Enzyme-based protein-tagging systems for site-specific labeling of proteins in living cells

IF 1.8 4区 工程技术
Microscopy Pub Date : 2019-11-01 DOI:10.1093/jmicro/dfaa011
Shinji Sueda
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引用次数: 2

Abstract

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.
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
已经提出了基于遗传编码标签和合成探针之间的特定相互作用的各种蛋白质标记方法来补充基于荧光蛋白质的标记。特别是,利用酶及其底物之间的高度特异性相互作用,已经深入开发了基于酶反应的标记方法。在这种方法中,肽或蛋白质作为标签在基因上附着在靶蛋白上,然后通过与携带这些标签的底物的酶反应将各种标签结合到标签中。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,通过用携带这些标签的酶进行翻译后修饰,然后在酶和底物蛋白之间紧密络合,将标签结合到标签中。在这篇综述中,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,然后描述了我们基于酶和底物蛋白质之间紧密络合的标记系统。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Microscopy
Microscopy 工程技术-显微镜技术
自引率
11.10%
发文量
0
审稿时长
>12 weeks
期刊介绍: Microscopy, previously Journal of Electron Microscopy, promotes research combined with any type of microscopy techniques, applied in life and material sciences. Microscopy is the official journal of the Japanese Society of Microscopy.
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