Kungulovski Goran, Kycia Ina, Mauser Rebekka, Jeltsch Albert
Institute of Biochemistry, Faculty of Chemistry, University Stuttgart, Pfaffenwaldring 55, 70569, Stuttgart, Germany.
The Jackson Laboratory for Genomic Medicine, Farmington, CT, USA.
Methods Mol Biol. 2015;1348:275-84. doi: 10.1007/978-1-4939-2999-3_24.
Histone posttranslational modifications (PTMs) have a crucial role in chromatin regulation and dynamics. They are specifically bound by so-called reading domains, which mediate the biological effects of histone PTMs. On a similar note, antibodies are invaluable reagents in chromatin biology for the detection, characterization, and mapping of histone PTMs. Despite these central roles in chromatin research and biology, the specificity of many antibodies and reading domains has been insufficiently characterized and documented. Here we describe in detail the application of the MODified™ Histone Peptide Array for the investigation of the binding specificity of histone binding antibodies or domains. The array contains 384 histone tail peptides carrying 59 posttranslational modifications in different combinations which can be used to study the primary binding specificity, but at the same time also allow to determine the combinatorial effect of secondary marks on antibody or reading domain binding.
组蛋白翻译后修饰（PTMs）在染色质调控和动态变化中起着关键作用。它们由所谓的阅读结构域特异性结合，这些阅读结构域介导组蛋白PTMs的生物学效应。同样，抗体是染色质生物学中用于检测、表征和定位组蛋白PTMs的宝贵试剂。尽管在染色质研究和生物学中具有这些核心作用，但许多抗体和阅读结构域的特异性尚未得到充分表征和记录。在这里，我们详细描述了MODified™组蛋白肽阵列在研究组蛋白结合抗体或结构域结合特异性方面的应用。该阵列包含384个组蛋白尾肽，这些肽带有59种不同组合的翻译后修饰，可用于研究主要结合特异性，但同时也能够确定二级标记对抗体或阅读结构域结合的组合效应。
