Brightness-gated two-color coincidence detection unravels two distinct mechanisms in bacterial protein translation initiation.
Biochemical assays
Single-molecule biophysics
Journal
Communications biology
ISSN: 2399-3642
Titre abrégé: Commun Biol
Pays: England
ID NLM: 101719179
Informations de publication
Date de publication:
2019
2019
Historique:
received:
20
12
2018
accepted:
22
11
2019
entrez:
17
12
2019
pubmed:
17
12
2019
medline:
7
7
2020
Statut:
epublish
Résumé
Life on the molecular scale is based on a complex interplay of biomolecules under which the ability of binding is crucial. Fluorescence based two-color coincidence detection (TCCD) is commonly used to characterize molecular binding, but suffers from an underestimation of coincident events. Here, we introduce a brightness-gated TCCD which overcomes this limitation and benchmark our approach with two custom-made calibration samples. Applied to a cell-free protein synthesis assay, brightness-gated TCCD unraveled a previously disregarded mode of translation initiation in bacteria.
Identifiants
pubmed: 31840104
doi: 10.1038/s42003-019-0709-7
pii: 709
pmc: PMC6897966
doi:
Substances chimiques
Bacterial Proteins
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
459Informations de copyright
© The Author(s) 2019.
Déclaration de conflit d'intérêts
Competing interestsThe authors declare no competing interests.
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