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Diversity in ATP concentrations in a single bacterial cell population revealed by quantitative single-cell imaging

Recent advances in quantitative single-cell analysis revealed large diversity in gene expression levels between individual cells, which could affect the physiology and/or fate of each cell. In contrast, for most metabolites, the concentrations were only measureable as ensemble averages of many cells...

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Autores principales: Yaginuma, Hideyuki, Kawai, Shinnosuke, Tabata, Kazuhito V., Tomiyama, Keisuke, Kakizuka, Akira, Komatsuzaki, Tamiki, Noji, Hiroyuki, Imamura, Hiromi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4185378/
https://www.ncbi.nlm.nih.gov/pubmed/25283467
http://dx.doi.org/10.1038/srep06522
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author Yaginuma, Hideyuki
Kawai, Shinnosuke
Tabata, Kazuhito V.
Tomiyama, Keisuke
Kakizuka, Akira
Komatsuzaki, Tamiki
Noji, Hiroyuki
Imamura, Hiromi
author_facet Yaginuma, Hideyuki
Kawai, Shinnosuke
Tabata, Kazuhito V.
Tomiyama, Keisuke
Kakizuka, Akira
Komatsuzaki, Tamiki
Noji, Hiroyuki
Imamura, Hiromi
author_sort Yaginuma, Hideyuki
collection PubMed
description Recent advances in quantitative single-cell analysis revealed large diversity in gene expression levels between individual cells, which could affect the physiology and/or fate of each cell. In contrast, for most metabolites, the concentrations were only measureable as ensemble averages of many cells. In living cells, adenosine triphosphate (ATP) is a critically important metabolite that powers many intracellular reactions. Quantitative measurement of the absolute ATP concentration in individual cells has not been achieved because of the lack of reliable methods. In this study, we developed a new genetically-encoded ratiometric fluorescent ATP indicator “QUEEN”, which is composed of a single circularly-permuted fluorescent protein and a bacterial ATP binding protein. Unlike previous FRET-based indicators, QUEEN was apparently insensitive to bacteria growth rate changes. Importantly, intracellular ATP concentrations of numbers of bacterial cells calculated from QUEEN fluorescence were almost equal to those from firefly luciferase assay. Thus, QUEEN is suitable for quantifying the absolute ATP concentration inside bacteria cells. Finally, we found that, even for a genetically-identical Escherichia coli cell population, absolute concentrations of intracellular ATP were significantly diverse between individual cells from the same culture, by imaging QUEEN signals from single cells.
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spelling pubmed-41853782014-10-17 Diversity in ATP concentrations in a single bacterial cell population revealed by quantitative single-cell imaging Yaginuma, Hideyuki Kawai, Shinnosuke Tabata, Kazuhito V. Tomiyama, Keisuke Kakizuka, Akira Komatsuzaki, Tamiki Noji, Hiroyuki Imamura, Hiromi Sci Rep Article Recent advances in quantitative single-cell analysis revealed large diversity in gene expression levels between individual cells, which could affect the physiology and/or fate of each cell. In contrast, for most metabolites, the concentrations were only measureable as ensemble averages of many cells. In living cells, adenosine triphosphate (ATP) is a critically important metabolite that powers many intracellular reactions. Quantitative measurement of the absolute ATP concentration in individual cells has not been achieved because of the lack of reliable methods. In this study, we developed a new genetically-encoded ratiometric fluorescent ATP indicator “QUEEN”, which is composed of a single circularly-permuted fluorescent protein and a bacterial ATP binding protein. Unlike previous FRET-based indicators, QUEEN was apparently insensitive to bacteria growth rate changes. Importantly, intracellular ATP concentrations of numbers of bacterial cells calculated from QUEEN fluorescence were almost equal to those from firefly luciferase assay. Thus, QUEEN is suitable for quantifying the absolute ATP concentration inside bacteria cells. Finally, we found that, even for a genetically-identical Escherichia coli cell population, absolute concentrations of intracellular ATP were significantly diverse between individual cells from the same culture, by imaging QUEEN signals from single cells. Nature Publishing Group 2014-10-06 /pmc/articles/PMC4185378/ /pubmed/25283467 http://dx.doi.org/10.1038/srep06522 Text en Copyright © 2014, Macmillan Publishers Limited. All rights reserved http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder in order to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Article
Yaginuma, Hideyuki
Kawai, Shinnosuke
Tabata, Kazuhito V.
Tomiyama, Keisuke
Kakizuka, Akira
Komatsuzaki, Tamiki
Noji, Hiroyuki
Imamura, Hiromi
Diversity in ATP concentrations in a single bacterial cell population revealed by quantitative single-cell imaging
title Diversity in ATP concentrations in a single bacterial cell population revealed by quantitative single-cell imaging
title_full Diversity in ATP concentrations in a single bacterial cell population revealed by quantitative single-cell imaging
title_fullStr Diversity in ATP concentrations in a single bacterial cell population revealed by quantitative single-cell imaging
title_full_unstemmed Diversity in ATP concentrations in a single bacterial cell population revealed by quantitative single-cell imaging
title_short Diversity in ATP concentrations in a single bacterial cell population revealed by quantitative single-cell imaging
title_sort diversity in atp concentrations in a single bacterial cell population revealed by quantitative single-cell imaging
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4185378/
https://www.ncbi.nlm.nih.gov/pubmed/25283467
http://dx.doi.org/10.1038/srep06522
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