TR&D Project 2. The Analysis Stage I: Tools for Analyzing the Composition and Stoichiometry of Macromolecular Assemblies

TR

• 批准号: 10621356
• 负责人: JOHN D. AITCHISON
• 金额: $ 16.94万
• 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/10621356
• 关键词: Address; Biological Process; Cell physiology; Cells; Communities; Complex; Constitution; Constitutional; Data; Freezing; Genomics; Goals; Hand; Macromolecular Complexes; Mediating; Methods; Microscope; Modeling; Molecular; Morphology; Nature; Neighborhoods; Process; Property; Proteins; Proteomics; Refractory; Research; Research Personnel; Resources; Sensitivity and Specificity; System; Techniques; Technology; Time; bone; empowerment; improved; interest; macromolecular assembly; macromolecule; molecular assembly/self assembly; nucleic acid quantitation; reconstruction; response; stoichiometry; three-dimensional modeling; tool

PROJECT SUMMARY TR&D Project 2. The Analysis Stage I: Tools for Analyzing the Composition and Stoichiometry of Macromolecular Assemblies The determination and quantitation of proteins within assemblies remain a significant challenge, as proteins carry much of the burden of the rapid dynamic responses of the cell and are incredibly diverse in their abundance and their physico-chemical properties, making them highly versatile for the dynamic tasks at hand but at the same time difficult to analyze. This TR&D will allow the researcher to quantitatively characterize the constitution of isolated assemblies, and achieve this objective with a fraction of the time, effort, and resources normally required. Thus, we will develop methods to faithfully and fully identify the bone fide components of any target macromolecular complex in the cell, as well as that of vicinal associators, with high accuracy and throughput. We will advance current quantitative approaches for determining complex composition. We will also provide the community with a suite of options for discriminating between specific interactors and contaminants.We will also develop methods to faithfully and fully identify with high accuracy the stoichiometry of any target macromolecular complex in the cell. We will provide this technology to the community in the form of a “Stoichiometry Package”. This package will incorporate a parallel suite of techniques that give complementary and cross-verifying information, collectively providing accurate, comprehensive sets of stoichiometry data (and dynamic changes) for an isolated assembly. Users can choose any or a combination of approaches, depending on need. We will also gather data that inform on the dynamics of macromolecular complexes, by isolating complexes in defined sequences in space and time; comparisons of how the complexes change with time or different assembly state in terms of composition and stoichiometry will allow reconstruction of these dynamic processes. The major challenge we are addressing here is to achieve these goals with high fidelity in a mode that is readily transportable to any researcher.

项目摘要 TR＆D项目2。分析阶段I：分析组成和化学计量的工具 大分子组件 由于蛋白质携带 细胞快速动态反应的许多负担，它们的丰富性非常多样 它们的物理化学特性，使其在手头的动态任务方面具有高度的用途 时间难以分析。该TR＆D将允许研究人员定量地表征 孤立的集会，并以通常需要的时间，精力和资源的一小部分来实现这一目标。 这，我们将开发忠实并充分识别任何目标的骨骼组成部分的方法 细胞中的大分子复合物，以及速溶副手的精度和吞吐量。 我们将推进当前的定量方法来确定复杂组成。我们还将提供 社区有一套选择以区分特定交互者和污染物的选择。我们还将 开发忠实和充分识别的方法，以高精度为准任何靶标大分子的化学计量 在细胞中复杂。我们将以“化学计量套件”的形式向社区提供这项技术。 该软件包将包含一套平行的技术，这些技术可提供完整和交叉验证 信息，共同提供准确，全面的化学计量数据（以及动态更改） 用于隔离组件。用户可以根据需要选择任何或组合方法。我们将 还通过隔离定义的复合物来收集有关大分子复合物动态的数据 时空序列；比较复合物如何随时间或不同的装配状态变化 就组成和化学计量计而言，将允许重建这些动态过程。专业 我们在这里解决的挑战是在很容易的模式下以高忠诚实现这些目标 可运输到任何研究人员。