Xue Bingbing, Xiao Kaijie, Tian Zhixin
School of Chemical Science and Engineering and Shanghai Key Laboratory of Chemical Assessment and Sustainability, Tongji University, Shanghai, 200092, China.
J Mass Spectrom. 2019 Mar;54(3):258-265. doi: 10.1002/jms.4339.
Histone post-translational modifications (PTMs) play various roles in chromatin-related cellular processes, and comprehensive analysis of these combinatorial PTMs at the intact protein level by top-down proteomics is the method of choice to reveal their crosstalk and biological functions. Here, we report our top-down characterization of the core histones from mouse fibroblasts cells NIH/3T3, which is a classic model used in many kinds of research. With nanoRPLC-MS/MS analysis and ProteinGoggle database search, 547 protein species were identified with spectrum-level FDR ≤ 1%, where PTMs in 51 protein species were unambiguously localized with PTM scores ≥1. High-resolution MS/MS data also allowed the unambiguous identification of acetylation instead of trimethylation. This study presents a general picture of combinatorial PTMs of mouse core histones, which serves as a basic reference for all future related biological studies.
组蛋白翻译后修饰（PTMs）在与染色质相关的细胞过程中发挥着多种作用，通过自上而下的蛋白质组学在完整蛋白质水平上对这些组合PTM进行全面分析，是揭示它们之间相互作用和生物学功能的首选方法。在此，我们报告了对小鼠成纤维细胞NIH/3T3核心组蛋白的自上而下表征，NIH/3T3是多种研究中使用的经典模型。通过纳升反相液相色谱-串联质谱（nanoRPLC-MS/MS）分析和ProteinGoggle数据库搜索，鉴定出547种蛋白质，其谱级错误发现率（FDR）≤1%，其中51种蛋白质中的PTM通过PTM得分≥1被明确定位。高分辨率MS/MS数据还能明确鉴定出乙酰化而非三甲基化。本研究展示了小鼠核心组蛋白组合PTM的总体情况，为所有未来相关生物学研究提供了基础参考。
