RUI: The Role of Protein Dynamics in Catalysis and Subunit Cooperativity

这项研究将促进我们对蛋白质的动态性质如何在涉及蛋白质、碳水化合物和脂肪代谢的整合的酶中发挥关键作用的理解。该项目将研究在代谢中起核心作用的三种酶：谷氨酸脱氢酶、苹果酸脱氢酶和3-磷酸甘油脱氢酶的催化和活性调节之间的联系。影响催化和底物结合的亚基之间的通讯调节这些酶中的每一个。个别侧链在催化或亚基之间的通讯中的作用将通过位点特异性突变来探索。突变的影响将通过功能分析、使用X射线结晶学和质谱学的结构分析以及计算方法来研究。催化活性和生物调节都涉及蛋白质动态性质的变化的假设将通过蛋白质不同区域的工程灵活性变化并将这些影响与活性和亚单位通信的变化相关联来直接检验。更广泛的影响：这项研究完全融入了本科生的教育。学生将从事个人研究项目，参加实验室课程，并将研究嵌入到课程中，或者为非理科专业的学生选择以研究为基础的课程。学生完整地参与到实验的设计、研究的进行以及数据分析和展示中。学生在第一年就开始参与研究，并经常跟踪研究2-3年。这项研究为参与研究的学生带来了对生物过程的定量和物理理解。他们还参加地区小学的外联活动，将科学的兴奋带给当地的儿童和他们的老师。参与该项目的学生来自里士满大学和弗吉尼亚州立大学。

This research will advance our understanding of how the dynamic properties of proteins play a critical role in the enzymes involved with the integration of protein, carbohydrate, and fat metabolism. The project will investigate the linkage between dynamic protein structure and both catalysis and the regulation of activity in three enzymes that play central roles in metabolism: glutamate dehydrogenase, malate dehydrogenase, and 3 phosphoglycerate dehydrogenase. Communication between subunits affecting catalysis and substrate binding regulate each of these enzymes. The roles of individual side chains in catalysis or in communication between subunits will be probed using site-specific mutations. The effects of mutations will be studied by functional assays, structural analysis using X ray crystallography and mass spectrometry, and computational approaches. The hypothesis that both catalytic activity and biological regulation involve changes in the dynamic properties of the protein will be directly tested by engineering flexibility changes in various regions of the protein and correlating the effects with alterations in both activity and subunit communication. Broader Impacts: The research is integrated fully into the education of undergraduates. Students will work on individual research projects, take laboratory courses with the research embedded into the course, or may take a research-based course for non-science majors. Students are integrally involved in the design of experiments, conduct of the research, and data analysis and presentation. Students become involved in their first year and often follow the research for 2-3 years. The research brings both a quantitative and physical understanding of biological processes to the participating students. They also participate in outreach to area elementary schools to bring the excitement of science to local children and their teachers. Students engaged in the project are drawn from the University of Richmond and Virginia State University.