Interaction of basic fibroblast growth factor with bovine growth plate chondrocytes.
The basic fibroblast growth factor (bFGF) family of peptides influences a wide range of cellular actions. To better understand the possible role of bFGF in the growth plate, we have characterized the interaction of this growth factor with isolated bovine growth plate chondrocytes. Basic FGF interacts with two classes of binding sites on these cells. One is consistent with high-affinity bFGF receptors and the other with low-affinity heparin-like binding sites on the chondrocyte surface. Radiolabeled bFGF binding studies revealed approximately 4 x 10(6) binding sites per cell, with a Kd of approximately 42 nM. Graded concentrations of heparin or NaCl competed with [125I]-labeled bFGF in a dose-dependent fashion, reducing [125I]-labeled bFGF binding by 75 and 97%, respectively. The data suggest the presence of a high-capacity, low-affinity class of binding sites with the properties of a heparin-like moiety. Affinity cross-linking of [125I]-labeled bFGF to chondrocytes labeled two principal species with apparent molecular masses of 135 and 160 kDa. Labeled bFGF was specifically displaced from both species by subnanomolar concentrations of unlabeled bFGF. These high-affinity, low-capacity binding sites are characteristic of classical bFGF receptors. Binding of [125I]-labeled bFGF to these sites was also influenced by heparin, consistent with coregulation of binding to the two classes of binding sites. The data suggest that bFGF participates in the regulation of skeletal growth at the growth plate and that this regulation may involve bFGF interaction with at least two distinct classes of binding sites.
Trippel
S B
SB
Department of Orthopaedics, Massachusetts General Hospital, Boston 02115.
Whelan
M C
MC
Klagsbrun
M
M
Doctrow
S R
SR
D000818:Animals; D001345:Autoradiography; D001667:Binding, Competitive; D002417:Cattle; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D016222:Fibroblast Growth Factor 2 / Q000378:metabolism* / Q000494:pharmacology; D006132:Growth Plate / Q000166:cytology* / Q000378:metabolism* / Q000648:ultrastructure; D006493:Heparin / Q000378:metabolism / Q000494:pharmacology; D007457:Iodine Radioisotopes; D011956:Receptors, Cell Surface / Q000032:analysis / Q000378:metabolism; D017468:Receptors, Fibroblast Growth Factor; D012965:Sodium Chloride / Q000378:metabolism / Q000494:pharmacology
D007457:Iodine Radioisotopes; D011956:Receptors, Cell Surface; D017468:Receptors, Fibroblast Growth Factor; D016222:Fibroblast Growth Factor 2; D012965:Sodium Chloride; D006493:Heparin