通过免疫沉淀技术分析组蛋白去乙酰化酶的SUMO化修饰

Analysis of Histone Deacetylases Sumoylation by Immunoprecipitation Techniques.

作者信息

Wagner Tobias, Godmann Maren, Heinzel Thorsten

机构信息

Department of Biochemistry, Institute of Biochemistry and Biophysics, CMB - Center for Molecular Biomedicine, Friedrich Schiller University Jena, Hans-Knöll-Str. 2, Jena, 07745, Germany.

出版信息

Methods Mol Biol. 2017;1510:339-351. doi: 10.1007/978-1-4939-6527-4_25.

DOI:10.1007/978-1-4939-6527-4_25

PMID:27761833

Abstract

Histone deacetylases (HDACs) are controlling dynamic protein acetylation by removing acetyl moieties from lysine. Histone deacetylases themselves are regulated on the posttranslational level, including modifications with small ubiquitin-like modifier (SUMO) proteins. Detecting SUMO modifications of deacetylases by immunoblotting is technically challenging due to the typically low ratio of the modified compared to the unmodified species. Here, we describe a set of methods for the detection of endogenous sumoylated HDACs by immunoprecipitation and immunoblotting techniques.

摘要

组蛋白去乙酰化酶（HDACs）通过去除赖氨酸上的乙酰基部分来控制动态蛋白质乙酰化。组蛋白去乙酰化酶自身在翻译后水平受到调控，包括被小泛素样修饰物（SUMO）蛋白修饰。由于与未修饰形式相比，修饰形式的比例通常较低，通过免疫印迹检测去乙酰化酶的SUMO修饰在技术上具有挑战性。在此，我们描述了一组通过免疫沉淀和免疫印迹技术检测内源性SUMO化HDACs的方法。