Linked Life Data

10713099

Source:http://linkedlifedata.com/resource/pubmed/id/10713099

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
11
pubmed:dateCreated
2000-4-12
pubmed:abstractText
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.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Mar
pubmed:issn
0021-9258
pubmed:author
pubmed:issnType
Print
pubmed:day
17
pubmed:volume
275
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
7839-53
pubmed:dateRevised
2011-9-7
pubmed:meshHeading
pubmed-meshheading:10713099-Amino Acid Sequence, pubmed-meshheading:10713099-Animals, pubmed-meshheading:10713099-Antigens, CD, pubmed-meshheading:10713099-Antigens, CD43, pubmed-meshheading:10713099-Biomechanics, pubmed-meshheading:10713099-CHO Cells, pubmed-meshheading:10713099-Carbohydrate Sequence, pubmed-meshheading:10713099-Cell Adhesion, pubmed-meshheading:10713099-Cell Membrane, pubmed-meshheading:10713099-Cricetinae, pubmed-meshheading:10713099-Dimerization, pubmed-meshheading:10713099-Glycosylation, pubmed-meshheading:10713099-HL-60 Cells, pubmed-meshheading:10713099-Humans, pubmed-meshheading:10713099-Ligands, pubmed-meshheading:10713099-Membrane Glycoproteins, pubmed-meshheading:10713099-Models, Molecular, pubmed-meshheading:10713099-Molecular Sequence Data, pubmed-meshheading:10713099-Mucins, pubmed-meshheading:10713099-P-Selectin, pubmed-meshheading:10713099-Physical Stimulation, pubmed-meshheading:10713099-Protein Binding, pubmed-meshheading:10713099-Protein Conformation, pubmed-meshheading:10713099-Recombinant Proteins, pubmed-meshheading:10713099-Repetitive Sequences, Amino Acid, pubmed-meshheading:10713099-Sialoglycoproteins
pubmed:year
2000
pubmed:articleTitle
Noncovalent association of P-selectin glycoprotein ligand-1 and minimal determinants for binding to P-selectin.
pubmed:affiliation
Department of Biochemistry and Molecular Biology, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73104, USA.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S., Research Support, U.S. Gov't, Non-P.H.S.