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

智力价值大多数酶都是以蛋白质为基础的催化剂，在催化效率和专一性方面都很突出。酶可以被认为是一种“纳米机器”，就像日常机器一样，酶的运动部分被预测为对其功能重要。这个项目的主要研究目标是了解内部蛋白质波动和酶催化之间的关系。更具体地说，将实施基于核磁共振的方法来严格量化吲哚-3-甘油磷酸合成酶的内部运动与其底物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(S)中所起的作用。对氨基酸的突变分析预计对内部蛋白质运动的调节和协调非常重要，将进一步揭示内部蛋白质波动是如何与化学相关的底物运动相耦合的。该模型酶体系的研究结果将有助于对蛋白质运动和酶催化的基本认识。这些结果还将使人们对新型酶催化剂的设计和开发有更深入的了解，例如那些用于许多工业过程和产品的催化剂，并可以为利用这里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指导。更广泛的影响本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，并激励代表不足的初中生继续STEM教育和职业生涯，特别是在化学和生物领域。学生将通过高等成就计划(Higher Achievement Program)和宾夕法尼亚州家庭科学之夜(Penn State Family Science Night)参加，该计划将把华盛顿特区地区的非大学学生和少数族裔家庭的学生带到宾夕法尼亚州，将科学示范和动手活动带到宾夕法尼亚州服务不足的农村学区。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STEM师范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个国家的教师都可以访问该领域。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来加强教育基础设施，并促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.