Diversity in ATP Concentrations in a Single Bacterial Cell Population Revealed by Quantitative Single-cell Imaging

Overview

Journal Sci Rep

Specialty Science

Date 2014 Oct 7

PMID 25283467

Citations 168

Authors

Hideyuki Yaginuma

Shinnosuke Kawai

Kazuhito V Tabata

Keisuke Tomiyama

Akira Kakizuka

Tamiki Komatsuzaki

Hiroyuki Noji

Hiromi Imamura

Affiliations

Soon will be listed here.

Abstract

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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