UNDERSTANDING SPECIFICITY OF SERINE PROTEASES

了解丝氨酸蛋白酶的特异性

• 批准号: 6119275
• 负责人: SANDRA W RUGGLES
• 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/6119275
• 关键词: biological products; biomedical resource; lipids

The relationship between the three dimensional structure and the function of an enzyme is still poorly understood. This is especially so in the context of predictably altering the function of an enzyme. The interactive computer graphics in the Computer Graphics Laboratory provide a resource with which the binding surfaces and catalytic active sites can be discovered and mapped. This project involves creating and analyzing an altered serine protease. Granzyme B is a serine protease that had been implicated in cytotoxic lymphocyte mediated cell death. Experimenta evidence shows that granzyme B had strict extended specificity with the unusual property of cleaving after an acidic residue. The well characterized protease trypsin will be used as a model system to explore the specificity of granzyme B and other proteases with strict extended sites. Through three dimensional and sequence comparison the chymotrypsin family of enzymes will be analyzed for common motifs in extended specificity. When common themes occur, they will be tested on a hybrid scaffold for changes in Km. A hybrid will be designed that takes into account differences in the granzyme B and trypsin active sites. Trypsin has both a characteristic loop region and a disulfide bridge that is absent in granzyme B and may account for the differences in specificity. Mutations will be made systematically to trypsin to convert it to a protease of granzyme B specificity. Molecular modeling with Midas Plus and computer visualization will play important roles in the experimental design towards understanding the differences between these two and other serine proteases.

三维结构与 酶的功能仍然鲜为人知。 尤其是 因此，在可以预测改变​​酶功能的背景下。 计算机图形实验室中的交互式计算机图形 提供结合表面和催化的资源 可以发现和映射活动地点。 这个项目涉及 创建和分析改变的丝氨酸蛋白酶。 Granzyme B是A 与细胞毒性淋巴细胞有关的丝氨酸蛋白酶 介导的细胞死亡。 实验证据表明颗粒酶B 严格的扩展特异性具有分裂的异常特性 酸性残留后。 特征良好的蛋白酶胰蛋白酶将 用作探索颗粒酶B和的特异性的模型系统 其他具有严格扩展位点的蛋白酶。 通过三维 和序列比较酶的辣椒蛋白酶家族将是 分析了扩展特异性的共同基序。 当常见时 主题发生，将在混合脚手架上进行测试以改变 公里。设计的混合动力车将考虑到 颗粒酶B和胰蛋白酶活性位点。 胰蛋白酶都有两个 特征环区域和一座二硫键不存在 粒酶B，可能解释了特异性的差异。 将系统地进行突变以将其转换为胰蛋白酶 粒状B特异性的蛋白酶。 MIDAS分子建模 加上计算机可视化将在 实验设计，以理解之间的差异 这两个丝氨酸蛋白酶。