Albrow Victoria E, Grimley Rachel L, Clulow James, Rose Colin R, Sun Jianmin, Warmus Joseph S, Tate Edward W, Jones Lyn H, Storer R Ian
Pfizer Ltd, The Portway Building, Granta Park, Great Abington, Cambridge, CB21 6GS, UK.
Department of Chemistry and Institute of Chemical Biology, Imperial College London, London, SW7 2AZ, UK.
Mol Biosyst. 2016 May 24;12(6):1781-9. doi: 10.1039/c6mb00109b.
Histone deacetylases (HDACs) contribute to regulation of gene expression by mediating higher-order chromatin structures. They assemble into large multiprotein complexes that regulate activity and specificity. We report the development of small molecule probes with class IIa and pan-HDAC activity that contain photoreactive crosslinking groups and either a biotin reporter, or a terminal alkyne handle for subsequent bioorthogonal ligation. The probes retained inhibitory activity against recombinant HDAC proteins and caused an accumulation of acetylated histone and tubulin following cell treatment. The versatility of the probes has been demonstrated by their ability to photoaffinity modify HDAC targets in vitro. An affinity enrichment probe was used in conjunction with mass spectrometry proteomics to isolate HDACs and their interacting proteins in a native proteome. The performance of the probes in recombinant versus cell-based systems highlights issues for the development of chemoproteomic technologies targeting class IIa HDACs in particular.
组蛋白去乙酰化酶（HDACs）通过介导高阶染色质结构来调控基因表达。它们组装成大型多蛋白复合物，从而调节活性和特异性。我们报道了具有IIa类和泛HDAC活性的小分子探针的开发，这些探针含有光反应性交联基团以及生物素报告基团或末端炔烃手柄，用于后续的生物正交连接。这些探针保留了对重组HDAC蛋白的抑制活性，并在细胞处理后导致乙酰化组蛋白和微管蛋白的积累。这些探针的多功能性已通过其在体外对HDAC靶点进行光亲和修饰的能力得到证明。一种亲和富集探针与质谱蛋白质组学结合使用，以在天然蛋白质组中分离HDACs及其相互作用蛋白。这些探针在重组系统与基于细胞的系统中的性能突出了特别是针对IIa类HDACs的化学蛋白质组学技术开发中存在的问题。
