TABLE 2.
Kinetic parameters for binding of bA2 variants and FIXa
Kinetic parameters were determined by SPR as described under “Experimental Procedures” using A2 in the mobile phase and bFPR-FIXa bound to a SA chip. Samples were run at least three times, and mean ± S.D. shown.
| A2 analyte | ka1 | kd1 | ka2 | kd2 | Kd | χ2 |
|---|---|---|---|---|---|---|
| m−1 s−1 | s−1 | s−1 | s−1 | nm | ||
| WT | (3.4 ± 0.3) × 105 | (1.5 ± 0.1) × 10−2 | (1.3 ± 0.1) × 10−2 | (6.5 ± 2.4) × 10−4 | 2.1 ± 0.02 | 4.2 |
| K707A | (2.6 ± 0.2) × 105 | (3.5 ± 0.05) × 10−2 | (2.5 ± 0.5) × 10−2 | (6.6 ± 0.4) × 10−5 | 0.41 ± 0.04 | 0.71 |
| S709A | (2.9 ± 0.3) × 105 | (0.8 ± 0.2) × 10−2 | (6.9 ± 0.2) × 10−3 | (5.7 ± 1.3) × 10−4 | 2.1 ± 0.1 | 1.5 |
| S710A | (1.6 ± 0.4) × 105 | (4.3 ± 0.2) × 10−2 | (1.9 ± 0.02) × 10−3 | (0.54 ± 0.04) × 10−3 | 8.0 ± 2.5 | 0.20 |
| D712A | (3.2 ± 0.5) × 105 | (1.8 ± 0.4) × 10−2 | (1.3 ± 0.1) × 10−2 | (8.3 ± 0.5) × 10−4 | 3.4 ± 0.1 | 0.20 |
| K713A | (2.7 ± 1.0) × 105 | (1.8 ± 0.3) × 10−2 | (1.2 ± 0.05) × 10−2 | (1.5 ± 0.1) × 10−3 | 7.3 ± 0.1 | 0.15 |
| N714A | (1.1 ± 0.2) × 105 | (3.3 ± 0.1) × 10−2 | (1.9 ± 0.1) × 10−2 | (2.9 ± 0.7) × 10−3 | 4.7 ± 0.2 | 1.0 |