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• W2567554261 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." @default.
• W2567554261 created "2017-01-06" @default.
• W2567554261 creator A5015613537 @default.
• W2567554261 creator A5055094058 @default.
• W2567554261 creator A5056908833 @default.
• W2567554261 date "2016-12-01" @default.
• W2567554261 modified "2023-10-18" @default.
• W2567554261 title "BTeam, a Novel BRET-based Biosensor for the Accurate Quantification of ATP Concentration within Living Cells" @default.
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• W2567554261 doi "https://doi.org/10.1038/srep39618" @default.
• W2567554261 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/5175186" @default.
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• W2567554261 hasPublicationYear "2016" @default.
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