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Brightness-gated two-color coincidence detection unravels two distinct mechanisms in bacterial protein translation initiation.

ABSTRACT: 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.

SUBMITTER: Hofig H

PROVIDER: S-EPMC6897966 | biostudies-literature | 2019

REPOSITORIES: biostudies-literature

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Brightness-gated two-color coincidence detection unravels two distinct mechanisms in bacterial protein translation initiation.

Höfig Henning H Yukhnovets Olessya O Remes Cristina C Kempf Noemie N Katranidis Alexandros A Kempe Daryan D Fitter Jörg J

Communications biology 20191206

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 ...[more]

PMID: 31840104

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