title: Expression cloning of a functional glycoprotein ligand for P-selectin.
abstract: The initial adhesive interactions between circulating leukocytes and endothelia are mediated, in part, by P-selectin. We now report the expression cloning of a functional ligand for P-selectin from an HL-60 cDNA library. The predicted amino acid sequence reveals a novel mucin-like transmembrane protein. Significant binding of transfected COS cells to P-selectin requires coexpression of both the protein ligand and a fucosyltransferase. This binding is calcium dependent and can be inhibited by a neutralizing monoclonal antibody to P-selectin. Cotransfected COS cells express the ligand as a homodimer of 220 kd. A soluble ligand construct, when coexpressed with fucosyltransferase in COS cells, also mediates P-selectin binding and is immunocrossreactive with the major HL-60 glycoprotein that specifically binds P-selectin.
authors.affiliation: Genetics Institute, Small Molecule Drug Discovery Group, Cambridge, Massachusetts 02140.
authors.forename: D
authors.initials: D
authors.lastname: Sako
authors.forename: X J
authors.initials: XJ
authors.lastname: Chang
authors.forename: K M
authors.initials: KM
authors.lastname: Barone
authors.forename: G
authors.initials: G
authors.lastname: Vachino
authors.forename: H M
authors.initials: HM
authors.lastname: White
authors.forename: G
authors.initials: G
authors.lastname: Shaw
authors.forename: G M
authors.initials: GM
authors.lastname: Veldman
authors.forename: K M
authors.initials: KM
authors.lastname: Bean
authors.forename: T J
authors.initials: TJ
authors.lastname: Ahern
authors.forename: B
authors.initials: B
authors.lastname: Furie
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D016466:CHO Cells; D002448:Cell Adhesion; D015815:Cell Adhesion Molecules / Q000378:metabolism; D002460:Cell Line; D003001:Cloning, Molecular; D006224:Cricetinae; D018076:DNA, Complementary / Q000378:metabolism; D004730:Endothelium, Vascular / Q000502:physiology; D015723:Gene Library; D006801:Humans; D015473:Leukemia, Promyelocytic, Acute; D007962:Leukocytes / Q000502:physiology; D046911:Macromolecular Substances; D008562:Membrane Glycoproteins / Q000096:biosynthesis* / Q000378:metabolism; D008969:Molecular Sequence Data; D019007:P-Selectin; D010980:Platelet Membrane Glycoproteins / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D014162:Transfection; D014407:Tumor Cells, Cultured
chemical_list: D015815:Cell Adhesion Molecules; D018076:DNA, Complementary; D046911:Macromolecular Substances; D008562:Membrane Glycoproteins; D019007:P-Selectin; C084882:P-selectin ligand protein; D010980:Platelet Membrane Glycoproteins