Overexpression of Merlin in B16F10 mouse melanoma cells reduces their metastatic activity: Role of the cell surface heparan sulfate glycosaminoglycans

Cell surface heparan sulfate proteoglycan (HSPG) from metastatic mouse melanoma cells initiates cell adhesion to the synthetic peptide FN-C/H II, a heparin-binding peptide from the 33-kD A chain-derived fragment of fibronectin. Mouse melanoma cell adhesion to FN-C/H II was sensitive to soluble heparin and pretreatment of mouse melanoma cells with heparitinase. In contrast, cell adhesion to the fibronectin synthetic peptide CS1 is mediated through an alpha 4 beta 1 integrin and was resistant to heparin or heparitinase treatment. Mouse melanoma cell HSPG was metabolically labeled with [35S]sulfate and extracted with detergent. After HPLC-DEAE purification, 35S-HSPG eluted from a dissociative CL-4B column with a Kav approximately 0.45, while 35S-heparan sulfate (HS) chains eluted with a Kav approximately 0.62. The HSPG contained a major 63-kD core protein after heparitinase digestion. Polyclonal antibodies generated against HSPG purified from mouse melanoma cells grown in vivo also identified a 63-kD core protein. This HSPG is an integral plasma membrane component by virtue of its binding to Octyl Sepharose affinity columns and that anti-HSPG antibody staining exhibited a cell surface localization. The HSPG is anchored to the cell surface through phosphatidylinositol (PI) linkages, as evidenced in part by the ability of PI-specific phospholipase C to eliminate binding of the detergent-extracted HSPG to Octyl Sepharose. Furthermore, the mouse melanoma HSPG core protein could be metabolically labeled with 3H-ethanolamine. The involvement of mouse melanoma cell surface HSPG in cell adhesion to fibronectin was also demonstrated by the ability of anti-HSPG antibodies and anti-HSPG IgG Fab monomers to inhibit mouse melanoma cell adhesion to FN-C/H II. 35S-HSPG and 35S-HS bind to FN-C/H II affinity columns and require 0.25 M NaCl for elution. However, heparitinase-treated 125I-labeled HSPG failed to bind FN-C/H II, suggesting that HS, and not HSPG core protein, binds FN-C/H II. These data support the hypothesis that a phosphatidylinositol-anchored HSPG on mouse melanoma cells (MPIHP-63) initiates recognition to FN-C/H II, and implicate PI-associated signal transduction pathways in mediating melanoma cell adhesion to this defined ligand.

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Genogroup II Noroviruses Efficiently Bind to Heparan Sulfate Proteoglycan Associated with the Cellular Membrane

Journal of Virology ◽

10.1128/jvi.78.8.3817-3826.2004 ◽

2004 ◽

Vol 78 (8) ◽

pp. 3817-3826 ◽

Cited By ~ 67

Author(s):

Masaru Tamura ◽

Katsuro Natori ◽

Masahiko Kobayashi ◽

Tatsuo Miyamura ◽

Naokazu Takeda

Keyword(s):

Cell Surface ◽

Heparan Sulfate ◽

Cellular Membrane ◽

Heparan Sulfate Proteoglycan ◽

Sulfate Proteoglycan ◽

Sulfated Glycosaminoglycan ◽

The Family ◽

Sulfated Derivative ◽

Causative Agents

ABSTRACT Norovirus (NV), a member of the family Caliciviridae, is one of the important causative agents of acute gastroenteritis. In the present study, we found that virus-like particles (VLPs) derived from genogroup II (GII) NV were bound to cell surface heparan sulfate proteoglycan. Interestingly, the VLPs derived from GII were more than ten times likelier to bind to cells than were those derived from genogroup I (GI). Heparin, a sulfated glycosaminoglycan, and suramin, a highly sulfated derivative of urea, efficiently blocked VLP binding to mammalian cell surfaces. The reagents known to bind to cell surface heparan sulfate, as well as the enzymes that specifically digest heparan sulfate, markedly reduced VLP binding to the cells. Treatment of the cells with chlorate revealed that sulfation of heparan sulfate plays an important role in the NV-heparan sulfate interaction. The binding efficiency of NV to undifferentiated Caco-2 (U-Caco-2) cells differed largely between GI NV and GII NV, whereas the efficiency of binding to differentiated Caco-2 (D-Caco-2) cells did not differ significantly between the two genogroups, although slight differences between strains were observed. Digestion with heparinase I resulted in a reduction of up to 90% in U-Caco-2 cells and a reduction of up to only 50% in D-Caco-2 cells, indicating that heparan sulfate is the major binding molecule for U-Caco-2 cells, while it contributed to only half of the binding in the case of D-Caco-2 cells. The other half of those VLPs was likely to be associated with H-type blood antigen, suggesting that GII NV has two separate binding sites. The present study is the first to address the possible role of cell surface glycosaminoglycans in the binding of recombinant VLPs of NV.

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Cell-cycle analysis of insulin binding and internalization on mouse melanoma cells.

The Journal of Cell Biology ◽

10.1083/jcb.88.1.241 ◽

1981 ◽

Vol 88 (1) ◽

pp. 241-244 ◽

Cited By ~ 20

Author(s):

N Shimizu ◽

Y Shimizu ◽

B B Fuller

Keyword(s):

Cell Cycle ◽

Cell Surface ◽

Insulin Binding ◽

Melanoma Cells ◽

Binding Activity ◽

S Phase ◽

Cell Cycle Analysis ◽

Surface Receptors ◽

Mouse Melanoma ◽

Melanocyte Stimulating Hormone

Binding of 125I-labeled insulin to the surface receptors of Cloudman S-91 mouse melanoma cells (CCL 53.1) was studied at various phases (M, G1, S, and G2) in the cell cycle. Insulin-binding activity was persistently present during the cell cycle but the highest activity was noted at the S-phase. The insulin once bound to the cell surface receptors at any phase of the cell cycle was internalized and degraded, presumably through a lysosomal pathway. Insulin-indexing activity of melanoma cells was not affected by melanocyte-stimulating hormone.

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Association of cell surface receptors for melanotropin with the Golgi region in mouse melanoma cells.

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.73.2.559 ◽

1976 ◽

Vol 73 (2) ◽

pp. 559-562 ◽

Cited By ~ 51

Author(s):

J. M. Varga ◽

G. Moellmann ◽

P. Fritsch ◽

E. Godawska ◽

A. B. Lerner

Keyword(s):

Cell Surface ◽

Melanoma Cells ◽

Cell Surface Receptors ◽

Surface Receptors ◽

Mouse Melanoma ◽

Golgi Region

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The role of membrane-bound heat shock Hsp90 proteins in the migration of tumor cells in vitro and the involvement of cell surface heparan sulfate proteoglycans in protein binding to the plasma membrane

BIOPHYSICS ◽

10.1134/s0006350916020196 ◽

2016 ◽

Vol 61 (2) ◽

pp. 277-283 ◽

Cited By ~ 2

Author(s):

A. V. Snigireva ◽

V. V. Vrublevskaya ◽

Y. Y. Skarga ◽

O. S. Morenkov

Keyword(s):

Plasma Membrane ◽

Heat Shock ◽

Cell Surface ◽

Protein Binding ◽

Heparan Sulfate ◽

Tumor Cells ◽

Heparan Sulfate Proteoglycans ◽

Membrane Bound

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Syntenin-knock out reduces exosome turnover and viral transduction

Scientific Reports ◽

10.1038/s41598-021-81697-4 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Rudra Kashyap ◽

Marielle Balzano ◽

Benoit Lechat ◽

Kathleen Lambaerts ◽

Antonio Luis Egea-Jimenez ◽

...

Keyword(s):

Cell Surface ◽

Heparan Sulfate ◽

Intercellular Communication ◽

Scaffold Protein ◽

Heparan Sulfate Proteoglycans ◽

Knock Out ◽

Major Class ◽

Endosomal Membranes ◽

Viral Transduction

AbstractExosomal transfers represent an important mode of intercellular communication. Syntenin is a small scaffold protein that, when binding ALIX, can direct endocytosed syndecans and syndecan cargo to budding endosomal membranes, supporting the formation of intraluminal vesicles that compose the source of a major class of exosomes. Syntenin, however, can also support the recycling of these same components to the cell surface. Here, by studying mice and cells with syntenin-knock out, we identify syntenin as part of dedicated machinery that integrates both the production and the uptake of secreted vesicles, supporting viral/exosomal exchanges. This study significantly extends the emerging role of heparan sulfate proteoglycans and syntenin as key components for macromolecular cargo internalization into cells.

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