Institute for Biology, Freie Universität Berlin, Berlin, Germany.
Methods Mol Biol. 2025;2873:167-184. doi: 10.1007/978-1-0716-4228-3_10.
Histone posttranslational modifications (PTMs) contribute to transcriptional regulation and generate in combination a "histone code," which is largely conserved among organismal kingdoms. By binding to specific PTMs, histone reader proteins act as molecular interpreters of the histone code. These proteins play a crucial role in gene regulation and chromatin structure by recruiting other proteins to the chromatin or blocking histone-modifying enzymes from accessing chromatin. Revealing the specificity of histone readers is, therefore, fundamental to understanding their function in gene regulation. Here, we describe a protocol to identify the binding specificity of histone readers to PTMs using histone peptide arrays and in vitro pull-down assays.
组蛋白翻译后修饰（PTMs）有助于转录调控，并结合生成“组蛋白密码”，该密码在生物界中广泛保守。通过与特定的 PTM 结合，组蛋白读码器蛋白充当组蛋白密码的分子解读器。这些蛋白通过将其他蛋白募集到染色质上或阻止组蛋白修饰酶接近染色质，在基因调控和染色质结构中发挥关键作用。因此，揭示组蛋白读码器的特异性对于理解其在基因调控中的功能至关重要。在这里，我们描述了一种使用组蛋白肽阵列和体外下拉测定来鉴定组蛋白读码器与 PTM 结合特异性的方案。
