Proteoheparan sulfates contribute to the binding of basic FGF to its high affinity receptors on bovine adrenocortical cells.
Bovine adrenocortical cells in primary culture express the basic fibroblast growth factor (bFGF) gene and their proliferation is stimulated by this growth factor. We report here the characterization of bFGF receptors on these cells. Binding studies revealed the presence of two bFGF receptor types: a limited number (4,300 sites/cell) of high affinity sites (Kd congruent to 2 pM) and a larger number (230,000 sites/cell) of lower affinity sites (Kd congruent to 400 pM). Cross-linking of 125I-bFGF to adrenocortical cells revealed two bands at 145 kDa and 125 kDa which are attributed to molecular complexes between the high affinity receptors and their ligand. These high affinity receptors possess N-linked carbohydrate chains that are important for proper cell surface expression but are devoid of glycosaminoglycan chains. The low-affinity (2 M NaCl-sensitive) binding sites are totally degraded by heparitinase treatment of adrenocortical cells indicating that low-affinity sites are borne by heparan sulfate proteoglycans. However, heparitinase treatment also reduced partially the binding of bFGF to high-affinity (2 M NaCl-resistant) sites. This argues for a contribution of heparan sulfate proteoglycans to the binding of bFGF to high-affinity receptors. Exogenous soluble heparin or heparan sulfate did not restore normal high affinity bFGF binding onto heparitinase-treated cells, suggesting that heparan sulfate proteoglycans either must be membrane-anchored or must contain specific structural features to enhance FGF binding to high-affinity receptors. Taken together with previous reports, this work supports the hypothesis that bFGF may act as an autocrine growth factor in the adrenal cortex.
Savona
C
C
Unité INSERM 244, Biochimie des Régulations Cellulaires Endocrines, DBMS, Centre d'Etudes Nucléaires, Grenoble, France.
Chambaz
E M
EM
Feige
J J
JJ
D000302:Adrenal Cortex / Q000166:cytology / Q000378:metabolism*; D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D004591:Electrophoresis, Polyacrylamide Gel; D016222:Fibroblast Growth Factor 2 / Q000378:metabolism*; D006031:Glycosylation; D019812:Heparan Sulfate Proteoglycans; D019763:Heparin Lyase; D006497:Heparan Sulfate / Q000378:metabolism*; D011133:Polysaccharide-Lyases / Q000378:metabolism; D011509:Proteoglycans / Q000378:metabolism*; D011956:Receptors, Cell Surface / Q000378:metabolism*; D017468:Receptors, Fibroblast Growth Factor
D016222:Fibroblast Growth Factor 2; D019812:Heparan Sulfate Proteoglycans; D019763:Heparin Lyase; D006497:Heparan Sulfate; D011133:Polysaccharide-Lyases; D011509:Proteoglycans; D011956:Receptors, Cell Surface; D017468:Receptors, Fibroblast Growth Factor