E-selectin and ICAM-1 are incorporated into detergent-insoluble membrane domains following clustering in endothelial cells.
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.
Tilghman
Robert W
RW
Department of Pathology, Vanderbilt University School of Medicine, U4202 Medical Center North, 1161 21st Avenue South, Nashville, TN 37232, USA.
Hoover
Richard L
RL
D000906:Antibodies / Q000494:pharmacology; D015153:Blotting, Western; D002448:Cell Adhesion / Q000187:drug effects / Q000502:physiology; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism*; D002478:Cells, Cultured; D002784:Cholesterol / Q000378:metabolism; D003902:Detergents / Q000737:chemistry / Q000494:pharmacology; D019040:E-Selectin / Q000737:chemistry / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D004730:Endothelium, Vascular / Q000737:chemistry / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000737:chemistry / Q000378:metabolism*; D021962:Membrane Microdomains / Q000737:chemistry / Q000378:metabolism*; D009000:Monocytes / Q000166:cytology; D011940:Receptor Aggregation / Q000187:drug effects; D012995:Solubility / Q000187:drug effects; D013347:Subcellular Fractions / Q000737:chemistry / Q000378:metabolism; D019061:src-Family Kinases / Q000378:metabolism
D000906:Antibodies; D003902:Detergents; D019040:E-Selectin; D018799:Intercellular Adhesion Molecule-1; D002784:Cholesterol; D019061:src-Family Kinases