Table 2.
Dissociation constants (KD) of the PDI-MPD interaction with 68% confidence interval (CI) obtained by fitting the data from at least 3 independent MST analyses each with sixteen dilutions (1:2 dilution series) of the respective PDI at 25 °C. KD’s were calculated from the concentration-dependent ligand induced fluorescence intensity changes according to Eq. 1. Additional parameters resulting from the fit: Response amplitude of the fluorescence intensity shift between bound and unbound state, standard error (SE) of regression, reduced χ² and signal to noise.
| Analyte | Ligand | KD ± 68% CI [nM] | Response Amplitude [counts] | n | SE of Regression | Reduced χ² | Signal to Noise |
|---|---|---|---|---|---|---|---|
| PDIA1 | opMPD | 158 ± 34 | 298.9 | 3 | 19.75 | 0.45 | 16.25 |
| clMPD | 211 ± 67 | 210.4 | 5* | 20.17 | 2.05 | 11.21 | |
| PDIA3 | opMPD | 90 ± 27 | 331.5 | 3 | 30.26 | 2.24 | 11.77 |
| clMPD | 55 ± 22 | 208.2 | 3 | 25.08 | 1.30 | 8.92 | |
| PDIA6 | opMPD | 93 ± 23 | 348.6 | 3 | 26.50 | 0.86 | 14.13 |
| clMPD | 61 ± 14 | 276.9 | 3 | 19.41 | 1.30 | 15.32 | |
| PDIA6 C58S | opMPD | 141 ± 38 | 411.6 | 3 | 34.58 | 3.73 | 12.79 |
| clMPD | 113 ± 31 | 198.2 | 3 | 16.87 | 0.69 | 12.62 | |
| PDIA6 C193S | opMPD | 142 ± 30 | 400.5 | 3 | 26.11 | 4.74 | 16.48 |
| clMPD | 91 ± 55 | 155.4 | 3 | 28.86 | 2.47 | 5.78 | |
| PDIA6 C58S C193S | opMPD | 192 ± 40 | 406.4 | 3 | 25.50 | 3.87 | 17.12 |
| clMPD | 151 ± 40 | 148.2 | 3 | 11.96 | 0.26 | 13.31 | |
| PDIA6 C55S C58S | opMPD | 138 ± 27 | 447.3 | 3 | 26.57 | 22.13 | 18.09 |
| clMPD | 191 ± 88 | 171.5 | 3 | 24.15 | 1.15 | 7.63 | |
| PDIA6 C55S C190S | opMPD | 115 ± 20 | 314.3 | 3 | 16.59 | 8.73 | 20.35 |
| clMPD | 116 ± 42 | 178.2 | 3 | 19.79 | 0.74 | 9.67 |
*Replicates increased to 5 due to variations at low concentration.