Ganglioside Modulates Ligand Binding to the Epidermal Growth Factor Receptor

Wang, Xiaoqi; Rahman, Zakia; Sun, Ping; Meuillet, Emmanuelle J.; George, David; Bremer, E.; Al-Qamari, Abbas; Paller, Amy S.

doi:10.1046/j.1523-1747.2001.00222.x

Cited by 46 publications

(67 citation statements)

References 40 publications

(47 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Three findings suggest, however, that gangliosides may work through additional mechanisms. First, we previously reported that GM3-treated keratinocyte-derived SCC12 cells decreased the number of EGF-binding sites even though EGFR protein levels remained constant (34). Our current finding that GM3 does not alter EGF binding to the ECD suggests that the availability of the EGFR at the cell surface may be altered in the presence of gangliosides.…”

Section: Fig 3 Solid-phase Ganglioside Binding Assay Detecting Ecd mentioning

confidence: 54%

Interaction of the Extracellular Domain of the Epidermal Growth Factor Receptor with Gangliosides

Miljan¹,

Meuillet²,

Mania-Farnell³

et al. 2002

Journal of Biological Chemistry

Self Cite

160

113

View full text Add to dashboard Cite

Ganglioside GM3 inhibits epidermal growth factor (EGF)-dependent cell proliferation in a variety of cell lines. Both in vitro and in vivo, this glycosphingolipid inhibits the kinase activity of the EGF receptor (EGFR). Furthermore, membrane preparations containing EGFR can bind to GM3-coated surfaces. These data suggest that GM3 may interact directly with the EGFR. In this study, the interaction of gangliosides with the extracellular domain (ECD) of the EGFR was investigated. The purified human recombinant ECD from insect cells bound directly to ganglioside GM3. The ganglioside interaction site appears to be distinct from the EGF-binding site. In agreement with previous reports on the effects of specific gangliosides on EGFR kinase activity, the ECD preferentially interacted with GM3. The order of relative binding of other gangliosides investigated was as follows: GM3 > > GM2, GD3, GM4 > GM1, GD1a, GD1b, GT1b, GD2, GQ1b > lactosylceramide. These data suggest that NeuAc-lactose is essential for binding and that any sugar substitution reduces binding. In agreement with the specificity of soluble ECD binding to gangliosides, GM3 specifically inhibited EGFR autophosphorylation. Identification of a ganglioside interaction site on the ECD of the EGFR is consistent with the hypothesis that endogenous GM3 may function as a direct modulator of EGFR activity.

show abstract

Section: Fig 3 Solid-phase Ganglioside Binding Assay Detecting Ecd mentioning

confidence: 54%

Interaction of the Extracellular Domain of the Epidermal Growth Factor Receptor with Gangliosides

Miljan¹,

Meuillet²,

Mania-Farnell³

et al. 2002

Journal of Biological Chemistry

Self Cite

160

113

View full text Add to dashboard Cite

show abstract

“…23) and was decreased by knocking down GM3S with 3 siRNAs of GM3S (Table 1) or by inhibiting the biosynthesis of GM3 with PPPP (P4, racemic threo-1-phenyl-2-hexadecanoylamino-3-pyrrolidinopropan-1-ol, HCl; Calbiochem), a chemical reagent that blocks the synthesis of GM3 without accumulating ceramide (24,25). The content of ganglioside was detected by ganglioside ELISA (24)(25)(26)(27), routine thinlayer chromatography (TLC), and TLC immunostaining (24,28,29) using antibodies directed against total GM3 (DH2, Glycotech) or d-GM3 (DH5, generous gift from Dr. Hakomori, Pacific Northwest Diabetes Research Institute, Seattle, WA; ref. 7).…”

Section: Modulating D-gm3 Expressionmentioning

confidence: 99%

“…For routine TLC and TLC immunostaining, total lipids were extracted from cells (5 Â 10 7 ) using 10 volume of chloroform/ methanol (2:1; ref. 28). The aqueous phase was separated and desalted, and the bands were separated by TLC in chloroform/methanol/water in 0.02% CaCl 2 , 55:45:10 (v/v/v).…”

Section: Modulating D-gm3 Expressionmentioning

confidence: 99%

See 1 more Smart Citation

Deacetylated GM3 Promotes uPAR-Associated Membrane Molecular Complex to Activate p38 MAPK in Metastatic Melanoma

Qiu

Bach

Gatla

et al. 2013

Molecular Cancer Research

Self Cite

View full text Add to dashboard Cite

GM3, the simplest ganglioside, regulates cell proliferation, migration, and invasion by influencing cell signaling at the membrane level. Although the classic N-acetylated form of GM3 (NeuAcLacCer) is commonly expressed and has been well studied, deacetylated GM3 (NeuNH 2 LacCer, d-GM3) has been poorly investigated, despite its presence in metastatic tumors but not in noninvasive melanomas or benign nevi. We have recently found that d-GM3 stimulates cell migration and invasion by activating urokinase plasminogen activator receptor (uPAR) signaling to augment matrix metalloproteinase-2 (MMP-2) function. However, the mechanisms by which d-GM3/ uPAR increase MMP-2 expression and activation are not clear. By modifying the expression of d-GM3 genetically and biochemically, we found that decreasing d-GM3 expression inhibits, whereas overexpressing d-GM3 stimulates, p38 mitogen-activated protein kinase (MAPK) activity to influence MMP-2 expression and activation. p38 MAPK (p38) activation requires the formation of a membrane complex that contains uPAR, caveolin-1, and integrin a5b1 in membrane lipid rafts. In addition, knocking down or inhibiting focal adhesion kinase (FAK), phosphoinositide 3-kinase (PI3K), or Src kinase significantly reduces d-GM3-induced p38 phosphorylation and activation. Taken together, these results suggest that d-GM3 enhances the metastatic phenotype by activating p38 signaling through uPAR/integrin signaling with FAK, PI3K, and Src kinase as intermediates. Elucidation of the mechanisms by which d-GM3, a newly discovered, potential biomarker of metastatic melanomas, promotes cell metastasis will help us to understand the function of d-GM3 in metastatic melanomas and may lead to novel GM3-based cancer therapies. Mol Cancer Res; 11(6); 665-75. Ó2013 AACR. IntroductionAlthough recent therapeutic progress for metastatic melanomas has significantly improved, including the more recent availability of small-molecule inhibitors targeting BRAF mutation or both BRAF mutation and MEK, or anti-CTLA4 antibody, survival is minimally prolonged and mortality rate of metastatic melanomas remains high (1-3). It is of utmost importance to discover specific molecular biomarkers that can distinguish melanomas with metastatic propensity from more indolent ones, which will improve prognostication, allow for earlier and more efficacious treatments, and provide additional targets for novel therapy.

show abstract

“…Gangliosides are sialylated membrane glycosphingolipids that modulate several biologic processes of keratinocytes and keratinocyte-derived cell lines in vitro, including cell proliferation, adhesion, migration, differentiation, and apoptosis, at least in part by affecting transmembrane signaling (23)(24)(25)(26)(27)(28)(29). Studies from our laboratory and others have shown that GM3, the predominant ganglioside of the membranes of both normal cultured keratinocytes and the keratinocyte-derived SCC12 cell line (26,30), inhibits cell proliferation by suppressing EGFR activation (31) and downstream signaling (26).…”

mentioning

confidence: 99%

Ganglioside Induces Caveolin-1 Redistribution and Interaction with the Epidermal Growth Factor Receptor

Wang

Sun

Paller

2002

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

Although caveolin-1 is thought to facilitate the interaction of receptors and signaling components, its role in epidermal growth factor receptor (EGFR) signaling remains poorly understood. Ganglioside GM3 inhibits EGFR autophosphorylation and may thus affect the interaction of caveolin-1 and the EGFR. We report here that endogenous overexpression of GM3 leads to the clustering of GM3 on the cell membrane of the keratinocyte-derived SCC12 cell line and promotes co-immunoprecipitation of caveolin-1 and GM3 with the EGFR. Overexpression of GM3 does not affect EGFR distribution but shifts caveolin-1 to the detergent-soluble, EGFR-containing region; consistently, caveolin-1 is retained in the detergent-insoluble membrane when ganglioside is depleted. GM3 overexpression inhibits EGFR tyrosine phosphorylation and receptor dimerization and concurrently increases both the content and tyrosine phosphorylation of EGFR-associated caveolin-1, providing evidence that tyrosine phosphorylation of caveolin-1 inhibits EGFR signaling. Consistently, depletion of ganglioside both increases EGFR phosphorylation and prevents the EGF-induced tyrosine phosphorylation of caveolin-1. GM3 also induces delayed serine phosphorylation of EGFR-unassociated caveolin-1, suggesting a role for serine phosphorylation of caveolin-1 in regulating EGFR signaling. These studies suggest that GM3 modulates the caveolin-1/EGFR association and is critical for the EGF-induced tyrosine phosphorylation of caveolin-1 that is associated with its inhibition of EGFR activation.The plasma membrane of eukaryotic cells is not uniform and instead is composed of microdomains or rafts, highly dynamic lateral assemblies composed of glycosphingolipids and cholesterol. One form of these rafts, caveolae, has been distinguished ultrastructurally by its invaginated shape and by the presence of caveolin-1, the major structural protein of caveolae. These caveolin-containing rafts are insoluble in both nonionic detergents, such as Triton X-100, and in sodium carbonate at alkaline pH and can be isolated as low density fractions by sucrose density gradient. Although caveolae have been implicated as sites at which receptors and signaling molecules interact (1, 2), signaling also occurs at noninvaginated rafts, devoid of caveolin-1 but enriched in glycosphingolipids (3-5). In addition, caveolin-1 may also exist in lipid rafts without caveolae (6). The distinction between lipid rafts and caveolae and the attribution of specific biologic functions and signaling pathways to one or the other remains confusing in the literature (7).Caveolae have been proposed to be the site of epidermal growth factor receptor (EGFR) 1 signaling, including EGFR autophosphorylation, based largely on the demonstration that caveolin-1 and EGFR both co-localize to low density, carbonateinsoluble membrane regions (8 -11). However, caveolin-1 and EGFR can be separated by differences in solubility in nonionic detergent, with EGFR restricted to the detergent-soluble high buoyancy, low density membrane regions (1...

show abstract

Ganglioside Modulates Ligand Binding to the Epidermal Growth Factor Receptor

Cited by 46 publications

References 40 publications

Interaction of the Extracellular Domain of the Epidermal Growth Factor Receptor with Gangliosides

Interaction of the Extracellular Domain of the Epidermal Growth Factor Receptor with Gangliosides

Deacetylated GM3 Promotes uPAR-Associated Membrane Molecular Complex to Activate p38 MAPK in Metastatic Melanoma

Ganglioside Induces Caveolin-1 Redistribution and Interaction with the Epidermal Growth Factor Receptor

Contact Info

Product

Resources

About