Interaction energies between residue pairs evaluated using GIA
| Site pairs | ΔΔGGIA | δΔΔGGIA |
| kcal | kcal | |
| R394-E395 | 1.05 | 0.58 |
| R394-Y485 | 3.14 | 0.58 |
| E395-Y485 | 5.27 | 0.58 |
| E395-V476 | 0.64 | 0.65 |
| V476-Y485 | 0.84 | 0.69 |
| R394-E395 (Y485A) | −3.03 | 0.76 |
| R394-Y485 (E395A) | −0.94 | 0.76 |
| E395-Y485 (R394A) | 1.15 | 0.76 |
| E395-Y485 (V476A) | 3.86 | 0.72 |
| Site pairs | ΔΔGGIA | δΔΔGGIA |
| kcal | kcal | |
| R394-E395 | 1.05 | 0.58 |
| R394-Y485 | 3.14 | 0.58 |
| E395-Y485 | 5.27 | 0.58 |
| E395-V476 | 0.64 | 0.65 |
| V476-Y485 | 0.84 | 0.69 |
| R394-E395 (Y485A) | −3.03 | 0.76 |
| R394-Y485 (E395A) | −0.94 | 0.76 |
| E395-Y485 (R394A) | 1.15 | 0.76 |
| E395-Y485 (V476A) | 3.86 | 0.72 |
ΔΔGGIA and its uncertainty (δΔΔGGIA) for each pair of sites were calculated as described in Materials and methods. The first five rows correspond to mutant cycles in which the WT channel was the reference channel. For the last four rows, the control/reference channel is a mutant indicated in parenthesis and reflects the interaction energy between two residues in the presence of a third mutation.