ArticleScientific reports2016
BTeam, a Novel BRET-based Biosensor for the Accurate Quantification of ATP Concentration within Living Cells.
Article in Scientific reports, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 47 papers.
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47 citing papers in PubMed, 89 citations in OpenAlex.
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- Autonomous multicolor bioluminescence imaging in bacteria, mammalian, and plant hosts.Proceedings of the National Academy of Sciences of the United States of America · 2024Article
- Toward a molecular mechanism for the interaction of ATP with alpha-synuclein.Chemical science · 2023Article
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- Biological and Molecular Components for Genetically Engineering Biosensors in Plants.Biodesign research · 2022Article
- Fueling Cell Invasion through Extracellular Matrix.Trends in cell biology · 2021Review
- Measuring and modeling energy and power consumption in living microbial cells with a synthetic ATP reporter.BMC biology · 2021Article
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Authors and funding
3 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
ATP levels may represent fundamental health conditions of cells. However, precise measurement of intracellular ATP levels in living cells is hindered by the lack of suitable methodologies. Here, we developed a novel ATP biosensor termed "BTeam". BTeam comprises a yellow fluorescent protein (YFP), the ATP binding domain of the ε subunit of the bacterial ATP synthase, and an ATP-nonconsuming luciferase (NLuc). To attain emission, BTeam simply required NLuc substrate. BTeam showed elevated bioluminescence resonance energy transfer efficiency upon ATP binding, resulted in the emission spectra changes correlating with ATP concentrations. By using values of YFP/NLuc emission ratio to represent ATP levels, BTeam achieved steady signal outputs even though emission intensities were altered. With this biosensor, we succeeded in the accurate quantification of intracellular ATP concentrations of a population of living cells, as demonstrated by detecting the slight distribution in the cytosol (3.7-4.1 mM) and mitochondrial matrix (2.4-2.7 mM) within some cultured cell lines. Furthermore, BTeam allowed continuous tracing of cytosolic ATP levels of the same cells, as well as bioluminescent imaging of cytosolic ATP dynamics within individual cells. This simple and accurate technique should be an effective method for quantitative measurement of intracellular ATP concentrations.
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