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Thrombin-induced release of active basic fibroblast growth factor- heparan
sulfate complexes from subendothelial extracellular matrix
M Benezra, I Vlodavsky, R Ishai-Michaeli, G Neufeld and R Bar-Shavit
Department of Oncology, Hadassah-Hebrew University Hospital, Jerusalem,
Israel.
The angiogenic factor, basic fibroblast growth factor (bFGF), is
sequestered and protected by binding to heparan sulfate proteoglycans
(HSPG) in the subendothelial extracellular matrix (ECM). Release of ECM-
bound bFGF provides a novel mechanism for regulation of cell proliferation
and neovascularization in normal and pathologic situations. Exposure of ECM
to thrombin, the final activation product of the clotting cascade, resulted
in release of high molecular weight HSPG-bFGF complex, as indicated by its
immunoprecipitation with anti- bFGF antibodies, susceptibility to
degradation by bacterial heparinase, and inhibition of its mitogenic
activity in the presence of neutralizing anti-bFGF antibodies. The
ECM-resident bFGF-HSPG complex was not released by thrombin in the presence
of hirudin or antithrombin III, or by catalytically blocked thrombin
preparations. A threefold to fivefold higher mitogenic activity was
released by thrombin from ECM that was preheated (1 hour, 80 degrees C), as
compared with native ECM. This difference is attributed to heat stable
bFGF-HSPG complexes that are more readily released after heat treatment of
the ECM and to activation and release of ECM-resident transforming growth
factor-beta (TGF-beta) activity. Our results indicate that the large
reservoir of proteolytic activity present in plasma in the form of
prothrombin may participate in release from the subendothelial ECM of
biologically active bFGF and TGF-beta, depending on the accessibility of
thrombin. Thrombin may gain access to the subendothelium on clot formation
after tissue injury and as a result of the conversion of prothrombin to
thrombin induced by the ECM itself.
Volume 81,
Issue 12,
pp. 3324-3331,
06/15/1993
Copyright © 1993 by The American Society of Hematology
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