ITC analysis of interactions between the SPSB2 SPRY domain and wild-type or mutant iNOS N-terminal peptides
| Peptide | Sequence | Kd |
| nM | ||
| A | Ac-KEEKDINNNVKKT-NH2 | 13.3 ± 3.0 |
| B | Ac-KEAKDINNNVKKT-NH2 | 14.0 ± 3.0 |
| C | Ac-KEEADINNNVKKT-NH2 | 127 ± 23 |
| D | Ac-KEAADINNNVKKT-NH2 | 65.4 ± 7.4 |
| E | Ac-KEEKAINNNVKKT-NH2 | 21,600 ± 750 |
| F | Ac-KEEKDANNNVKKT-NH2 | 23.5 ± 9.9 |
| G | Ac-KEEKDIANNVKKT-NH2 | 17,200 ± 7,400 |
| H | Ac-KEEKDIQNNVKKT-NH2 | 40,500 ± 7,200 |
| I | Ac-KEEKDINANVKKT-NH2 | 826 ± 20 |
| J | Ac-KEEKDINNAVKKT-NH2 | NDa |
| K | Ac-KEEKDINNQVKKT-NH2 | NDa |
| L | Ac-KEEKDINNNAKKT-NH2 | 56.8 ± 11.2 |
| M | Ac-KEEKDINNNVKAT-NH2 | 29.8 ± 12.2 |
| N | Ac-KEEKDINNNAKAT-NH2 | 180 ± 39 |
| O | Ac-KEEADINNNAKAT-NH2 | 311 ± 37 |
| Peptide | Sequence | Kd |
| nM | ||
| A | Ac-KEEKDINNNVKKT-NH2 | 13.3 ± 3.0 |
| B | Ac-KEAKDINNNVKKT-NH2 | 14.0 ± 3.0 |
| C | Ac-KEEADINNNVKKT-NH2 | 127 ± 23 |
| D | Ac-KEAADINNNVKKT-NH2 | 65.4 ± 7.4 |
| E | Ac-KEEKAINNNVKKT-NH2 | 21,600 ± 750 |
| F | Ac-KEEKDANNNVKKT-NH2 | 23.5 ± 9.9 |
| G | Ac-KEEKDIANNVKKT-NH2 | 17,200 ± 7,400 |
| H | Ac-KEEKDIQNNVKKT-NH2 | 40,500 ± 7,200 |
| I | Ac-KEEKDINANVKKT-NH2 | 826 ± 20 |
| J | Ac-KEEKDINNAVKKT-NH2 | NDa |
| K | Ac-KEEKDINNQVKKT-NH2 | NDa |
| L | Ac-KEEKDINNNAKKT-NH2 | 56.8 ± 11.2 |
| M | Ac-KEEKDINNNVKAT-NH2 | 29.8 ± 12.2 |
| N | Ac-KEEKDINNNAKAT-NH2 | 180 ± 39 |
| O | Ac-KEEADINNNAKAT-NH2 | 311 ± 37 |
Typical ITC raw data and titration curves are shown in Fig. S1. Bold letters indicate the amino acid substitution. The Kd column shows dissociation constants with errors from the Origin-calculated association constants transferred as the same fractions of primary values.
Binding affinity was too low to be determined under these conditions.