Extracellular matrix heparan sulfate proteoglycans modulate the mitogenic capacity of acidic fibroblast growth factor

P. B. Gordon, H. U. Choi, G. Conn, A. Ahmed, B. Ehrmann, L. Rosenberg, Victor Bernard Hatcher

Research output: Contribution to journalArticle

40 Citations (Scopus)

Abstract

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 HSPG(P), 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 HSPG(I), inhibited aFGF-stimulated proliferation in both sparse and subconfluent endothelial cell cultures. The majority of the biological activity of both aFGF-potentiating HSPG(P) and aFGF-inhibitory HSPG1 was contained in the glycosaminoglycan chains released by alkaline borohydride treatment of intact HSPG(P) or HSPG(I), respectively. 3H-Core protein derived from HSPG(P) or HSPG(I) 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.

Original languageEnglish (US)
Pages (from-to)584-592
Number of pages9
JournalJournal of Cellular Physiology
Volume140
Issue number3
StatePublished - 1989

Fingerprint

Fibroblast Growth Factor 1
Heparan Sulfate Proteoglycans
Extracellular Matrix
Endothelial cells
Bioactivity
Endothelial Cells
Column chromatography
heparitinsulfate lyase
Cell culture
Chromatography
Syndecan-2
Heparin Lyase
Nitrous Acid
Chondroitin ABC Lyase
Flavobacterium
Borohydrides
Glycosaminoglycans
Sepharose

ASJC Scopus subject areas

  • Cell Biology
  • Clinical Biochemistry
  • Physiology

Cite this

Extracellular matrix heparan sulfate proteoglycans modulate the mitogenic capacity of acidic fibroblast growth factor. / Gordon, P. B.; Choi, H. U.; Conn, G.; Ahmed, A.; Ehrmann, B.; Rosenberg, L.; Hatcher, Victor Bernard.

In: Journal of Cellular Physiology, Vol. 140, No. 3, 1989, p. 584-592.

Research output: Contribution to journalArticle

Gordon, P. B. ; Choi, H. U. ; Conn, G. ; Ahmed, A. ; Ehrmann, B. ; Rosenberg, L. ; Hatcher, Victor Bernard. / Extracellular matrix heparan sulfate proteoglycans modulate the mitogenic capacity of acidic fibroblast growth factor. In: Journal of Cellular Physiology. 1989 ; Vol. 140, No. 3. pp. 584-592.
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abstract = "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 HSPG(P), 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 HSPG(I), inhibited aFGF-stimulated proliferation in both sparse and subconfluent endothelial cell cultures. The majority of the biological activity of both aFGF-potentiating HSPG(P) and aFGF-inhibitory HSPG1 was contained in the glycosaminoglycan chains released by alkaline borohydride treatment of intact HSPG(P) or HSPG(I), respectively. 3H-Core protein derived from HSPG(P) or HSPG(I) 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.",
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