Department of Medicinal Chemistry and Pharmacognosy, College of Pharmacy, University of Illinois at Chicago, 833 South Wood Street, Chicago, IL, 60612, USA.
Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Tanta University, Tanta, 31527, Egypt.
ChemMedChem. 2019 Jun 5;14(11):1096-1107. doi: 10.1002/cmdc.201900114. Epub 2019 Apr 10.
Histone deacetylase (HDAC) activity is modulated in vivo by post-translational modifications and formation of multiprotein complexes. Novel chemical tools to study how these factors affect engagement of HDAC isoforms by HDAC inhibitors (HDACi) in cells and tissues are needed. In this study, a synthetic strategy to access chemically diverse photoreactive probes (PRPs) was developed and used to prepare seven novel HDAC PRPs 9-15. The class I HDAC isoform engagement by PRPs was determined in biochemical assays and photolabeling experiments in live SET-2, HepG2, HuH7, and HEK293T cell lines and in mouse liver tissue. Unlike the HDAC protein abundance and biochemical activity against recombinant HDACs, the chemotype of the PRPs and the type of cells were key in defining the engagement of HDAC isoforms in live cells. Our findings suggest that engagement of HDAC isoforms by HDACi in vivo may be substantially modulated in a cell- and tissue-type-dependent manner.
组蛋白去乙酰化酶 (HDAC) 的活性在体内受到翻译后修饰和多蛋白复合物形成的调节。需要新的化学工具来研究这些因素如何影响 HDAC 抑制剂 (HDACi) 与细胞和组织中 HDAC 同工型的结合。在这项研究中，开发了一种用于获得化学多样性光反应探针 (PRP) 的合成策略，并用于制备七种新型 HDAC PRP 9-15。在生化测定和活 SET-2、HepG2、HuH7 和 HEK293T 细胞系以及小鼠肝组织中的光标记实验中，确定了 PRP 对 I 类 HDAC 同工型的结合。与 HDAC 蛋白丰度和针对重组 HDAC 的生化活性不同，PRP 的化学类型和细胞类型是定义活细胞中 HDAC 同工型结合的关键。我们的研究结果表明，HDACi 在体内与 HDAC 同工型的结合可能在细胞和组织类型依赖性方式下受到实质性调节。
