Center for Biopharmaceuticals & Department of Drug Design and Pharmacology, University of Copenhagen, Universitetsparken 2, DK-2100, Copenhagen, Denmark.
Institute of Applied Biosciences & Department of Food Chemistry and Toxicology, Karlsruhe Institute of Technology, Adenauerring 20a, D-76131, Karlsruhe, Germany.
Nat Commun. 2021 Jan 4;12(1):62. doi: 10.1038/s41467-020-20250-9.
Histones control gene expression by regulating chromatin structure and function. The posttranslational modifications (PTMs) on the side chains of histones form the epigenetic landscape, which is tightly controlled by epigenetic modulator enzymes and further recognized by so-called reader domains. Histone microarrays have been widely applied to investigate histone-reader interactions, but not the transient interactions of Zn-dependent histone deacetylase (HDAC) eraser enzymes. Here, we synthesize hydroxamic acid-modified histone peptides and use them in femtomolar microarrays for the direct capture and detection of the four class I HDAC isozymes. Follow-up functional assays in solution provide insights into their suitability to discover HDAC substrates and inhibitors with nanomolar potency and activity in cellular assays. We conclude that similar hydroxamic acid-modified histone peptide microarrays and libraries could find broad application to identify class I HDAC isozyme-specific substrates and facilitate the development of isozyme-selective HDAC inhibitors and probes.
组蛋白通过调节染色质结构和功能来控制基因表达。组蛋白侧链上的翻译后修饰（PTMs）形成表观遗传景观，由表观遗传修饰酶严格控制，并进一步被所谓的阅读器结构域识别。组蛋白微阵列已广泛应用于研究组蛋白-阅读器相互作用，但不能用于研究 Zn 依赖性组蛋白去乙酰化酶（HDAC）擦除酶的瞬时相互作用。在这里，我们合成了羟肟酸修饰的组蛋白肽，并将其用于飞摩尔微阵列中，用于直接捕获和检测四种 I 类 HDAC 同工酶。随后在溶液中的功能测定提供了关于其发现具有纳摩尔效力和细胞测定中活性的 HDAC 底物和抑制剂的适用性的见解。我们得出结论，类似的羟肟酸修饰的组蛋白肽微阵列和文库可以广泛应用于鉴定 I 类 HDAC 同工酶特异性底物，并促进同工酶选择性 HDAC 抑制剂和探针的开发。
