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@ARTICLE{Katranidis:842811,
author = {Katranidis, Alexandros and Sadoine, Mayuri and Cerminara,
M. and Gerrits, M. and Fitter, Jörg},
title = {{C}o-translational incorporation into proteins of a
fluorophore suitable for sm{FRET} studies},
journal = {ACS synthetic biology},
volume = {7},
number = {2},
issn = {2161-5063},
address = {Washington, DC},
publisher = {ACS},
reportid = {FZJ-2018-01007},
pages = {405–411},
year = {2018},
abstract = {Single-molecule FRET (smFRET) is a powerful tool to
investigate conformational changes of biological molecules.
In general, smFRET studies require protein samples that are
site-specifically double-labeled with a pair of donor and
acceptor fluorophores. The common approaches to produce such
samples cannot be applied when studying the synthesis and
folding of the polypeptide chain on the ribosome. The best
strategy is to incorporate two fluorescent amino acids
cotranslationally using cell-free protein synthesis systems.
Here, we demonstrate the cotranslational site-specific
incorporation into a model protein of Atto633, a dye with
excellent photophysical properties, suitable for single
molecule spectroscopy, together with a second dye using a
combination of the sense cysteine and the nonsense amber
codon. In this work we show that cotranslational
incorporation of good fluorophores into proteins is a viable
strategy to produce suitable samples for smFRET studies.},
cin = {ICS-5},
ddc = {540},
cid = {I:(DE-Juel1)ICS-5-20110106},
pnm = {551 - Functional Macromolecules and Complexes (POF3-551)},
pid = {G:(DE-HGF)POF3-551},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:29370697},
UT = {WOS:000426012600013},
doi = {10.1021/acssynbio.7b00433},
url = {https://juser.fz-juelich.de/record/842811},
}