Enzyme-based protein-tagging systems for site-specific labeling of proteins in living cells
Enzyme-based protein-tagging systems for site-specific labeling of proteins in living cells
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基于酶的蛋白质标记系统，用于活细胞中蛋白质的位点特异性标记
DOI：
10.1093/jmicro/dfaa011
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发表时间：
2020
期刊：
影响因子：
1.8
通讯作者：
Sueda Shinji
中科院分区：
文献类型：
--
作者：
○山澤隆史;高橋浩三;作花哲夫;堀田弘樹;花岡健二郎;西 直哉;Sueda Shinji
各种基于基因编码标签和合成探针之间特异性相互作用的蛋白质标记方法已经被提出，以补充基于荧光蛋白的标记。特别是，基于酶反应的标记方法已经通过利用酶与其底物之间高度特异性的相互作用而得到了广泛的发展。在这种方法中，肽或蛋白质作为标签遗传地附着在目标蛋白质上，然后通过酶反应将各种标签与携带这些标签的底物结合到标签中。另一方面，我们一直在开发一种不同于现有的基于酶的蛋白质标记系统。在我们的系统中，底物蛋白作为标签附着在靶蛋白上，通过携带这些标签的酶的翻译后修饰，然后酶和底物蛋白之间紧密络合，将标签纳入标签中。在本文中，我总结了基于酶的蛋白质标记系统，重点介绍了几种典型的方法，然后介绍了基于酶与底物蛋白紧密络合的标记系统。
Various protein-labeling methods based on the specific interactions between genetically encoded tags and synthetic probes have been proposed to complement fluorescent protein-based labeling. In particular, labeling methods based on enzyme reactions have been intensively developed by taking advantage of the highly specific interactions between enzymes and their substrates. In this approach, the peptides or proteins are genetically attached to the target proteins as a tag, and the various labels are then incorporated into the tags by enzyme reactions with the substrates carrying those labels. On the other hand, we have been developing an enzyme-based protein-labeling system distinct from the existing ones. In our system, the substrate protein is attached to the target proteins as a tag, and the labels are incorporated into the tag by post-translational modification with an enzyme carrying those labels followed by tight complexation between the enzyme and the substrate protein. In this review, I summarize the enzyme-based protein-labeling systems with a focus on several typical methods and then describe our labeling system based on tight complexation between the enzyme and the substrate protein.