Collaborative Research: Expanding the repertoire of chemical tools to study and characterize bacterial Gcn5-related N-acetyltransferase functions

合作研究：扩展化学工具库来研究和表征细菌 Gcn5 相关的 N-乙酰转移酶功能

• 批准号: 1708927
• 负责人: Daniel Becker
• 金额: $ 5.4万
• 依托单位: Loyola University of Chicago
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-01-01 至 2021-12-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1708927&HistoricalAwards=false
• 关键词: Collaborative; Research; Expanding; repertoire; chemical

This National Science Foundation Research in Undergraduate Institutions award from the Chemistry of Life Processes Program funds Dr. Misty L. Kuhn from San Francisco State University (SFSU) and Dr. Daniel P. Becker from Loyola University Chicago to create biochemical tools to study enzymes in the family of Gcn5-related N-acetyltransferase (GNAT) proteins of unknown function from the soil microorganism, Pseudomonas aeruginosa. Specifically, these tools are used in combination with structure determinations via X-ray crystallography to gain a greater understanding of how molecules bind to GNATs. Since relatively little is known about the structure/function relationship of GNATs from P. aeruginosa, this work fills this gap in knowledge. Determining these functions is important because human welfare depends on healthy agriculture. This project enhances the research infrastructure at SFSU by establishing its first protein crystallization resource and increasing the repertoire of sophisticated biochemistry skills of underrepresented minority and disadvantaged undergraduate students. The goal of this project is to improve the functional annotation of P. aeruginosa GNATs of unknown function using a combination of protein X-ray crystallography, molecular modeling, organic synthesis, and enzymology. The architecture of the acceptor site of several GNAT proteins are probed using biochemical tools and X-ray crystallography. Active site residues that are critical for catalysis and substrate specificity are identified and tested using molecular modeling, site-directed mutagenesis, and enzyme kinetics. The results of this project provides the foundational knowledge to categorize experimentally uncharacterized members of the GNAT family and improve computational functional annotation of GNATs of unknown function.

这个国家科学基金会研究本科院校奖从生命过程计划的化学基金博士米斯蒂L。来自旧金山弗朗西斯科州立大学（SFSU）的Kuhn和来自洛约拉大学芝加哥的丹尼尔P.贝克尔博士创建了生物化学工具，用于研究来自土壤微生物铜绿假单胞菌的Gcn 5相关N-乙酰转移酶（GNAT）蛋白家族中的酶，该蛋白的功能未知。具体而言，这些工具与通过X射线晶体学进行的结构测定结合使用，以更好地了解分子如何与GNAT结合。由于对铜绿假单胞菌GNAT的结构/功能关系知之甚少，因此这项工作填补了这一知识空白。确定这些功能很重要，因为人类福利取决于健康的农业。 本项目旨在通过建立第一个蛋白质结晶资源库和提高少数民族和弱势本科生的复杂生物化学技能，加强SFSU的研究基础设施。本项目的目标是利用蛋白质X射线晶体学、分子模拟、有机合成和酶学相结合的方法，改进功能未知的铜绿假单胞菌GNAT的功能注释。几个GNAT蛋白质的受体位点的架构探测使用生化工具和X射线晶体学。活性位点的残基是催化和底物特异性的关键，确定和测试使用分子建模，定点诱变，和酶动力学。该项目的结果提供了基础知识来分类实验未表征的GNAT家族成员，并改进未知功能的GNAT的计算功能注释。