NH2-Terminal Sequence of Human Factor Va-Derived Fragments After HNE Digestion
| Cycle No. . | 76 kD* . | 56 kD . | 53 kD . | 48 kD . | 35 kD . | 28 kD . | 22 kD . |
|---|---|---|---|---|---|---|---|
| 1 | A(8.0)/S(31.0) | A(20.0) | A(6.0) | A(4.0)/S(13.0) | S(13.0) | N(10.0) | Q(17.0) |
| 2 | Q(4.0)/N(6.0) | Q(18.0) | Q(8.0) | Q(4.0)/S(9.0) | N(10.0) | M(5.0) | R(21.0) |
| 3 | L(4.0)/N(4.0) | L(20.0) | L(9.0) | L(2.0)/E(5.0) | N(6.0) | D(9.0) | A(16.0) |
| 4 | R(3.0)/G(4.0) | R(20.0) | R(7.0) | R(2.0)/M(3.0) | G(4.0) | K(3.0) | A(14.0) |
| 5 | Q(2.0)/N(2.0) | Q(20.0) | Q(6.0) | Q(4.0)/K(3.0) | N(4.0) | K(3.0) | D(15.0) |
| 6 | F(14.0) | F(5.0) | F(4.0)/K(3.0) | R(5.0) | Y(6.0) | I(9.0) | |
| 7 | Y(15.0) | Y(7.0) | Y(4.0)/S(4.0) | R(4.0) |
| Cycle No. . | 76 kD* . | 56 kD . | 53 kD . | 48 kD . | 35 kD . | 28 kD . | 22 kD . |
|---|---|---|---|---|---|---|---|
| 1 | A(8.0)/S(31.0) | A(20.0) | A(6.0) | A(4.0)/S(13.0) | S(13.0) | N(10.0) | Q(17.0) |
| 2 | Q(4.0)/N(6.0) | Q(18.0) | Q(8.0) | Q(4.0)/S(9.0) | N(10.0) | M(5.0) | R(21.0) |
| 3 | L(4.0)/N(4.0) | L(20.0) | L(9.0) | L(2.0)/E(5.0) | N(6.0) | D(9.0) | A(16.0) |
| 4 | R(3.0)/G(4.0) | R(20.0) | R(7.0) | R(2.0)/M(3.0) | G(4.0) | K(3.0) | A(14.0) |
| 5 | Q(2.0)/N(2.0) | Q(20.0) | Q(6.0) | Q(4.0)/K(3.0) | N(4.0) | K(3.0) | D(15.0) |
| 6 | F(14.0) | F(5.0) | F(4.0)/K(3.0) | R(5.0) | Y(6.0) | I(9.0) | |
| 7 | Y(15.0) | Y(7.0) | Y(4.0)/S(4.0) | R(4.0) |
Human F.Va (600 nmol/L) was incubated with 60 nmol/L HNE for 30 minutes in HBS, pH 7.4 with 5 mmol/L CaCl2 as described in Materials and Methods. Approximately 900 pmol was used for the F.Va reaction. The apparent molecular mass of the specific fragments is shown along with the pmol of the amino acid identified (in parentheses) after each number of cycles performed.
Fragments 76 kD; 56 kD; 53 kD; 48 kD; 35 kD; 28 kD; 22 kD represent the HNE-derived cleavage products of F.Va.