NATURE OF THE INTERACTION OF HEPARIN WITH ACIDIC FIBROBLAST GROWTH-FACTOR

The binding of human acidic fibroblast growth factor (aFGF) to heparin has been analyzed by a variety of different approaches to better eluc idate the nature of this protein/sulfated polysaccharide interaction. Static and dynamic light scattering as well as analytical ultracentrif ugation analyses indicates that 14-15 molecules of aFGF can bind to a 16-kDa heparin chain, with approximately 10 of these bound relatively uniformly to high-affinity sites. The dissociation constants of these latter sites are estimated to be approximately 50-140 nM on the basis of surface plasmon resonance experiments in which the association and dissociation rates of aFGF interaction with immobilized heparin were m easured. The size of the binding site of aFGF on heparin was also dete rmined by heparin lyase digestion of aFGF/heparin complexes followed b y isolation and characterization of protected oligosaccharides. The sm allest aFGF-protected oligosaccharide comigrated with S(1-->4)-alpha-L -IdoAp-2S(1-->4)-alpha-D-GlcNp2S6S (where DELTAUA represents 4-deoxy-a lpha-L-threo-hex-4-enopyranosyluronic acid and S is sulfate). Thus, a FGF appears to bind at high density (one molecule every 4-5 polysaccha ride units) and with high affinity to heparin. This potentially provid es a concentrated, stabilized storage form of the growth factor that c an be released for receptor-mediated cellular activation in response t o the proper stimuli. It is also possible that close proximity of aFGF molecules on the highly sulfated regions of heparan chains may be inv olved in the induction of receptor aggregation as suggested by Ornitz et al.