NH2-Terminal Sequence of Human F. IX-Derived Fragments After HNE Digestion
| Cycle No. . | 44 kD-150 . | 34 kD . | 30 kD . | 20 kD . | 17 kD . |
|---|---|---|---|---|---|
| 1 | S (15.0)/R (7.0) | L (30.0)/Y (8.0) | V (42.0)/Q (12.0) | Y (59.0)/V (11.0) | V (4.0) |
| 2 | K (29.0)/A (9.0) | D (28.0)/N (10.0) | G (43.0)/S (16.0) | N (62.0)/G (9.0) | G (4.0) |
| 3 | L (17.0)/E (8.0) | N (4.0)/S (6.0) | G (35.0)/F (14.0) | S (37.0)/G (10.0) | G (5.0) |
| 4 | T (23.0)/A (30.0) | I (25.0)/G (8.0) | E (34.0)/N (19.0) | G (15.0)/E (8.0) | E (4.0) |
| 5 | R (23.0)/V (10.0) | T (14.0)/K (13.0) | D (49.0) | K (36.0)/D (10.0) | D (8.0) |
| 6 | A (12.0)/F (8.0) | Q (11.0)/L (8.0) | A (30.0)/F (14.0) | L (28.0)/A (8.0) | |
| 7 | E (11.0)/P (6.0) | S (12.0)/E (4.0) |
| Cycle No. . | 44 kD-150 . | 34 kD . | 30 kD . | 20 kD . | 17 kD . |
|---|---|---|---|---|---|
| 1 | S (15.0)/R (7.0) | L (30.0)/Y (8.0) | V (42.0)/Q (12.0) | Y (59.0)/V (11.0) | V (4.0) |
| 2 | K (29.0)/A (9.0) | D (28.0)/N (10.0) | G (43.0)/S (16.0) | N (62.0)/G (9.0) | G (4.0) |
| 3 | L (17.0)/E (8.0) | N (4.0)/S (6.0) | G (35.0)/F (14.0) | S (37.0)/G (10.0) | G (5.0) |
| 4 | T (23.0)/A (30.0) | I (25.0)/G (8.0) | E (34.0)/N (19.0) | G (15.0)/E (8.0) | E (4.0) |
| 5 | R (23.0)/V (10.0) | T (14.0)/K (13.0) | D (49.0) | K (36.0)/D (10.0) | D (8.0) |
| 6 | A (12.0)/F (8.0) | Q (11.0)/L (8.0) | A (30.0)/F (14.0) | L (28.0)/A (8.0) | |
| 7 | E (11.0)/P (6.0) | S (12.0)/E (4.0) |
NH2-terminal sequencing of human F.IX-derived fragments after HNE cleavage. Human F.IX (4 μmol/L) was incubated with HNE (400 nmol/L) for 2 and 30 minutes at 37°C in HBS, pH 7.4, with 5.0 mmol/L CaCl2 as described in the Materials and Methods. Approximately 100 μg (1,750 pmol) was used for each sample reaction. The apparent molecular mass of the specific fragments is shown along with the picomoles of the amino acid identified (in parentheses) after each number of cycles performed.
Fragments 44 kD, 34 kD, 30 kD, 20 kD, and 17 kD represent the HNE-derived cleavage products of F.IX.