Mass spectrometry identifies and quantifies 74 unique histone H4 isoforms in differentiating human embryonic stem cells
Mass spectrometry identifies and quantifies 74 unique histone H4 isoforms in differentiating human embryonic stem cells
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DOI：
10.1073/pnas.0710515105
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发表时间：
2008-03-18
影响因子：
11.1
通讯作者：
Coon, Joshua J.
中科院分区：
文献类型：
--
作者：
Phanstiel, Doug;Brumbaugh, Justin;Coon, Joshua J.
通过染色质进行的表观遗传调控被认为在多能性的建立和维持中起着关键作用。传统上，基于抗体的技术用于探测组蛋白尾部存在的特定翻译后修饰（ptm），但这些方法通常不能揭示单个组蛋白尾部存在的多重修饰（组合代码）。在这里，我们描述了基于色谱法和质谱法的组蛋白组合密码的发现和定量技术。我们应用这种方法破译了人类胚胎干细胞组蛋白H4尾部的74个离散组合密码。最后，我们量化了人类胚胎干细胞分化过程中这些编码的丰度，揭示了甲基化和乙酰化模式的显著变化。例如，仅在存在H4K20二甲基化的情况下才观察到H4R3甲基化；这种特定于上下文的模式说明了这种技术的强大功能。
Epigenetic regulation through chromatin is thought to play a critical role in the establishment and maintenance of pluripotency. Traditionally, antibody-based technologies were used to probe for specific posttranslational modifications (PTMs) present on histone tails, but these methods do not generally reveal the presence of multiple modifications on a single-histone tail (combinatorial codes). Here, we describe technology for the discovery and quantification of histone combinatorial codes that is based on chromatography and mass spectrometry. We applied this methodology to decipher 74 discrete combinatorial codes on the tail of histone H4 from human embryonic stem (ES) cells. Finally, we quantified the abundances of these codes as human ES cells undergo differentiation to reveal striking changes in methylation and acetylation patterns. For example, H4R3 methylation was observed only in the presence of H4K20 dimethylation; such context-specific patterning exemplifies the power of this technique.