TR&D Project 3. The Analysis Stage II: Tools for Analyzing the Connectivity and Morphology of Macromolecular Assemblies

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• 批准号: 10621359
• 负责人: Brian T Chait
• 金额: $ 6.99万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-01 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10621359
• 关键词: Architecture; Cell physiology; Cells; Chemicals; Complex; Data; Dimensions; Electron Microscopy; Freezing; Location; Macromolecular Complexes; Maps; Mass Spectrum Analysis; Methodology; Methods; Microscope; Modeling; Molecular; Morphology; Neighborhoods; Positioning Attribute; Proteins; Research; Resolution; Role; Shapes; Structural Models; Structure; Time; crosslink; empowerment; interest; macromolecular assembly; macromolecule; spatial relationship; temporal measurement; three-dimensional modeling; tool

PROJECT SUMMARY TR&D Project 3. The Analysis Stage II: Tools for Analyzing the Connectivity and Morphology of Macromolecular Assemblies As the structure of an assembly is directly related to its functional role, we aim to define the architecture of isolated native macromolecular complexes of interest. For this, our molecular microscope pipeline needs information on the shape and connectivity of an assembly’s components, accurately representing their arrangement at the highest spatial and temporal resolution. Thus, we will develop and refine methodologies to determine the morphologies and spatial relationships between components within complexes at several distance scales, ranging from a description of the overall subunit shape and arrangement to defining atomic resolution contacts between pairs of macromolecules. Our strategy entails using orthologous methods, in order to provide complementary data and to cover a wide range of resolutions, to inform us about the shape, dimensions and connectivity of single proteins and macromolecular assemblies. We will focus on methods that have already proven particularly empowering, but which have significant scope for further advancement. These include electron microscopy (EM) and chemical cross-linking with mass spectrometry (XL-MS): through the former, we can produce morphological maps with sufficient detail to resolve the shapes and locations of complexes, proteins, domains and folds; in parallel, through the latter, we will obtain information on how each component of the assembly is positioned relative to all other components, and the entire structure. These data when combined with data from complementary, well-established methods, will be used to generate structural models of assemblies. To elucidate cellular functions, we propose to gather and interpret dynamic data about the changing morphologies of assemblies, and the changing interactions within these assemblies. Our molecular microscope pipeline thus seeks to build concrete high precision 3D models, and 4D models that change in time.

项目概要 TR&D 项目 3. 分析阶段 II：分析连通性和形态的工具 高分子组装体 由于组件的结构与其功能角色直接相关，因此我们的目标是定义组件的架构 分离出感兴趣的天然大分子复合物。为此，我们的分子显微镜管道需要 有关装配体组件的形状和连接性的信息，准确地表示它们 以最高的空间和时间分辨率排列。因此，我们将开发和完善方法 确定多个距离处复合物内组分之间的形态和空间关系 尺度，范围从整体子单元形状和排列的描述到定义原子分辨率 大分子对之间的接触。我们的策略需要使用直系同源方法，以便提供 补充数据并涵盖广泛的分辨率，让我们了解形状、尺寸和 单个蛋白质和大分子组装体的连接性。我们将重点关注已经存在的方法 事实证明特别具有赋权作用，但还有很大的进一步发展空间。这些包括 电子显微镜 (EM) 和化学交联质谱 (XL-MS)：通过前者，我们 可以生成具有足够细节的形态图来解析复合物的形状和位置， 蛋白质、结构域和折叠；同时，通过后者，我们将获得有关每个组件如何 该组件相对于所有其他组件和整个结构进行定位。这些数据合并后 与来自补充的、行之有效的方法的数据一起，将被用来生成结构模型 组件。为了阐明细胞功能，我们建议收集和解释有关变化的动态数据 程序集的形态，以及这些程序集内不断变化的交互。我们的分子显微镜 因此，pipeline 致力于构建具体的高精度 3D 模型和随时间变化的 4D 模型。