Use of Rab GTPases to study lipid trafficking in normal and sphingolipid storage disease fibroblasts

Methods Enzymol. 2005;403:166-82. doi: 10.1016/S0076-6879(05)03014-4.

Abstract

We describe methods for studying lipid transport in normal and sphingolipid storage disease fibroblasts. These techniques include endocytic assays with fluorescent sphingolipid analogs, expression of dominant negative (DN) Rab GTPases, and methods of manipulating cholesterol levels in intact cells and isolated cell membranes. These methods should be useful in future studies of lipid trafficking in normal and disease cell types.

MeSH terms

  • Biological Transport
  • Cell Membrane / metabolism
  • Cholesterol / metabolism
  • Fibroblasts / metabolism
  • Glutathione Transferase / metabolism
  • Golgi Apparatus / metabolism
  • Guanine Nucleotide Dissociation Inhibitors / metabolism
  • Humans
  • Lipid Metabolism*
  • Mutation
  • Skin / metabolism
  • Skin / pathology
  • Sphingolipidoses / metabolism*
  • Sphingolipidoses / pathology
  • rab GTP-Binding Proteins / genetics
  • rab GTP-Binding Proteins / metabolism*
  • rho-Specific Guanine Nucleotide Dissociation Inhibitors

Substances

  • Guanine Nucleotide Dissociation Inhibitors
  • rho-Specific Guanine Nucleotide Dissociation Inhibitors
  • Cholesterol
  • Glutathione Transferase
  • rab GTP-Binding Proteins