[Heterocomplex formation between high and low affinity FGF receptors is mediated by the formation of a FGF dimer].
Interactions between the two classes of fibroblast growth factor receptors 1) the high affinity receptors (HAR) a membrane glycoprotein containing an intrinsic tyrosine kinase activity, 2) low affinity receptors (LAR) cell surface proteoglycans containing heparan sulfate side chains (HSPG), and aFGF (MW: 15.5 kDa) were studied in bovine lens epithelial (BEL) cells. By Scatchard analysis of the aFGF binding to the BEL cell surface, we show that heparin at 10 micrograms/ml abolishes completely aFGF binding to LAR and reduces by half the number of aFGF HAR. By using cross-linking experiments, aFGF-HAR complexes are present in two forms (150 kDa and 135 kDa). Addition of heparin at 10 micrograms/ml abolishes the formation of the 150 kDa complex and does not affect the 135 kDa complex. Furthermore, binding of aFGF to LAR induces the spontaneous formation of a 31 kDa aFGF dimer. The dimerization process of aFGF on LAR is abolished by addition of heparin. During aFGF internalization at 37 degrees C, we have shown that aFGF-dimer is internalized, accumulated and degraded in the cell as is the 15.5 kDa native form. Heparin at 10 micrograms/ml suppresses specifically aFGF dimer internalization and reduces by half the total amount of internalized aFGF native form. Moreover, after aFGF binding and internalization, the affinity of HAR for aFGF increases concomitantly with its downregulation. Heparin does not seem to affect this phenomenon. All these results strongly suggest that an heteroreceptor dimer-aFGF complex (150 kDa) is formed by one molecule of HAR associated to one molecule of LAR through their respective interaction with a very stable homodimer of aFGF. Such a three component receptor complex induced by FGF dimerization may be a general process of FGF receptor activation which could explain the diversity of the biological response to FGF of different cell type expressing different HAR and LAR or HSPG.
Mascarelli
F
F
INSERM U 118, Association Claude-Bernard, Paris, France.
Courtois
Y
Y
D000818:Animals; D001667:Binding, Competitive; D002417:Cattle; D002478:Cells, Cultured; D016220:Fibroblast Growth Factor 1 / Q000378:metabolism*; D006493:Heparin / Q000494:pharmacology; D007908:Lens, Crystalline / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D008024:Ligands; D017468:Receptors, Fibroblast Growth Factor / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D013498:Suramin / Q000494:pharmacology
D008024:Ligands; D017468:Receptors, Fibroblast Growth Factor; D016220:Fibroblast Growth Factor 1; D013498:Suramin; D006493:Heparin