Heat-stable antigen (CD24) as ligand for mouse P-selectin.
Heat-stable antigen (HSA)/CD24 is a cell surface molecule expressed by many cell types in the mouse. The molecule has an unusual structure because of its small protein core and extensive glycosylation. In order to study the functional role of the HSA-associated glycoconjugates we have isolated different forms of HSA. Using lectin analysis we provide evidence for extensive heterogeneity in carbohydrate composition and sialic acid linkage. Several HSA forms were recognized by mouse P-selectin-IgG but not E-selectin-IgG in ELISA. As expected, P-selectin-IgG also bound to L2/HNK-1-positive neural glycoproteins (L2-glycoproteins) and sulfatides but not to gangliosides and other control glycoproteins. The binding of P-selectin-IgG to L2-glycoproteins and HSA required bivalent cations. The reactivity to HSA was sensitive to sialidase treatment whereas the binding to L2-glycoproteins was not. Studies with alpha 2-6 sialytransferase indicated that alpha 2-6 linked sialic acid was not involved in the P-selectin binding to HSA. Surprisingly, an L2/HNK-1 specific antibody was found to cross-react with some HSA glycoforms and its binding correlated with P-selectin-IgG reactivity. L2/HNK-1-positive or L2/HNK-1-negative HSA glycoforms were also analyzed after coating to polystyrene beads. Only the L2/HNK-1-positive HSA coated beads were reactive with P-selectin-IgG and could bind to activated bend3 endothelioma cells expressing P-selectin whereas the L2/HNK-1-negative HSA beads did not. It is suggested that in its L2/HNK-1 modified form the HSA molecule on leukocytes could represent a ligand for P-selectin on endothelial cells or platelets.
Sammar
M
M
Tumor Immunology Programme, German Cancer Research Center, Heidelberg.
Aigner
S
S
Hubbe
M
M
Schirrmacher
V
V
Schachner
M
M
Vestweber
D
D
Altevogt
P
P
D000818:Animals; D015703:Antigens, CD / Q000378:metabolism*; D001665:Binding Sites / Q000187:drug effects; D051927:CD24 Antigen; D015815:Cell Adhesion Molecules / Q000378:metabolism; D019040:E-Selectin; D004727:Endothelium / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D007074:Immunoglobulin G / Q000378:metabolism; D008024:Ligands; D008562:Membrane Glycoproteins; D051379:Mice; D008807:Mice, Inbred BALB C; D008811:Mice, Inbred DBA; D008863:Microspheres; D009439:Neuraminidase / Q000494:pharmacology; D019007:P-Selectin; D010980:Platelet Membrane Glycoproteins / Q000378:metabolism*; D012799:Sialyltransferases / Q000494:pharmacology; D014407:Tumor Cells, Cultured; D000094802:beta-D-Galactoside alpha 2-6-Sialyltransferase
D015703:Antigens, CD; D051927:CD24 Antigen; C497890:Cd24a protein, mouse; D015815:Cell Adhesion Molecules; D019040:E-Selectin; D007074:Immunoglobulin G; D008024:Ligands; D008562:Membrane Glycoproteins; D019007:P-Selectin; D010980:Platelet Membrane Glycoproteins; D012799:Sialyltransferases; D009439:Neuraminidase; D000094802:beta-D-Galactoside alpha 2-6-Sialyltransferase