PROTEIN ENGINEERING OF TRYPSIN & ECOTIN

胰蛋白酶的蛋白质工程

• 批准号: 6119229
• 负责人: Qing X Yang
• 金额: $ 0.54万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-07-01 至 2000-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6119229
• 关键词: bioengineering /biomedical engineering; biological products; biomedical resource; enzymes

We are investigating the macromolecular recognition and inhibition between serine proteases and their inhibitors. In particular, we are studying the origin of the broad inhibitory specificity of ecotin, an E. coli protease inhibitor. The goal of this study is to understand the molecular basis of ecotin's inhibitor specificity and introduce novel specificity into ecotin to target disease-related serine proteases. Based on the high resolution crystal structure of trypsin-ecotin complex, we have designed a series of ecotin variants at the two loops of ecotin and tested these mutants against different proteases. The result suggests a unique mode of inhibition of ecotin through a secondary binding site that does not occur in other serine protease inhibitors. Using phage display techniques, we have isolated several potent inhibitors against a serine protease, urokinase-type plasminogen activator, that plays an important role in cancer. This experiment validates a novel approach to design specific protease inhibitors. The molecular modeling of ecotin loop mutants is accomplished w ith MidasPlus software available in the Computer Graphics Laboratory.

我们正在研究大分子的识别和抑制 丝氨酸蛋白酶及其抑制剂之间的关系。 我们尤其 研究ecotin广泛抑制特异性的起源， E.大肠杆菌蛋白酶抑制剂。 这项研究的目的是了解 ecotin抑制剂特异性的分子基础，并介绍了 靶向疾病相关丝氨酸的ecotin的新特异性 蛋白酶 基于高分辨率的晶体结构， 胰蛋白酶-ecotin复合物，我们设计了一系列ecotin变体 在两个环的ecotin和测试这些突变体对不同的 蛋白酶 这一结果提示了ecotin的一种独特的抑制模式 通过在其他丝氨酸中不存在的二级结合位点 蛋白酶抑制剂 利用噬菌体展示技术，我们分离了 几种有效的丝氨酸蛋白酶抑制剂，尿激酶型 纤溶酶原激活剂，在癌症中起重要作用。 本实验验证了一种新的方法，设计特定的 蛋白酶抑制剂 ecotin环突变体的分子建模是 使用计算机中的MidasPlus软件完成 图形实验室