Multiplexed Imaging of Posttranslational Modifications of Endogenous Proteins in Live Cells
Multiplexed Imaging of Posttranslational Modifications of Endogenous Proteins in Live Cells
复制标题
活细胞内源蛋白翻译后修饰的多重成像
DOI：
10.1007/978-1-0716-1593-5_3
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发表时间：
2021
影响因子：
--
通讯作者：
Kimura Hiroshi
中科院分区：
文献类型：
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作者：
Sato Yuko;Kimura Hiroshi
翻译后组蛋白修饰与基因组功能的调节有关。一些修饰非常稳定，可以维持染色质的表观基因组状态，而另一些修饰可以响应内部和外部刺激而表现出动态变化。为了追踪组蛋白修饰的局部和整体变化，活细胞中的多重成像是有益的。在用于检测组蛋白修饰的活细胞探针中，基因编码的修饰特异性胞内抗体或薄荷体是用于此目的的方便且合适的工具。我们在这里描述了基于 mintbody 的方法来监测组蛋白甲基转移酶和脱乙酰酶抑制剂诱导的组蛋白修饰水平的变化。通过测量活细胞中薄荷体的核与细胞质强度比，检测到组蛋白 H4 赖氨酸 20 甲基化状态的变化和组蛋白 H3 乙酰化的增加。
Posttranslational histone modifications are associated with the regulation of genome function. Some modifications are quite stable to maintain epigenome states of chromatin, and others can exhibit dynamic changes in response to internal and external stimuli. To track the local and global changes in histone modifications, multiplexed imaging in living cells is beneficial. Among live cell probes for detecting histone modifications, genetically encoded modification-specific intracellular antibodies, or mintbodies, are convenient and suitable tools for this purpose. We here describe the mintbody-based methods to monitor the changes in histone modification levels induced by histone methyltransferase and deacetylase inhibitors. By measuring the nuclear to cytoplasmic intensity ratios of mintbodies in living cells, changes in histone H4 lysine 20 methylation states and the increase in histone H3 acetylation were detected.