A new chromatographic approach to analyze methylproteome with enhanced lysine methylation identification performance
A new chromatographic approach to analyze methylproteome with enhanced lysine methylation identification performance
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一种分析甲基蛋白质组的新色谱方法，具有增强的赖氨酸甲基化鉴定性能
DOI：
10.1016/j.aca.2019.03.042
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发表时间：
2019
影响因子：
6.2
通讯作者：
Ye Mingliang
中科院分区：
文献类型：
--
作者：
Wang Qi;Liu Zhen;Wang Keyun;Wang Yan;Ye Mingliang
精氨酸/赖氨酸甲基化是一种重要的翻译后修饰（PTM），参与DNA修复、转录调控等。免疫亲和富集是目前应用最广泛的甲基化蛋白质组分析方法。精氨酸甲基化的大规模分析已经通过使用泛R-甲基抗体实现。不幸的是，靶向所有三种赖氨酸甲基化形式的泛特异性抗体不可用。在这项研究中，我们提出了一种新的基于色谱的富集方法的全球甲基蛋白质组分析。将强阳离子交换色谱（SCX）、固定化金属离子亲和色谱（IMAC）和高pH反相色谱（high-pH RP）相结合的离线多维串联色谱应用于甲基蛋白质组的大规模分析。从BEL细胞中总共鉴定了765个位点上的860种形式，涵盖了所有五种精氨酸/赖氨酸甲基化形式。其中27.21%为赖氨酸甲基化形式。该技术允许同时分析精氨酸和赖氨酸甲基化，同时其具有改进的鉴定赖氨酸甲基化的性能。因此，这是一个很有前途的策略，为研究与甲基化相关的生物学功能。
Arginine/lysine methylation is an important post-translational modification (PTM) involved in DNA repairing, transcriptional regulation, etc. Immunoaffinity enrichment is currently the most widely used methods for the methylproteome analysis. Large-scale analysis of arginine methylation has been realized by using pan-R-methyl antibodies. Unfortunately, pan specific antibodies targeting all three lysine methylation forms are not available. In this study, we presented a novel chromatography-based enrichment method for global methylproteome analysis. The offline multidimensional tandem chromatography combining strong cation exchange (SCX) chromatography, immobilized metal ion affinity chromatography (IMAC) and high-pH reversed-phase chromatography (high-pH RP) was applied in the large-scale analysis of methylproteome. Totally, 860 forms on 765 sites were identified from BEL cells, covering all five arginine/lysine methylation forms. Among them, 27.21% were lysine methylation forms. This technique allows the simultaneous analysis of both arginine and lysine methylation while it has improved performance for the identification of lysine methylation. Therefore, it is a promising strategy for the investigation of biological functions related to methylation.