Specific binding of basic fibroblast growth factor to basement membrane-like structures and to purified heparan sulfate proteoglycan of the EHS tumor.
The binding of iodinated basic fibroblast growth factor (bFGF) to low-density heparan sulfate proteoglycan purified from the Engelbreth Holm Swarm (EHS) sarcoma was investigated using different techniques. The tumor clearly contained bFGF, the level being comparable to that found in other tissues such as human or bovine brain. 125I bFGF strongly bound to the basement membrane-like matrix of EHS frozen sections as revealed by autoradiography. Iodinated bFGF bound to purified heparan sulfate proteoglycan but not to laminin or collagen type IV, three components isolated from the same tumor. In contrast, acidic fibroblast growth factor (aFGF) displayed negligible binding to heparan sulfate proteoglycan. Binding of bFGF to frozen sections and to purified proteoglycan could be strongly inhibited by heparin and was displaced by an excess of unlabeled factor and completely suppressed after heparitinase and heparinase treatments. Binding was a function of the salt concentration and was abolished at 0.6 M NaCl. Scatchard analysis indicated the affinity site had a Kd of about 30 nM, a value 10-15 higher than that recently reported by Moscatelli (J. Cell. Physiol., 131:123-130, 1987) in the case of the low-affinity binding sites present on the surface of baby hamster kidney (BHK) cells.
Vigny
M
M
INSERM U. 118, CNRS U.A. 630, Paris, France.
Ollier-Hartmann
M P
MP
Lavigne
M
M
Fayein
N
N
Jeanny
J C
JC
Laurent
M
M
Courtois
Y
Y
D000818:Animals; D001345:Autoradiography; D001485:Basement Membrane / Q000378:metabolism; D002417:Cattle; D011508:Chondroitin Sulfate Proteoglycans / Q000378:metabolism*; D005346:Fibroblast Growth Factors / Q000378:metabolism*; D006025:Glycosaminoglycans / Q000378:metabolism*; D019812:Heparan Sulfate Proteoglycans; D006497:Heparan Sulfate / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D011509:Proteoglycans / Q000378:metabolism*; D012513:Sarcoma, Experimental / Q000378:metabolism*
D011508:Chondroitin Sulfate Proteoglycans; D005346:Fibroblast Growth Factors; D006025:Glycosaminoglycans; D019812:Heparan Sulfate Proteoglycans; D006497:Heparan Sulfate; D011509:Proteoglycans