Basic fibroblast growth factor binds to heparan sulfate in the extracellular matrix of rat growth plate chondrocytes.
Recent studies have demonstrated that basic fibroblast growth factor (bFGF) plays a key role in the terminal differentiation of growth plate chondrocytes during endochondral ossification. We therefore examined the binding of [125I]bFGF to an extract of extracellular matrix (ECM) of rat growth plate. Using a solid phase binding assay, binding of [125I]bFGF to ECM was demonstrated to be specific and saturable at 0.5-1 microgram/ml of bFGF. Scatchard analysis demonstrated the presence of a single class of binding sites with an apparent Kd of 14 nM. The binding of [125I]bFGF to rat growth plate ECM was inhibited by the addition of heparin, heparan sulfate, and dermatan sulfate. The binding was reversible as these glycosaminoglycans were also effective at displacement of bound [125I]bFGF from rat growth plate ECM. Chondroitin 4- or 6-sulfate had no effect on either binding or displacement when added at the same concentrations. Preincubation of the rat growth plate ECM with heparinase or heparitinase resulted in a reduction of the binding of [125I]bFGF to the ECM. Furthermore, these enzymes were able to significantly displace bound growth factor. In contrast, chondroitinase ABC or AC failed to displace bound [125I]bFGF from the ECM. Similar results were obtained when matrix derived from rat growth plate tissue was used for the binding studies. The results demonstrate that bFGF binds to a heparan sulfate in matrix produced by rat growth plate chondrocytes and matrix extracted from rat growth plate and suggest that this glycosaminoglycan may serve as a storage depot for this growth factor during endochondral ossification.
Chintala
S K
SK
Department of Biomedical Engineering, Cleveland Clinic Foundation Research Institute, Ohio 44195-5254.
Miller
R R
RR
McDevitt
C A
CA
D000469:Alkaline Phosphatase / Q000032:analysis; D000818:Animals; D002356:Cartilage / Q000166:cytology / Q000201:enzymology / Q000378:metabolism*; D005109:Extracellular Matrix / Q000378:metabolism*; D005269:Femur / Q000166:cytology; D016222:Fibroblast Growth Factor 2 / Q000378:metabolism*; D006025:Glycosaminoglycans / Q000378:metabolism / Q000494:pharmacology; D006132:Growth Plate / Q000201:enzymology / Q000378:metabolism*; D006497:Heparan Sulfate / Q000378:metabolism*; D011133:Polysaccharide-Lyases / Q000494:pharmacology; D011485:Protein Binding / Q000187:drug effects; D051381:Rats; D017207:Rats, Sprague-Dawley
D000469:Alkaline Phosphatase; D016222:Fibroblast Growth Factor 2; D006025:Glycosaminoglycans; D006497:Heparan Sulfate; D011133:Polysaccharide-Lyases