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http://purl.uniprot.org/citations/10713099http://www.w3.org/1999/02/22-rdf-syntax-ns#typehttp://purl.uniprot.org/core/Journal_Citation
http://purl.uniprot.org/citations/10713099http://www.w3.org/1999/02/22-rdf-syntax-ns#typehttp://purl.uniprot.org/core/Journal_Citation
http://purl.uniprot.org/citations/10713099http://www.w3.org/2000/01/rdf-schema#comment"P-selectin glycoprotein ligand-1 (PSGL-1) is a disulfide-bonded, homodimeric mucin ( approximately 250 kDa) on leukocytes that binds to P-selectin on platelets and endothelial cells during the initial steps in inflammation. Because it has been proposed that only covalently dimerized PSGL-1 can bind P-selectin, we investigated the factors controlling dimerization of PSGL-1 and re-examined whether covalent dimers are required for binding its P-selectin. Recombinant forms of PSGL-1 were created in which the single extracellular Cys (Cys(320)) was replaced with either Ser (C320S-PSGL-1) or Ala (C320A-PSGL-1). Both recombinants migrated as monomeric species of approximately 120 kDa under both nonreducing and reducing conditions on SDS-polyacrylamide gel electrophoresis. P-selectin bound similarly to cells expressing either wild type or mutated forms of PSGL-1 in both flow cytometric and rolling adhesion assays. Unexpectedly, chemical cross-linking studies revealed that both C320S- and C320A-PSGL-1 noncovalently associate in the plasma membrane and cross-linking generates dimeric species. Chimeric recombinants of PSGL-1 in which the transmembrane domain in PSGL-1 was replaced with the transmembrane domain of CD43 (CD43TMD-PSGL-1) could not be chemically cross-linked, suggesting that residues within the transmembrane domain of PSGL-1 are required for noncovalent association. Cells expressing CD43TMD-PSGL-1 bound P-selectin. To further address the ability of P-selectin to bind monomeric derivatives of PSGL-1, intact HL-60 cells were trypsin-treated, which generated a soluble approximately 25-kDa NH(2)-terminal fragment of PSGL-1 that bound to immobilized P-selectin. Because N-glycosylation of PSGL-1 hinders trypsin cleavage, a recombinant form of PSGL-1 was generated in which all three potential N-glycosylation sites were mutated (DeltaN-PSGL-1). Cells expressing DeltaN-PSGL-1 bound P-selectin, and trypsin treatment of the cells generated NH(2)-terminal monomeric fragments (<10 kDa) of PSGL-1 that bound to P-selectin. These results demonstrate that Cys(320)-dependent dimerization of PSGL-1 is not required for binding to P-selectin and that a small monomeric fragment of PSGL-1 is sufficient for P-selectin recognition."xsd:string
http://purl.uniprot.org/citations/10713099http://purl.org/dc/terms/identifier"doi:10.1074/jbc.275.11.7839"xsd:string
http://purl.uniprot.org/citations/10713099http://purl.org/dc/terms/identifier"doi:10.1074/jbc.275.11.7839"xsd:string
http://purl.uniprot.org/citations/10713099http://purl.uniprot.org/core/author"Cummings R.D."xsd:string
http://purl.uniprot.org/citations/10713099http://purl.uniprot.org/core/author"Cummings R.D."xsd:string
http://purl.uniprot.org/citations/10713099http://purl.uniprot.org/core/author"McEver R.P."xsd:string
http://purl.uniprot.org/citations/10713099http://purl.uniprot.org/core/author"McEver R.P."xsd:string
http://purl.uniprot.org/citations/10713099http://purl.uniprot.org/core/author"Patel K.D."xsd:string
http://purl.uniprot.org/citations/10713099http://purl.uniprot.org/core/author"Patel K.D."xsd:string
http://purl.uniprot.org/citations/10713099http://purl.uniprot.org/core/author"Epperson T.K."xsd:string
http://purl.uniprot.org/citations/10713099http://purl.uniprot.org/core/author"Epperson T.K."xsd:string
http://purl.uniprot.org/citations/10713099http://purl.uniprot.org/core/date"2000"xsd:gYear
http://purl.uniprot.org/citations/10713099http://purl.uniprot.org/core/date"2000"xsd:gYear
http://purl.uniprot.org/citations/10713099http://purl.uniprot.org/core/name"J. Biol. Chem."xsd:string
http://purl.uniprot.org/citations/10713099http://purl.uniprot.org/core/name"J. Biol. Chem."xsd:string
http://purl.uniprot.org/citations/10713099http://purl.uniprot.org/core/pages"7839-7853"xsd:string
http://purl.uniprot.org/citations/10713099http://purl.uniprot.org/core/pages"7839-7853"xsd:string
http://purl.uniprot.org/citations/10713099http://purl.uniprot.org/core/title"Noncovalent association of P-selectin glycoprotein ligand-1 and minimal determinants for binding to P-selectin."xsd:string
http://purl.uniprot.org/citations/10713099http://purl.uniprot.org/core/title"Noncovalent association of P-selectin glycoprotein ligand-1 and minimal determinants for binding to P-selectin."xsd:string
http://purl.uniprot.org/citations/10713099http://purl.uniprot.org/core/volume"275"xsd:string
http://purl.uniprot.org/citations/10713099http://purl.uniprot.org/core/volume"275"xsd:string
http://purl.uniprot.org/citations/10713099http://www.w3.org/2004/02/skos/core#exactMatchhttp://purl.uniprot.org/pubmed/10713099
http://purl.uniprot.org/citations/10713099http://www.w3.org/2004/02/skos/core#exactMatchhttp://purl.uniprot.org/pubmed/10713099
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