Identification and verification of lysine propionylation and butyrylation in yeast core histones using PTMap software.

Identification and verification of lysine propionylation and butyrylation in yeast core histones using PTMap software.
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DOI:
10.1021/pr8005155
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发表时间:
2009-02
影响因子:
4.4
通讯作者:
Zhao Y
Zhao Y
中科院分区:
生物学2区
文献类型:
--
作者:
Zhang K;Chen Y;Zhang Z;Zhao Y

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核心组蛋白中描述了已知对多种细胞功能至关重要的十种翻译后修饰 (PTM)。然而,目前尚不清楚组蛋白中是否存在其他 PTM,如果存在,这些未发现的信号在表观遗传现象中发挥什么作用。在这里,我们报告了通过质谱法结合使用我们最近开发的计算机程序 PTMap 的蛋白质序列比对对酵母组蛋白 PTM 进行的系统分析。我们首次在酵母组蛋白 H2B、H3 和 H4 中鉴定出赖氨酸丙酰化和丁酰化的多个位点。我们通过使用修饰特异性抗体进行蛋白质印迹、合成肽的 MS/MS 以及来自 HPLC 柱的合成肽和体内衍生肽的共洗脱来确认这些修饰。几种酵母组蛋白中存在多个修饰位点表明这两个 PTM 是真核生物中进化保守的组蛋白标记。此外,我们还发现了 14 个与任何已知 PTM 都不匹配的新质量转移,表明存在先前未描述的组蛋白修饰。这些修饰的化学性质仍有待确定。因此,我们的研究扩展了“组蛋白密码”的当前知识。
Ten types of post-translational modifications (PTMs) known to be critical to diverse cellular functions have been described in core histone proteins. However, it remains unclear whether additional PTMs exist in histones, and if so, what roles these undiscovered signals play in epigenetic phenomena. Here, we report a systematic analysis of yeast histone PTMs by mass spectrometry in combination with protein sequence alignment using PTMap, a computer program we recently developed. We have identified, for the first time, multiple sites of lysine propionylation and butyrylation in yeast histones H2B, H3, and H4. We confirmed these modifications by Western blotting using modification-specific antibodies, MS/MS of synthetic peptides, and coelution of synthetic and in vivo-derived peptides from an HPLC column. The presence of multiple modification sites in several yeast histones suggests that these two PTMs are histone marks that are evolutionarily conserved among eukaryotes. In addition, we identified 14 novel mass shifts that do not match any known PTM, suggesting the presence of previously undescribed histone modifications. The chemical natures of these modifications remain to be determined. Our studies therefore expand current knowledge of the “histone code”.
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