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T‐CrAsH: A Heterologous Chemical Crosslinker
Author(s) -
Rutkowska Anna,
Plass Tilman,
Hoffmann JanErik,
Yushchenko Dmytro A.,
Feng Suihan,
Schultz Carsten
Publication year - 2014
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201402189
Subject(s) - click chemistry , cycloaddition , combinatorial chemistry , heterologous , in vitro , chemistry , membrane , chemical synthesis , strain (injury) , chemical biology , cell , nanotechnology , biophysics , biochemistry , materials science , biology , gene , anatomy , catalysis
Copper‐free click chemistry is currently the most promising and most rapidly developing technology for performing tailored chemical reactions inside intact living cells and animals. Its potential is particularly intensely explored in the field of live cell imaging, for both proteins and metabolites. Here we expand the application spectrum of click reactions to the chemical crosslinking of two proteins of choice in living cells. By combining strain‐promoted Diels–Alder cycloaddition with FlAsH‐based labeling of peptidic tetracysteine motifs, we developed the membrane‐permeating reversible crosslinker T‐CrAsH. We demonstrate the feasibility of the method both in vitro and inside cells. The biggest advantage of this new tool is the small size of the crosslinkable groups; this significantly decreases the risk of functional interference.