Table 1. Table of Measured Biophysical Constants.
| Construct | WT RIG-I | S411A | S411L |
|---|---|---|---|
| Steady state ATPase activity | |||
| 5΄ppp10L: | |||
| k cat (ATP/RIG-I•s) | 18.2 ± 0.3 | 5.1 ± 0.4 | Inactive |
| K M,RNA (nM) | 10.0 ± 1.0 | 5.0 ± 1 | Inactive |
| Maximum IFN-β Production | |||
| 5΄ppp10L (Firefly Luc/Renilla Luc) | 20.1 ± 1.7 | 56.6 ± 10.8 | 7.8 ± 1.5 |
| 5΄OH10L (Firefly Luc/Renilla Luc) | 3.8 ± 0.3 | 19.5 ± 3.8 | 0.2 ± 0..2 |
| PIC (Firefly Luc/Renilla Luc) | 59.2 ± 15 | 209.8 ± 17 | 1.8 ± 0.4 |
| Mock (Firefly Luc/Renilla Luc) | 0.99 ± 0.4 | 3.5 ± 0.5 | 0.2 ± 0.01 |
| MANT-ATP binding | |||
| 5΄ppp10L (μM) | 72 ± 3 | 82 ± 6 | 110 ± 8 |
| OH10L (μM) | 136 ± 6 | 75 ± 5 | – |
| Equilibrium RNA binding | |||
| OH10L (nM) | 17 ± 1.0 | 23 ± 1.8 | 41 ± 1.8 |
Biophysical constants for binding, catalysis and cell-based signaling assays performed for wild type, S411A and S411L constructs of RIG-I. Values represent mean ± SD (n = 3). See Materials and Methods for details of the experimental setups and data analysis used in each assay.