Table 1.
Comparison of the affinity measurements between NCOA4 and FTH1-WT and its mutants obtained by ITC, immunoprecipitation and colocalization assays
| Mutant | Thermodynamic parameters from ITC | Relative interaction by immunoprecipitation | Cells with colocalized mRuby3-FTH1/FTL:mGFP-NCOA4 puncta | ||||||
|---|---|---|---|---|---|---|---|---|---|
| ΔG (kJ/mol) | ΔH (kJ/mol) | ΔS J/(K*mol) | Kd (µM) | n | % | p value* | % | p value* | |
| FTH1-WT | −32.2 ± 1.1 | −23.0 ± 1.8 | 30.9 ± 8.2 | 2.4 ± 1.1 | 2.5 ± 0.6 | 100 | n.a. | 100 | n.a. |
| FTH1-A19Y | – | – | – | – | 8.86 | <0.0001 | 0 | <0.0001 | |
| FTH1-N22A | No Binding | 3.72 | <0.0001 | 0.89 | <0.0001 | ||||
| FTH1-R23A | No Binding | 5.35 | <0.0001 | 7.04 | <0.0001 | ||||
| FTH1-R23I | – | – | – | – | 13.0 | <0.0001 | 23.7 | <0.0001 | |
| FTH1-R23L | No Binding | 86.0 | 0.4614 | 33.7 | 0.0005 | ||||
| FTH1-S114D | - 31.2 ± 0.3 | - 19.5 ± 2.1 | 39.1 ± 5.9 | 3.5 ± 0.5 | 3.0 ± 0.1 | 111 | 0.6897 | 23.9 | <0.0001 |
| FTH1-S114E | No Binding | 113 | 0.5198 | 24.9 | <0.0001 | ||||
| FTL-WT | – | – | – | – | 14.0 | <0.0001 | 0 | <0.0001 | |
| L2H | −30.1 ± 0.3 | −22.0 ± 0.4 | 26.9 ± 2.4 | 5.6 ± 0.7 | 1.9 ± 0.1 | 147 | <0.0001 | 15.5 | <0.0001 |
*Dunnett’s multiple comparison.
Source data are provided as a Source Data file.