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

• Published in: FEBS letters 2002-08, Vol.525 (1), p.83-87
• Main Authors: Tilghman, Robert W, Hoover, Richard L
• Format: Article
• Language: English
• Subjects: Adhesion; Antibodies - pharmacology; BCD, β-cyclodextrin; 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; Endothelial cell; Endothelium, Vascular - chemistry; Endothelium, Vascular - cytology; Endothelium, Vascular - metabolism; Humans; HUVEC, human umbilical vein endothelial cell; ICAM-1, intercellular adhesion molecule-1; Intercellular Adhesion Molecule-1 - chemistry; Intercellular Adhesion Molecule-1 - metabolism; Lipid raft; Membrane Microdomains - chemistry; Membrane Microdomains - metabolism; Monocytes - cytology; Receptor Aggregation - drug effects; Solubility - drug effects; src-Family Kinases - metabolism; Subcellular Fractions - chemistry; Subcellular Fractions - metabolism; TNFα, tumor necrosis factor-α; Tyrosine kinase
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/S0014-5793(02)03070-3

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.