Proteomic Analysis of Histone Variants and Their PTMs: Strategies and Pitfalls.
Proteomic Analysis of Histone Variants and Their PTMs: Strategies and Pitfalls.
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DOI：
10.3390/proteomes6030029
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发表时间：
2018-06-21
期刊：
影响因子：
3.3
通讯作者：
Pflieger D
中科院分区：
文献类型：
--
作者：
El Kennani S;Crespo M;Govin J;Pflieger D
表观遗传修饰有助于决定细胞的命运和分化。组蛋白变异和翻译后修饰（PTMs）的分子机制已经在发育、分化和疾病的背景下进行了研究。基于抗体的检测通常用于靶向ptm，但这些方法无法揭示修饰的组合模式。此外，一些组蛋白变体与标准组蛋白非常相似，抗体很难区分这些同工型。质谱（MS）已逐渐发展成为研究组蛋白变异及其PTMs的有力技术。事实上，质谱分析强调了PTM的复杂组合，表明它们之间存在“串扰”，并且还揭示了PTM模式通常是变异特异性的。尽管随着用于多种PTMs研究的计算工具的发展，质谱仪器的灵敏度和采集速度大大提高，但正确描述组蛋白PTMs的结构仍然具有挑战性，特别是自信地将修饰分配到特定的氨基酸上。在这里，我们提供了一份基于ms的策略清单，以及组蛋白PTM和变异表征固有的陷阱，同时强调了PTM和组蛋白序列变异之间复杂的相互作用。我们将特别说明基于质谱的分析在识别和量化组蛋白变异和修饰方面所起的作用。
Epigenetic modifications contribute to the determination of cell fate and differentiation. The molecular mechanisms underlying histone variants and post-translational modifications (PTMs) have been studied in the contexts of development, differentiation, and disease. Antibody-based assays have classically been used to target PTMs, but these approaches fail to reveal combinatorial patterns of modifications. In addition, some histone variants are so similar to canonical histones that antibodies have difficulty distinguishing between these isoforms. Mass spectrometry (MS) has progressively developed as a powerful technology for the study of histone variants and their PTMs. Indeed, MS analyses highlighted exquisitely complex combinations of PTMs, suggesting “crosstalk” between them, and also revealed that PTM patterns are often variant-specific. Even though the sensitivity and acquisition speed of MS instruments have considerably increased alongside the development of computational tools for the study of multiple PTMs, it remains challenging to correctly describe the landscape of histone PTMs, and in particular to confidently assign modifications to specific amino acids. Here, we provide an inventory of MS-based strategies and of the pitfalls inherent to histone PTM and variant characterization, while stressing the complex interplay between PTMs and histone sequence variations. We will particularly illustrate the roles played by MS-based analyses in identifying and quantifying histone variants and modifications.