- Policy & Practices
It was determined that the quality of each mutation set depended on two characteristics, namely:
- The ability of the scaffold protein to bind with its complementary binding partner. For example, CT52(I947H) should have the ability to bind to the T14-3-3 (S71L) binding pocket. The magnitude of this binding interaction should be comparable with the existing mutation set, that functions as a control (T14-3-3 (E19R) and CT52 (K943D)).
- Orthogonality of the designed mutation set. For example, the T14-3-3 (S71L) binding pocket should only have a strong interaction with its complementary binding partner CT52 (I947H). This also goes vice versa. The binding protein CT52, for instance CT52 (I947H) should only have a strong binding interaction with its complementary binding pocket T14-3-3 (S71L). Ideally the scaffolds have no other interaction other than their complementary interaction. If this is not completely the case this does not mean the mutation is not usable. It is important that there is a significant difference between the interaction which is expected to be the highest and the interaction which should not show activity.
The evaluation of criterion 1 can be found on the Functionality page, the evaluation of criterion 2 can be found on the Orthogonality page. The conclusion about the quality of each mutation set can be found at the Conclusion page. An overview of the mutation sets can be seen in figure 1.