Biochemistry Laboratory, School of Engineering and Science, Jacobs University Bremen, Campus Ring 1, 28759 Bremen, Germany.
BMC Biochem. 2011 Aug 31;12:48. doi: 10.1186/1471-2091-12-48.
Epigenetic reading domains are involved in the regulation of gene expression and chromatin state by interacting with histones in a post-translational modification specific manner. A detailed knowledge of the target modifications of reading domains, including enhancing and inhibiting secondary modifications, will lead to a better understanding of the biological signaling processes mediated by reading domains.
We describe the application of Celluspots peptide arrays which contain 384 histone peptides carrying 59 post translational modifications in different combinations as an inexpensive, reliable and fast method for initial screening for specific interactions of reading domains with modified histone peptides. To validate the method, we tested the binding specificities of seven known epigenetic reading domains on Celluspots peptide arrays, viz. the HP1ß and MPP8 Chromo domains, JMJD2A and 53BP1 Tudor domains, Dnmt3a PWWP domain, Rag2 PHD domain and BRD2 Bromo domain. In general, the binding results agreed with literature data with respect to the primary specificity of the reading domains, but in almost all cases we obtained additional new information concerning the influence of secondary modifications surrounding the target modification.
We conclude that Celluspots peptide arrays are powerful screening tools for studying the specificity of putative reading domains binding to modified histone peptides.
表观遗传阅读结构域通过与组蛋白发生特定的翻译后修饰相互作用，参与基因表达和染色质状态的调控。详细了解阅读结构域的靶标修饰，包括增强和抑制二级修饰，将有助于更好地理解阅读结构域介导的生物学信号转导过程。
我们描述了 Celluspots 肽阵列的应用，该阵列包含 384 个带有 59 种不同组合翻译后修饰的组蛋白肽，作为一种廉价、可靠和快速的方法，用于初始筛选阅读结构域与修饰组蛋白肽的特异性相互作用。为了验证该方法，我们在 Celluspots 肽阵列上测试了七个已知的表观遗传阅读结构域的结合特异性，即 HP1β 和 MPP8 Chromo 结构域、JMJD2A 和 53BP1 Tudor 结构域、Dnmt3a PWWP 结构域、Rag2 PHD 结构域和 BRD2 Bromo 结构域。一般来说，结合结果与文献数据在阅读结构域的主要特异性方面一致，但在几乎所有情况下，我们获得了关于靶标修饰周围二级修饰影响的额外新信息。
我们得出结论，Celluspots 肽阵列是研究假定的阅读结构域与修饰组蛋白肽结合特异性的强大筛选工具。
