Binding, internalization, and degradation of basic fibroblast growth factor in human microvascular endothelial cells

Bikfalvi, Andréas; Dupuy, Évelyne; Inyang, A. L.; Fayein, N.A.; Lesèche, Guy; Courtois, Yves; Tobelem, Gérard

doi:10.1016/0014-4827(89)90183-3

Cited by 42 publications

(11 citation statements)

References 16 publications

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“…We do not know if these subfragments of FGF2 have a biological activity of their own. However, they were not seen in earlier studies with '"I-FGF2 (Moscatelli, 1988;Bikfalvi et al, 1989;Moenner et al, 1989;Mascarelli et al, 1991;Walicke and Baird, 1991;Rusnati et al, 1993). Several reasons could be envisioned for the absence of the 14-kDa and 7-kDa catabolic fragments in the case of Iz5I-FGF2: (a) these fragments corresponded to sequences of FGF2 which did not contain "'1-fixing tyrosine residues ; (b) these fragments had lost their radiolabelling agent during the catabolism of FGF2 which rendered them undetectable ; (c) the presence of iodine influenced the FGF2-processing pathways.…”

Section: Discussionmentioning

confidence: 56%

Comparative Study In Vivo and In Vitro of Uniformly ¹⁴C‐Labelled and ¹²⁵I‐Labelled Recombinant Fibroblast Growth Factor 2

Colin

Mascarelli

Jeanny

et al. 1997

European Journal of Biochemistry

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Recombinant bovine fibroblast growth factor (FGF2), uniformly labelled with 14C ([14C]FGF2), was purified and showed to be highly stable and to retain full biological activity. Organ distribution of [14C]FGF2 after intravenous injection of young rats was assessed by autoradiography of whole body sections and compared with those obtained with [125I]iodinated FGF2 (125I‐FGF2). Thyroid, stomach, intestine, bladder and skin were radioactively labelled only in the case of 125I‐FGF2. This tissue‐labelling is artefactual, probably due to free iodide binding not observed when using [14C]FGF2. High‐resolution autoradiography showed a complex tissue distribution of [14C]FGF2 in kidney and adrenal organs. Incubation of frozen eye sections with [14C]FGF2 showed a specific and high‐resolution labelling pattern of ocular tissues. After cellular internalization, [14C]FGF2 was processed into five distinct polypeptides of 16, 14, 8, 7, and 5.5 kDa. The 14‐kDa and 7‐kDa polypeptides are novel catabolic fragments not detected with radioiodinated FGF2. In terms of stability, tissue distribution specificity, and autoradiographic resolution, [14C]FGF2 proved to have more advantages than 125I‐FGF2 for pharmacokinetic and catabolism studies.

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Section: Discussionmentioning

confidence: 56%

Comparative Study In Vivo and In Vitro of Uniformly ¹⁴C‐Labelled and ¹²⁵I‐Labelled Recombinant Fibroblast Growth Factor 2

Colin

Mascarelli

Jeanny

et al. 1997

European Journal of Biochemistry

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“…Internalization of VASiVEGF reaches a maximum of 55% after 30 min. Internalization of bFGF has been reported to be slower than for VASNEGF (Moscatelli, 1988;Bikfalvi et al, 1989). These differences might be due to the absence of binding of VASIVEGF to slow internalizing low-affinity binding sites as has been demonstrated for bFGF (Moscatelli, 1988).…”

Section: 5*mentioning

confidence: 88%

Interaction of vasculotropin/vascular endothelial cell growth factor with human umbilical vein endothelial cells: Binding, internalization, degradation, and biological effects

Bikfalvi

Sauzeau

Moukadiri

et al. 1991

Journal Cellular Physiology

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114

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Vasculotropin/vascular endothelial cell growth factor (VAS/VEGF) is a newly purified growth factor with a unique specificity for vascular endothelial cells. We have investigated the interactions of VAS/VEGF with human umbilical vein endothelial cells (HUVE cells). 125I-VAS/VEGF was found to HUVE cells in a saturable manner with a half-maximum binding at 2.8 ng/ml. Scatchard analysis did show two classes of high-affinity binding sites. The first class displayed a dissociation constant of 9 pM with 500 sites/cell. The dissociation constant and the number of binding sites of the second binding class were variable for different HUVE cell cultures (KD = 179 +/- 101 pM, 5,850 +/- 2,950 sites/cell). Half-maximal inhibition of 125I-VAS/VEGF occurred with a threefold excess of unlabeled ligand. Basic fibroblast growth factor (bFGF) and heparin did not compete with 125I-VAS/VEGF binding. In contrast, suramin and protamin sulfate completely displaced 125I-VAS/VEGF binding from HUVE cells. VAS/VEGF was shown to be internalized in HUVE cells. Maximum internalization (55% of total cell-associated radioactivity) was observed after 30 min. 125I-VAS/VEGF was completely degraded 2-3 hr after binding. At 3 hr, the trichloroacetic acid (TCA)-soluble radioactivity accumulated in the medium was 60% of the total radioactivity released by HUVE cells. No degradation fragment of 125I-VAS/VEGF was observed. Chloroquine completely inhibited degradation. VAS/VEGF was able to induce angiogenesis in vitro in HUVE cells. However, it did not significantly modulate urokinase-type plasminogen activator (u-PA), tissue-type plasminogen activator (t-PA), plasminogen activator inhibitor (PAI-1), and tissue factor (TF). Prostacyclin production was only stimulated at very high VAS/VEGF concentrations. Taken together, these results indicate that VAS/VEGF might be a potent inducer of neovascularization resulting from a direct interaction with endothelial cells. The angiogenic activity seems to be independent of the plasminogen activator or inhibitor system.

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“…Maximum degradation was observed between 12 and 24 h. Interestingly, at 24 h, 18-kDa FGF-2 was still present in significant amounts in LECs. Together, our biochemical study provides detailed information on binding kinetics of FGF-2 to its receptors and subsequent internalization and degradation patterns of the ligand in lymphatic endothelial cells, which are largely comparable with those of vascular endothelial cells as described previously (Bikfalvi et al, 1989).…”

Section: Fgf-2 Binds Directly To Low-and High-affinity Receptors In Lmentioning

confidence: 90%

Prox1 Promotes Lineage-specific Expression of Fibroblast Growth Factor (FGF) Receptor-3 in Lymphatic Endothelium: A Role for FGF Signaling in Lymphangiogenesis

Shin

Min

Larrieu-Lahargue³

et al. 2006

MBoC

168

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Fibroblast growth factors play important roles in angiogenesis, but their functions in lymphangiogenesis remain poorly understood. The homeodomain transcription factor Prox1 is essential for development of the lymphatic system by specifying lymphatic endothelial cell (LEC) fate. Here, we identify fibroblast growth factor (FGF) receptor (FGFR)-3 as a novel Prox1 target gene. Ectopic overexpression of Prox1 in blood vascular endothelial cells up-regulates FGFR-3. Prox1 induces the expression of the IIIc isoform, which we also found to be the major isoform of FGFR-3 expressed in LECs. This transcriptional activation is mediated by a direct binding of Prox1 to newly identified Prox1-response elements in the FGFR-3 promoter. Consistently, FGFR-3 is up-regulated in Prox1-positive newly formed lymphatic vessels during embryogenesis and its lymphatic-specific expression is maintained throughout development. We also found that FGF-1 and FGF-2 promote proliferation, migration, and survival of cultured LECs without involvement of vascular endothelial cell growth factor receptor-3. We show that FGF-2 binds to low-and high-affinity receptors on LECs and is efficiently internalized and processed. Moreover, functional inhibition of FGFR-3 using small interfering RNA represses LEC proliferation. Together, these results indicate that FGFR-3 is an initial target of Prox1 during the lymphatic cell fate specification and that FGF signaling may play an important role in lymphatic vessel development.

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Binding, internalization, and degradation of basic fibroblast growth factor in human microvascular endothelial cells

Cited by 42 publications

References 16 publications

Comparative Study In Vivo and In Vitro of Uniformly ¹⁴C‐Labelled and ¹²⁵I‐Labelled Recombinant Fibroblast Growth Factor 2

Comparative Study In Vivo and In Vitro of Uniformly ¹⁴C‐Labelled and ¹²⁵I‐Labelled Recombinant Fibroblast Growth Factor 2

Interaction of vasculotropin/vascular endothelial cell growth factor with human umbilical vein endothelial cells: Binding, internalization, degradation, and biological effects

Prox1 Promotes Lineage-specific Expression of Fibroblast Growth Factor (FGF) Receptor-3 in Lymphatic Endothelium: A Role for FGF Signaling in Lymphangiogenesis

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Binding, internalization, and degradation of basic fibroblast growth factor in human microvascular endothelial cells

Cited by 42 publications

References 16 publications

Comparative Study In Vivo and In Vitro of Uniformly 14C‐Labelled and 125I‐Labelled Recombinant Fibroblast Growth Factor 2

Comparative Study In Vivo and In Vitro of Uniformly 14C‐Labelled and 125I‐Labelled Recombinant Fibroblast Growth Factor 2

Interaction of vasculotropin/vascular endothelial cell growth factor with human umbilical vein endothelial cells: Binding, internalization, degradation, and biological effects

Prox1 Promotes Lineage-specific Expression of Fibroblast Growth Factor (FGF) Receptor-3 in Lymphatic Endothelium: A Role for FGF Signaling in Lymphangiogenesis

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Comparative Study In Vivo and In Vitro of Uniformly ¹⁴C‐Labelled and ¹²⁵I‐Labelled Recombinant Fibroblast Growth Factor 2

Comparative Study In Vivo and In Vitro of Uniformly ¹⁴C‐Labelled and ¹²⁵I‐Labelled Recombinant Fibroblast Growth Factor 2