Table 1.
ITC-derived thermodynamic parameters for the association of HHARI, HOIP, TRIAD3A and RNF144A RBR and Ring1 domains with UbcH7, UbcH5A and their ubiquitin conjugates. Each experiment was repeated multiple times and the average values reported with their standard deviation.
| Interactions | n | Kd (uM) | ΔH Kcal/mol | −TΔS Kcal/mol | ΔG Kcal/mol |
|---|---|---|---|---|---|
| HHARI-RBR/UbcH7 | 0.9 ± 0.1 | 0.41 ± 0.05 | 1.8 ± 0.2 | −10 ± 0.6 | −8.2 ± 0.8 |
| HHARI-RBR/UbcH7-Ub | 0.9 ± 0.1 | 0.20 ± 0.01 | −2.3 ± 0.2 | −6.6 ± 0.7 | −8.9 ± 0.9 |
| HHARI-Ring1/UbcH7 | 0.8 ± 0.1 | 0.21 ± 0.01 | 4.2 ± 0.5 | −13.2 ± 0.4 | −9.0 ± 0.9 |
| HHARI-Ring1/UbcH7-Ub | 0.8 ± 0.2 | 0.12 ± 0.01 | 3.9 ± 0.4 | −13.3 ± 0.6 | −9.4 ± 0.9 |
| HHARI-RBR/UbcH5A | No binding detected | ||||
| HHARI-RBR/UbcH5A-Ub | 0.8 ± 0.1 | 4.5 ± 0.4 | −3.7 ± 0.4 | −3.4 ± 0.2 | −7.1 ± 0.6 |
| HHARI-Ring1/UbcH5A-Ub | No binding detected | ||||
| HOIP-RBR/UbcH7 | 0.8 ± 0.2 | 2.3 ± 0.2 | 0.8 ± 0.1 | −8.3 ± 0.4 | −7.5 ± 0.6 |
| HOIP-RBR/UbcH7-Ub | 0.9 ± 0.1 | 0.9 ± 0.1 | 0.8 ± 0.1 | −8.9 ± 0.6 | −8.1 ± 0.8 |
| HOIP-Ring1/UbcH7 | 1.0 ± 0.1 | 4.8 ± 0.5 | 1.4 ± 0.2 | −8.5 ± 0.5 | −7.1 ± 0.7 |
| HOIP-Ring1/UbcH7-Ub | 0.9 ± 0.2 | 4.3 ± 0.4 | 1.1 ± 0.2 | −8.3 ± 0.5 | −7.2 ± 0.7 |
| HOIP-RBR/UbcH5A | No binding detected | ||||
| HOIP-RBR/UbcH5A-Ub | 0.9 ± 0.1 | 4.3 ± 0.3 | −8.2 ± 0.5 | 1.0 ± 0.1 | −7.2 ± 0.7 |
| HOIP-Ring1/UbcH5A-Ub | No binding detected | ||||
| TRIAD3A-RBR/UbcH7 | 1.0 ± 0.1 | 2.0 ± 0.1 | 1.8 ± 0.7 | −9.4 ± 0.1 | −7.6 ± 0.8 |
| TRIAD3A-RBR/UbcH7-Ub | 0.9 ± 0.1 | 2.4 ± 0.1 | 3.7 ± 0.3 | −11.2 ± 0.5 | −7.5 ± 0.9 |
| TRIAD3A-Ring1/UbcH7 | 0.8 ± 0.2 | 3.2 ± 0.2 | 4.5 ± 0.3 | −11.8 ± 0.6 | −7.3 ± 0.9 |
| TRIAD3A-Ring1/UbcH7-Ub | 0.8 ± 0.2 | 4.3 ± 0.3 | 3.6 ± 0.4 | −10.8 ± 0.2 | −7.1 ± 0.7 |
| TRIAD3A-RBR/UbcH5A | No binding detected | ||||
| TRIAD3A-RBR/UbcH5A-Ub | No binding detected | ||||
| TRIAD3A-Ring1/UbcH5A-Ub | No binding detected | ||||
| RNF144A-RBR/UbcH7 | 0.9 ± 0.1 | 1.0 ± 0.1 | 1.7 ± 0.2 | −9.7 ± 0.5 | −8.0 ± 0.8 |
| RNF144A-RBR/UbcH7-Ub | 0.8 ± 0.2 | 0.52 ± 0.04 | −0.9 ± 0.1 | −7.5 ± 0.7 | −8.4 ± 0.9 |
| RNF144A-Ring1/UbcH7 | 0.8 ± 0.2 | 2.2 ± 0.2 | 3.5 ± 0.4 | −11.1 ± 0.6 | −7.5 ± 0.9 |
| RNF144A-Ring1/UbcH7-Ub | 0.8 ± 0.1 | 1.0 ± 0.1 | 2.7 ± 0.4 | −10.7 ± 0.5 | −8.0 ± 0.8 |
| RNF144A-RBR/UbcH5A | No binding detected | ||||
| RNF144A-RBR/UbcH5A-Ub | No binding detected | ||||
| RNF144A-Ring1/UbcH5A-Ub | No binding detected | ||||