Proteomics systems for integrative structural biology

综合结构生物学的蛋白质组学系统

• 批准号: RGPIN-2017-04879
• 负责人: Schriemer, David
• 金额: $ 9.03万
• 依托单位: University of Calgary
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2017
• 资助国家: 加拿大
• 起止时间: 2017-01-01 至 2018-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=632612
• 关键词: Proteomics; systems; integrative; structural; biology

We present a program of research designed to provide new tools for characterizing and modeling the structure of complex protein systems. Our strategy for generating structure-rich biophysical data involves condensed-phase protein chemistries that code protein properties into quantities measurable by mass spectrometry, at all organizational scales (structural proteomics). There are four interrelated themes in the proposed program. First, we will investigate new protein bioconjugation strategies that involve fast-acting, light-triggered chemical reactions, and develop next-generation reagents for both covalent labeling mass spectrometry (CL-MS) and crosslinking mass spectrometry (XL-MS). We propose that separating the equilibrium protein solvation properties of these reagents from the photolytic process will provide unprecedented opportunities to increase the structural value of modeling restraints derived from both CL-MS (topographical mapping) and XL-MS (distance measurements). Second, we present concepts for implementing protein bioconjugation methods in complex mixtures and cells, to avoid reconstituting protein complexes. Reconstitution is a laborious and failure-prone process, thus overcoming this current requirement is critical step towards extending structural methods into a cellular context. The work in this theme will involve the production fast-isolation methods for removing complexes from

我们提出了一个研究计划，旨在提供新的工具，复杂的蛋白质系统的结构表征和建模。我们生成结构丰富的生物物理数据的策略涉及凝聚相蛋白质化学，该化学将蛋白质特性编码为可通过质谱测量的数量，在所有组织尺度上（结构蛋白质组学）。在拟议的方案中有四个相互关联的主题。首先，我们将研究新的蛋白质生物缀合策略，涉及快速作用，光触发化学反应，并开发下一代共价标记质谱（CL-MS）和交联质谱（XL-MS）试剂。我们建议，从光解过程中分离这些试剂的平衡蛋白质溶剂化特性将提供前所未有的机会，以增加来自CL-MS（地形图）和XL-MS（距离测量）的建模约束的结构价值。其次，我们提出了在复杂的混合物和细胞中实施蛋白质生物缀合方法的概念，以避免重构蛋白质复合物。重组是一个费力且容易失败的过程，因此克服当前的要求是将结构方法扩展到细胞环境中的关键一步。本课题的工作将涉及生产快速分离方法，