Mendel Centre for Plant Genomics and Proteomics, Central European Institute of Technology, Masaryk University, Brno, Czech Republic.
Phytophthora Research Centre, Department of Molecular Biology and Radiobiology, Faculty of AgriSciences, Mendel University in Brno, Brno, Czech Republic.
Methods Mol Biol. 2022;2526:241-257. doi: 10.1007/978-1-0716-2469-2_18.
Reshaping of the chromatin landscape under oxidative stress is of paramount importance for mounting an effective stress response. Unbiased systemic identification and quantification of histone marks is crucial for understanding the epigenetic component of plant responses to adverse environmental conditions. We describe a detailed method for isolation of plant histones and subsequent bottom-up proteomics approach for characterization of acetylation and methylation status. By performing label-free quantitative mass spectrometry analysis, relative abundances of histone marks can be statistically compared between experimental conditions.
在氧化应激下重塑染色质景观对于引发有效的应激反应至关重要。无偏系统地鉴定和定量组蛋白标记对于理解植物对不利环境条件的反应的表观遗传成分至关重要。我们描述了一种详细的植物组蛋白分离方法,以及随后用于鉴定乙酰化和甲基化状态的自上而下的蛋白质组学方法。通过进行无标记定量质谱分析,可以在实验条件之间对组蛋白标记的相对丰度进行统计学比较。