Journal Article

FZJ-2018-01007

http://join2-wiki.gsi.de/foswiki/pub/Main/Artwork/join2_logo100x88.png Co-translational incorporation into proteins of a fluorophore suitable for smFRET studies

Katranidis, A. (Corresponding author)FZJ* ; Sadoine, M. ; Cerminara, M. ; Gerrits, M. ; Fitter, J. (Corresponding author)FZJ*

2018
ACS Washington, DC

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Please use a persistent id in citations: doi:10.1021/acssynbio.7b00433

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.

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Contributing Institute(s):

1. Molekulare Biophysik (ICS-5)

Research Program(s):

1. 551 - Functional Macromolecules and Complexes (POF3-551) (POF3-551)

Appears in the scientific report 2018

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Database coverage:
; BIOSIS Previews ; IF >= 5 ; JCR ; NCBI Molecular Biology Database ; SCOPUS ; Science Citation Index Expanded ; Thomson Reuters Master Journal List ; Web of Science Core Collection

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