Rapid Purification of a New P-I Class Metalloproteinase from Bothrops moojeni Venom with Antiplatelet Activity
Figure 3
Proteolysis of bovine fibrinogen by BmooMPα-II. Line 1: negative control-fibrinogen incubated with enzyme for 0 minutes. (a) Proteolysis of bovine fibrinogen by BmooMPα-II time-dependent. Lanes 2–5: fibrinogen incubated with enzyme for 15, 30, 60 and 120 minutes, respectively. (b) Proteolysis of bovine fibrinogen by BmooMPα-II and effect of inhibitors. Lanes 2: positive control-fibrinogen incubated with enzyme for 60 minutes, 3–8: fibrinogen incubated with enzyme for 60 minutes after preincubation of BmooMPα-II with 5 mmol/L EDTA, 5 mmol/L β-mercaptoethanol, 5 mmol/L 1,10-phenanthroline, 5 mmol/L benzamidine, 5 mmol/L leupeptin, and 5 mmol/L aprotinin for 15 minutes, respectively. (c) Effect of the pH on the stability of BmooMPα-II. Lanes 2–8: fibrinogen incubated with enzyme for 60 minutes after preincubation of BmooMPα-II in pH 4.0, 5.0, 6.0, 7.0, 8.0, 9.0, and 10.0, respectively. (d) Effect of temperature on the stability of the BmooMPα-II. Lanes 2–8: fibrinogen incubated with enzyme for 60 minutes after preincubation of BmooMPα-II for 15 minutes at 30, 40, 50, 60, 70, 80, and 90°C, respectively.
