Proteomics systems for integrative structural biology

我们提出了一个研究计划，旨在为描述和模拟复杂蛋白质系统的结构提供新的工具。我们生成结构丰富的生物物理数据的策略涉及浓缩相蛋白质化学，它将蛋白质属性编码成所有组织规模的可通过质谱学测量的数量(结构蛋白质组学)。拟议的计划有四个相互关联的主题。首先，我们将研究涉及快速反应、光触发化学反应的蛋白质生物偶联新策略，并开发用于共价标记质谱学(CL-MS)和交联质谱学(XL-MS)的新一代试剂。我们认为，将这些试剂的平衡蛋白质溶剂化性质从光解过程中分离出来，将提供前所未有的机会来增加来自CL-MS(地形测绘)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现蛋白质生物结合方法的概念，以避免重组蛋白质复合体。重建是一个费力且容易失败的过程，因此，克服目前的要求是将结构方法扩展到细胞环境中的关键一步。本主题的工作将涉及生产快速分离方法，用于从

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