Analysis of the substrate specificity of the dim-5 histone lysine methyltransferase using peptide arrays
Analysis of the substrate specificity of the dim-5 histone lysine methyltransferase using peptide arrays
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DOI：
10.1016/j.chembiol.2007.11.013
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发表时间：
2008-01-01
影响因子：
--
通讯作者：
Jeltsch, Albert
中科院分区：
文献类型：
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作者：
Rathert, Philipp;Zhang, Xing;Jeltsch, Albert
组蛋白甲基化是基因调控和发育所必需的表观遗传标记。我们引入肽 SPOT 合成来研究 Dim-5 组蛋白 3 赖氨酸 9 甲基转移酶的序列特异性。 Dim-5 识别 H3 尾部的 R8-G12，其中 T11 和 G12 是最重要的特异性决定因素。 E 交换 H3 尾残基 S10 和 T11 强烈降低了 Dim-5 的甲基化，表明 S10 或 T11 的磷酸化可能调节 Dim-5 的活性。在 Dim-5/肽结构中，E227 与 H3R8 相互作用，D209 与 H3-S10 相互作用。 E227 或 D209 的突变导致底物偏好发生可预测的变化，这表明组蛋白甲基转移酶的肽识别可以通过蛋白质设计来改变。因此，肽阵列与野生型和突变型酶的比较分析非常适合研究蛋白质甲基转移酶的靶标特异性和研究表观遗传串扰。
Histone methylation is an epigenetic mark essential for gene regulation and development. We introduce peptide SPOT synthesis to study sequence specificity of the Dim-5 histone-3 lysine-9 methyltransferase. Dim-5 recognizes R8-G12 of the H3 tail with T11 and G12 being the most important specificity determinants. Exchange of H3 tail residue S10 and T11 by E strongly reduced methylation by Dim-5, suggesting that phosphorylation of S10 or T11 may regulate the activity of Dim-5. In the Dim-5/peptide structure, E227 interacts with H3R8 and D209 with H3-S10. Mutations of E227 or D209 caused predictable changes in the substrate preference, illustrating that peptide recognition of histone methyltransferases can be altered by protein design. Comparative analyses of peptide arrays with wild-type and mutant enzymes, therefore, are well suited to investigate the target specificity of protein methyltransferases and study epigenetid crosstalk.