Table 2. Effects of membrane-impermeable and -permeable protease inhibitors on zymogen activation.
Inhibitor in Cyt | ZG | ZG/Cyt | ZG/Cyt/ATP | Mic | Mic/Cyt | Mic/Cyt/ATP |
---|---|---|---|---|---|---|
Trypsin activity | ||||||
−STI/PIC | 0 | 101±12 | 100 | 0 | 110±10 | 100 |
+STI | 0 | 114±18 | 111±4 | 0 | 107±4 | 91±2 |
+PIC | 0 | 44±14* | 57±10* | 0 | 54±3* | 58±8* |
Chymotrypsin activity | ||||||
−STI/PIC | 0 | 39±8 | 100 | 0 | 67±10 | 100 |
+STI | 0 | 42±5 | 119±10 | 0 | 69±13 | 97±4 |
+PIC | 0 | 13±5* | 41 ±12* | 0 | 22±13* | 44±6* |
Values are means of %activity ± SE. For ZG data, maximal activity (100%) is set as ZG/Cyt/ATP using −STI/PIC Cyt. For Mic data, maximal activity (100%) is set as Mic/Cyt/ATP using −STI/PIC Cyt. In the absence or presence of membrane-impermeable protease inhibitor in the Cyt, no major difference in activity is seen, indicating that free enzyme is not contributing to activity. When PIC (containing membrane-permeant inhibitors) is present in Cyt, there is a significant reduction in activity (*P < 0.05). STI, soybean trypsin inhibitor; PIC, protease inhibitor cocktail.