Cell membrane GM1 ganglioside is a functional coreceptor for fibroblast growth factor 2

Free gangliosides bind fibroblast growth factor 2 (FGF2), thus preventing cell interaction and biological activity of the growth factor in endothelial cells. Here we investigated the role of cell-associated gangliosides in mediating the biological activity of FGF2. Treatment of endothelial cells of different origin with the ganglioside biosynthesis inhibitors fumonisin B1, D-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol or D-1-threo-1-phenyl-2-hexa-decanoylamino-3-pyrrolidino-1-propanol-HCl, impairs their capacity to proliferate when exposed to FGF2. Also, the mitogenic activity of FGF2 is inhibited by the GM1-binding cholera toxin B subunit (CTB). Conversely, overloading of endothelial GM 7373 cell membranes with exogenous GM1 causes a 10-fold increase of the mitogenic potency of FGF2. 125I-FGF2 binds to cell membrane GM1 (Kd = 3 nM) in complex ganglioside/heparan sulfate-deficient Chinese hamster ovary (CHO)-K1-pgsA745 cell mutants that were overloaded with exogenous GM1. Moreover, FGF2 competes with FITC-CTB for the binding to cell membrane GM1 in different CHO cell lines independently of their capacity to express heparan sulfate proteoglycans. Conversely, CTB inhibits cell proliferation triggered by FGF2 in CHO cells overexpressing the tyrosine kinase FGF receptor 1. Finally, GM1-overloading confers to FGF receptor 1-transfected, complex ganglioside-deficient CHO-K1 cell mutants the capacity to proliferate when stimulated by FGF2. This proliferation is inhibited by CTB. Cell proliferation triggered by serum or by phorbol 12-myristate 13-acetate is instead independent of the cell membrane ganglioside milieu. In conclusion, cell membrane GM1 binds FGF2 and is required for the mitogenic activity of the growth factor. Our data indicate that cell-associated gangliosides may act as functional FGF2 co-receptors in different cell types.

[1]  M. Hollenberg,et al.  Interferon action: role of membrane gangliosides. , 1976, Virology.

[2]  D. Moscatelli,et al.  The FGF family of growth factors and oncogenes. , 1992, Advances in cancer research.

[3]  S. Tzeng,et al.  Myelin Basic Protein and Myelin Basic Protein Peptides Induce the Proliferation of Schwann Cells Via Ganglioside GM1 and the FGF Receptor , 1999, Neurochemical Research.

[4]  G. Schwarzmann,et al.  Incorporation and metabolism of ganglioside GM2 in skin fibroblasts from normal and GM2 gangliosidosis subjects. , 1985, European journal of biochemistry.

[5]  Marco Presta,et al.  αvβ3 Integrin Mediates the Cell-adhesive Capacity and Biological Activity of Basic Fibroblast Growth Factor (FGF-2) in Cultured Endothelial Cells , 1997 .

[6]  J. Peter-Katalinic,et al.  Isolation and structural characterization of glycosphingolipids of in vitro propagated human umbilical vein endothelial cells. , 1999, Glycobiology.

[7]  R. Esnouf,et al.  Modulation of fibroblast growth factor-2 receptor binding, signaling, and mitogenic activity by heparin-mimicking polysulfonated compounds. , 1999, Molecular pharmacology.

[8]  P. Vermeulen,et al.  Elevated levels of the angiogenic cytokines basic fibroblast growth factor and vascular endothelial growth factor in sera of cancer patients. , 1997, British Journal of Cancer.

[9]  Dudley H. Williams,et al.  A vesicle capture sensor chip for kinetic analysis of interactions with membrane-bound receptors. , 2000, Analytical biochemistry.

[10]  V. Georgiev.  Virology , 1955, Nature.

[11]  L. Culp,et al.  Ganglioside-dependent adhesion events of human neuroblastoma cells regulated by the RGDS-dependent fibronectin receptor and proteoglycans. , 1988, Experimental cell research.

[12]  A. Isacchi,et al.  A six-amino acid deletion in basic fibroblast growth factor dissociates its mitogenic activity from its plasminogen activator-inducing capacity. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[13]  F. Young.  Biochemistry , 1955, The Indian Medical Gazette.

[14]  E. Estey,et al.  Angiogenesis in acute and chronic leukemias and myelodysplastic syndromes. , 2000, Blood.

[15]  A. Merrill,et al.  Inhibition of sphingolipid biosynthesis by fumonisins. Implications for diseases associated with Fusarium moniliforme. , 1991, The Journal of biological chemistry.

[16]  A. Yates,et al.  Gangliosides Inhibit Platelet‐Derived Growth Factor‐Stimulated Growth, Receptor Phosphorylation, and Dimerization in Neuroblastoma SH‐SY5Y Cells , 1995, Journal of neurochemistry.

[17]  H. Maccioni,et al.  Chinese hamster ovary cells lacking GM1 and GD1a synthesize gangliosides upon transfection with human GM2 synthase. , 1997, Biochimica et biophysica acta.

[18]  A. Abe,et al.  Improved Inhibitors of Glucosylceramide Synthase* , 1999, The Journal of Biological Chemistry.

[19]  Jeffrey D. Esko,et al.  Cell surface, heparin-like molecules are required for binding of basic fibroblast growth factor to its high affinity receptor , 1991, Cell.

[20]  C. Powers,et al.  Fibroblast growth factors, their receptors and signaling. , 2000, Endocrine-related cancer.

[21]  A. Futerman,et al.  Up-regulation of Glucosylceramide Synthesis upon Stimulation of Axonal Growth by Basic Fibroblast Growth Factor , 2000, The Journal of Biological Chemistry.

[22]  M. Ziche,et al.  Interaction of fibroblast growth factor-2 (FGF-2) with free gangliosides: biochemical characterization and biological consequences in endothelial cell cultures. , 1999, Molecular biology of the cell.

[23]  Ruixiang Li,et al.  Cellular Gangliosides Promote Growth Factor-induced Proliferation of Fibroblasts* , 2000, The Journal of Biological Chemistry.

[24]  J. Esko,et al.  Microbial adherence to and invasion through proteoglycans , 1997, Infection and immunity.

[25]  M. Rusnati,et al.  Interaction of angiogenic basic fibroblast growth factor with endothelial cell heparan sulfate proteoglycans , 1996, International journal of clinical & laboratory research.

[26]  R C Stevens,et al.  Cholera toxin binding affinity and specificity for gangliosides determined by surface plasmon resonance. , 1996, Biochemistry.

[27]  D. Gospodarowicz,et al.  The identification and partial characterization of the fibroblast growth factor receptor of baby hamster kidney cells. , 1985, The Journal of biological chemistry.

[28]  L. Riboni,et al.  Basic Fibroblast Growth Factor-induced Proliferation of Primary Astrocytes , 2001, The Journal of Biological Chemistry.

[29]  R. Burry,et al.  Gangliosides inhibit growth factor‐stimulated neurite outgrowth in SH‐SY5Y human neuroblastoma cells , 1997, Journal of neuroscience research.

[30]  E. Levine,et al.  Bovine endothelial cells transformed in vitro by benzo(a)pyrene , 1983, Journal of cellular physiology.

[31]  M. Presta,et al.  Role of endothelial cell extracellular signal-regulated kinase1/2 in urokinase-type plasminogen activator upregulation and in vitro angiogenesis by fibroblast growth factor-2. , 1999, Journal of cell science.

[32]  K. Fuxe,et al.  Codistribution of choleratoxin binding sites and tyrosine hydroxylase/FGF-2 immunoreactive nigral nerve cells. , 1993, Neuroreport.

[33]  S. Milstien,et al.  The role of calcium influx in cellular proliferation induced by interaction of endogenous ganglioside GM1 with the B subunit of cholera toxin. , 1995, Biochimica et biophysica acta.

[34]  N. Radin,et al.  Preparation of the active isomer of 1-phenyl-2-decanoylamino-3-morpholino-1-propanol, inhibitor of murine glucocerebroside synthetase. , 1987, Journal of lipid research.

[35]  Y. Nagamine,et al.  A distinct basic fibroblast growth factor (FGF-2)/FGF receptor interaction distinguishes urokinase-type plasminogen activator induction from mitogenicity in endothelial cells. , 1996, Molecular biology of the cell.

[36]  E. Merritt,et al.  A mutant cholera toxin B subunit that binds GM1- ganglioside but lacks immunomodulatory or toxic activity , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[37]  S. Ladisch,et al.  Shedding of gangliosides by human medulloblastoma cells. , 1997, Experimental cell research.

[38]  F. Besançon,et al.  Binding of interferon to gangliosides , 1974, Nature.

[39]  R. Dwek,et al.  Glycobiology , 2018, Biochimie.