Reference: Bermejo C, et al. (2010) Dynamic analysis of cytosolic glucose and ATP levels in yeast using optical sensors. Biochem J 432(2):399-406

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Abstract

Precise and dynamic measurement of intracellular metabolite levels has been hampered by difficulties in differentiating between adsorbed and imported fractions and the subcellular distribution between cytosol, endomembrane compartments and mitochondria. Here, genetically encoded Forster Resonance Energy Transfer (FRET)-based sensors were deployed for dynamic measurements of free cytosolic glucose and ATP at varying external supply and in glucose transport mutants. Moreover, by using the FRET sensors in a microfluidic platform, we were able to monitor in vivo changes in intracellular free glucose in individual yeast cells. We demonstrate the suitability of the FRET sensors for gaining physiological insight by demonstrating that free intracellular glucose and ATP levels are reduced in an hxt5 hexose transporter mutant compared to wild type and to other hxt strains.

Reference Type
Journal Article
Authors
Bermejo C, Haerizadeh F, Takanaga H, Chermak D, Frommer WB
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