Identification of a glycoprotein ligand for E-selectin on mouse myeloid cells.
E-selectin is an inducible endothelial cell adhesion molecule for neutrophils which functions as a Ca(2+)-dependent lectin. Using a recombinant, antibody-like form of mouse E-selectin, we have searched for glycoprotein ligands on mouse neutrophils and the neutrophil progenitor cell line 32D cl 3. We have identified a 150-kD glycoprotein as the only protein which could be affinity-isolated with soluble E-selectin from [35S]methionine/[35S]cysteine-labeled 32D cl 3 cells. Binding of this protein was strictly Ca(2+)-dependent, was blocked by a cell adhesion-blocking mAb against mouse E-selectin, and required the presence of sialic acid on the 150-kD ligand. This glycoprotein was also affinity-isolated from mature neutrophils, in addition to a minor component at 250 kD, but could not be isolated from several other non-myeloid cell lines. The 150-kD glycoprotein was the only protein from 32D cl 3 cells, which was detectable by silver-staining after a one-step affinity-isolation.
Levinovitz
A
A
Hans Spemann Laboratory, Max-Planck-Institute of Immunology, Freiburg, FRG.
Mühlhoff
J
J
Isenmann
S
S
Vestweber
D
D
D000818:Animals; D000906:Antibodies / Q000276:immunology; D001665:Binding Sites; D002118:Calcium / Q000494:pharmacology; D015815:Cell Adhesion Molecules / Q000276:immunology / Q000378:metabolism*; D002460:Cell Line / Q000737:chemistry; D019040:E-Selectin; D006023:Glycoproteins / Q000737:chemistry* / Q000302:isolation & purification; D008024:Ligands; D051379:Mice; D008970:Molecular Weight; D019158:N-Acetylneuraminic Acid; D009504:Neutrophils / Q000378:metabolism*; D011993:Recombinant Fusion Proteins; D012794:Sialic Acids / Q000032:analysis / Q000378:metabolism; D014407:Tumor Cells, Cultured
D000906:Antibodies; D015815:Cell Adhesion Molecules; D019040:E-Selectin; D006023:Glycoproteins; D008024:Ligands; D011993:Recombinant Fusion Proteins; D012794:Sialic Acids; D019158:N-Acetylneuraminic Acid; D002118:Calcium