Extracellular matrix heparan sulfate proteoglycans modulate the mitogenic capacity of acidic fibroblast growth factor.
Confluent cultures of human endothelial cells deposit into extracellular matrix (ECM) distinct heparan sulfate proteoglycans (HSPG) which modulate acidic fibroblast growth factor's (aFGF) ability to stimulate human endothelial cell mitogenic capacity. Extracellular matrix 35S-HSPG were isolated from cultures metabolically labelled with Na235SO4 by DEAE-Sepharose, Sepharose CL-4B, and aFGF-Affi-Gel 15 column chromatography and identified by resistance to chondroitinase ABC and sensitivity to nitrous acid. Fifty to sixty percent of the 35S-HSPG deposited into ECM do not bind aFGF. The bound 35S-HSGP (40-50% of the total counts applied) eluted from the aFGF-Affi-Gel column after the addition of buffer containing 2 M NaCl. aFGF-binding and aFGF-nonbinding 35S-HSPG were individually pooled and further purified by Sepharose CL-4B column chromatography. 35S-HSPG which bind aFGF, designated HSPGP, were 100-fold superior to heparin in augmenting the mitogenic efficacy of aFGF in sparse proliferating cultures. In contrast, however, 35S-HSPG, which did not bind aFGF, designated HSPG1, inhibited aFGF-stimulated proliferation in both sparse and subconfluent endothelial cell cultures. The majority of the biological activity of both aFGF-potentiating HSPGP and aFGF-inhibitory HSPG1 was contained in the glycosaminoglycan chains released by alkaline borohydride treatment of intact HSPGP or HSPG1, respectively. 3H-Core protein derived from HSPGP or HSPG1 contained only minor biological activity. The ability of heparitinase or heparinase (Flavobacterium heparinum) to abolish biological activity differed, depending upon the HSPG tested, also suggested that these are two distinct HSPGs.
Gordon
P B
PB
Department of Medicine, Albert Einstein College of Medicine, Bronx, New York 10467.
Choi
H U
HU
Conn
G
G
Ahmed
A
A
Ehrmann
B
B
Rosenberg
L
L
Hatcher
V B
VB
D002455:Cell Division / Q000187:drug effects; D011508:Chondroitin Sulfate Proteoglycans / Q000145:classification / Q000494:pharmacology / Q000502:physiology*; D002846:Chromatography, Affinity; D004357:Drug Synergism; D004730:Endothelium, Vascular / Q000166:cytology*; D005109:Extracellular Matrix / Q000502:physiology*; D005346:Fibroblast Growth Factors / Q000378:metabolism / Q000494:pharmacology*; D006025:Glycosaminoglycans / Q000502:physiology*; D006131:Growth Inhibitors; D019812:Heparan Sulfate Proteoglycans; D006497:Heparan Sulfate / Q000145:classification / Q000494:pharmacology / Q000502:physiology*; D006801:Humans; D011509:Proteoglycans / Q000502:physiology*; D013329:Structure-Activity Relationship; D014758:Viral Core Proteins / Q000032:analysis
D011508:Chondroitin Sulfate Proteoglycans; D005346:Fibroblast Growth Factors; D006025:Glycosaminoglycans; D006131:Growth Inhibitors; D019812:Heparan Sulfate Proteoglycans; D006497:Heparan Sulfate; D011509:Proteoglycans; D014758:Viral Core Proteins