The E-selectin-ligand ESL-1 is located in the Golgi as well as on microvilli on the cell surface.
Neutrophils and subsets of lymphocytes bind to E-selectin, a cytokine inducible adhesion molecule on endothelial cells. The E-selectin-ligand-1 (ESL-1) is a high affinity glycoprotein ligand which participates in the binding of mouse myeloid cells to E-selectin. The sequence of mouse ESL-1 is highly homologous to the cysteine rich FGF receptor (CFR) in chicken and the rat Golgi protein MG160. We have analysed the subcellular distribution of ESL-1 by indirect immunofluorescence, flow cytometry, various biochemical techniques and by immunogold scanning electron microscopy. We could localize ESL-1 in the Golgi as well as on the cell surface of 32Dc13 cells and neutrophils. Cell surface staining was confirmed by cell surface biotinylation and by cell surface immunoprecipitations in which antibodies only had access to surface proteins on intact cells. In addition, ESL-1(high) and ESL-1(low) expressing cells, sorted by flow cytometry, gave rise to high and low immunoprecipitation signals for ESL-1, respectively. Based on immunogold labeling of intact cells, we localized ESL-1 on microvilli of 32Dc13 cells and of the lymphoma cell line K46. Quantitative evaluation determined 80% of the total labeling for ESL-1 on microvilli of K46 cells while 69% of the labeling for the control antigen B220 was found on the planar cell surface. These data indicate that ESL-1 occurs at sites on the leukocyte cell surface which are destined for the initiation of cell contacts to the endothelium.
Steegmaier
M
M
Institute of Cell Biology, ZMBE, University of Münster, Germany.
Borges
E
E
Berger
J
J
Schwarz
H
H
Vestweber
D
D
D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D002645:Chickens; D019084:Fluorescent Antibody Technique, Indirect; D006056:Golgi Apparatus / Q000378:metabolism* / Q000648:ultrastructure; D007962:Leukocytes / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000378:metabolism*; D051379:Mice; D008855:Microscopy, Electron, Scanning; D008871:Microvilli / Q000378:metabolism* / Q000648:ultrastructure; D051381:Rats; D017468:Receptors, Fibroblast Growth Factor / Q000378:metabolism*; D012795:Sialoglycoproteins; D013347:Subcellular Fractions
D008562:Membrane Glycoproteins; D017468:Receptors, Fibroblast Growth Factor; D012795:Sialoglycoproteins; C086711:cysteine-rich fibroblast growth factor receptor