Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Kyoto 615-8510, Japan.
Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Kyoto 615-8510, Japan; JST-ERATO, Hamachi Innovative Molecular Technology for Neuroscience, Kyoto University, Kyoto 615-8530, Japan.
Cell Chem Biol. 2020 Aug 20;27(8):970-985. doi: 10.1016/j.chembiol.2020.06.016. Epub 2020 Jul 16.
Protein analysis under biological conditions is now regarded as indispensable for understanding the structure and function of proteins, in addition to in vitro studies using purified target proteins. Because there are many molecules other than the protein-of-interest (POI) under live cell conditions, selective labeling of a POI is critical to distinguish the POI from other proteins for precise analysis. Protein labeling strategies utilizing genetically encoded tags have been used in POI modification in the complex environment of live cells. However, genetic manipulation may often induce overexpression of the POI and/or perturb the cellular context, resulting in unexpected artifacts in the protein analysis. Alternatively, recent progress in chemical biology has produced two major chemical approaches for analyzing endogenous proteins under native conditions. In this review, we summarize these techniques that utilize either protein-selective chemical labeling or proteome-directed chemical modification.
在生物条件下进行蛋白质分析，除了使用纯化的靶蛋白进行体外研究之外，现在被认为对于理解蛋白质的结构和功能也是必不可少的。由于在活细胞条件下存在许多除目标蛋白（POI）之外的分子，因此选择性标记 POI 对于将 POI 与其他蛋白质区分开来以进行精确分析至关重要。在活细胞复杂环境中，利用遗传编码标签的蛋白质标记策略已被用于 POI 修饰。然而，遗传操作通常会导致 POI 的过表达和/或破坏细胞环境，从而导致蛋白质分析中出现意想不到的假象。或者，化学生物学的最新进展已经产生了两种用于在天然条件下分析内源性蛋白质的主要化学方法。在这篇综述中，我们总结了这些利用蛋白质选择性化学标记或蛋白质组导向的化学修饰的技术。
