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Ras diffusion is sensitive to plasma membrane viscosity.

J Shawn Goodwin1, Kimberly R Drake, Catha L Remmert

  • 1Department of Molecular Physiology & Biophysics, Vanderbilt University School of Medicine, Nashville, Tennessee 37232, USA.

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|June 1, 2005
PubMed
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This summary is machine-generated.

The small GTPase Ras

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Area of Science:

  • Cell biology
  • Membrane biophysics

Background:

  • Protein diffusion on the cell surface is hindered by membrane barriers.
  • Regulation of inner leaflet protein diffusion remains largely unknown.

Purpose of the Study:

  • To investigate if the diffusion of the small GTPase Ras is influenced by plasma membrane viscosity.
  • To explore the role of cholesterol in modulating Ras protein diffusion.

Main Methods:

  • Confocal fluorescence recovery after photobleaching (FRAP).
  • Utilized green fluorescent protein (GFP)-tagged HRas, NRas, and KRas.
  • Manipulated cellular cholesterol levels in COS-7 cells.

Main Results:

  • Increased cellular cholesterol significantly reduced the diffusion of GFP-tagged Ras proteins.
  • Ras protein diffusion changes mirrored those of viscosity-sensitive lipid probes.
  • Cholesterol depletion effects varied by method: methyl-beta-cyclodextrin slowed diffusion via a non-viscosity mechanism, while overnight depletion increased diffusion.

Conclusions:

  • Ras protein diffusion is sensitive to plasma membrane viscosity.
  • Cholesterol content directly impacts Ras mobility.
  • Viscosity sensing may be a general property of cytoplasmic face proteins.