Ellis Danielle J P, Yuan Zhigang, Seto Edward
H. Lee Moffitt Cancer Center & Research Institute, Tampa, Florida.
Curr Protoc Protein Sci. 2008 Nov;Chapter 14:14.12.1-14.12.14. doi: 10.1002/0471140864.ps1412s54.
Histone deacetylases (HDACs) are members of a diverse family of enzymes that catalyze the removal of an acetyl moiety from an acetyl-lysine-containing substrate. HDACs target a variety of substrates, including histone and nonhistone proteins, to mediate alterations in protein localization, stability, and activity. In addition, HDACs have been shown to modulate changes in gene expression, primarily through the recruitment of transcriptional cofactors to promoter regions. Mammalian HDACs are organized into distinct classes based on their homology to yeast HDACs. Classes I, II and IV HDACs are structurally and catalytically similar, whereas, class III HDACs require NAD(+) as a cofactor in the deacetylation reaction. This unit provides guidance for choosing and preparing a substrate suitable for assaying an HDAC of interest and describes key protocols necessary for assaying HDAC activity.
组蛋白脱乙酰酶（HDACs）是一类多样的酶家族成员，可催化从含乙酰赖氨酸的底物上去除乙酰基部分。HDACs作用于多种底物，包括组蛋白和非组蛋白，以介导蛋白质定位、稳定性和活性的改变。此外，HDACs已被证明主要通过将转录辅因子募集到启动子区域来调节基因表达的变化。哺乳动物HDACs根据其与酵母HDACs的同源性分为不同类别。I类、II类和IV类HDACs在结构和催化方面相似，而III类HDACs在脱乙酰化反应中需要NAD(+)作为辅因子。本单元为选择和制备适合检测目标HDAC的底物提供指导，并描述检测HDAC活性所需的关键方案。
