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首页> 外文期刊>The FEBS journal >Using unnatural amino acids to selectively label proteins for cellular imaging: a cell biologist viewpoint
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Using unnatural amino acids to selectively label proteins for cellular imaging: a cell biologist viewpoint

机译:使用非天然氨基酸对细胞成像的选择性标记蛋白质:细胞生物学家观点

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摘要

Twenty-five years ago, GFP revolutionized the field of cell biology by enabling scientists to visualize, for the first time, proteins in living cells. However, when it comes to current, state-of-the-art imaging technologies, fluorescent proteins (such as GFP) have several limitations that result from their size and photophysics. Over the past decade, an elegant, alternative approach, which is based on the direct labeling of proteins with fluorescent dyes and is compatible with live-cell and super-resolution imaging applications, has been introduced. In this approach, an unnatural amino acid that can covalently bind a fluorescent dye is incorporated into the coding sequence of a protein. The protein of interest is thereby site-specifically fluorescently labeled inside the cell, eliminating the need for protein- or peptide-labeling tags. Whether this labeling approach will change cell biology research is currently unclear, but it clearly has the potential to do so. In this short review, a general overview of this approach is provided, focusing on the imaging of site-specifically labeled proteins in mammalian tissue culture cells, and highlighting its advantages and limitations for cellular imaging.
机译:25年前,绿色荧光蛋白(GFP)通过使科学家首次可视化活细胞中的蛋白质,彻底改变了细胞生物学领域。然而,就目前最先进的成像技术而言,荧光蛋白(如GFP)因其大小和光物理性而存在一些局限性。在过去的十年里,一种优雅的替代方法被引入,它基于荧光染料直接标记蛋白质,并与活细胞和超分辨率成像应用兼容。在这种方法中,可以共价结合荧光染料的非天然氨基酸被并入蛋白质的编码序列。因此,感兴趣的蛋白质在细胞内位置特异性地荧光标记,消除了对蛋白质或肽标记标签的需要。目前尚不清楚这种标记方法是否会改变细胞生物学研究,但它显然有这样做的潜力。在这篇简短的综述中,对这种方法进行了概述,重点介绍了哺乳动物组织培养细胞中位点特异性标记蛋白质的成像,并强调了其在细胞成像方面的优势和局限性。

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