛭素， 标准化α-凝血酶活性（没有简单的、绝对的方法来 日期），并评估与合成和基因 工程化的类似物，iv）开发生物活性标记的凝血酶 (e.g.,荧光α-凝血酶、生物素缀合物）用于生物 探测器，和V）继续主要的合作研究（例如，x射线 细胞生物学结晶学）。我们的整体方法具有成本效益 科学，在许多学科工作，以最大限度地提高我们的 贡献（例如，前三年共出版42篇）。

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).