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Extracellular matrix heparan sulfate proteoglycans modulate the mitogenic capacity of acidic fibroblast growth factor
Author(s) -
Gordon Portia B.,
Choi Haing U.,
Conn Greg,
Ahmed Asra,
Ehrmann Bettina,
Rosenberg Lawrence,
Hatcher Victor B.
Publication year - 1989
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.1041400325
Subject(s) - extracellular matrix , fibroblast growth factor , heparan sulfate , microbiology and biotechnology , chemistry , perlecan , fibroblast , extracellular , fibroblast growth factor receptor 3 , fibroblast growth factor receptor 2 , biochemistry , proteoglycan , glycosaminoglycan , biology , in vitro , receptor
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 35 S‐HSPG were isolated from cultures metabolically labelled with Na 2 35 SO 4 by DEAE‐Sepharose, Sepharose CL‐4B, and aFGF‐Affi‐Gel 15 column chromatography and identified by resistance to chon‐droitinase ABC and sensitivity to nitrous acid. Fifty to sixty percent of the 35 S‐HSPG deposited into ECM do not bind aFGF. The bound 35 S‐HSGP (40‐50% of the total counts applied) eluted from the aFGF‐Affi‐Gel column after the addition of buffer containing 2 M NaCI. aFGF‐binding and aFGF‐nonbinding 35 S‐HSPG were individually pooled and further purified by Sepharose CL‐4B column chromatography. 35 S‐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, 35 S‐HSPG, which did not bind aFGF, designated HSPG 1 , 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 HSPG 1 was contained in the glycosaminoglycan chains released by alkaline borohydride treatment of intact HSPG P or HSPG 1 , respectively. 3 H‐Core protein derived from HSPG P or HSPG 1 contained only minor biological activity. The ability of heparitinase or hepnrinase (Flavobacterium heparinum) to abolish biological activity differed, depending upon the HSPG tested, also suggested that these are two distinct HSPGs.