INVESTIGATIONS OF MACROMOLECULAR STRUCTURES AND DYNAMICS IN SOLUTION BY NMR

通过核磁共振研究溶液中的大分子结构和动力学

• 批准号: 6105208
• 负责人: ANGELA M. GRONENBORN
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6105208
• 关键词: DNA binding protein; bacterial proteins; human immunodeficiency virus 1; human tissue; integrase; method development; molecular dynamics; nuclear magnetic resonance spectroscopy; protein folding; protein structure function; solutions; structural biology; thioredoxin; transposon /insertion element

The objective of the overall research in this laboratory is centered on achieving as complete a description as possible for the structures of peptides, proteins, nucleic acids and their complexes in solution, principally by NMR spectroscopy. At present particular emphasis is being placed on developing approaches which allow the investigation of larger and complex systems as well as increase the precision with which these solution structures can be obtained. Studies aimed at correlating structure and function, and experiments aimed at investigating protein folding are conducted. Structural studies for several proteins have been carried out. These include the DNA binding domains of Mu Transposase and the N-terminal domain of HIV-1 integrase . In addition, work was also carried out on a number of protein nucleic acid complexes, including those of GAGA, Are A and HMG-I/Y. A novel method was developed for large scale preparation of isotopically labelled DNA for structural studies. Also, mutant core libraries of streptococcal Protein G have been prepared and characterized structurally as well as with respect to stability. One of the largest systems whose structure we determined by NMR is the N-terminal domain of Enzyme I of the PTS pathway. In addition, the binding surface for HPr on EI was determined and structural implications of phosphorylation investigated. Finally, the structure of EI-N complexed to HPr was determined

本研究的总体目标是 实验室的中心是实现作为一个完整的描述， 可能的肽，蛋白质，核酸和 它们的络合物在溶液中，主要通过NMR光谱。在 目前特别强调发展 这些方法允许调查更大和更复杂的 系统，并提高这些解决方案的精度 可以得到结构。旨在关联结构的研究 和功能，以及旨在研究蛋白质折叠的实验 进行。对几种蛋白质的结构研究已经 贯彻这些包括Mu的DNA结合结构域 转座酶和HIV-1整合酶的N-末端结构域。在 此外，还对一些蛋白质核酸进行了研究， 酸络合物，包括GAGA、Are A和HMG-I/Y的酸络合物。一 发展了一种新的大规模制备 同位素标记的DNA用于结构研究。还有，变异核心 已经制备了链球菌蛋白G的文库， 在结构上和稳定性方面都有特征。之一 我们通过核磁共振确定的最大的系统是 PTS途径酶I的N-末端结构域。此外，本发明还提供了一种方法， 测定了HPr在EI上的结合表面， 研究磷酸化的影响。最后，结构 测定了与HPr络合的EI-N的量