Application of Celluspots peptide arrays for the analysis of the binding specificity of epigenetic reading domains to modified histone tails.
Application of Celluspots peptide arrays for the analysis of the binding specificity of epigenetic reading domains to modified histone tails.
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DOI：
10.1186/1471-2091-12-48
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发表时间：
2011-08-31
期刊：
影响因子：
--
通讯作者：
Jeltsch A
中科院分区：
文献类型：
--
作者：
Bock I;Kudithipudi S;Tamas R;Kungulovski G;Dhayalan A;Jeltsch A
表观遗传阅读结构域通过与组蛋白以翻译后修饰的特异性方式相互作用，参与基因表达和染色质状态的调节。详细了解阅读结构域的靶向修饰，包括加强和抑制二次修饰，将有助于更好地理解阅读结构域介导的生物信号过程。我们描述了Celluspots多肽阵列的应用，它包含384个组蛋白多肽，在不同的组合中携带59个翻译后修饰，作为一种廉价、可靠和快速的方法来初步筛选阅读结构域与修饰的组蛋白多肽的特异性相互作用。为了验证该方法，我们测试了Celluspots多肽阵列上已知的七个表观遗传读取域的结合特异性，即。HP1？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。总的来说，结合结果与文献数据在阅读结构域的主要特异性方面是一致的，但在几乎所有的情况下，我们都获得了关于靶标修饰周围的二级修饰的影响的额外的新信息。我们的结论是，Celluspots多肽阵列是研究与修饰的组蛋白多肽结合的假定阅读结构域的特异性的强有力的筛选工具。
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.
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影响因子：
16.8
作者：
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作者：
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