The identification and partial characterization of the fibroblast growth factor receptor of baby hamster kidney cells.
The binding of biologically active, 125I-labeled basic fibroblast growth factor (FGF) to baby hamster kidney-derived cell line cells (BHK-21) was studied at 4 degrees C. Unlabeled FGF displaced cell surface bound 125I-FGF, but platelet-derived growth factor, epidermal growth factor, insulin, or transferrin did not. Binding was saturable both as a function of time and as a function of increasing 125I-FGF concentrations. Scatchard analysis of the binding data revealed the presence of about 1.2 X 10(5) binding sites/cell with an apparent KD of 270 pM. The number of the binding sites was down-regulated following preincubation of the cells with FGF. The density of binding sites/cell also decreased as an inverse function of cell density. When 125I-FGF binding was studied in a BHK-21 cell membrane preparation, it was found that the membranal binding site displayed a lower KD of 21 pM. 125I-FGF was covalently cross-linked to its cell surface receptor on intact BHK-21 cells using the homobifunctional agent disuccinimidyl suberate. Two macromolecular species with an apparent molecular weight of 145,000 and 125,000, respectively, were labeled under both reducing and nonreducing conditions. Unlabeled FGF competed with 125I-FGF for binding to both macromolecular species. The labeling of the macromolecules was also inhibited by heparin. No labeling was observed in the absence of the cross-linkers or when heat-inactivated 125I-FGF was used instead of radiolabeled, biologically active FGF.
D000818:Animals; D002452:Cell Count; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D006224:Cricetinae; D005346:Fibroblast Growth Factors / Q000378:metabolism*; D007457:Iodine Radioisotopes; D007668:Kidney / Q000378:metabolism; D008970:Molecular Weight; D010766:Phosphorylation; D011956:Receptors, Cell Surface / Q000032:analysis*; D017468:Receptors, Fibroblast Growth Factor
D007457:Iodine Radioisotopes; D011956:Receptors, Cell Surface; D017468:Receptors, Fibroblast Growth Factor; D005346:Fibroblast Growth Factors