Optical Manipulation of Subcellular Protein Translocation Using a Photoactivatable Covalent Labeling System
Optical Manipulation of Subcellular Protein Translocation Using a Photoactivatable Covalent Labeling System
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使用光激活共价标记系统对亚细胞蛋白质易位进行光学操作
DOI：
10.1002/anie.202016684
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发表时间：
2021
期刊：
影响因子：
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通讯作者：
Mizukami Shin
中科院分区：
文献类型：
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作者：
Kowada Toshiyuki;Arai Keisuke;Yoshimura Akimasa;Matsui Toshitaka;Kikuchi Kazuya;Mizukami Shin
用于标签融合蛋白的光活化化学诱导二聚化（photo-CID）技术是调节亚细胞蛋白质易位和蛋白质-蛋白质相互作用的最有前途的方法之一。然而，光诱导的共价蛋白质二聚化在活细胞中尚未建立，尽管其具有各种优点。在此，我们通过将笼状配体应用于BL-标签系统（一种使用突变型β-内酰胺酶的共价蛋白质标记系统）开发了一种可光活化的共价蛋白质标记技术。我们进一步开发了CBHD，一种笼状蛋白二聚体，使用笼状BL-标签和HaloTag配体，并实现了光诱导的蛋白质从细胞质到亚细胞区域的易位。此外，这种共价pho-CID系统能够使蛋白质快速转移到激光照射的微区。这些结果表明，共价pho-CID系统将扩大CID在细胞功能的光学操纵中的应用范围。
The photoactivatable chemically induced dimerization (photo‐CID) technique for tag‐fused proteins is one of the most promising methods for regulating subcellular protein translocations and protein–protein interactions. However, light‐induced covalent protein dimerization in living cells has yet to be established, despite its various advantages. Herein, we developed a photoactivatable covalent protein‐labeling technology by applying a caged ligand to the BL‐tag system, a covalent protein labeling system that uses mutant β‐lactamase. We further developed CBHD, a caged protein dimerizer, using caged BL‐tag and HaloTag ligands, and achieved light‐induced protein translocation from the cytoplasm to subcellular regions. In addition, this covalent photo‐CID system enabled quick protein translocation to a laser‐illuminated microregion. These results indicate that the covalent photo‐CID system will expand the scope of CID applications in the optical manipulation of cellular functions.