CAREER: The Role of Protein Motions in Directing Substrate Conformational Dynamics in Enzyme Catalysis

知识优势大多数酶是蛋白质催化剂，在催化效率和特异性方面非常出色。酶可以被认为是一种“纳米机器”，就像每天的机器一样，酶有运动部件，预计对它们的功能很重要。本项目的主要研究目标是了解内部蛋白质波动与酶催化之间的关系。更具体地说，将实施基于NMR的方法来严格量化吲哚-3-甘油磷酸合酶的内部运动与其底物的构象动力学之间的关系，以更深入地了解蛋白质运动在指导酶催化的化学步骤中所起的作用。预测在内部蛋白质运动的调节和协调中重要的氨基酸的突变分析将进一步揭示内部蛋白质波动如何与化学相关的底物运动耦合。该模型酶系统的结果将适用于蛋白质运动和酶催化的基本理解。这些结果也将提供对新型酶催化剂的设计和开发的更透彻的理解，例如在许多工业过程和产品中使用的那些，并且可以为利用本文概述的基本催化原理的小分子仿生催化剂的设计提供进一步的方向。 该项目的综合教育目标是开发，指导和传播基于酶的演示和基于探究的活动，以教育不同的受众了解酶技术，并激励代表性不足的初中和高中学生追求STEM教育和职业，特别是在化学和生物学领域。学生将通过更高成就计划参与，该计划将华盛顿，华盛顿特区地区的非大学少数民族家庭的学生带到宾夕法尼亚州立大学，以及宾夕法尼亚州立大学家庭科学之夜，为宾夕法尼亚州服务不足的农村学区带来科学演示和实践活动。农村教师（6-12年级）将被带到宾夕法尼亚州立大学，通过宾夕法尼亚州立大学STEM教师学院参加与酶有关的演示。演示和相关材料将通过教师领域提供，美国和其他163个国家的教师可以访问该领域。这些活动的最终目标将是增加代表性不足的学生群体的参与，通过创建数字学习对象加强教育基础设施，并推进初中和高中科学教师的培训和发展。

Intellectual MeritThe majority of enzymes are protein-based catalysts that are remarkable in terms of their catalytic efficiency and specificity. Enzymes can be considered to be a type of 'nanomachine', and just like every day machines, enzymes have moving parts that are predicted to be important for their function. The main research goal of this project is to understand the relationships between internal protein fluctuations and enzyme catalysis. More specifically, NMR-based methodology will be implemented to rigorously quantify the relationship between the internal motions of the enzyme indole-3-glycerol phosphate synthase and the conformational dynamics of its substrate to provide deeper insight into the role protein motions play in directing the chemical step(s) of enzyme catalysis. Mutational analysis of amino acids predicted to be important in the regulation and coordination of internal protein motions will further reveal how internal protein fluctuations couple to chemically relevant substrate motions. The results for this model enzyme system will be applicable to the basic understanding of protein motion and enzyme catalysis. The results will also provide a more thorough understanding of the design and development of novel enzyme catalysts, such as those used in many industrial processes and products, and can provide further direction to the design of small molecule biomimetic catalysts that take advantage of the basic catalytic principles outlined here. Broader ImpactThe integrated education objectives of this project are to develop, direct and disseminate enzyme-based demonstrations and inquiry-based activities to educate diverse audiences about enzyme technology, and to inspire underrepresented middle and high school students to pursue STEM education and careers, especially in the fields of chemistry and biology. Students will participate through the Higher Achievement Program that brings students from non-college, minority families in the Washington, D.C. area to Penn State, and Penn State Family Science Night that brings science demonstrations and hands-on activities to underserved rural school districts in Pennsylvania. Rural-based teachers (grades 6-12) will be brought to Penn State to participate in enzyme-related demonstrations through the Penn State STEM Teacher Academy. Demonstration and related materials will be made available through Teacher's Domain that is accessed by teachers across the U.S. and in 163 other countries. The ultimate goals of these activities will be to increase participation of underrepresented student populations, enhance educational infrastructure through the creation of digital learning objects, and advance the training and development of middle and high school science teachers.