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

TR

• 批准号: 10401762
• 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 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10401762
• 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; Tertiary Protein Structure; Time; crosslink; empowered; 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.

项目摘要 TR＆D项目3。分析阶段II：分析连通性和形态的工具 大分子组件 由于组件的结构与其功能作用直接相关，因此我们旨在定义 分离的本地大分子络合物。为此，我们的分子显微镜管道需求 有关组件组件的形状和连通性的信息，准确地表示他们 以最高的空间和临时分辨率排列。那我们将开发并完善 确定几个距离的复合物中组件之间的形态和空间关系 量表，从对整体亚基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，以提供 完全数据并涵盖各种决议，以告知我们形状，尺寸和 单蛋白和大分子组件的连通性。我们将专注于已经 被证明是特别授权的，但它们具有进一步发展的显着范围。这些包括 电子显微镜（EM）和与质谱法（XL-MS）的化学交联：通过前者，我们 可以产生足够细节的形态图以解决复合物的形状和位置， 蛋白质，域和褶皱；并行，通过后者，我们将获得有关如何的每个组成部分的信息 组件与所有其他组件以及整个结构相对定位。这些数据合并 有了来自完成的数据，将使用良好的方法来生成结构模型 集会。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 组件的形态以及这些组件内的不断变化的相互作用。我们的分子显微镜 因此，管道试图构建混凝土高精度3D模型和时间变化的4D模型。