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Related Experiment Videos

Development of a fluorescent nanosensor for ribose.

Ida Lager1, Marcus Fehr, Wolf B Frommer

  • 1ZMBP Tübingen, Plant Physiology, Tübingen, Germany.

FEBS Letters
|October 11, 2003
PubMed
Summary

Researchers developed nanosensors to track ribose sugar in cells. They found ribose enters cells via glucose transporters and is slowly metabolized, with uptake appearing irreversible.

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Area of Science:

  • Biochemistry
  • Cell Biology
  • Molecular Imaging

Background:

  • Ribose is a fundamental sugar in cellular processes.
  • Understanding ribose transport and metabolism is crucial for cell biology.
  • Existing methods for tracking intracellular ribose are limited.

Purpose of the Study:

  • To develop and utilize a novel nanosensor for real-time analysis of ribose uptake and metabolism in living cells.
  • To investigate the mechanism and kinetics of ribose transport across the cell membrane.
  • To explore the metabolic fate of intracellular ribose.

Main Methods:

  • Engineering of nanosensors by fusing Escherichia coli ribose binding protein with green fluorescent protein variants.
  • Utilizing Förster Resonance Energy Transfer (FRET) to quantify ribose concentration.
  • Generating and characterizing five affinity mutants of the nanosensor.
  • Performing experiments in COS-7 cells, including inhibitor studies targeting monosaccharide transporters.

Main Results:

  • The nanosensor demonstrated a decrease in FRET efficiency with increasing ribose concentration.
  • Analysis in COS-7 cells revealed intracellular accumulation of free ribose.
  • Ribose was observed to be slowly metabolized within the cells.
  • Inhibitor studies implicated monosaccharide transporters of the GLUT family in ribose uptake.
  • Uptake of ribose into the cells appeared to be irreversible or showed very slow release.

Conclusions:

  • The developed nanosensor is effective for monitoring intracellular ribose dynamics.
  • Ribose uptake in COS-7 cells is mediated by GLUT family transporters.
  • Intracellular ribose accumulates and is slowly metabolized, suggesting a potential bottleneck in its utilization or release.

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