Reactive group-embedded affinity labeling reagent for efficient intracellular protein labeling
Reactive group-embedded affinity labeling reagent for efficient intracellular protein labeling
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嵌入反应基团的亲和标记试剂，可有效标记细胞内蛋白质
DOI：
10.1016/j.bmc.2017.02.059
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发表时间：
2017
期刊：
影响因子：
--
通讯作者：
M. Ueda
中科院分区：
文献类型：
--
作者：
Y. Takaoka;Y. Nukadzuka;M. Ueda
目的蛋白的亲和标记是化学生物学研究的有力手段。然而，在活细胞中有效地标记细胞内蛋白质仍然是困难的。我们提出了一种用于高效蛋白质标记的反应性基团包埋亲和标记剂的新设计策略。以FKBP12为模型靶蛋白，配基结合口袋导向标记剂可以标记细胞内蛋白质，而蛋白质表面导向标记剂不能有效地标记活细胞，部分原因是大分子拥挤作用下细胞内蛋白质的波动。这些结果为有效的细胞内蛋白质标记提供了新的见解。
Affinity labeling of a target protein is a powerful method for chemical biology studies. However, it is still difficult to label intracellular proteins efficiently in living cells. We propose the novel design strategy of a reactive group-embedded affinity labeling reagent for efficient protein labeling. With FKBP12 as the model target protein, the ligand binding pocket-oriented labeling reagent could label intracellular protein, whereas protein surface-oriented reagent was ineffective for labeling in living cells, partially because of the intracellular protein fluctuation under the macromolecular crowding effects. These results provide new insight for efficient intracellular protein labeling.