Table 1.
PDA analysis of oxidized (top) and reduced (bottom) PDI 88/467 WT and variants used in this study
| Rates (ms−1) | WT | AA/AA | CC/AA | AA/CC | R300H | W396A |
|---|---|---|---|---|---|---|
| k1,2 | 4.50 ± 0.81 | 2.14 ± 0.14 | 4.43 ± 0.05 | 2.73 ± 0.36 | 1.97 ± 0.41 | 1.36 ± 0.14 |
| k1,3 | 0.32 ± 0.31 | 0.63 ± 0.23 | 0.34 ± 0.13 | 0.26 ± 0.28 | 0.51 ± 0.47 | 1.38 ± 0.09 |
| k2,1 | 2.38 ± 0.36 | 1.59 ± 0.13 | 2.39 ± 0.09 | 1.51 ± 0.11 | 1.35 ± 0.22 | 1.51 ± 0.39 |
| k2,3 | 0.78 ± 0.26 | 1.03 ± 0.09 | 0.67 ± 0.06 | 0.68 ± 0.12 | 0.70 ± 0.55 | 0.80 ± 0.11 |
| k3,1 | 0.15 ± 0.11 | 0.66 ± 0.15 | 0.12 ± 0.02 | 0.48 ± 0.24 | 0.09 ± 0.11 | 0.13 ± 0.04 |
| k3,2 | 1.07 ± 0.24 | 0.95 ± 0.11 | 0.94 ± 0.06 | 0.66 ± 0.24 | 1.20 ± 0.16 | 0.93 ± 0.16 |
| % state | ||||||
| O1(80 Å) | 23 ± 1 | 29 ± 1 | 22 ± 1 | 26 ± 1 | 22 ± 2 | 20 ± 1 |
| O2(57 Å) | 44 ± 3 | 36 ± 2 | 43 ± 1 | 42 ± 2 | 39 ± 4 | 32 ± 2 |
| C (42 Å) | 34 ± 1 | 35 ± 2 | 35 ± 1 | 32 ± 1 | 39 ± 2 | 49 ± 1 |
| χ2(global) | 5.5 | 6.2 | 5.2 | 4.9 | 6.7 | 4.7 |
| Rates (ms−1) | WT | AA/AA | CC/AA | AA/CC | R300H | W396A |
|---|---|---|---|---|---|---|
| k1,2 | 8.49 ± 1.95 | 2.12 ± 0.15 | 9.35 ± 0.30 | 4.33 ± 1.29 | 6.64 ± 0.21 | 4.49 ± 0.62 |
| k1,3 | 0.36 ± 0.33 | 0.39 ± 0.34 | 0.99 ± 0.66 | 1.03 ± 1.04 | 0.94 ± 1.29 | 0.78 ± 0.09 |
| k2,1 | 6.28 ± 1.07 | 1.45 ± 0.39 | 7.50 ± 1.95 | 3.52 ± 0.32 | 4.09 ± 0.18 | 4.72 ± 0.40 |
| k2,3 | 0.11 ± 0.06 | 0.75 ± 0.59 | 0.75 ± 1.19 | 0.27 ± 0.09 | 0.17 ± 0.13 | 0.16 ± 0.05 |
| k3,1 | 1.39 ± 0.84 | 0.48 ± 0.17 | 3.95 ± 4.47 | 0.79 ± 0.19 | 0.36 ± 0.55 | 0.25 ± 0.22 |
| k3,2 | 0.40 ± 0.21 | 0.97 ± 0.23 | 2.05 ± 2.72 | 0.50 ± 0.50 | 0.68 ± 0.60 | 1.30 ± 0.11 |
| % state | ||||||
| O1(80 Å) | 39 ± 1 | 28 ± 1 | 41 ± 1 | 37 ± 3 | 33 ± 1 | 37 ± 1 |
| O2(57 Å) | 51 ± 3 | 37 ± 2 | 49 ± 3 | 45 ± 6 | 53 ± 1 | 40 ± 2 |
| C (42 Å) | 9 ± 2 | 35 ± 1 | 10 ± 2 | 18 ± 3 | 14 ± 1 | 23 ± 1 |
| χ2(global) | 4.5 | 5.1 | 4.2 | 5.2 | 6.1 | 5.1 |
PDA was performed on datasets binned at 0.25, 0.5, 0.75, and 1 ms. To assess robustness of the fit, the PDA fit was repeated by systematically varying the initial value of the rate constants to 1, 0.5, and 0.75 ms−1 (min 0, max 10) while keeping the other settings identical. The results in the tables above represent the average of the three independent determinations.