Imaging and manipulating proteins in live cells through covalent labeling
Imaging and manipulating proteins in live cells through covalent labeling
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DOI：
10.1038/nchembio.1959
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发表时间：
2015-12-01
影响因子：
14.8
通讯作者：
Johnsson, Kai
中科院分区：
文献类型：
--
作者：
Xue, Lin;Karpenko, Iuliia A.;Johnsson, Kai
在过去的20年里，已经见证了许多技术的出现，特异性和共价标记的蛋白质在细胞内和体内的合成探针。这些技术的范围从自我标记蛋白质标签到非天然氨基酸，问题不再是我们如何特异性地标记给定的蛋白质，而是我们希望为其配备什么额外的功能。在诸如超级、分辨率显微镜和基因组编辑为用先进的合成探针标记蛋白质提供了额外的动力，或者消除了进行蛋白质标记的一些困难。这样的实验。通过关注两个特定的应用，活细胞成像和可逆蛋白质开关的产生，我们概述了该领域的机遇和挑战，以及合成化学和蛋白质工程之间的协同作用如何使人们能够进行传统方法不可行的实验。
The past 20 years have witnessed the advent of numerous technologies to specifically and covalently label proteins in cellulo and in vivo with synthetic probes. These technologies range from self-labeling proteins tags to non-natural amino acids, and the question is no longer how we can specifically label a given protein but rather with what additional functionality we wish to equip it. In addition, progress in fields such as super-resolution microscopy and genome editing have either provided additional motivation to label proteins with advanced synthetic probes or removed some of the difficulties of conducting such experiments. By focusing on two particular applications, live-cell imaging and the generation of reversible protein switches, we outline the opportunities and challenges of the field and how the synergy between synthetic chemistry and protein engineering will make it possible to conduct experiments that are not feasible with conventional approaches.