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首页> 外文期刊>Molecular & cellular proteomics: MCP >Characterization of histone H2A and H2B variants and their post-translational modifications by mass spectrometry.
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Characterization of histone H2A and H2B variants and their post-translational modifications by mass spectrometry.

机译：通过质谱对组蛋白H2A和H2B变体及其翻译后修饰进行表征。

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摘要

The nucleosome, the fundamental structural unit of chromatin, contains an octamer of core histones H3, H4, H2A, and H2B. Incorporation of histone variants alters the functional properties of chromatin. To understand the global dynamics of chromatin structure and function, analysis of histone variants incorporated into the nucleosome and their covalent modifications is required. Here we report the first global mass spectrometric analysis of histone H2A and H2B variants derived from Jurkat cells. A combination of mass spectrometric techniques, HPLC separations, and enzymatic digestions using endoproteinase Glu-C, endoproteinase Arg-C, and trypsin were used to identify histone H2A and H2B subtypes and their modifications. We identified nine histone H2A and 11 histone H2B subtypes, among them proteins that only had been postulated at the gene level. The two main H2A variants, H2AO and H2AC, as well as H2AL were either acetylated at Lys-5 or phosphorylated at Ser-1. For the replacement histone H2AZ, acetylation at Lys-4 and Lys-7 was found. The main histone H2B variant, H2BA, was acetylated at Lys-12, -15, and -20. The analysis of core histone subtypes with their modifications provides a first step toward an understanding of the functional significance of the diversity of histone structures.
机译：核小体是染色质的基本结构单元，包含核心组蛋白H3，H4，H2A和H2B的八聚体。组蛋白变体的掺入会改变染色质的功能特性。为了了解染色质结构和功能的整体动力学，需要分析掺入核小体的组蛋白变体及其共价修饰。在这里，我们报告了源自Jurkat细胞的组蛋白H2A和H2B变体的首次全球质谱分析。质谱技术，HPLC分离和使用内切蛋白酶Glu-C，内切蛋白酶Arg-C和胰蛋白酶的酶消化法的组合用于鉴定组蛋白H2A和H2B亚型及其修饰。我们鉴定了9个组蛋白H2A和11个组蛋白H2B亚型，其中只有在基因水平上才假定的蛋白质。两个主要的H2A变体H2AO和H2AC以及H2AL在Lys-5处被乙酰化或在Ser-1处被磷酸化。对于替换的组蛋白H2AZ，发现在Lys-4和Lys-7的乙酰化。主要的组蛋白H2B变体H2BA在Lys-12，-15和-20处被乙酰化。核心组蛋白亚型及其修饰的分析为了解组蛋白结构多样性的功能重要性提供了第一步。

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