Equilibrium binding affinities for VcGluP wildtype and binding site mutants
| Variant . | Glu Kd (µM) . | FC . | n . | Gln Kd (mM) . | FC . | n . | Pyroglu Kd (mM) . | FC . | n . |
|---|---|---|---|---|---|---|---|---|---|
| WT | 0.065 ± 0.031 | - | 5 | 0.011 ± 0.002 | - | 5 | 0.050 ± 0.016 | - | 10 |
| Y45A | 13.88 ± 1.576 | 213 | 5 | 5.135 ± 0.864 | 466 | 6 | 2.000 ± 0.094 | 40 | 5 |
| S75A | 0.915 ± 0.126 | 14 | 5 | 0.073 ± 0.028 | 6.5 | 7 | 0.499 ± 0.050 | 10 | 5 |
| Q93A | 64.39 ± 13.74 | 990 | 10 | ND | ND | 5 | 18.62 ± 4.566 | 372 | 5 |
| E125A | 0.083 ± 0.023 | 1.3 | 5 | 0.025 ± 0.008 | 2.3 | 9 | 0.147 ± 0.010 | 2.9 | 6 |
| Y200A | 0.228 ± 0.088 | 3.5 | 6 | 0.022 ± 0.008 | 2 | 8 | 0.104 ± 0.022 | 2.1 | 3 |
| M201A | 0.139 ± 0.032 | 2.1 | 4 | 0.040 ± 0.009 | 3.6 | 9 | 0.302 ± 0.010 | 6 | 4 |
| Variant . | Glu Kd (µM) . | FC . | n . | Gln Kd (mM) . | FC . | n . | Pyroglu Kd (mM) . | FC . | n . |
|---|---|---|---|---|---|---|---|---|---|
| WT | 0.065 ± 0.031 | - | 5 | 0.011 ± 0.002 | - | 5 | 0.050 ± 0.016 | - | 10 |
| Y45A | 13.88 ± 1.576 | 213 | 5 | 5.135 ± 0.864 | 466 | 6 | 2.000 ± 0.094 | 40 | 5 |
| S75A | 0.915 ± 0.126 | 14 | 5 | 0.073 ± 0.028 | 6.5 | 7 | 0.499 ± 0.050 | 10 | 5 |
| Q93A | 64.39 ± 13.74 | 990 | 10 | ND | ND | 5 | 18.62 ± 4.566 | 372 | 5 |
| E125A | 0.083 ± 0.023 | 1.3 | 5 | 0.025 ± 0.008 | 2.3 | 9 | 0.147 ± 0.010 | 2.9 | 6 |
| Y200A | 0.228 ± 0.088 | 3.5 | 6 | 0.022 ± 0.008 | 2 | 8 | 0.104 ± 0.022 | 2.1 | 3 |
| M201A | 0.139 ± 0.032 | 2.1 | 4 | 0.040 ± 0.009 | 3.6 | 9 | 0.302 ± 0.010 | 6 | 4 |
FC is the fold change in Kd compared to wildtype, n indicates the number of technical replicates performed, and error indicated is the standard deviation. Values are color coded to indicate the extent of Kd change for a mutant compared to wildtype (red being most deleterious, blue being a mild effect, and orange being middling).