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蛋白质和蛋白质与其他大分子或小配体的络合物的方法。这一点意义重大，因为蛋白质相互作用是许多生物过程的关键，可能是抗击人类疾病的治疗干预的切入点。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，这种方法由P0-1开创，已被证明显著扩展了与现代核磁共振硬件可实现的分辨率相匹配的多维核磁共振实验的分辨率。特别令人感兴趣的将是对弱相互作用蛋白质复合体的关注，这些蛋白质复合体很难结晶，不能很好地表现出核磁共振谱，并且在用标准方法研究时会表现出严重的线宽。因此，P0-1将解决一个只有通过新的核磁共振方法才能解决的严重的结构生物学问题。拟议的研究包括五个研究组成部分和三个核心。组成部分1(瓦格纳)将为大型蛋白质和蛋白质复合体的结构研究开发实验和计算方法。组件2(Wagner)建议在实验约束稀疏的情况下开发和应用蛋白质与小分子和其他大分子相互作用的联合计算和实验方法。构成部分3(HOCH)建议进一步开发和应用蛋白质光谱的非均匀采样和处理非均匀采集的数据的方法。组分4(Walsh)针对非核糖体多肽合成酶Entf和EntB的结构和模块之间的相互作用。总体目标是了解细菌产生不寻常多肽的机制，其中一些具有重要的药理学特性。组件5(Chou)的重点是开发新的基于DNA的比对介质，用于测量剩余的偶极耦合，以及获得定义水溶液和洗涤剂胶束中的蛋白质复合体的新限制条件的策略。这项研究将得到管理(核心A)、核磁共振仪器(核心B)和计算(核心C)三个核心的支持。这一P0-1已经并将对该地区的生物科学产生广泛影响。它促进了与其他生物学家的许多合作，并产生了许多受益于这笔赠款所开发的专门知识的出版物。

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.