NMR and Computational Studies of Biomolecules

描述（由申请人提供）：拟议研究的总体目标是开发和应用表征大分子蛋白质和与其他大分子或小配体的蛋白质复合物的方法。这是非常重要的，因为蛋白质相互作用是许多生物过程的关键，可能是治疗性干预对抗人类疾病的切入点。计划中的研究将包括核磁共振、计算和生化方法。核磁共振方法将在很大程度上依赖于非均匀采样方法，这是由这个P0-1开创的，并已被证明可以显着扩展多维核磁共振实验的分辨率，与现代核磁共振硬件可以实现的分辨率相匹配。特别感兴趣的将是对弱相互作用的蛋白质复合物的关注，这些蛋白质复合物难以结晶，在用标准方法研究时不能表现出很好的核磁共振光谱，并且表现出严重的谱线展宽。因此，P0-1将解决结构生物学的一个严重问题，这个问题只能用新的核磁共振方法来解决。本研究包括五个研究组成部分和三个核心。组件1 （Wagner）将为大型蛋白质和蛋白质复合物的结构研究开发实验和计算方法。组件2 （Wagner）提出在实验约束较少的情况下，开发和应用计算和实验相结合的方法来研究蛋白质与小分子和其他大分子的相互作用。组件3 （Hoch）提出进一步开发和应用蛋白质光谱非均匀采样和非均匀采集数据处理的方法。组分4 （Walsh）针对非核糖体多肽合成酶EntF和EntB模块的结构和相互作用。总体目标是了解细菌制造不寻常肽的机制，其中一些具有重要的药理特性。组分5 （Chou）专注于开发用于测量残余偶极偶联的新的基于dna的比对介质，以及在水溶液和洗涤剂胶束中获得定义蛋白质复合物的新约束的策略。该研究将由三个核心支持：管理（核心A），核磁共振仪器（核心B）和计算（核心C）。这一p -1已经并将对该地区的生物科学产生广泛影响。它激发了与其他生物学家的大量合作，并产生了许多出版物，这些出版物受益于这项资助所开发的专业知识。

DESCRIPTION (provided by applicant): The overall goal of the proposed research is to develop and apply methods for characterizing large proteins and protein complexes with other macromolecules or small ligands. This is of high significance since protein interactions are key to many biological processes and may be entry points for therapeutic interventions to fight human diseases. The planned research will include NMR, computational and biochemical approaches. NMR methods will heavily bank on non-uniform sampling approaches, which have been pioneered by this P0-1 and have been shown to dramatically extend the resolution of multidimensional NMR experiments matching the resolution achievable with modern NMR hardware. Of particular interest will be the focus on weakly interacting protein complexes that are difficult to crystallize, do not exhibit well-resolved NMR spectra and exhibit severe line broadening when studied with standard approaches. Thus, the P0-1 will tackle a serious problem of structural biology that can only be solved with new NMR approaches. The proposed research contains five research components and three cores. Component 1 (Wagner) will develop experimental and computational approaches for structural studies of large proteins and protein complexes. Component 2 (Wagner) proposes development and application of combined computational and experimental approaches for protein interactions with small molecules and other macromolecules for situations where experimental constraints are sparse. Component 3 (Hoch) proposes to further develop and apply methods for non-uniform sampling of protein spectra and processing of the non-uniformly acquired data. Component 4 (Walsh) targets structures and interactions of modules of the non-ribosomal polypeptide synthetases EntF and EntB. The overall goal is to understand the mechanisms by which bacteria make unusual peptides, some of which have important pharmacological properties. Component 5 (Chou) is focused on the development of new DNA-based alignment media for measuring residual dipolar couplings and strategies for obtaining new restraints for defining protein complexes both in aqueous solution and detergent micelles. The research will be supported by three cores for Administration (Core A), NMR instrumentation (Core B) and Computation (Core C). This P0-1 has had and will have a wide impact on biological science in the region. It stimulated numerous collaborations with other biologists and resulting in many publications that benefited from the expertise developed in this grant.