Work

Dynamic analysis of cytosolic glucose and ATP levels in yeast using optical sensors

Year: 2010

Type: article

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. In the present study, genetically encoded FRET (Förster resonance energy transfer)-based sensors were deployed for dynamic measurements of free cytosolic glucose and ATP with 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 of 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 a hxt5Δ hexose-transporter mutant compared with wild-type and other hxtΔ strains.

Source: Biochemical Journal

DOI: 10.1042/bj20100946

Authors: Clara Bermejo, Farzad Haerizadeh, Hitomi Takanaga, Diane Chermak, Wolf B. Frommer

Institutions: Carnegie Institution for Science, Carnegie Department of Plant Biology

Language: English

Topic: Advanced Fluorescence Microscopy Techniques

Subfield: Biophysics

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: green

Funder: National Institute of Diabetes and Digestive and Kidney Diseases

Award: Sugar signaling networks detected by high content fluxomicx