Structural and mechanistic studies of Ser/Lys dyad proteases

Ser/Lys 二元蛋白酶的结构和机制研究

• 批准号: 250131-2007
• 负责人: Paetzel, Mark
• 金额: $ 2.7万
• 依托单位: Simon Fraser University
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2007
• 资助国家: 加拿大
• 起止时间: 2007-01-01 至 2008-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=365241
• 关键词: Structural; mechanistic; studies; Ser; Lys

The objectives of the proposed research program is to investigate the structural and mechanistic details of two unrelated families of proteolytic enzymes (one bacterial and one viral) that have evolved the same unusual catalytic mechanism. These proteases; bacterial signal peptidase and viral VP4 protease both utilize an essential lysine and serine in their hydrolytic reactions. The deprotonated lysine is thought to serve as the general-base by abstracting the proton from the serine hydroxyl thereby activating it for nucleophilic attack on the scissile carbonyl of the substrate.Signal peptidase is an essential bacterial enzyme that functions to release secreted proteins from the cell membrane and is being investigated by a number of pharmaceutical companies as a novel antibacterial target.  VP4 protease is involved in the proteolytic processing of viral coat proteins in birnavirus, the infectious agent that costs the fish farming and agricultural industry billions of dollars of lost revenue each year. We will use; molecular biology, protein chemistry, X-ray crystallography, enzyme kinetics, and Nucleur Magnetic Resonance to investigate the structural and mechanistic similarities and difference in these two enzymes families in order to provide a more detailed understanding of how the Ser/Lys dyad mechanism works, and how enzymes work in general.Canada will benefit socially and financially from the commercialized drugs that result from the structure-based rational antibacterial and antiviral drug design efforts on these enzymes.  Canada will also benefit from the training of highly qualifies personnel in our lab. Students and post-doctoral fellows are becoming structural biologist (highly sought after in both academics and industry) with skills in molecular biology, protein chemistry and X-ray crystallography.

拟议的研究计划的目标是调查的结构和机制的细节两个不相关的蛋白水解酶（一个细菌和一个病毒），已经演变出相同的不寻常的催化机制。这些蛋白酶;细菌信号肽酶和病毒VP 4蛋白酶在它们的水解反应中都利用必需的赖氨酸和丝氨酸。去质子化赖氨酸被认为是一般的-信号肽酶是一种重要的细菌酶，其功能是从细胞膜上释放分泌的蛋白质，许多制药公司正在研究其作为一种新的抗菌靶标。VP 4蛋白酶参与了信号肽酶的作用。双RNA病毒是一种传染性病原体，每年使养鱼业和农业损失数十亿美元的收入。我们将使用;分子生物学、蛋白质化学、X射线晶体学、酶动力学和核磁共振，研究这两种酶家族的结构和机制相似性和差异，以便更详细地了解Ser/Lys二联体机制如何工作，以及酶一般是如何工作的。加拿大将从由这种结构产生的商业化药物中获得社会和经济利益-基于这些酶的合理的抗菌和抗病毒药物设计工作。加拿大也将受益于我们实验室高素质人员的培训。学生和博士后研究员正在成为结构生物学家（在学术界和工业界都备受追捧），具有分子生物学，蛋白质化学和X射线晶体学方面的技能。