Table 2.
Kinetic Constants for Hydrolysis of FXIII (28–41) F34, W34, Y34, V34, and L34 Activation Peptides with Different Thrombin (IIa) Mutantsa. L34 is a Common Polymorphism
| FXIII (28–41) F34 AP | Km (µM) | kcat (s−1) | kcat/Km (µM−1 s−1) |
| WT IIab | 442 ± 93 | 6.27 ± 0.54 | 1.4×10−2 ± 3.0×10−3 |
| W60dA IIab | 435 ± 98 | 1.46 ± 0.15 | 3.4×10−3 ± 8.0×10−4 |
| Y60aA IIab | NA | NA | NA |
| L99A IIab | 570 ± 125 | 2.71 ± 0.3 | 4.8×10−3 ± 1.2×10−4 |
| I174A IIa | 920 ± 88 | 6.08 ± 0.25 | 6.6×10−3 ± 6.9×10−4 |
| W215A IIb | 437 ± 81 | 1.97 ± 0.14 | 4.5×10−3 ± 9.0×10−4 |
| WE (W215A/E217A) IIab | 190 ± 43 | 0.07 ± 0.007 | 3.6×10−4 ± 8.0×10−5 |
| FXIII (28–41) W34 AP | Km (µM) | kcat (s−1) | kcat/Km (µM−1 s−1) |
| WT IIa | 637 ± 56 | 7.62 ± 0.3 | 1.2×10−2 ± 1.2×10−3 |
| W60dA IIa | 976 ± 89 | 1.21 ± 0.05 | 1.2×10−3 ± 1.2×10−4 |
| Y60aA IIa | NA | NA | NA |
| L99A IIa | 719 ± 97 | 2.3 ± 0.17 | 3.2×10−3 ± 4.9×10−4 |
| I174A IIa | 256 ± 24 | 13.9 ± 0.52 | 5.4×10−2 ± 5.5×10−3 |
| W215A IIa | 1030 ± 57 | 5.76 ± 0.14 | 5.6×10−3 ± 3.4×10−4 |
| WE (W215A/E217A) IIa | 1281 ± 161 | 0.35 ± 0.03 | 2.7×10−4 ± 3.5×10−5 |
| FXIII (28–41) Y34 AP | Km (µM) | kcat (s−1) | kcat/Km (µM−1 s−1) |
| WT IIa | 484 ± 88 | 2.23 ± 0.18 | 4.6×10−3 ± 9.0×10−4 |
| W60dA IIa | NA | NA | NA |
| Y60aA IIa | NA | NA | NA |
| L99A IIa | 347 ± 80 | 13.5 ± 1.4 | 3.9×10−2 ± 9.8×10−3 |
| I174A IIa | 349 ± 120 | 1.17 ± 0.16 | 3.3×10−3 ± 2.0×10−3 |
| W215A IIa | NA | NA | NA |
| WE (W215A/E217A) IIa | NA | NA | NA |
| FXIII (28–41) V34 AP | Km (µM) | kcat (s−1) | kcat/Km (µM−1 s−1) |
| WT IIac | 298 ± 42 | 2.57 ± 0.005 | 8.6×10−3 ± 1.0×10−3 |
| W60dA IIab | 1375 ± 422 | 0.86 ± 0.046 | 6.3×10−4 ± 1.9×10−4 |
| Y60aA IIab | 678 ± 120 | 0.048 ± 0.0032 | 7.1×10−5 ± 1.3×10−5 |
| L99A IIac | 2074 ± 414 | 0.89 ± 0.07 | 4.3×10−4 ± 1.0×10−4 |
| I174A IIac | 204 ± 36 | 5.37 ± 0.01 | 2.6×10−2 ± 1.0×10−4 |
| W215A IIac | 2374 ± 372 | 5.53 ± 0.017 | 2.3×10−3 ± 7.0×10−4 |
| WE (W215A/E217A) IIac | 745 ± 98 | 0.117 ± 0.008 | 1.6×10−4 ± 2.0×10−5 |
| FXIII (28–41) L34 AP [common polymorphism] | Km (µM) | kcat (s−1) | kcat/Km (µM−1 s−1) |
| WT IIac | 315 ± 42 | 22.9 ± 0.003 | 7.3×10−2 ± 1.2×10−3 |
| W60dA IIab | 1000 ± 201 | 5.07 ± 0.021 | 5.1×10−3 ± 1.0×10−3 |
| Y60aA IIab | 696 ± 116 | 0.37 ± 0.012 | 5.3×10−4 ± 1.2×10−4 |
| L99A IIac | 1930 ± 354 | 5.94 ± 0.02 | 3.1×10−3 ± 2.0×10−4 |
| I174A IIc | 1109 ± 247 | 10.2 ± 0.04 | 9.1×10−3 ± 2.0×10−3 |
| W215A IIac | 3619 ± 470 | 5.31 ± 0.002 | 1.5×10−3 ± 2.0×10−4 |
| WE (W215A/E217A) IIac | 142 ± 28 | 0.052 ± 0.003 | 3.7×10−4 ± 1.0×10−4 |
a
The results shown here represent averages of at least three independent experiments. Kinetic values were calculated using nonlinear regression analysis methods using SigmaPlot or Kaleidagraph. The error values correspond to standard error of the mean (SEM).
b
Jadhav, M. et al. [40]
c
Isetti, G. et al [36]