Degradation of fibrinogen by Tuf-bound plasmin.

Tuf proteins, BBA70 and gelatin (5 μg/ml) were immobilized on microtiter plates, blocked and incubated with plasminogen (10 μg/ml). Following several wash steps, a reaction mixture containing the plasminogen activator uPA (0.16 μg/ml) and fibrinogen (20 μg/ml) was added and plates were incubated at 37°C. Samples were taken at the indicated time intervals and separated via SDS-PAGE. Upon transfer to nitrocellulose membranes, fibrinogen or its degradation products were detected in a Western blot analysis using a polyclonal fibrinogen antiserum. Controls included the lysine analog tranexamic acid (+T) and omission of plasminogen (-Plg). Fg, fibrinogen. Shown are representative results from several independent experiments.