Fibroblast growth factor-2 (FGF-2) binds to fibrin-(ogen) with high affinity, and fibrinogen potentiates FGF-2-stimulated proliferation of endothelial cells. Because plasmin degrades fibrin(ogen) physiologically and could liberate growth factor from fibrin deposits or alter its activity, we have now investigated the effect of plasmic degradation on the activity of fibrin(ogen)-bound FGF-2. Fibrinogen with bound FGF-2 was incubated with plasmin, the products characterized by SDS-PAGE, and the proliferative activity determined by 3Hthymidine incorporation into endothelial cells. Before plasmin exposure, proliferation was increased 3.7±0.6-fold with fibrinogen-bound FGF-2 compared with medium alone (P<0.005). Plasmic degradation resulted in progressive decrease in the proliferative capacity, with the 60-min digest showing predominantly fragment D1 and E and 3H-thymidine uptake of only 1.2±0.2-fold, significantly less than the activity of an equal concentration of free FGF-2 (P<0.02). However, further degradation increased activity, and proliferation with a 90-min digest increased to 2.6±0.5-fold, significantly greater than the 60-min digest (P<0.02). Plasmic degradation in the presence of 10mM calcium chloride prevented degradation of D1 to D2 and D3, and the activity did not increase with extended degradation. Immunoprecipitation of the digests with antifibrinogen antibody showed 70±8% of fibrinogen-bound FGF-2 in the presence of calcium but only 15±4% in its absence, indicating that cleavage of D1 to D2 and D3 is critical in binding. Fragment D1 and D2, but not D3, bound to a column containing immobilized FGF-2, indicating that a binding site is lost upon degradation to D3. The results demonstrate that plasmic degradation of fibrinogen modulates the activity and binding of FGF-2 that involves a site near the carboxyl terminus of the γ chain.
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