Bioanalytics, Institute of Biotechnology, Technische Universität Berlin, 13355, Berlin, Germany.
Bioanalytics, Institute of Biotechnology, Technische Universität Berlin, 13355, Berlin, Germany; Wellcome Centre for Cell Biology, University of Edinburgh, Edinburgh, EH9 3BF, UK.
Curr Opin Chem Biol. 2021 Feb;60:39-46. doi: 10.1016/j.cbpa.2020.07.008. Epub 2020 Aug 21.
Crosslinking mass spectrometry has become a core technology in structural biology and is expanding its reach towards systems biology. Its appeal lies in a rapid workflow, high sensitivity and the ability to provide data on proteins in complex systems, even in whole cells. The technology depends heavily on crosslinking reagents. The anatomy of crosslinkers can be modular, sometimes comprising combinations of functional groups. These groups are defined by concepts including: reaction selectivity to increase information density, enrichability to improve detection, cleavability to enhance the identification process and isotope-labelling for quantification. Here, we argue that both concepts and functional groups need more thorough experimental evaluation, so that we can show exactly how and where they are useful when applied to crosslinkers. Crosslinker design should be driven by data, not only concepts. We focus on two crosslinker concepts with large consequences for the technology, namely reactive group reaction kinetics and enrichment groups.
交联质谱已成为结构生物学的核心技术，并正在向系统生物学扩展。它的吸引力在于快速的工作流程、高灵敏度以及提供复杂系统中蛋白质数据的能力，甚至是整个细胞中的蛋白质数据。该技术严重依赖于交联试剂。交联剂的结构可以是模块化的，有时包括功能基团的组合。这些基团的定义包括：增加信息密度的反应选择性、提高检测效率的富集性、增强鉴定过程的可切割性以及用于定量的同位素标记。在这里，我们认为概念和功能基团都需要更彻底的实验评估，以便我们能够准确展示它们在应用于交联剂时的有用性和作用位置。交联剂的设计应该由数据驱动，而不仅仅是概念。我们重点关注两个对该技术有重大影响的交联剂概念，即反应基团反应动力学和富集基团。
