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Targeted fluorescent probes in peroxisome function.

T B Dansen1, Pap EHW, R J Wanders

  • 1Institute of Biomembranes, Centre for Biomembranes and Lipid Enzymology, Department of Biochemistry of Lipids, Utrecht University, The Netherlands.

The Histochemical Journal
|July 4, 2001
PubMed
Summary
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New fluorescent peptides target peroxisomes, enabling visualization of cellular processes. These probes reveal peroxisomal pH in living cells and aid in understanding import defects in genetic disorders.

Area of Science:

  • Cell Biology
  • Biochemistry
  • Molecular Imaging

Background:

  • Fluorescent peptides are emerging as advanced tools for visualizing intracellular dynamics.
  • They combine sensitive fluorescence detection with specific amino acid sequence information.
  • Targeting these peptides to specific organelles like peroxisomes allows for detailed cellular analysis.

Purpose of the Study:

  • To develop and utilize fluorescent peptides for targeting peroxisomes in live cells.
  • To investigate the mechanism of peptide uptake and translocation into peroxisomes.
  • To measure the internal pH of peroxisomes and study pH alterations in genetic disorders.

Main Methods:

  • Synthesis of fluorescent peptides conjugated with peroxisomal targeting signal 1 (PTS1).

Related Experiment Videos

  • Incubation of peptides with intact cells at various temperatures and ATP levels.
  • Confocal microscopy and ratio-imaging techniques for fluorescence detection and pH measurement.
  • Analysis of peptide uptake and peroxisomal localization in healthy and patient-derived fibroblasts.

    • Main Results:

    • Fluorescent PTS1-peptides were successfully targeted to peroxisomes in intact cells within 15 minutes.
    • Cellular uptake was diffusion-driven and not energy-dependent, while translocation to peroxisomes required active processes.
    • Peroxisomes in human fibroblasts were found to have an internal pH of 8.2.
    • Fibroblasts from patients with peroxisomal import defects (chondrodysplasia punctata) showed altered peroxisomal pH (6.8).

    Conclusions:

    • Fluorescent peptides are effective tools for studying peroxisome function and dynamics in living cells.
    • The study established a method for measuring peroxisomal pH using pH-sensitive fluorescent probes.
    • These findings provide insights into peroxisomal pH homeostasis and its disruption in specific genetic conditions.