TY  - JOUR
AU  - Katranidis, Alexandros
AU  - Sadoine, Mayuri
AU  - Cerminara, M.
AU  - Gerrits, M.
AU  - Fitter, Jörg
TI  - Co-translational incorporation into proteins of a fluorophore suitable for smFRET studies
JO  - ACS synthetic biology
VL  - 7
IS  - 2
SN  - 2161-5063
CY  - Washington, DC
PB  - ACS
M1  - FZJ-2018-01007
SP  - 405–411
PY  - 2018
AB  - 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.
LB  - PUB:(DE-HGF)16
C6  - pmid:29370697
UR  - <Go to ISI:>//WOS:000426012600013
DO  - DOI:10.1021/acssynbio.7b00433
UR  - https://juser.fz-juelich.de/record/842811
ER  -