TY - JOUR
T1 - Optical Manipulation of Subcellular Protein Translocation Using a Photoactivatable Covalent Labeling System
AU - Kowada, Toshiyuki
AU - Arai, Keisuke
AU - Yoshimura, Akimasa
AU - Matsui, Toshitaka
AU - Kikuchi, Kazuya
AU - Mizukami, Shin
N1 - Funding Information:
This work was supported by MEXT KAKENHI Grant Numbers JP18H04605 and JP19H05284, by JSPS KAKENHI Grant Numbers JP17K05921, JP18H02102, JP18F18340, JP19K22241, and JP20K05702, and by the Uehara Memorial Foundation, the Takeda Science Foundation, the Astellas Foundation for Research on Metabolic Disorders, the Tokyo Biochemical Research Foundation, the Kowa Life Science Foundation, the Nakatani Foundation, and the “Dynamic Alliance for Open Innovation Bridging Human, Environment and Materials” Research Program in the “Network Joint Research Center for Materials and Devices”. We appreciate the experimental assistance of Hiroto Takahashi, Naoki Honzawa, and Kashfia Ahamed. We thank Tagen Central Analytical Facility for providing the NMR and MS instruments.
Publisher Copyright:
© 2021 Wiley-VCH GmbH
PY - 2021/5/10
Y1 - 2021/5/10
N2 - 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.
AB - 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.
KW - caged compound
KW - dimerization
KW - photochemistry
KW - proteins
KW - protein–protein interactions
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U2 - 10.1002/anie.202016684
DO - 10.1002/anie.202016684
M3 - Article
C2 - 33644979
AN - SCOPUS:85103549305
SN - 1433-7851
VL - 60
SP - 11378
EP - 11383
JO - Angewandte Chemie - International Edition
JF - Angewandte Chemie - International Edition
IS - 20
ER -