TY - JOUR
T1 - Reactive group-embedded affinity labeling reagent for efficient intracellular protein labeling
AU - Takaoka, Yousuke
AU - Nukadzuka, Yuuki
AU - Ueda, Minoru
N1 - Funding Information:
Plasmid for expression of FKBP12 fused with GST-tag in E. coli (pET41a-G3-FKBP12) was kindly gifted from Prof. I. Hamachi (Kyoto Univ) and Prof. S. Tsukiji (Nagoya Institute of Technology). We also thank Dr. T. Tamura (Kyoto Univ) for helpful discussion of this project. This work was supported in part by a Grant-in-Aid for Scientific Research (No. 26282207 to M.U. and No. 25708026 to Y.T.) on Innovative Areas “Chemical Biology of Natural Products” from MEXT, Japan, and by JST (Y.T.).
Publisher Copyright:
© 2017 Elsevier Ltd
PY - 2017
Y1 - 2017
N2 - 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.
AB - 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.
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U2 - 10.1016/j.bmc.2017.02.059
DO - 10.1016/j.bmc.2017.02.059
M3 - Article
C2 - 28283334
AN - SCOPUS:85014513811
SN - 0968-0896
VL - 25
SP - 2888
EP - 2894
JO - Bioorganic and Medicinal Chemistry
JF - Bioorganic and Medicinal Chemistry
IS - 11
ER -