Oligomerization reduces heparin affinity but enhances receptor binding of fibroblast growth factor 2.
The biological response of cells to fibroblast growth factors (FGFs) depends on heparan sulphate glycosaminoglycans sharing particular structural motifs. Heparin induced FGF dimerization has been suggested to mediate receptor dimerization and activation. Here we demonstrate that heparin-derived oligosaccharides that promote receptor binding and activation specifically induce the dimerization of basic FGF (FGF2). These heparin-induced dimers of FGF2 acquire high affinity for receptor binding and are biologically active. Using biotinylated FGF2 bound to immobilized streptavidin gradually saturated with biotin, enabled a quantitative analysis of heparin-dependent and heparin-independent FGF2 monomers and oligomers. Streptavidin induced FGF2 dimers bind and activate FGF receptors only in the presence of heparin. An excess of streptavidin, forcing biotin-FGF2 into monomers, reduces receptor binding and blocks FGF-dependent cell proliferation. All these suggest predominant receptor binding and activation by heparin associated FGF2 oligomers. Unexpectedly, heparin induced dimers and higher order oligomers lose most of their affinity towards heparin. Direct binding of soluble FGF receptors (FGFRs) to either monomers or dimers of FGF2, immobilized on heparin, confirm the preferred association of FGFRs with dimers of FGF2. Computerized molecular docking predicts a cis-oriented FGF2 dimer, stabilized by heparin, which complies with all the experimental data.
Safran
M
M
Department of Molecular Cell Biology, Weizmann Institute of Science, Rehovot 76100, Israel.
Eisenstein
M
M
Aviezer
D
D
Yayon
A
A
D000818:Animals; D001665:Binding Sites; D001710:Biotin; D002455:Cell Division / Q000187:drug effects / Q000502:physiology; D002460:Cell Line; D003198:Computer Simulation; D019281:Dimerization; D016222:Fibroblast Growth Factor 2 / Q000737:chemistry* / Q000378:metabolism*; D006493:Heparin / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D007700:Kinetics; D051379:Mice; D008958:Models, Molecular; D009844:Oligosaccharides / Q000737:chemistry / Q000494:pharmacology; D020836:Protein Structure, Quaternary / Q000187:drug effects; D020794:Receptor Protein-Tyrosine Kinases / Q000378:metabolism*; D051497:Receptor, Fibroblast Growth Factor, Type 2; D017468:Receptors, Fibroblast Growth Factor / Q000378:metabolism*; D019809:Streptavidin
D009844:Oligosaccharides; D017468:Receptors, Fibroblast Growth Factor; D016222:Fibroblast Growth Factor 2; D001710:Biotin; D006493:Heparin; D019809:Streptavidin; C496354:Fgfr2 protein, mouse; D020794:Receptor Protein-Tyrosine Kinases; D051497:Receptor, Fibroblast Growth Factor, Type 2