TY - JOUR
T1 - Fluorescent "Turn-on" system utilizing a quencher-conjugated peptide for specific protein labeling of living cells
AU - Arai, Satoshi
AU - Yoon, Su In
AU - Murata, Atsushi
AU - Takabayashi, Masao
AU - Wu, Xiaoyu
AU - Lu, Yixin
AU - Takeoka, Shinji
AU - Ozaki, Miwako
N1 - Funding Information:
This work was supported by special coordination funds for promoting science and technology, global COE, High-Tech Research Center project and grant-aid for scientific research challenging exploratory research from MEXT and Singapore Economic Development Board.
Copyright:
Copyright 2011 Elsevier B.V., All rights reserved.
PY - 2011/1/7
Y1 - 2011/1/7
N2 - A specific protein fluorescent labeling method has been used as a tool for bio-imaging in living cells. We developed a novel system of switching " fluorescent turn on" by the recognition of a fluorescent probe to a hexahistidine-tagged (His-tag) protein. The tetramethyl rhodamine bearing three nitrilotriacetic acids, which was used as a fluorescent probe to target a His-tagged protein, formed a reversible complex with the quencher, (Dabcyl)-conjugated oligohistidines, in the homogeneous solution, causing fluorescence of the fluorophore to be quenched. The complex when applied to living cells (COS-7) expressing His-tagged proteins on the cell surface caused the quencher-conjugated oligohistidines to be dissociated from the complex by specific binding of the fluorescent probe to the tagged protein, resulting in the fluorescent emission. The complex that did not participate in the binding event remained in the quenched state to maintain a low level of background fluorescence.
AB - A specific protein fluorescent labeling method has been used as a tool for bio-imaging in living cells. We developed a novel system of switching " fluorescent turn on" by the recognition of a fluorescent probe to a hexahistidine-tagged (His-tag) protein. The tetramethyl rhodamine bearing three nitrilotriacetic acids, which was used as a fluorescent probe to target a His-tagged protein, formed a reversible complex with the quencher, (Dabcyl)-conjugated oligohistidines, in the homogeneous solution, causing fluorescence of the fluorophore to be quenched. The complex when applied to living cells (COS-7) expressing His-tagged proteins on the cell surface caused the quencher-conjugated oligohistidines to be dissociated from the complex by specific binding of the fluorescent probe to the tagged protein, resulting in the fluorescent emission. The complex that did not participate in the binding event remained in the quenched state to maintain a low level of background fluorescence.
KW - Fluorescent imaging
KW - His-tag
KW - Small chemical probe
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U2 - 10.1016/j.bbrc.2010.11.095
DO - 10.1016/j.bbrc.2010.11.095
M3 - Article
C2 - 21110945
AN - SCOPUS:78650887464
SN - 0006-291X
VL - 404
SP - 211
EP - 216
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 1
ER -