Substrate binding to histone deacetylases as shown by the crystal structure of the HDAC8-substrate complex
Substrate binding to histone deacetylases as shown by the crystal structure of the HDAC8-substrate complex
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DOI：
10.1038/sj.embor.7401047
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发表时间：
2007-09-01
期刊：
影响因子：
7.7
通讯作者：
Di Marco, Stefania
中科院分区：
文献类型：
--
作者：
Vannini, Alessandro;Volpari, Cinzia;Di Marco, Stefania
组蛋白去乙酰化酶（HDAC）-一种使组蛋白和非组蛋白蛋白去乙酰化的酶家族-与癌症等人类疾病有关，第一代HDAC抑制剂现已进入临床试验。在这里，我们报告的2.0埃分辨率的晶体结构的催化失活HDAC 8活性位点突变体，Tyr 306 Phe，结合到乙酰化的肽底物。该结构阐明了活性位点残基在脱乙酰化反应和底物识别中的作用。值得注意的是，该结构显示了活性位点边缘的保守残基Asp 101在通过直接与肽骨架相互作用并施加受约束的顺式构象来定位底物中的意想不到的作用。在我们也解决的新的异羟肟酸盐聚集体-HDAC 8结构中观察到类似的相互作用。野生型HDAC 8和Asp 101 Ala、Tyr 306 Phe和Asp 101 Ala/Tyr 306 Phe突变体的活性和结合试验证实了Asp 101在底物和抑制剂识别中的关键作用。
Histone deacetylases ( HDACs)-an enzyme family that deacetylates histones and non-histone proteins-are implicated in human diseases such as cancer, and the first-generation of HDAC inhibitors are now in clinical trials. Here, we report the 2.0 angstrom resolution crystal structure of a catalytically inactive HDAC8 active-site mutant, Tyr306Phe, bound to an acetylated peptidic substrate. The structure clarifies the role of active-site residues in the deacetylation reaction and substrate recognition. Notably, the structure shows the unexpected role of a conserved residue at the active-site rim, Asp 101, in positioning the substrate by directly interacting with the peptidic backbone and imposing a constrained cis-conformation. A similar interaction is observed in a new hydroxamate inhibitor-HDAC8 structure that we also solved. The crucial role of Asp 101 in substrate and inhibitor recognition was confirmed by activity and binding assays of wild-type HDAC8 and Asp101Ala, Tyr306Phe and Asp101Ala/Tyr306Phe mutants.