The Study of Isotropic and Anisotropic Motions in Proteins involving 13CO and 15ND

13CO和15ND蛋白质各向同性和各向异性运动的研究

• 批准号: 9513355
• 负责人: Erik Zuiderweg
• 金额: $ 31.5万
• 依托单位: Regents of the University of Michigan - Ann Arbor
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-02-01 至 2000-01-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9513355&HistoricalAwards=false
• 关键词: Study; Isotropic; Anisotropic; Motions; Proteins

9513355 Zuiderweg Dr. Zuiderweg will develop new NMR spectroscopy methodologies for the study of dynamics in biological macromolecules by measuring 13 CO and 15ND relaxation in 13C/15N labeled proteins. These experiments will yield information on the dynamics of the protein backbone and nitrogen-containing sidechains, complementary to the information obtained from the commonly performed 15NH NMR relaxation experiments. Efforts will go into extending above methods to allow the detection of the presence or absence of cross correlation between relaxation mechanisms. Such measurements will help the development of models for dynamical phenomena in proteins. The cross correlation (interference) between the heteronuclear relaxation mechanisms is determined with new two- and three-dimensional NMR methods. The methods will be mostly developed with the proteins T4-lysozyme (19 kDa) and Binase (12 kDa), and using a peptide loop on the protein Hsc70. Preliminary data shows that the extent of relaxation cross correlation is non-uniform over the backbone of T4-lysozyme, indicating that the dynamical characteristics measured as such are non-uniform and therefore of high informational content. The methods will be extended for determining the extent of motional correlation between relaxation vectors in protein surface loops, using the Hsc70 protein. These methods will be well-suited to distinguish between disjointed-chain motions and concerted motions involving only a few hinge-points. This information should be of importance for the understanding of the role of those loops and peptides that are involved in intermolecular interactions. %%% The research is aimed at developing new nuclear magnetic resonance (NMR) spectroscopy methodologies for the study of dynamics in biological macromolecules. The methods will concentrate on the NMR relaxation behavior of carbonyl moieties and will yield information on the dynamics of the protein backbone, complementary to the information obtained from the commonly performed NMR relaxation experiments focusing on amide moieties. The methods will be mostly developed with the proteins T4-lysozyme and Binase and will be extended for determining the extent of motional correlation between relaxation vectors in protein surface loops. This information should be of importance for the understanding of the dynamical characteristics of loops and peptides that are involved in intermolecular interactions. ***

Zuiderweg博士将通过测量13C/15N标记蛋白质中的13co和15ND弛豫，为生物大分子动力学研究开发新的核磁共振波谱方法。这些实验将得到蛋白质主链和含氮侧链的动力学信息，与通常进行的15NH NMR弛豫实验获得的信息相补充。将努力扩展上述方法，以允许检测松弛机制之间存在或不存在相互关系。这样的测量将有助于蛋白质动力学现象模型的发展。用新的二维和三维核磁共振方法确定了异核弛豫机制之间的相互关系（干涉）。该方法将主要利用蛋白t4 -溶菌酶（19 kDa）和Binase （12 kDa）开发，并在蛋白Hsc70上使用肽环。初步数据表明，在t4 -溶菌酶的主干上，弛豫互相关的程度是不均匀的，这表明用这种方法测量的动力学特性是不均匀的，因此具有很高的信息量。使用Hsc70蛋白，将扩展方法以确定蛋白表面环中松弛向量之间的运动相关程度。这些方法将非常适合于区分断链运动和仅涉及几个铰点的协调运动。这些信息对于理解那些参与分子间相互作用的环和肽的作用是很重要的。本研究旨在发展新的核磁共振（NMR）光谱方法，用于研究生物大分子的动力学。该方法将集中于羰基部分的核磁共振弛豫行为，并将获得有关蛋白质骨架动力学的信息，补充从通常进行的核磁共振弛豫实验中获得的信息，重点是酰胺部分。这些方法将主要与蛋白质t4溶菌酶和Binase一起开发，并将扩展到确定蛋白质表面环中松弛向量之间运动相关的程度。这一信息对于理解参与分子间相互作用的环和肽的动力学特性是很重要的。＊＊＊