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

合作研究：RUI：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

基本信息

批准号：
1708863
负责人：
Misty Kuhn
金额：
$ 24万
依托单位：
San Francisco State University
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=1708863&HistoricalAwards=false
关键词：
Collaborative Research RUI Expanding repertoire

项目摘要

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.

这项来自生命过程化学计划的国家科学基金会本科院校研究奖资助了旧金山州立大学（SFSU）的Misty L. Kuhn博士和芝加哥洛约拉大学的Daniel P. Becker博士创建生化工具，研究来自土壤微生物铜绿假单胞菌的gcn5相关n -乙酰转移酶（GNAT）蛋白家族中功能未知的酶。具体来说，这些工具与x射线晶体学的结构测定相结合，以更好地了解分子如何与gnat结合。由于对铜绿假单胞菌GNATs的结构/功能关系知之甚少，本研究填补了这一知识空白。确定这些功能很重要，因为人类的福祉取决于健康的农业。该项目通过建立首个蛋白质结晶资源，增强SFSU的研究基础设施，并增加少数民族和弱势本科生的复杂生物化学技能。本项目的目的是利用蛋白质x射线晶体学、分子建模、有机合成和酶学相结合的方法，改进P. aeruginosa gnat未知功能的功能注释。利用生化工具和x射线晶体学对几种GNAT蛋白受体位点的结构进行了探测。对催化和底物特异性至关重要的活性位点残基通过分子建模、位点定向诱变和酶动力学进行鉴定和测试。本项目的研究结果为GNAT家族实验中未表征成员的分类和改进未知功能GNAT的计算函数注释提供了基础知识。