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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...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group
2014
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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. |
format | Online Article Text |
id | pubmed-4185378 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
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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