Institute of Clinical Biochemistry, Biocenter, Innsbruck Medical University, Innsbruck, Austria.
Institute of Clinical Biochemistry, Biocenter, Innsbruck Medical University, Innsbruck, Austria.
Methods. 2020 Dec 1;184:125-134. doi: 10.1016/j.ymeth.2020.01.017. Epub 2020 Jan 31.
The analysis, identification and quantification of histones and their post-translational modifications plays a central role in chromatin research and in studying epigenetic regulations during physiological processes. In the last decade analytical strategies based on mass spectrometry have been greatly improved for providing a global view of single modification abundances or to determine combinatorial patterns of modifications. Presented here is a newly developed strategy for histone protein analysis and a number of applications are illustrated with an emphasis on PTM characterization. Capillary electrophoresis is coupled to mass spectrometry (CE-MS) and has proven to be a very promising concept as it enables to study intact histones (top-down proteomics) as well as the analysis of enzymatically digested proteins (bottom-up proteomics). This technology combines highly efficient low-flow CE separations with ionization in a single device and offers an orthogonal separation principle to conventional LC-MS analysis, thus expanding the existing analytical repertoire in a perfect manner.
组蛋白及其翻译后修饰的分析、鉴定和定量在染色质研究以及研究生理过程中的表观遗传调控中起着核心作用。在过去的十年中,基于质谱的分析策略得到了极大的改进,可提供单个修饰丰度的全局视图,或确定修饰的组合模式。本文介绍了一种新开发的组蛋白分析策略,并通过强调 PTM 表征来说明了许多应用。毛细管电泳与质谱(CE-MS)相结合,已被证明是一种非常有前途的概念,因为它能够研究完整的组蛋白(自上而下的蛋白质组学)以及酶解蛋白质的分析(自下而上的蛋白质组学)。该技术将高效的低流速 CE 分离与单个设备中的离子化相结合,并提供了与传统 LC-MS 分析正交的分离原理,从而以完美的方式扩展了现有的分析手段。