Table 2.
Thermodynamic parameters from ITC experiments between Cas8e and AcrIE4-F7 mutantsa
| AcrIE4-F7 | Cas8e | K D (nM) | N | ΔG (kcal/mol) | ΔH (kcal/mol) | −TΔS (kcal/mol) |
|---|---|---|---|---|---|---|
| WT | WT | 200 ± 28 | 1.0 ± 0.0 | −9.2 ± 0.1 | −14.7 ± 1.4 | 5.5 ± 1.5 |
| E12K/D13K | WT | 400 ± 120 | 0.8 ± 0.0 | −8.8 ± 0.2 | −22.8 ± 1.8 | 14.0 ± 1.9 |
| E19K/D22K | WT | N.B.b | - | - | - | - |
| E19K | WT | N.B. | - | - | - | - |
| D22K | WT | N.B. | - | - | - | - |
| Y20A | WT | 3500 ± 1300 | 1.1 ± 0.2 | −7.6 ± 0.3 | −5.6 ± 1.5 | −2.0 ± 1.5 |
| D30K/E31K | WT | 4800 ± 690 | 0.8 ± 0.1 | −7.3 ± 0.1 | −11.1 ± 1.8 | 3.8 ± 1.7 |
| E38K/D39K | WT | 190 ± 15 | 0.9 ± 0.1 | −9.2 ± 0.0 | −12.8 ± 2.2 | 3.6 ± 2.2 |
| E46K | WT | 430 ± 30 | 1.0 ± 0.1 | −8.7 ± 0.0 | −11.7 ± 1.5 | 3.0 ± 1.5 |
| AcrIE4-F7NTD | WT | 140 ± 40 | 0.8 ± 0.1 | −9.4 ± 0.2 | −18.1 ± 1.7 | 8.7 ± 1.5 |
aITC experiments were performed in triplicate, and their thermodynamic parameters are reported as average values with standard errors.
bNo binding: Integrated heats from the measurement were insufficient to constrain the least squares fit derived from a single-site binding model for the titration.