J Biophotonics. 2011 Jun;4(6):391-402. doi: 10.1002/jbio.201100018. Epub 2011 May 12.
Technologies to visualize cellular structures and dynamics enable cell biologists to gain insight into complex biological processes. Currently, fluorescent proteins are used routinely to investigate the behavior of proteins in live cells. Chemical biology techniques for selective labeling of proteins with fluorescent labels have become an attractive alternative to fluorescent protein labeling. In the last ten years the progress in the development of chemical tagging methods have been substantial offering a broad palette of applications for live cell fluorescent microscopy. Several methods for protein labeling have been established, using protein tags, peptide tags and enzyme mediated tagging. This review focuses on the different strategies to achieve the attachment of fluorophores to proteins in live cells and cast light on the advantages and disadvantages of each individual method. Selected experiments in which chemical tags have been successfully applied to live cell imaging will be discussed and evaluated.
可视化细胞结构和动态的技术使细胞生物学家能够深入了解复杂的生物学过程。目前，荧光蛋白被常规用于研究活细胞中蛋白质的行为。用荧光标记物选择性标记蛋白质的化学生物学技术已成为荧光蛋白标记的一种有吸引力的替代方法。在过去十年中，化学标记方法的发展取得了实质性的进展，为活细胞荧光显微镜提供了广泛的应用。已经建立了几种蛋白质标记方法，使用蛋白质标签、肽标签和酶介导的标记。本综述重点介绍了在活细胞中实现荧光团与蛋白质连接的不同策略，并阐明了每种方法的优缺点。将讨论和评估已成功应用于活细胞成像的化学标记的选定实验。
