title: The HNK-1 reactive sulfoglucuronyl glycolipids are ligands for L-selectin and P-selectin but not E-selectin.
abstract: E-selectin, L-selectin, and P-selectin are related cell adhesion molecules that bind via their lectin domains to sialyl Lewis x and related carbohydrate determinants. Reports have indicated that sulfated glycolipids and polysaccharides also bind selectins. To extend these findings, we compared binding of selectin-IgG chimeras to immobilized sulfated and sialylated glycosphingolipids. E-, L-, and P-selectin chimeras all bound to surfaces absorbed with 2,3-sialyl Lewis x glycolipid or sulfatide (galactosylceramide I3-sulfate) but not to surfaces adsorbed with control sulfated lipids (octadecyl sulfate, sphingosine sulfate). Notably, the L- and P-selectin chimeras but not E-selectin chimera bound to surfaces adsorbed with sulfoglucuronyl glycosphingolipids (SGNL lipids; e.g., IV3 glucuronylneolactotetraosylceramide V3-sulfate). These unusual lipids have been reported as antigenic determinants for monoclonal IgM antibodies produced in patients with neuropathy associated with paraproteinemia and react with the mouse monoclonal antibody HNK-1. Binding of L- and P-selectin chimeras to SGNL lipids was specifically inhibited by appropriate anti-selectin antibodies. While binding of all three selectin chimeras to sialyl Lewis x was blocked by removal of calcium, binding to SGNL lipid was only modestly reduced by EDTA. Chemically desulfated SGNL lipid retained binding activity for L- and P-selectin chimeras, while methyl esterification of the glucuronic acid eliminated binding. We conclude that SGNL lipids, unlike sialyl Lewis x and sulfatides, selectively support L- and P-selectin but not E-selectin chimera binding. The presence of SGNL lipids on brain microvascular endothelium (and other endothelia) may implicate these molecules in leukocyte trafficking to the nervous system and elsewhere.
authors.affiliation: Department of Pharmacology, Johns Hopkins University School of Medicine, Baltimore, MD 21205.
authors.forename: L K
authors.initials: LK
authors.lastname: Needham
authors.forename: R L
authors.initials: RL
authors.lastname: Schnaar
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D000943:Antigens, Differentiation / Q000276:immunology / Q000378:metabolism*; D018959:CD57 Antigens; D002417:Cattle; D002420:Cauda Equina / Q000737:chemistry; D015815:Cell Adhesion Molecules / Q000378:metabolism*; D019040:E-Selectin; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis / Q000378:metabolism; D006017:Glycolipids / Q000378:metabolism*; D006028:Glycosphingolipids / Q000494:pharmacology; D006801:Humans; D007074:Immunoglobulin G / Q000378:metabolism; D007075:Immunoglobulin M / Q000378:metabolism; D007700:Kinetics; D019041:L-Selectin; D008024:Ligands; D008562:Membrane Glycoproteins / Q000378:metabolism; D019007:P-Selectin; D010980:Platelet Membrane Glycoproteins / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013329:Structure-Activity Relationship; D013433:Sulfoglycosphingolipids / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology
chemical_list: D000911:Antibodies, Monoclonal; D000943:Antigens, Differentiation; D018959:CD57 Antigens; D015815:Cell Adhesion Molecules; D019040:E-Selectin; D000939:Epitopes; D006017:Glycolipids; D006028:Glycosphingolipids; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D008024:Ligands; D008562:Membrane Glycoproteins; D019007:P-Selectin; D010980:Platelet Membrane Glycoproteins; D011993:Recombinant Fusion Proteins; D013433:Sulfoglycosphingolipids; D019041:L-Selectin