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Related Experiment Videos

GFP associates with microfilaments in fixed cells.

H D Schmitz1, J Bereiter-Hahn

  • 1Kinematic Cell Research Group, Biocentre, Goethe University of Frankfurt/Main, Marie-Curie-Strasse 9, 60439 Frankfurt, Germany. d.schmitz@em.uni-frankfurt.de

Histochemistry and Cell Biology
|August 2, 2001
PubMed
Summary
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Green fluorescent protein (GFP) binds to microfilaments in fixed cells but not in living cells. This actin-binding property necessitates careful controls when using GFP-tagged proteins to study cellular structures in vivo.

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • Green fluorescent protein (GFP) is a widely used marker in cell biology.
  • It is typically considered inert and homogeneously distributed in the cytoplasm.
  • Applications include protein localization and studying cytoskeleton dynamics.

Purpose of the Study:

  • To investigate the intracellular distribution of GFP.
  • To determine if GFP interacts with cellular structures.
  • To assess the reliability of GFP as a marker in different cell types.

Main Methods:

  • Confocal laser scanning microscopy was used.
  • Studies were conducted on amphibian and various mammalian cell lines (XTH2, CHO-K1, HaCaT, MDCK, NIH-3T3).
  • Experiments involved both fixed and living cells, with and without detergent treatment.

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Main Results:

  • Fixed cells showed GFP associated with microfilaments after paraformaldehyde fixation.
  • This association persisted after detergent treatment.
  • In contrast, GFP association with stress fibers was not observed in living cells.

Conclusions:

  • GFP exhibits actin-binding properties in fixed cells.
  • This interaction is not detectable in living cells.
  • Careful controls are essential when using GFP-tagged proteins, and in vivo monitoring is recommended.