Structural Dynamics and Domain Reorientation of a Phosphohexomutase: Long Range Effects and Catalysis

磷酸己糖变位酶的结构动力学和结构域重新定向：长程效应和催化

• 批准号: 1409898
• 负责人: Lesa Beamer
• 金额: $ 50万
• 依托单位: University of Missouri-Columbia
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-01 至 2018-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1409898&HistoricalAwards=false
• 关键词: Structural; Dynamics; Domain; Reorientation; Phosphohexomutase

Structural dynamics and domain reorientation of a phosphohexomutase: long range effects and catalysis The investigators will address fundamental questions about the functions of enzymes, the intricate molecular machines found in all living things. In particular, this work will define pathways used for transmitting information across the three-dimensional structures of enzymes. The program will train young scientists in powerful, state-of-the-art biophysical methods and their applications for unraveling the mysteries of biological macromolecules. The research and associated training activities in this project will benefit the chemical and biotechnology industries in the U.S., as well as the research communities in protein science and biophysics. The PI will expand her Science ARTreach program at a local elementary school, using art to improve science literacy of a diverse student population. The co-PI will launch Scoutreach to introduce molecules of living cells to adolescent youth and will enhance teaching of biophysical methods in the graduate student curricula, to provide highly skilled employees for the work force. Catalytically relevant motions among domains and in internal networks will be investigated in the enzyme phosphomannomutase/phosphoglucomutase (PMM/PGM). PMM/PGM is a member of a large enzyme superfamily whose members have important roles in carbohydrate biosynthesis. Previous studies showed that both phosphorylation and ligand binding affect the structure and flexibility of this enzyme. As phosphorylation and allostery are topics of wide interest, the mechanisms involved in long-range phospho-regulation of catalysis have implications for understanding many enzyme systems. Specifically, this will be investigated at the levels of fluctuating domain orientations and networks of communicating and mobile residues, using a combination of cutting-edge X-ray crystallography, NMR, and protein chemistry. Multiple enzyme states (phosphorylated, dephosphorylated, etc.) will be characterized in terms of the nature and populations of their side chain and domain conformers, their dynamics, and residue networks of transmission and dynamics. How these characteristics are affected by covalent phosphorylation vs. noncovalent ligand binding will be compared and contrasted. This will provide a comprehensive case study of long-range communication in an enzyme, and its tuning by phosphorylation and ligand binding.

磷酸己变位酶的结构动力学和结构域重定向：长程效应和催化研究人员将解决有关酶功能的基本问题，酶是所有生物中发现的复杂分子机器。 特别是，这项工作将定义用于在酶的三维结构中传输信息的途径。 该计划将培养年轻科学家强大的，最先进的生物物理方法及其应用，以解开生物大分子的奥秘。 该项目的研究和相关培训活动将使美国的化学和生物技术行业受益，以及蛋白质科学和生物物理学的研究团体。 PI将在当地一所小学扩大她的科学ARTreach计划，利用艺术来提高不同学生群体的科学素养。 共同主要研究者将启动Scoutreach，向青少年介绍活细胞分子，并将加强研究生课程中的生物物理方法教学，为劳动力提供高技能员工。 在磷酸甘露变位酶/磷酸葡萄糖变位酶（PMM/PGM）中，将研究结构域之间和内部网络中的催化相关运动。 PMM/PGM是一个大的酶超家族的成员，其成员在碳水化合物的生物合成中具有重要作用。 以前的研究表明，磷酸化和配体结合都会影响这种酶的结构和灵活性。 由于磷酸化和变构是广泛关注的主题，参与催化的长程磷酸调节的机制对理解许多酶系统具有意义。 具体而言，这将在波动的结构域方向和网络的通信和移动的残基的水平上进行研究，使用尖端的X射线晶体学，NMR和蛋白质化学的组合。 多种酶状态（磷酸化、去磷酸化等）将其特征在于其侧链和结构域构象的性质和人口，他们的动力学，和残留网络的传输和动力学。将比较和对比共价磷酸化与非共价配体结合如何影响这些特征。这将提供一个全面的案例研究的远程通信的酶，其调整磷酸化和配体结合。