如何通过蛋白质免疫印迹法区分HDAC1 - 3和HDAC6的活性
How to Distinguish Between the Activity of HDAC1-3 and HDAC6 with Western Blot.
作者信息
Beyer Mandy, Kiweler Nicole, Mahboobi Siavosh, Krämer Oliver H
机构信息
Institute of Toxicology, University Medical Center Mainz, Mainz, 55131, Germany.
Faculty of Chemistry and Pharmacy, Institute of Pharmacy, University of Regensburg, Regensburg, 93040, Germany.
出版信息
Methods Mol Biol. 2017;1510:355-364. doi: 10.1007/978-1-4939-6527-4_26.
Histone deacetylases (HDACs) catalyze the deacetylation of lysine residues in their target proteins. This biochemical modification can have profound effects on the functions of these proteins and a dysregulation of HDAC activity contributes to severe diseases, including neoplastic transformation. In the following chapter, we present a strategy that allows to distinguish between the inhibition of the class I HDACs HDAC1, 2, and 3 and of the class IIb HDAC HDAC6. This method is based on Western blot and relies on the detection of hyperacetylated substrates of class I or class IIb HDACs in lysates from cells that were treated with histone deacetylase inhibitors (HDACi).
组蛋白去乙酰化酶（HDACs）催化其靶蛋白中赖氨酸残基的去乙酰化反应。这种生化修饰可对这些蛋白质的功能产生深远影响，而HDAC活性的失调会导致包括肿瘤转化在内的严重疾病。在接下来的章节中，我们将介绍一种策略，该策略能够区分I类HDACs（HDAC1、2和3）和IIb类HDAC（HDAC6）的抑制作用。此方法基于蛋白质免疫印迹法，依赖于在经组蛋白去乙酰化酶抑制剂（HDACi）处理的细胞裂解物中检测I类或IIb类HDACs的高乙酰化底物。
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