BTeam, a Novel BRET-based Biosensor for the Accurate Quantification of ATP Concentration within Living Cells.

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 th

SUBMITTER: Yoshida T 

PROVIDER: S-EPMC5175186 | biostudies-literature | 2016 Dec

REPOSITORIES: biostudies-literature