Determination Of 3 Dimensional Structures Of Macromolecu

该实验室的研究主要集中在使用多维NMR光谱对蛋白质，蛋白质-蛋白质复合物和蛋白质-核酸复合物的结构和动力学进行溶液研究，以及开发和应用新型NMR和计算方法来帮助这些研究。特别强调的是参与信号转导和转录调节的复合物，以及艾滋病和艾滋病相关蛋白。最近的成就包括将NMR方法的适用性扩展到大于40 kDa的结构。在过去一年中解决的结构包括酶IIA（甘露醇）+ HPr的磷酰基转移复合物，和酶IIA（葡萄糖）酶IIB（葡萄糖）的PTS途径，和一个42 kDa的三元Oct 1-Sox 2-DNA复合物。方法学发展的例子包括NIH开发的全套3D和4D异频NMR实验，这些实验对于研究较大的蛋白质至关重要，这些蛋白质的重叠共振构成了一个可怕的问题;利用排列张量各向异性的方法（例如，在溶解于稀液晶介质中的大分子上测量的残余偶极耦合，例如棒状液晶的双相，形状的病毒颗粒）或扩散张量（对于高度非球形的分子）以提供从完全依赖于原子的紧密空间接近度的其他NMR参数无法获得的长程取向信息;和快速和有效的算法的NMR光谱分析和计算的三个发展-基于所有可用的实验NMR约束的三维结构。最近的发展包括使用刚性mody最小化和约束/约束模拟退火，以准确对接蛋白质-蛋白质复合物的分子间NOE和偶极耦合数据的基础上，以及化学位移扰动数据。

Research in this laboratory is centered around solution studies on the structure and dynamics of proteins, protein-protein complexes and protein-nucleic acid complexes using multidimensional NMR spectroscopy, and on the development and application of novel NMR and computational methods to aid in these studies. Particular emphasis is being placed on complexes involved in signal transduction and transcriptional regulation, and on AIDS and AIDS-related proteins. Recent accomplishments include the extension of the applicability of the NMR method to structures larger than 40 kDa. Structures solved in the last year include phosphoryl transfer complexes of enzyme IIA(mannitol) + HPr, and enzyme IIA(glucose)enzyme IIB(glucose) of the PTS pathway, and a 42 kDa ternary Oct1-Sox2-DNA complex. Examples of methodological developments include the panoply of 3D and 4D heteronuclear NMR experiments that have been developed at the NIH and are essential for studying larger proteins whose overlapping resonances pose a formidable problem; methods that make use of anisotropy of the alignment tensor (e.g. residual dipolar couplings measured on macromolecules dissolved in dilute liquid crystalline media such as the nematic phases of rod-shaped virus particles) or the diffusion tensor (for highly non-spherical molecules) to provide long-range orientational information that is not available from other NMR parameters that rely entirely on close spatial proximity of atoms; and the development of fast and efficient algorithms for the analysis of NMR spectra and for the computation of three-dimensional structures based on all available experimental NMR restraints. Very recent developments include the use of rigid mody minimization and constrained/restrained simulated annealing to accurately dock protein-protein complexes on the basis of intermolecular NOE and dipolar coupling data, as well as chemical shift perturbation data.