PSGL-1 recognition of P-selectin is controlled by a tyrosine sulfation consensus at the PSGL-1 amino terminus.
P-selectin binding to neutrophils requires a specific protein, P-selectin glycoprotein ligand 1 (PSGL-1), as well as sialyl-Lewis X (sLex) glycan determinants. We have found that a short segment near the amino terminus of PSGL-1 that contains a tyrosine sulfation consensus is essential for P-selectin adhesion and that addition of the amino-terminal segment to some but not all mucin-like molecules confers on those molecules the ability to bind P-selectin. PSGL-1 synthesized in the presence of sulfation inhibitors binds P-selectin weakly, and within the amino-terminal 20 residues, mutation of the tyrosines to phenylalanine abolishes binding. Rolling of HL-60 cells on P-selectin-coated coverslips is strongly attenuated by treatment of cells with an inhibitor of sulfation.
Pouyani
T
T
Department of Genetics, Harvard Medical School, Massachusetts General Hospital, Boston 02114, USA.
Seed
B
B
D000595:Amino Acid Sequence; D000818:Animals; D002448:Cell Adhesion / Q000502:physiology; D002478:Cells, Cultured; D002704:Chlorates / Q000494:pharmacology; D016384:Consensus Sequence; D004252:DNA Mutational Analysis; D004730:Endothelium, Vascular / Q000502:physiology; D018922:HL-60 Cells; D006801:Humans; D007962:Leukocytes / Q000502:physiology; D008024:Ligands; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D009077:Mucins / Q000235:genetics / Q000378:metabolism*; D009844:Oligosaccharides; D019007:P-Selectin / Q000378:metabolism*; D010446:Peptide Fragments / Q000378:metabolism; D011485:Protein Binding; D011499:Protein Processing, Post-Translational / Q000187:drug effects; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D017384:Sequence Deletion; D000080506:Sialyl Lewis X Antigen; D014162:Transfection; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000096:biosynthesis
D002704:Chlorates; D008024:Ligands; D008562:Membrane Glycoproteins; D009077:Mucins; D009844:Oligosaccharides; D019007:P-Selectin; C084882:P-selectin ligand protein; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D000080506:Sialyl Lewis X Antigen; C005662:tyrosine O-sulfate; D014443:Tyrosine