Multiple cellular proteins modulate the dynamics of K-ras association with the plasma membrane

Biophys J. 2010 Nov 17;99(10):3327-35. doi: 10.1016/j.bpj.2010.10.001.

Abstract

Although specific proteins have been identified that regulate the membrane association and facilitate intracellular transport of prenylated Rho- and Rab-family proteins, it is not known whether cellular proteins fulfill similar roles for other prenylated species, such as Ras-family proteins. We used a previously described method to evaluate how several cellular proteins, previously identified as potential binding partners (but not effectors) of K-ras4B, influence the dynamics of K-ras association with the plasma membrane. Overexpression of either PDEδ or PRA1 enhances, whereas knockdown of either protein reduces, the rate of dissociation of K-ras from the plasma membrane. Inhibition of calmodulin likewise reduces the rate of K-ras dissociation from the plasma membrane, in this case in a manner specific for the activated form of K-ras. By contrast, galectin-3 specifically reduces the rate of plasma membrane dissociation of activated K-ras, an effect that is blocked by the K-ras antagonist farnesylthiosalicylic acid (salirasib). Multiple cellular proteins thus control the dynamics of membrane association and intercompartmental movement of K-ras to an important degree even under basal cellular conditions.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Calmodulin / metabolism
  • Cell Membrane / drug effects
  • Cell Membrane / metabolism*
  • Cyclic Nucleotide Phosphodiesterases, Type 6 / metabolism
  • Farnesol / analogs & derivatives
  • Farnesol / pharmacology
  • GTP-Binding Proteins / metabolism
  • Galectin 3 / metabolism
  • HeLa Cells
  • Humans
  • Intracellular Space / drug effects
  • Intracellular Space / metabolism
  • Kinetics
  • Microscopy, Confocal
  • Molecular Sequence Data
  • Peptides / chemistry
  • Peptides / metabolism
  • Prenylation / drug effects
  • Protein Binding / drug effects
  • Proto-Oncogene Proteins p21(ras) / metabolism*
  • Salicylates / pharmacology
  • Sirolimus / pharmacology
  • Vesicular Transport Proteins / metabolism

Substances

  • Calmodulin
  • Galectin 3
  • Peptides
  • Salicylates
  • Vesicular Transport Proteins
  • farnesylthiosalicylic acid
  • Farnesol
  • Cyclic Nucleotide Phosphodiesterases, Type 6
  • GTP-Binding Proteins
  • K-Ras4B protein, human
  • RABAC1 protein, human
  • Proto-Oncogene Proteins p21(ras)
  • Sirolimus