The EMARS Reaction for Proximity Labeling
The EMARS Reaction for Proximity Labeling
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近距离标签的 EMARS 反应
DOI：
10.1007/978-1-4939-9537-0_1
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发表时间：
2019
影响因子：
--
通讯作者：
Kotani Norihiro
中科院分区：
文献类型：
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作者：
Honke Koichi;Miyagawa-Yamaguchi Arisa;Kotani Norihiro
为了在分子水平上理解细胞过程，需要阐明在活细胞中特定位置发生的蛋白质-蛋白质相互作用。我们已经开发了一种由酶介导的自由基源激活（EMARS）反应介导的邻近标记方法，其特征在于由辣根过氧化物酶（HRP）设置在感兴趣的分子（探测分子）上的标记试剂形成自由基。近端分子用与标记试剂缀合的标签共价标记。在这里，我们描述了用于制备标记试剂、在活细胞中标记被探测分子的邻近蛋白质以及鉴定标记蛋白质的方案。
To understand cellular processes at molecular levels, elucidation of protein-protein interactions occurring at a specific location in living cells is required. We have developed a proximity labeling method mediated by the enzyme-mediated activation of radical source (EMARS) reaction, which features a radical formation from labeling reagents by horseradish peroxidase (HRP) set on a molecule of interest (probed molecule). Proximal molecules are covalently labeled with a tag conjugated with the labeling reagent. Here we describe protocols for preparation of a labeling reagent, labeling of neighboring proteins of the probed molecule in living cells, and identification of the labeled proteins.