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: 6289752
• 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/6289752
• 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 N-terminal domain of HIV-1 integrase, the chemokine SDF-1, and cyanovirin-N. In addition, work was also carried out on a number of protein nucleic acid complexes, including those of the wild-type SRY and a sex-reversal mutant of this protein, Are A and Mef-2. Methodology for the large scale preparation of isotopically labelled DNA for structural studies was refined and optimized. In addition, from the mutant core libraries of streptococcal Protein G an unusual, tertameric variant has been characterized. As one of the largest systems to date, the NMR structure of EI-N complexed to HPr was determined. New media for partially aligning molecules in the magnetic field were developed and exploited for measuring residual dipolar couplings. - NMR structures, DNA binding domains, protein/DNA complexes, protein/protein complexes,isotope labelling of DNA, heteronuclear NMR.

该实验室的总体研究目标集中在尽可能完整地描述肽，蛋白质，核酸及其复合物在溶液中的结构，主要是通过NMR光谱。目前，特别强调的是发展的方法，使调查更大和复杂的系统，以及增加精度，这些解决方案的结构可以得到。进行了旨在关联结构和功能的研究，以及旨在研究蛋白质折叠的实验。已经对几种蛋白质进行了结构研究。这些包括HIV-1整合酶的N-末端结构域、趋化因子SDF-1和cyanovirin-N。此外，还对一些蛋白质核酸复合物进行了研究，包括野生型SRY和这种蛋白质的性别逆转突变体Are A和Mef-2的蛋白质核酸复合物。改进和优化了用于结构研究的同位素标记DNA的大规模制备方法。此外，从链球菌蛋白G的突变体核心文库中，一种不寻常的四聚体变体已被表征。作为迄今为止最大的体系之一，确定了EI-N与HPr络合的NMR结构。开发了在磁场中部分排列分子的新介质，并用于测量残余偶极耦合。- NMR结构，DNA结合域，蛋白质/DNA复合物，蛋白质/蛋白质复合物，DNA的同位素标记，杂原子NMR。