Highly selective inhibition of histone demethylases by de novo macrocyclic peptides.
Highly selective inhibition of histone demethylases by de novo macrocyclic peptides.
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DOI：
10.1038/ncomms14773
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发表时间：
2017-04-06
影响因子：
16.6
通讯作者：
Schofield CJ
中科院分区：
文献类型：
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作者：
Kawamura A;Münzel M;Kojima T;Yapp C;Bhushan B;Goto Y;Tumber A;Katoh T;King ON;Passioura T;Walport LJ;Hatch SB;Madden S;Müller S;Brennan PE;Chowdhury R;Hopkinson RJ;Suga H;Schofield CJ
JmjC 组蛋白去甲基酶 (KDM) 与肿瘤细胞增殖有关，是当前的癌症靶点；然而，很少有针对这些的高选择性抑制剂。在此，我们报告了 KDM4A-C 环肽抑制剂，其选择性优于其他 KDM/2OG 加氧酶，包括密切相关的 KDM4D/E 亚型。对其中一种抑制剂 (CP2) 与 KDM4A 的晶体结构和生化分析表明，CP2 与活性位点的组蛋白底物的结合方式不同，但与之竞争。将 CP2 的活性位点结合精氨酸替换为 N-ɛ-三甲基赖氨酸或甲基化精氨酸会产生环肽底物，表明 KDM4 可能作用于非组蛋白底物。基于晶体学和质谱分析对 CP2 进行靶向修饰，产生具有更强蛋白水解稳健性的变体。细胞中的肽剂量表现出 KDM4A 靶标的稳定性。尽管需要进一步开发来优化细胞活性，但结果揭示了 JmjC KDM 的高选择性非金属螯合、底物竞争性抑制剂的可行性。 JmjC 组蛋白去甲基酶 (KDM) 由于与细胞增殖相关而成为癌症靶标，但选择性抑制仍然是一个挑战。在此，作者通过从大量环肽库中进行体外选择，鉴定出 KDM4A-C 的有效抑制剂，这些抑制剂显示出优于其他 KDM 的选择性。
The JmjC histone demethylases (KDMs) are linked to tumour cell proliferation and are current cancer targets; however, very few highly selective inhibitors for these are available. Here we report cyclic peptide inhibitors of the KDM4A-C with selectivity over other KDMs/2OG oxygenases, including closely related KDM4D/E isoforms. Crystal structures and biochemical analyses of one of the inhibitors (CP2) with KDM4A reveals that CP2 binds differently to, but competes with, histone substrates in the active site. Substitution of the active site binding arginine of CP2 to N-ɛ-trimethyl-lysine or methylated arginine results in cyclic peptide substrates, indicating that KDM4s may act on non-histone substrates. Targeted modifications to CP2 based on crystallographic and mass spectrometry analyses results in variants with greater proteolytic robustness. Peptide dosing in cells manifests KDM4A target stabilization. Although further development is required to optimize cellular activity, the results reveal the feasibility of highly selective non-metal chelating, substrate-competitive inhibitors of the JmjC KDMs. JmjC histone demethylases (KDMs) are cancer targets due to their links to cell proliferation, but selective inhibition remains a challenge. Here the authors identify potent inhibitors of KDM4A-C—via in vitro selection from a vast library of cyclic peptides—that show selectivity over other KDMs.