DETERMINATION OF 3 DIMENSIONAL STRUCTURES OF MACROMOLECULES IN SOLUTION BY NMR

核磁共振法测定溶液中大分子的三维结构

• 批准号: 6289751
• 负责人: G. MARIUS CLORE
• 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/6289751
• 关键词: DNA; DNA binding protein; Streptococcus; bacterial proteins; human immunodeficiency virus; integrase; ligands; method development; nuclear magnetic resonance spectroscopy; protein structure; solutions; structural biology; thioredoxin; transcription factor; transposon /insertion element; virus protein

Work in this laboratory has been focused on the determination of three- dimensional structures of larger proteins in solution by NMR, with a particular emphasis on protein-protein, protein-ligand and protein-DNA complexes. A considerable effort has been placed on the development of three- and four-dimensional heteronuclear NMR to extend the application of NMR as a method for determining three-dimensional structures of proteins in solution beyond the limits of conventional two-dimensional NMR (-100 residues) to molecules in the 150- to 400- residue range, and to develop new NMR methods for determining long range order apriori, including the dependence of heteronuclear relexation data on diffusion anisotropy and the use of residual dipolar couplings. Solution structures of a number of proteins have been determined. These include the complexes of the transcription factors GAGA, AreA and HMG-I/Y with DNA, the DNA MuA binding domains of the Mu transposase, the 30 kDa (259 residue) N terminal domain of Enzyme I of the PTS pathway, the 40 kDa complex of the N-terminal domain of enzyme I with the histidine containing protein HPr (40 kDa),the complete 44 kDa trimeric ectodomain of gp41, theN-terminal domain of HIV-1 integrase, and the barrier-to- autointegration factor BAF. - Structural Biology / Proteins / Solution Structure / NMR / protein-DNA complexes / protein-protein complexes

该实验室的工作重点是通过NMR测定溶液中较大蛋白质的三维结构，特别强调蛋白质-蛋白质、蛋白质-配体和蛋白质-DNA复合物。为了将NMR作为一种确定溶液中蛋白质三维结构的方法的应用扩展到传统二维NMR的极限之外，人们已经在三维和四维异构NMR的发展上投入了相当大的努力（~ 100个残基）到150- 400-残基范围内的分子，并开发新的NMR方法来确定长程有序的先验，包括异质结数据对扩散各向异性的依赖性和残余偶极耦合的使用。一些蛋白质的溶液结构已被确定。这些包括转录因子GAGA、AreA和HMG-I/Y与DNA的复合物，Mu转座酶的DNA MuA结合结构域，30 kDa的DNA结合结构域，以及DNA结合结构域。（259个残基）PTS途径的酶I的N-末端结构域，酶I的N-末端结构域与含组氨酸的蛋白HPr（40 kDa）的40 kDa复合物，gp 41的完整的44 kDa三聚体胞外域，HIV-1整合酶的N-末端结构域和自整合屏障因子BAF。- 结构生物学/蛋白质/溶液结构/ NMR /蛋白质-DNA复合物/蛋白质-蛋白质复合物