Partial characterization of the N-linked oligosaccharide structures on P-selectin glycoprotein ligand-1 (PSGL-1)

Selectins, a family of cell adhesion molecules, bind to sialylated and fucosylated carbohydrates, such as sialyl Lewisx (SLex) and its derivatives, as their minimal recognition motif. Here we report that P-selectin bound to human malignant melanoma A375 cells and mediated their adhesion under flow. However, probing with a specific Ab failed to detect any apparent expression of SLex. This finding was bolstered by reduced expression of α-1,3-fucosyltransferase VII mRNA and by absence of the cell surface expression of P-selectin glycoprotein ligand-1. Instead, they expressed heparan sulfate-like proteoglycans on their cell surfaces. Treatment with β-d-xyloside (a proteoglycan biosynthesis inhibitor) or heparinases could reduce the binding of these cells to P-selectin. In the competition assays, heparin, but not other proteoglycans, could abolish the P-selectin recognition. Further, we found that P-selectin could bind specifically to human tongue squamous cancer Tca-8113 cells, which had negative staining of SLex but positive staining of heparan sulfates. Both β-d-xyloside and heparinases could reduce the binding of P-selectin to Tca-8113 cells. Our results thus indicate that heparan sulfate-like proteoglycans can mediate adhesion of certain types of non-blood borne, “epithelial-like” human cancer cells to P-selectin.

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“…The existence of SLe x -like moieties on O-linked glycans of PSGL-1 further supports this notion (5,6).…”

Section: Heparan Sulfate-like Proteoglycans Mediate Adhesion Of Humansupporting

confidence: 63%

Heparan Sulfate-Like Proteoglycans Mediate Adhesion of Human Malignant Melanoma A375 Cells to P-Selectin Under Flow

Qing

Geng

2000

The Journal of Immunology

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“…CVA24v appears to bind to both ␣2,3-linked and ␣2,6-linked sialic acids, and this may at least partially explain why CVA24v causes respiratory disease more frequently than EV70, which mainly binds ␣2,3-linked sialic acid and almost exclusively causes ocular disease (50,53,76). On PSGL-1, the 3Ј-sialyl-TF trisaccharide is linked to either 3ЈSLN or sLe X , thus creating a disialic acid-containing glycan (2,72), which may provide even better binding to CVA24v. In agreement with this suggestion, we demonstrated recently that branched, disialylated glycans are functional receptors for another human ocular pathogen, Ad37, and showed that this soluble glycan was 200-fold more efficient in inhibiting Ad37 binding to corneal cells (51).…”

Section: Discussionmentioning

confidence: 99%

“…PSGL-1 is a heavily O-glycosylated, mucinlike dimeric glycoprotein that is rich in serine/threonine repeat regions, with no less than 70 potential sites for O-glycosylation and only 3 potential sites for N-glycosylation on each subunit (17,63). The majority of the glycans on PSGL-1 are nonfucosylated, nonsialylated core 2 O-glycans, and thus, only a few of these structures contain sLe X (2,72). Here, we demonstrate that sLe X is present mainly on corneal cells but also on conjunctival cells and that soluble sLe X is a better inhibitor of CVA24v binding to corneal cells than sialic acid monosaccharides.…”

Section: Discussionmentioning

confidence: 99%

Coxsackievirus A24 Variant Uses Sialic Acid-Containing O -Linked Glycoconjugates as Cellular Receptors on Human Ocular Cells

Mistry¹,

Inoue²,

Jamshidi³

et al. 2011

J Virol

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Coxsackievirus A24 variant (CVA24v) is a main causative agent of acute hemorrhagic conjunctivitis (AHC), which is a highly contagious eye infection. Previously it has been suggested that CVA24v uses sialic acidcontaining glycoconjugates as attachment receptors on corneal cells, but the nature of these receptors is poorly described. Here, we set out to characterize and identify the cellular components serving as receptors for CVA24v. Binding and infection experiments using corneal cells treated with deglycosylating enzymes or metabolic inhibitors of de novo glycosylation suggested that the receptor(s) used by CVA24v are constituted by sialylated O-linked glycans that are linked to one or more cell surface proteins but not to lipids. CVA24v bound better to mouse L929 cells overexpressing human P-selectin glycoprotein ligand-1 (PSGL-1) than to mocktransfected cells, suggesting that PSGL-1 is a candidate receptor for CVA24v. Finally, binding competition experiments using a library of mono-and oligosaccharides mimicking known PSGL-1 glycans suggested that CVA24v binds to Neu5Ac␣2,3Gal disaccharides (Neu5Ac is N-acetylneuraminic acid). These results provide further insights into the early steps of the CVA24v life cycle.

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“…Dynamic rolling assays conducted in the presence of a blocking anti-PSGL-1 mAb (KPL-1) indicated that PSGL-1-coated microspheres are still capable of tethering on E-but not P-selectin substrates (Goetz et al, 1997), supporting the notion that PSGL-1 has binding sites for E-selectin other than the crucial 19 amino acid sequence recognized by KPL-1. Along these lines, E-selectin appears to be capable of binding to N-linked oligosaccharides, including those found on PSGL-1 (Aeed et al, 2001;Patel et al, 1994;Vestweber, 1996). Taken altogether, E-selectin mediated tethering and rolling appears to involve additional as-yet-unidentified endogenous ligands distinct from PSGL-1.…”

Section: Introductionmentioning

confidence: 97%

Distinct kinetic and mechanical properties govern selectin-leukocyte interactions

Hanley

Wirtz

Κωνσταντόπουλος

2004

Journal of Cell Science

113

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Leukocytes are recruited from the bloodstream to sites of inflammation by the selectin family of adhesion receptors. In vivo and in vitro studies reveal distinctive rolling velocities of polymorphonuclear leukocytes over E-, P- and L-selectin substrates. The kinetic and mechanical properties of the selectin-ligand bonds responsible for these differences at the single-molecule level are not well understood. Using single-molecule force spectroscopy, we probe in situ the rupture force, unstressed off-rate and reactive compliance of single selectin receptors to single ligands on whole human polymorphonuclear leukocytes (PMNs) under conditions that preserve the proper orientation and post-translational modifications of the selectin ligands. Single L-selectin bonds to PMNs were more labile than either E- or P-selectin in the presence of an applied force. This outcome, along with a higher unstressed off-rate and a higher reactive compliance, explain the faster L-selectin-mediated rolling. By quantifying binding frequency in the presence of a specific blocking monoclonal antibody or following enzyme treatment, we determined that P-selectin glycoprotein ligand-1 is a high-affinity ligand for E-selectin on PMNs under force. The rupture force spectra and corresponding unstressed off-rate and reactive compliance of selectin-ligand bonds provide mechanistic insights that might help to explain the variable rolling of leukocytes over different selectin substrates.

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Partial characterization of the N-linked oligosaccharide structures on P-selectin glycoprotein ligand-1 (PSGL-1)

Cited by 22 publications

References 32 publications

Heparan Sulfate-Like Proteoglycans Mediate Adhesion of Human Malignant Melanoma A375 Cells to P-Selectin Under Flow

Heparan Sulfate-Like Proteoglycans Mediate Adhesion of Human Malignant Melanoma A375 Cells to P-Selectin Under Flow

Coxsackievirus A24 Variant Uses Sialic Acid-Containing O -Linked Glycoconjugates as Cellular Receptors on Human Ocular Cells

Distinct kinetic and mechanical properties govern selectin-leukocyte interactions

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