NH2-terminal sequence of human factor IX-derived fragments following plasmin digestion
| Cycle No. . | Plasmin Cleavage Products of Human F.IX . | |||||
|---|---|---|---|---|---|---|
| 45 kd . | 30 kd . | 20 kd . | 18 kd . | 16 kd . | 14 kd . | |
| 1 | A (20.0) | A (45.0)/Y (26.0)/V (20.0) | Y (142.0) | Q (29.0) | V (36.0)/G (4.0) | G (67.0)/S (10.0) |
| 2 | E (20.0) | E (36.0)/N (20.0)/V (10.0) | N (116.0) | Y (35.0) | V (35.0)/R (5.0) | R (90.0)/A (4.0) |
| 3 | T (10.0)/A (19.0) | T (30.0)/A (30.0)/S (15.0)/G (9.0) | S (66.0) | V (24.0) | G (36.0)/S (5.0) | S (48.0)/L (6.0) |
| 4 | V (14.0) | V (27.0)/G (7.0) | G (32.0) | D (27.0) | G (33.0)/A (6.0) | A (70.0)/V (4.0) |
| 5 | F (14.0) | F (33.0)/K (8.0)/E (5.0) | K (60.0) | G (17.0) | E (21.0)/L (3.0) | L (70.0) |
| 6 | P (10.0) | P (25.0)/L (12.0)/D (9.0) | L (56.0) | D (16.0) | D (27.0)/V (8.0) | V (50.0)/Q (8.0) |
| 7 | D (35.0)/E (7.0)/A (8.0) | Q (8.0) | A (30.0)/L (5.0) | |||
| 8 | K (40.0)/Q (6.0) | |||||
| Cycle No. . | Plasmin Cleavage Products of Human F.IX . | |||||
|---|---|---|---|---|---|---|
| 45 kd . | 30 kd . | 20 kd . | 18 kd . | 16 kd . | 14 kd . | |
| 1 | A (20.0) | A (45.0)/Y (26.0)/V (20.0) | Y (142.0) | Q (29.0) | V (36.0)/G (4.0) | G (67.0)/S (10.0) |
| 2 | E (20.0) | E (36.0)/N (20.0)/V (10.0) | N (116.0) | Y (35.0) | V (35.0)/R (5.0) | R (90.0)/A (4.0) |
| 3 | T (10.0)/A (19.0) | T (30.0)/A (30.0)/S (15.0)/G (9.0) | S (66.0) | V (24.0) | G (36.0)/S (5.0) | S (48.0)/L (6.0) |
| 4 | V (14.0) | V (27.0)/G (7.0) | G (32.0) | D (27.0) | G (33.0)/A (6.0) | A (70.0)/V (4.0) |
| 5 | F (14.0) | F (33.0)/K (8.0)/E (5.0) | K (60.0) | G (17.0) | E (21.0)/L (3.0) | L (70.0) |
| 6 | P (10.0) | P (25.0)/L (12.0)/D (9.0) | L (56.0) | D (16.0) | D (27.0)/V (8.0) | V (50.0)/Q (8.0) |
| 7 | D (35.0)/E (7.0)/A (8.0) | Q (8.0) | A (30.0)/L (5.0) | |||
| 8 | K (40.0)/Q (6.0) | |||||
Human F.IX (4 μM) was treated with plasmin (50 nM) for 2 and 30 minutes at 37°C in HBS/Ca, as described in “Materials and Methods.” Approximately 100 μg (1750 pmol) of F.IX was used for each reaction. The apparent molecular mass of the protein fragments is indicated with the amount of each amino acid (in pmol) after the number of cycles performed. Fragments of 45, 30, 20, 18, 16, and 14 kd represent the plasmin cleavage products of human F.IX.