INVESTIGATIONS OF MACROMOLECULAR STRUCTURES AND DYNAMICS IN SOLUTION BY NMR

本实验室的整体研究目标是以核磁共振波谱为主要手段，对多肽、蛋白质、核酸及其络合物在溶液中的结构进行尽可能完整的描述。目前，正在特别重视开发方法，以便能够调查较大和复杂的系统，并提高获得这些解结构的精度。旨在将结构和功能联系起来的研究，以及旨在研究蛋白质折叠的实验。对几种蛋白质的结构进行了研究。其中包括HIV-1整合酶的N-末端结构域、趋化因子SDF-1和氰基韦林-N。此外，还对一些蛋白质核酸复合体进行了研究，包括野生型SRY和该蛋白质的性反转突变体ARE A和MEF-2。改进和优化了用于结构研究的大规模制备同位素标记DNA的方法学。此外，从链球菌蛋白G的突变核心文库中还发现了一种不寻常的变种。作为迄今最大的体系之一，测定了与HPR络合的EI-N的核磁共振结构。开发了在磁场中部分排列分子的新介质，并用于测量剩余偶极耦合。-核磁共振结构，DNA结合结构域，蛋白质/DNA复合体，蛋白质/蛋白质复合体，DNA的同位素标记，异核核磁共振。

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.