Brightness-gated two-color coincidence detection unravels two distinct mechanisms in bacterial protein translation initiation
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...
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| Veröffentlicht in: | Communications biology 2019-12, Vol.2 (1), p.459-459, Article 459 |
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| creator | Höfig, Henning Yukhnovets, Olessya Remes, Cristina Kempf, Noemie Katranidis, Alexandros Kempe, Daryan Fitter, Jörg |
| description | 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.
Henning Höfig et al. present a brightness-gated two-color coincidence detection method that overcomes the limitations of confocal detection. The method adds a selection criterion for the brightness of single fluorescence bursts, substantially improving the counting accuracy for single fluorophores. |
| doi_str_mv | 10.1038/s42003-019-0709-7 |
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Henning Höfig et al. present a brightness-gated two-color coincidence detection method that overcomes the limitations of confocal detection. The method adds a selection criterion for the brightness of single fluorescence bursts, substantially improving the counting accuracy for single fluorophores.</description><identifier>ISSN: 2399-3642</identifier><identifier>EISSN: 2399-3642</identifier><identifier>DOI: 10.1038/s42003-019-0709-7</identifier><identifier>PMID: 31840104</identifier><language>eng</language><publisher>London: Nature Publishing Group UK</publisher><subject>14/19 ; 14/35 ; 631/1647/2017/2197 ; 631/57/2265 ; Bacteria - genetics ; Bacteria - metabolism ; Bacterial Proteins - genetics ; Bacterial Proteins - metabolism ; Biology ; Biomedical and Life Sciences ; Brightness ; Color ; Fluorescence Resonance Energy Transfer ; Fluorophores ; Life Sciences ; Molecular Imaging - methods ; Peptide Chain Initiation, Translational ; Protein biosynthesis ; Spectrometry, Fluorescence - methods ; Translation ; Translation initiation</subject><ispartof>Communications biology, 2019-12, Vol.2 (1), p.459-459, Article 459</ispartof><rights>The Author(s) 2019</rights><rights>The Author(s) 2019.</rights><rights>The Author(s) 2019. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c470t-61d202e77c5d9c5917db41ff4e5cb5c6977c98799bd2f3d5e75f2ee04ba578463</citedby><cites>FETCH-LOGICAL-c470t-61d202e77c5d9c5917db41ff4e5cb5c6977c98799bd2f3d5e75f2ee04ba578463</cites><orcidid>0000-0003-0506-783X ; 0000-0001-9443-929X ; 0000-0002-1785-1659 ; 0000-0002-4503-2079</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC6897966/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC6897966/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,723,776,780,860,881,27901,27902,41096,42165,51551,53766,53768</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/31840104$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Höfig, Henning</creatorcontrib><creatorcontrib>Yukhnovets, Olessya</creatorcontrib><creatorcontrib>Remes, Cristina</creatorcontrib><creatorcontrib>Kempf, Noemie</creatorcontrib><creatorcontrib>Katranidis, Alexandros</creatorcontrib><creatorcontrib>Kempe, Daryan</creatorcontrib><creatorcontrib>Fitter, Jörg</creatorcontrib><title>Brightness-gated two-color coincidence detection unravels two distinct mechanisms in bacterial protein translation initiation</title><title>Communications biology</title><addtitle>Commun Biol</addtitle><addtitle>Commun Biol</addtitle><description>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.
Henning Höfig et al. present a brightness-gated two-color coincidence detection method that overcomes the limitations of confocal detection. 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Henning Höfig et al. present a brightness-gated two-color coincidence detection method that overcomes the limitations of confocal detection. The method adds a selection criterion for the brightness of single fluorescence bursts, substantially improving the counting accuracy for single fluorophores.</abstract><cop>London</cop><pub>Nature Publishing Group UK</pub><pmid>31840104</pmid><doi>10.1038/s42003-019-0709-7</doi><tpages>1</tpages><orcidid>https://orcid.org/0000-0003-0506-783X</orcidid><orcidid>https://orcid.org/0000-0001-9443-929X</orcidid><orcidid>https://orcid.org/0000-0002-1785-1659</orcidid><orcidid>https://orcid.org/0000-0002-4503-2079</orcidid><oa>free_for_read</oa></addata></record> |
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| subjects | 14/19 14/35 631/1647/2017/2197 631/57/2265 Bacteria - genetics Bacteria - metabolism Bacterial Proteins - genetics Bacterial Proteins - metabolism Biology Biomedical and Life Sciences Brightness Color Fluorescence Resonance Energy Transfer Fluorophores Life Sciences Molecular Imaging - methods Peptide Chain Initiation, Translational Protein biosynthesis Spectrometry, Fluorescence - methods Translation Translation initiation |
| title | Brightness-gated two-color coincidence detection unravels two distinct mechanisms in bacterial protein translation initiation |
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