TABLE 2.
Interaction of Pol IV with the Pol IIIα catalytic and Pol IIIεθ proofreading subunitsa
| Interactionb | Expt | KD (nM) | Avg KD, nM (range) |
|
|---|---|---|---|---|
| KD 1 | KD 2 | |||
| Pol IV-Pol IIIαHis | 1 | KD1 = 11.8 | 13.9 (4.10) | 52.6 (4.80) |
| KD2 = 50.2 | ||||
| 2 | KD1 = 15.9 | |||
| KD2 = 55.0 | ||||
| Pol IV-T120P–Pol IIIαHis | 1 | KD1 = 5.97 | 4.88 (2.18) | 62.1 (29.9) |
| KD2 = 77.0 | ||||
| 2 | KD1 = 3.79 | |||
| KD2 = 47.1 | ||||
| Pol IVCD-Pol IIIαHis | 1 | KD1 = 68.7 | 59.0 (19.5) | 819 (362) |
| KD2 = 1,000 | ||||
| 2 | KD1 = 49.2 | |||
| KD2 = 638 | ||||
| Pol IVCD-T120P–Pol IIIαHis | 1 | KD1 = 215 | 216 (2) | 3,790 (40) |
| KD2 = 3,810 | ||||
| 2 | KD1 = 217 | |||
| KD2 = 3,770 | ||||
| Pol IVLF-Pol IIIαHis | 1 | KD1 = 4.59 | 5.02 (0.85) | 15.7 (8.20) |
| KD2 = 19.8 | ||||
| 2 | KD1 = 5.44 | |||
| KD2 = 11.6 | ||||
a
See Table S2 in the supplemental material for the ka (on rate), kd (off rate), χ2 (likelihood of no relationship), and R2 (goodness of fit) values for these interactions.
b
The indicated His-tagged protein (ligand) was captured on a HIS1K biosensor, and interactions were measured using BLI. Interactions were identified between Pol IIIεθHis and Pol IV, Pol IVCD, and Pol IVLF, but we were unable to obtain reliable numbers describing the kinetics of their interactions.