E-selectin and ICAM-1 are incorporated into detergent-insoluble membrane domains following clustering in endothelial cells

FEBS Lett. 2002 Aug 14;525(1-3):83-7. doi: 10.1016/s0014-5793(02)03070-3.

Abstract

Here we present data supporting the role of lipid rafts in endothelial cells during leukocyte adhesion. Following adhesion of THP-1 cells or antibody-mediated clustering, both E-selectin and intercellular adhesion molecule-1 (ICAM-1) partitioned into the detergent-insoluble portion of the endothelial cellular lysate. Sucrose gradient centrifugation revealed the partitioning of clustered E-selectin and ICAM-1 with the low-density fraction where they co-fractionated with src family kinases, markers of lipid rafts. Depleting the plasma membrane of cholesterol inhibited clustering of adhesion molecules following their antibody-induced crosslinking and inhibited their association with src kinases. Thus, our data suggest that E-selectin and ICAM-1 associate with lipid rafts in human endothelial cells following leukocyte adhesion.

MeSH terms

  • Antibodies / pharmacology
  • Blotting, Western
  • Cell Adhesion / drug effects
  • Cell Adhesion / physiology
  • Cell Membrane / chemistry
  • Cell Membrane / metabolism*
  • Cells, Cultured
  • Cholesterol / metabolism
  • Detergents / chemistry
  • Detergents / pharmacology
  • E-Selectin / chemistry
  • E-Selectin / metabolism*
  • Electrophoresis, Polyacrylamide Gel
  • Endothelium, Vascular / chemistry
  • Endothelium, Vascular / cytology
  • Endothelium, Vascular / metabolism*
  • Humans
  • Intercellular Adhesion Molecule-1 / chemistry
  • Intercellular Adhesion Molecule-1 / metabolism*
  • Membrane Microdomains / chemistry
  • Membrane Microdomains / metabolism*
  • Monocytes / cytology
  • Receptor Aggregation / drug effects
  • Solubility / drug effects
  • Subcellular Fractions / chemistry
  • Subcellular Fractions / metabolism
  • src-Family Kinases / metabolism

Substances

  • Antibodies
  • Detergents
  • E-Selectin
  • Intercellular Adhesion Molecule-1
  • Cholesterol
  • src-Family Kinases