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用于蛋白质组学分析的特定染色质位点的纯化

Purification of specific chromatin loci for proteomic analysis.

作者信息

Byrum Stephanie D, Taverna Sean D, Tackett Alan J

机构信息

Department of Biochemistry and Molecular Biology, University of Arkansas for Medical Sciences, 4301 West Markham St, Little Rock, AR, 72205, USA.

出版信息

Methods Mol Biol. 2015;1228:83-92. doi: 10.1007/978-1-4939-1680-1_8.

DOI:10.1007/978-1-4939-1680-1_8
PMID:25311124
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4324170/
Abstract

Purification of small, native chromatin regions for proteomic identification of specifically bound proteins and histone posttranslational modifications is a powerful approach for studying mechanisms of chromosome metabolism. Here we detail a Chromatin Affinity Purification with Mass Spectrometry (ChAP-MS) approach for affinity purification of 1 kb regions of chromatin for targeted proteomic analysis. This approach utilizes quantitative, high resolution mass spectrometry to categorize proteins and histone posttranslational modifications co-enriched with the given chromatin region as either "specific" to the targeted chromatin or "nonspecific" contamination. In this way, the ChAP-MS approach can help define and redefine mechanisms of chromatin-templated activities.

摘要

纯化小的天然染色质区域以用于蛋白质组学鉴定特异性结合蛋白和组蛋白翻译后修饰，是研究染色体代谢机制的一种强大方法。在此，我们详细介绍一种用于亲和纯化1 kb染色质区域以进行靶向蛋白质组分析的染色质亲和纯化与质谱联用（ChAP-MS）方法。该方法利用定量、高分辨率质谱将与给定染色质区域共富集的蛋白质和组蛋白翻译后修饰分类为对靶向染色质“特异性”或“非特异性”污染。通过这种方式，ChAP-MS方法有助于定义和重新定义染色质模板化活动的机制。

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