Fluorescent Labeling of Proteins of Interest in Live Cells: Beyond Fluorescent Proteins.,Bioconjugate Chemistry

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Fluorescent Labeling of Proteins of Interest in Live Cells: Beyond Fluorescent Proteins.
Bioconjugate Chemistry ( IF 4.0 ) Pub Date : 2020-05-07 , DOI: 10.1021/acs.bioconjchem.0c00181
Ji Liu _{1,

2} , Zongqiang Cui _{1,

2}

Affiliation

Live cell imaging brings us into a new era of direct visualization of biological processes and molecular dynamics in real time. To visualize dynamic cellular processes and virus–host interactions, fluorescent labeling of proteins of interest is often necessary. Fluorescent proteins are widely used for protein imaging, but they have some intrinsic deficiencies such as big size, photobleaching, and spectrum restriction. Thus, a variety of labeling strategies have been established and continuously developed. To protect the natural biological function(s) of the protein of interest, especially in viral life cycle, in vivo labeling requires smaller-sized tags, more specificity, and lower cytotoxicity. Here, we briefly summarized the principles, development, and their applications mainly in the virology field of three strategies for fluorescent labeling of proteins of interest including self-labeling enzyme derivatives, stainable peptide tags, and non-canonical amino acid incorporation. These labeling techniques greatly expand the fluorescent labeling toolbox and provide new opportunities for imaging biological processes.

中文翻译：

在活细胞中感兴趣的蛋白质的荧光标记：超越荧光蛋白质。

活细胞成像使我们进入了直接实时可视化生物过程和分子动力学的新时代。为了使动态细胞过程和病毒-宿主相互作用可视化，通常需要对目标蛋白进行荧光标记。荧光蛋白被广泛用于蛋白成像，但是它们具有一些固有的缺陷，例如大尺寸，光致漂白和光谱限制。因此，已经建立并不断发展各种标记策略。在体内保护目标蛋白质的天然生物学功能，尤其是在病毒生命周期中标记需要较小尺寸的标签，更高的特异性和更低的细胞毒性。在这里，我们简要地概述了原理，发展及其在主要用于三种荧光蛋白标记策略的病毒学领域中的应用，这些策略包括自标记酶衍生物，可染肽标签和非规范氨基酸掺入。这些标记技术极大地扩展了荧光标记工具箱，并为生物过程成像提供了新的机会。

更新日期：2020-05-07

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