Department of Chemistry and the State Key Laboratory of Synthetic Chemistry, The University of Hong Kong, Laboratory for Synthetic Chemistry and Chemical Biology of Health@InnoHK, Pokfulam Road, Hong Kong SAR, China.
Department of Chemistry, Southern University of Science and Technology China, 1088 Xueyuan Road, Shenzhen, China.
Angew Chem Int Ed Engl. 2020 Sep 28;59(40):17525-17532. doi: 10.1002/anie.202001205. Epub 2020 Aug 11.
Histone deacetylase (HDAC) is a major class of deacetylation enzymes. Many HDACs exist in large protein complexes in cells and their functions strongly depend on the complex composition. The identification of HDAC-associated proteins is highly important in understanding their molecular mechanisms. Although affinity probes have been developed to study HDACs, they were mostly targeting the direct binder HDAC, while other proteins in the complex remain underexplored. We report a DNA-based affinity labeling method capable of presenting different probe configurations without the need for preparing multiple probes. Using one binding probe, 9 probe configurations were created to profile HDAC complexes. Notably, this method identified indirect HDAC binders that may be inaccessible to traditional affinity probes, and it also revealed new biological implications for HDAC-associated proteins. This study provided a simple and broadly applicable method for characterizing protein-protein interactions.
组蛋白去乙酰化酶 (HDAC) 是去乙酰化酶的主要类别。许多 HDAC 存在于细胞中的大型蛋白质复合物中，其功能强烈依赖于复合物的组成。鉴定与 HDAC 相关的蛋白质对于理解其分子机制非常重要。尽管已经开发了亲和探针来研究 HDAC，但它们大多针对直接结合 HDAC 的探针，而复合物中的其他蛋白质仍未得到充分探索。我们报告了一种基于 DNA 的亲和标记方法，该方法能够在不制备多个探针的情况下呈现不同的探针构型。使用一个结合探针，我们创建了 9 种探针构型来分析 HDAC 复合物。值得注意的是，该方法鉴定了间接的 HDAC 结合物，这些结合物可能无法被传统的亲和探针所接近，并且它还揭示了与 HDAC 相关的蛋白质的新的生物学意义。这项研究为蛋白质-蛋白质相互作用的表征提供了一种简单且广泛适用的方法。
