For Fibroblast Growth Factor Research Articles

Affinity of fibroblast growth factors for β-cyclodextrin tetradecasulfate

β-Cyclodextrin tetradecasulfate was found to have a very strong affinity for fibroblast growth factor (FGF) and could substitute for heparin in FGF purification. Basic FGF was purified about 200,000-fold from a rat chondrosarcoma using a method of biaffinity chromatography in which the β-cyclodextrin tetradecasulfate polymer was mixed with copper-Sepharose. This method takes advantage of the strong affinity of FGF for both β-cyclodextrin tetradecasulfate and copper.

Fibroblast growth factor receptor levels decrease during chick embryogenesis.

Two putative receptors for fibroblast growth factor (FGF) of approximately 150 and 200 kD were identified in membrane preparations from chick embryos. Specific binding (femtomoles/milligram) of 125I-aFGF to whole chick embryonic membranes was relatively constant from day 2 to 7, then decreased fivefold between days 7 and 13. Day-19 chick embryos retained 125I-aFGF binding at low levels to brain, eye, and liver tissues but not to skeletal muscle or cardiac tissues. The 200-kD FGF receptor began to decline between day 4.5 and 7 and was barely detectable by day 9, whereas the 150-kD FGF receptor began to decline by day 7 but was still detectable in day-9 embryonic membranes. It is not known whether the two FGF-binding proteins represent altered forms of one polypeptide, but it is clear that their levels undergo differential changes during development. Because endogenous chick FGF may remain bound to FGF receptor in membrane preparations, membranes were treated with acidic (pH 4.0) buffers to release bound FGF; such treatment did not affect 125I-aFGF binding and moderately increased the number of binding sites in day-7 and -19 embryos. Consequently, the observed loss of high affinity 125I-aFGF binding sites and FGF-binding polypeptides most likely represents a loss of FGF receptor protein. These experiments provide in vivo evidence to support the hypothesis that regulation of FGF receptor levels may function as a mechanism for controlling FGF-dependent processes during embryonic development.

Identification of a new heparin‐binding protein localized within chick basement membranes

A protein with a molecular mass of 19 kDa has been purified to homogeneity from 11-day-old chick embryos using a procedure involving chromatography on heparin - Sepharose, immunoaffinity resin and C4 reversed-phase. Indirect immunofluorescence studies, using polyclonal and monoclonal antibodies raised against this protein, indicate that it is essentially localized within the basement membranes in early embryonic tissues. After the 18th day of embryonic life and in post-hatched chicken, this protein could only be detected in some eye basement membranes. It appears to be bound to heparan sulfate chains of the proteoglycan present in these structures. Thus, the protein exhibits similar properties to those previously described for fibroblast growth factors (FGF), such as heparin affinity, molecular mass and localization in the basement membranes. In contrast, this protein is present in much larger amounts than FGFs, at least in 11-day-old embryos. Furthermore, the first 17 amino acid residues of the N-terminal sequence show that it does not strictly correspond to any previously described protein.

Radioimmunoassay for fibroblast growth factor (FGF): release by the bovine anterior pituitary in vitro.

A radioimmunoassay (RIA) was developed to measure fibroblast growth factor (FGF) using antiserum generated against a synthetic replicate of [Tyr10]FGF(1-10). The antisera, previously shown to be capable of inhibiting the biological action of FGF on bovine aortic arch endothelial cells in vitro, are highly specific for the amino-terminus of FGF. In the RIA, the antisera recognize the decapeptide antigen [Tyr10]FGF(1-10) and the intact mitogen on an equimolar basis and show less than 0.01% cross-reactivity with N-acetyl-[Tyr10]FGF(1-10). Bovine adenohypophysial cells maintained in primary monolayer culture release and ir-FGF which is indistinguishable from the intact mitogen in as much as it is retained on heparin-Sepharose affinity columns and shows a dose-dependent and parallel displacement in RIA. The release of ir-FGF by the bovine adenohypophysis can be increased with forskolin (10(-5) M) or KCl (50 mM). Preincubation of pituitary cells with 17 beta-estradiol has no measurable effects on basal ir-FGF, but increases the release after KCl treatment 2-3-fold. These results show that ir-FGF can be released by the bovine adenohypophysis in vitro and lend credence to the hypothesis that FGF plays a physiological role in the homeostatic mechanisms regulating mesoderm-derived cell growth.