Interaction of Heparan Sulfate from Mammary Cells with Acidic Fibroblast Growth Factor (FGF) and Basic FGF

Rahmoune, Hassan; Chen, Hailan; Gallagher, John T.; Rudland, Philip S.; Fernig, David G.

doi:10.1074/jbc.273.13.7303

Cited by 116 publications

(80 citation statements)

References 45 publications

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“…The 2 M NaCl fractions were desalted on a 75-ml G-25 Sephadex (Amersham Pharmacia Biotech) column equilibrated in 100 mM NH 4 HCO 3 . After freezedrying, the proteoglycans were treated sequentially with sialidases, chondroitinase ABC, ␣-fucosidase, endo-␤-galactosidase, and nucleases to remove anionic contaminants (9). The proteoglycans were concentrated on a 1-ml DEAE Fast Flow column and the 2M NaCl fraction was desalted using a 10 ml G-25 Sephadex column, as described above.…”

Section: Methodsmentioning

confidence: 99%

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Stimulation of DNA Synthesis and Cell Proliferation of Human Mammary Myoepithelial-like Cells by Hepatocyte Growth Factor/Scatter Factor Depends on Heparan Sulfate Proteoglycans and Sustained Phosphorylation of Mitogen-activated Protein Kinases p42/44

Sergeant

Lyon

Rudland

et al. 2000

Journal of Biological Chemistry

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Section: Methodsmentioning

confidence: 99%

“…After 4 h, the wells were rinsed twice with PBS before seeding the cells. Sulfated glycosaminoglycan-deficient Huma 109 cells were prepared as described previously (5,9,25). Cells were incubated for 4 h in sulfate-free Dulbecco's modified Eagle's medium supplemented with 10% dialyzed fetal calf serum (v/v) and 15 mM NaClO 3 .…”

Section: Methodsmentioning

confidence: 99%

Stimulation of DNA Synthesis and Cell Proliferation of Human Mammary Myoepithelial-like Cells by Hepatocyte Growth Factor/Scatter Factor Depends on Heparan Sulfate Proteoglycans and Sustained Phosphorylation of Mitogen-activated Protein Kinases p42/44

Sergeant

Lyon

Rudland

et al. 2000

Journal of Biological Chemistry

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“…The distribution of the immobilized heparin, DS, and of the bound HARP on the surface of the biosensor cuvette was inspected by examination of the resonance scan, which showed that at all times these molecules were distributed uniformly on the sensor surface and therefore were not microaggregated. Binding assays were as described (27), and the amount of bound HARP is reported in arc s (1 arc s ϭ 1/3600°. Briefly, the ligate, HARP, was added at a known concentration in 100 l of PBS-T (PBS supplemented with 0.02% (v/v) Tween 20), and then the association reaction was followed over a set time, usually 150 s. The cuvette was then washed twice with 200 l of PBS-T, and the dissociation of bound ligate into the bulk PBS-T was followed over time.…”

Section: Activation Of Harp By Heparin In Heparitinase-treated Bel Cementioning

confidence: 99%

“…To remove residual bound ligate and thus regenerate the immobilized ligand, the cuvette was washed twice with 200 l of 2 M NaCl, 10 mM Na 2 HPO 4 , pH 7.2. Binding parameters were calculated from the association and dissociation rate constants, k ass and k diss , respectively, using the nonlinear curve-fitting FastFit software (Affinity Sensors) provided with the instrument, as described (27). HARP did not itself bind to streptavidin-derivatized surfaces.…”

Section: Activation Of Harp By Heparin In Heparitinase-treated Bel Cementioning

confidence: 99%

Glycosaminoglycans Differentially Bind HARP and Modulate Its Biological Activity

Vacherot

Delbé

Héroult

et al. 1999

Journal of Biological Chemistry

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Heparin affin regulatory peptide (HARP) is a polypep-) and a lower affinity (K d ‫؍‬ 51 nM). Exogenous heparin, heparan sulfate, and dermatan sulfate potentiated the growth-stimulatory activity of HARP, suggesting that corresponding proteoglycans could be involved in the regulation of the mitogenic activity of HARP. Heparin affin regulatory peptide (HARP)1 belongs to a growing group of heparin binding extracellular regulatory molecules, and it has mitogenic and neurite outgrowth activities.Independently purified from perinatal rat brain as a polypeptide that induces neurite outgrowth of embryonic neurons (1), this protein, also named pleiotrophin (2), has been purified from uterus (3) and adult brain (4) as a growth factor for fibroblastic and endothelial cells. Expression of HARP mRNA is developmentally regulated, and major expression is found during the perinatal growth period in rat brain (5, 6), pointing to the possible function of this molecule in the maturation of nerve cells. Moreover, HARP mRNA is found in several adult tissues, and HARP has been implicated in tumor growth (7).HARP cDNA has been cloned and sequenced from several species. The predicted amino acid sequence has been determined in humans, mice, and rats (2, 6, 8, 9) and shows 98% homology among the three species. In addition, the HARP amino acid sequence shares 55% homology with the midkine gene product (10). In contrast to their neurite outgrowth activity, initial studies differed in their results about the mitogenic activity of midkine and HARP (3, 9, 11-15), possibly because of differences in the cell type used or to the isolation procedure.More recent studies clearly demonstrate that this family of polypeptides have growth-stimulatory activity (16).The high affinity of HARP for heparin suggests that HARP may bind to heparan sulfate proteoglycans (HSPGs) present in extracellular compartments defined as cell surface and extracellular matrix (ECM). Recent studies have demonstrated that syndecan-1, syndecan-3 (N-syndecan), and syndecan-4 (ryudocan) bind HARP with high affinity (13,17,18). In addition, biochemical and cell biological studies have pointed to syndecan-3 as the HARP receptor involved in neurite outgrowth activity (19). Despite the correlation derived from previous studies, there has been no direct biochemical demonstration that HARP is trapped in the extracellular compartment as a mitogenic molecule bound to HSPGs. In this study we investigated whether HARP is present in the extracellular compartment as well as the role of glycosaminoglycans (GAGs) in controlling HARP mitogenic activity. (Proteus vulgaris; EC 4.2.2.4), leupeptin, pepstatin, phenylmethylsulfonyl fluoride, heparan sulfate (HS) from bovine intestinal mucosa, keratan sulfate (KS) from bovine cornea, chondroitin sulfate A (CS-A) from bovine trachea, dermatan sulfate (DS) from porcine skin, and chondroitin sulfate C (CS-C) from shark cartilage were purchased from Sigma. Horseradish peroxidase-conjugated goat anti-rabbit immunoglobulins were obtained from Diagnostics-...

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“…The HS chain mediates a second extracellular regulatory input. Specific sequences in HS can allow only a restricted subset of FGF-FGFR interactions to lead to cell proliferation (13)(14)(15)(16) and this is likely to occur in vivo (17,18). A third extracellular regulatory input is likely to be the assembly of the FGF receptor ligand system into different complexes that have different signaling potentials.…”

mentioning

confidence: 99%

N-Glycosylation of Fibroblast Growth Factor Receptor 1 Regulates Ligand and Heparan Sulfate Co-receptor Binding

Duchesne

Tissot

Rudd

et al. 2006

Journal of Biological Chemistry

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The regulation of cell function by fibroblast growth factors (FGF) occurs through a dual receptor system consisting of a receptor-tyrosine kinase, FGFR and the glycosaminoglycan heparan sulfate (HS). Mutations of some potential N-glycosylation sites in human fgfr lead to phenotypes characteristic of receptor overactivation. To establish how N-glycosylation may affect FGFR function, soluble-and membrane-bound recombinant receptors corresponding to the extracellular ligand binding domain of FGFR1-IIIc were produced in Chinese Hamster Ovary cells. Both forms of FGFR1-IIIc were observed to be heavily N-glycosylated and migrated on SDS-PAGE as a series of multiple bands between 50 and 75 kDa, whereas the deglycosylated receptors migrated at 32 kDa, corresponding to the expected molecular weight of the polypeptides. Optical biosensor and quartz crystal microbalance-dissipation binding assays show that the removal of the N-glycans from FGFR1-IIIc caused an increase in the binding of the receptor to FGF-2 and to heparin-derived oligosaccharides, a proxy for cellular HS. This effect is mediated by N-glycosylation reducing the association rate constant of the receptor for FGF-2 and heparin oligosaccharides. N-Glycans were analyzed by mass spectrometry, which demonstrates a predominance of bi-and tri-antennary core-fucosylated complex type structures carrying one, two, and/or three sialic acids. Modeling of such glycan structures on the receptor protein suggests that at least some may be strategically positioned to interfere with interactions of the receptor with FGF ligand and/or the HS co-receptor. Thus, the N-glycans of the receptor represent an additional pathway for the regulation of the activity of FGFs.The fibroblast growth factors (FGFs) 5 and their receptors co-evolved with metazoans (1, 2), a reflection of their central role in mediating cell-cell communication. FGFs regulate the specification of stem cell fate, organogenesis, skeletal growth (3), tissue repair (4) and phosphate metabolism. FGFs are involved in numerous pathologies such as genetic skeletal defects in children (5) and carcinomas in adults (6, 7).In humans and rodents there are 22 genes encoding more than 30 different FGF proteins and the fgfr 1-4 genes encode over 48 major isoforms of the cognate receptor tyrosine kinase (FGFR) (1, 2, 8). FGFs possess a glycosaminoglycan co-receptor, usually heparan sulfate (HS). The interaction of FGFs with HS is required for the stimulation of cell proliferation, which is initiated by the formation of a complex of FGF ligand, HS co-receptor and FGFR (9 -11). Not surprisingly for a family of key regulatory ligands, the activation of FGF signaling is subjected to multiple regulatory inputs, some of which operate at the level of the assembly of the receptor-ligand complex, whereas others operate inside the cell, on the active receptor complex. Regulatory inputs that operate extracellularly include partial selectivity in the choice of ligand by certain receptor isoforms. FGF-7 is the most specific FGF ligand, beca...

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Interaction of Heparan Sulfate from Mammary Cells with Acidic Fibroblast Growth Factor (FGF) and Basic FGF

Cited by 116 publications

References 45 publications

Stimulation of DNA Synthesis and Cell Proliferation of Human Mammary Myoepithelial-like Cells by Hepatocyte Growth Factor/Scatter Factor Depends on Heparan Sulfate Proteoglycans and Sustained Phosphorylation of Mitogen-activated Protein Kinases p42/44

Stimulation of DNA Synthesis and Cell Proliferation of Human Mammary Myoepithelial-like Cells by Hepatocyte Growth Factor/Scatter Factor Depends on Heparan Sulfate Proteoglycans and Sustained Phosphorylation of Mitogen-activated Protein Kinases p42/44

Glycosaminoglycans Differentially Bind HARP and Modulate Its Biological Activity

N-Glycosylation of Fibroblast Growth Factor Receptor 1 Regulates Ligand and Heparan Sulfate Co-receptor Binding

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