A Novel Glycosulfopeptide Binds to P-selectin and Inhibits Leukocyte Adhesion to P-selectin

Leppänen, Anne; Mehta, Padmaja; Ouyang, Ying-Bin; Ju, Tongzhong; Helin, Jari; Moore, Kevin L.; Die, Irma van; Canfield, William M.; McEver, Rodger P.; Cummings, Richard D.

doi:10.1074/jbc.274.35.24838

Cited by 199 publications

(230 citation statements)

References 67 publications

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“…Cells expressing type 2 glycosynapse are capable of binding to cells expressing P-selectin, E-selectin, or siglecs. A specific structure with three tyrosine phosphates and O-linked glycan to define P-selectin-dependent adhesion in PSGL-1 was recently elucidated (33). (C) Type 3 glycosynapse with integrin receptor (ITR) having ␣-and ␤-subunits and tetraspanin (Tsp.).…”

Section: Type 2 Glycosynapse: O-linked Mucin-type Adhesion Epitopes Omentioning

confidence: 99%

The glycosynapse

Hakomori

2002

Proc. Natl. Acad. Sci. U.S.A.

467

215

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Physically distinguishable microdomains associated with various functional membrane proteins are one of the major current topics in cell biology. Glycosphingolipids present in such microdomains have been used as “markers;” however, the functional role of glycosyl epitopes in microdomains has received little attention. In this review, I have tried to summarize the evidence that glycosyl epitopes in microdomains mediate cell adhesion and signal transduction events that affect cellular phenotypes. Molecular assemblies that perform such functions are hereby termed “glycosynapse” in analogy to “immunological synapse,” the membrane assembly of immunocyte adhesion and signaling. Three types of glycosynapses are so far distinguishable: (i) Glycosphingolipids organized with cytoplasmic signal transducers and proteolipid tetraspanin with or without growth factor receptors; (ii) transmembrane mucin-type glycoproteins with clustered O-linked glycoepitopes for cell adhesion and associated signal transducers at lipid domain; and (iii) N-glycosylated transmembrane adhesion receptors complexed with tetraspanin and gangliosides, as typically seen with the integrin–tetraspanin–ganglioside complex. The possibility is discussed that glycosynapses give rise to a high degree of diversity and complexity of phenotypes

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Section: Type 2 Glycosynapse: O-linked Mucin-type Adhesion Epitopes Omentioning

confidence: 99%

The glycosynapse

Hakomori

2002

Proc. Natl. Acad. Sci. U.S.A.

467

215

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“…This region contains the epitope for PL1, a mAb that blocks binding of PSGL-1 to P-selectin (37). 2-GSP-6, a glycosulfopeptide modeled after this region, binds with equivalent affinity to P-selectin (20). 2-GP-6, which lacks sulfate on the tyrosines, binds with much lower affinity to P-selectin.…”

Section: A Phagocytophilum Binds To a Glycopeptide Modeled After Thementioning

confidence: 99%

Structurally Distinct Requirements for Binding of P-selectin Glycoprotein Ligand-1 and Sialyl Lewis x to Anaplasma phagocytophilum and P-selectin

Yago

Leppänen

Carlyon

et al. 2003

Journal of Biological Chemistry

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Colonization of neutrophils by the bacterium Anaplasma phagocytophilum causes the disease human granulocytic ehrlichiosis. The pathogen also infects mice, its natural host. Like binding of P-selectin, binding of A. phagocytophilum to human neutrophils requires expression of P-selectin glycoprotein ligand-1 (PSGL-1) and ␣1-3-fucosyltransferases that construct the glycan determinant sialyl Lewis x (sLe x ). Binding of A. phagocytophilum to murine neutrophils, however, requires expression of ␣1-3-fucosyltransferases but not PSGL-1. To further characterize the molecular features that A. phagocytophilum recognizes, we measured bacterial binding to microspheres bearing specific glycoconjugates or to cells expressing human PSGL-1 and particular glycosyltransferases. Like P-selectin, A. phagocytophilum bound to purified human PSGL-1 and to glycopeptides modeled after the N terminus of human PSGL-1 that presented sLe x on an O-glycan. Unlike P-selectin, A. phagocytophilum bound to glycopeptides that contained sLe x but lacked tyrosine sulfation or a specific core-2 orientation of sLe x on the O-glycan. A. phagocytophilum bound only to glycopeptides that contained a short amino acid sequence found in the N-terminal region of human but not murine PSGL-1. Unlike P-selectin, A. phagocytophilum bound to cells expressing PSGL-1 in cooperation with sLe x on both N-and O-glycans. Moreover, bacteria bound to microspheres coupled independently with glycopeptide lacking sLe x and with sLe x lacking peptide. These results demonstrate that, unlike P-selectin, A. phagocytophilum binds cooperatively to a nonsulfated N-terminal peptide in human PSGL-1 and to sLe x expressed on PSGL-1 or other glycoproteins. Distinct bacterial adhesins may mediate these cooperative interactions.Human granulocytic ehrlichiosis is a tick-transmitted disease caused by a bacterium recently named Anaplasma phagocytophilum (1-3). Clinical manifestations include fever, headache, myalgia, leukopenia, thrombocytopenia, and impaired host defenses that occasionally lead to fatal complications (4 -7). A. phagocytophilum is maintained in a zoonotic cycle between the arthropod vector, Ixodes scapularis, and mice that serve as reservoir hosts (8 -10). Human infection is inadvertent and is not an obligate component of the life cycle of the bacterium.A hallmark of infection in both mice and humans is the colonization of neutrophils (1, 2). This tropism for a particular cell type suggests that the bacteria recognize specific molecular determinants on the neutrophil surface. Current information suggests that these determinants are related to the cell-surface ligands for selectins, a family of cell adhesion molecules that mediate tethering and rolling of leukocytes on vascular surfaces during the earliest steps of inflammation (11, 12). P-, E-, and L-selectin bind to ␣2-3-sialylated and ␣1-3-fucosylated glycans such as sialyl Lewis x (sLe x ), 1 which are expressed on most leukocytes. Targeted disruption of the genes encoding FTVII and FTIV, the ␣1-3-fucosyltransferases exp...

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“…Each of these two ligands requires sLe x for binding, with the difference that ESL-1 requires sLe x on N-glycans (18), whereas PSGL-1 requires it on O-glycans that carry a core-2 branch (21,24). In addition, PSGL-1 requires sulfation of the tyrosine residues within its N terminus for binding to P-selectin (25)(26)(27)(28)(29). The physiological relevance of PSGL-1 in leukocyte extravasation is well established (23).…”

mentioning

confidence: 99%

The α(1,3)-Fucosyltransferase Fuc-TIV, but Not Fuc-TVII, Generates Sialyl Lewis X-like Epitopes Preferentially on Glycolipids

Huang

Laskowska²,

Vestweber

et al. 2002

Journal of Biological Chemistry

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Fuc-TIV and Fuc-TVII are the two ␣(1, 3)-fucosyltransferases in myeloid cells responsible for the biosynthesis of sialyl Lewis X (sLe x ), the minimal ligand structure for the selectins. We have compared the ability of Fuc-TIV and Fuc-TVII to generate sLe x -like epitopes in transfected Chinese hamster ovary (CHO)-Pro ؊ 5 cells expressing the P-selectin glycoprotein ligand-1 and the core-2 branching enzyme C2GnT. We found that mouse Fuc-TIV and Fuc-TVII can generate similar levels of cell surface sLe x . Surprisingly however, Fuc-TIV-generated sLe x was resistant to proteinase K and trypsin treatment and could be removed from cells by delipidation with chloroform/methanol, whereas 80 -90% of Fuc-TVIIgenerated sLe x was protease-sensitive, and most of it resistant to delipidation. Despite similar levels of sLe x on the cell surface, Fuc-TVII transfectants adhered to immobilized E-selectin-IgG under static and flow conditions better than Fuc-TIV transfectants. Binding was mainly protease sensitive, indicating that glycoproteins were more efficient ligands than glycolipids. In summary, we conclude that the two fucosyltransferases differ in their in vivo specificity for acceptor substrates with Fuc-TVII generating sLe x preferentially on glycoproteins, whereas most of the Fuc-TIV-generated sLe x is found on glycolipids. Interestingly, the non-catalytic portion of Fuc-TIV in a Fuc-TIV/VII chimeric enzyme mediated the specificity for glycolipid substrates.The three known selectins, L-, E-, and P-selectin are cell adhesion molecules that initiate interactions between leukocytes and endothelial cells during leukocyte extravasation (1). The minimal ligand structure for all three selectins is the tetrasaccharide sialyl Lewis X (sLe x ). 1

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A Novel Glycosulfopeptide Binds to P-selectin and Inhibits Leukocyte Adhesion to P-selectin

Cited by 199 publications

References 67 publications

The glycosynapse

The glycosynapse

Structurally Distinct Requirements for Binding of P-selectin Glycoprotein Ligand-1 and Sialyl Lewis x to Anaplasma phagocytophilum and P-selectin

The α(1,3)-Fucosyltransferase Fuc-TIV, but Not Fuc-TVII, Generates Sialyl Lewis X-like Epitopes Preferentially on Glycolipids

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