使用不同染色质底物测量组蛋白甲基转移酶活性的体外分析

In Vitro Assays to Measure Histone Methyltransferase Activity Using Different Chromatin Substrates.

作者信息

Jacob Yannick, Voigt Philipp

机构信息

Department of Molecular, Cellular and Developmental Biology, Yale University, 165 Prospect St., New Haven, CT, 06511-2106, USA.

Wellcome Trust Centre for Cell Biology, The University of Edinburgh, Michael Swann Building, Max Born Crescent, Edinburgh, EH9 3BF, UK.

出版信息

Methods Mol Biol. 2018;1675:345-360. doi: 10.1007/978-1-4939-7318-7_20.

DOI:10.1007/978-1-4939-7318-7_20

PMID:29052201

Abstract

In vitro histone modification (HM) assays are used to characterize the activity of chromatin-modifying enzymes. These assays provide information regarding the modification sites on histones, the product specificity, and the impact of other histone or nucleotide modifications on enzyme activity. In particular, histone methyltransferase (HMT) assays have been instrumental in elucidating the activity and site specificity of many plant HMT enzymes. In this chapter, we describe a general protocol that can be used to perform HMT assays using different chromatin substrates, detection methods, and enzymes directly purified from plant material or heterologous sources.

摘要

体外组蛋白修饰（HM）分析用于表征染色质修饰酶的活性。这些分析提供了有关组蛋白上修饰位点、产物特异性以及其他组蛋白或核苷酸修饰对酶活性影响的信息。特别是，组蛋白甲基转移酶（HMT）分析在阐明许多植物HMT酶的活性和位点特异性方面发挥了重要作用。在本章中，我们描述了一种通用方案，可用于使用不同的染色质底物、检测方法以及直接从植物材料或异源来源纯化的酶来进行HMT分析。