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Blood, 15 February 2008, Vol. 111, No. 4, pp. 1989-1998. Prepublished online as a Blood First Edition Paper on November 20, 2007; DOI 10.1182/blood-2007-09-113423. This Article Full Text Full Text (PDF) All Versions of this Article: blood-2007-09-113423v1 111/4/1989    most recent Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Rights and Permissions Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Setiadi, H. Articles by McEver, R. P. Search for Related Content PubMed PubMed Citation Articles by Setiadi, H. Articles by McEver, R. P. Related Collections Hemostasis, Thrombosis, and Vascular Biology Social Bookmarking                   What's this?  Previous Article  |  Table of Contents  |  Next Article  HEMOSTASIS, THROMBOSIS, AND VASCULAR BIOLOGY Clustering endothelial E-selectin in clathrin-coated pits and lipid rafts enhances leukocyte adhesion under flow Hendra Setiadi1, and Rodger P. McEver1,2 1 Cardiovascular Biology Research Program, Oklahoma Medical Research Foundation, and 2 Department of Biochemistry and Molecular Biology, Oklahoma Center for Medical Glycobiology, University of Oklahoma Health Sciences Center, Oklahoma City During inflammation, E-selectin expressed on cytokine-activated endothelial cells mediates leukocyte rolling under flow. E-selectin undergoes endocytosis and may associate with lipid rafts. We asked whether distribution of E-selectin in membrane domains affects its functions. E-selectin was internalized in transfected CHO cells or cytokine-activated human umbilical vein endothelial cells (HUVECs). Confocal microscopy demonstrated colocalization of E-selectin with -adaptin, a clathrin-associated protein. Deleting the cytoplasmic domain of E-selectin or disrupting clathrin-coated pits with hypertonic medium blocked internalization of E-selectin, reduced colocalization of E-selectin with -adaptin, and inhibited E-selectin-mediated neutrophil rolling under flow. Unlike CHO cells, HUVECs expressed a small percentage of E-selectin in lipid rafts. Even fewer neutrophils rolled on E-selectin in HUVECs treated with hypertonic medium and with methyl-β-cyclodextrin, which disrupts lipid rafts. These data demonstrate that E-selectin clusters in both clathrin-coated pits and lipid rafts of endothelial cells but is internalized in clathrin-coated pits. Distribution in both domains markedly enhances E-selectin's ability to mediate leukocyte rolling under flow. CiteULike    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this? This article has been cited by other articles: S. Bournazos, S. P. Hart, L. H. Chamberlain, M. J. Glennie, and I. Dransfield Association of Fc{gamma}RIIa (CD32a) with Lipid Rafts Regulates Ligand Binding Activity J. Immunol., June 15, 2009; 182(12): 8026 - 8036. [Abstract] [Full Text] [PDF] J. J. Miner, L. Xia, T. Yago, J. Kappelmayer, Z. Liu, A. G. Klopocki, B. Shao, J. M. McDaniel, H. Setiadi, D. W. Schmidtke, et al. Separable requirements for cytoplasmic domain of PSGL-1 in leukocyte rolling and signaling under flow Blood, September 1, 2008; 112(5): 2035 - 2045. [Abstract] [Full Text] [PDF] T. M. Randis, N. Diacovo, J. Calafat, and T. G. Diacovo Shedding light on class I phosphoinositide 3-kinase activity in endothelium Blood, May 1, 2008; 111(9): 4827 - 4828. [Full Text] [PDF]

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