Determination Of 3 Dimensional Structures Of Macromolecu

该实验室的研究主要集中在利用多维核磁共振波谱对蛋白质、蛋白质-蛋白质复合体和蛋白质-核酸复合体的结构和动力学进行溶液研究，以及开发和应用新的核磁共振和计算方法来辅助这些研究。目前正在特别重视信号转导和转录调控的复合体，以及艾滋病和艾滋病相关蛋白。最近的成就包括将核磁共振方法的适用性扩展到大于40 kDa的结构。方法学发展的例子包括美国国立卫生研究院开发的全套3D和4D异核核磁共振实验，这些实验对于研究较大的蛋白质来说是必不可少的，其重叠的共振构成了一个可怕的问题；方法利用排列张量的各向异性(例如，在溶解于稀释型液晶介质中的大分子上测量的残余偶极耦合)或扩散张量(对于高度非球形的分子)来提供远距离取向信息，这是其他完全依赖于原子空间接近的核磁共振参数所不能获得的；基于顺磁松弛增强的方法，以获得结构信息并检测和表征参与大分子缔合的低密度中间体；以及根据所有可用的实验核磁共振限制条件，开发快速有效的算法来分析核磁共振谱和计算三维结构。最近的进展包括第一次直接展示和结构表征了蛋白质-DNA和蛋白质-蛋白质结合中的瞬时低群体中间体。

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. 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; methods based on paramagnetic relaxation enhancement to obtain both structural information and to detect and characterize low population intermediates involved in macromolecular association; 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. Recent developments include the first direct demonstration and structural characterization of transient low population intermediates iolved in both protein-DNA and protein-protein association.