Summary of Affinity and Charge Mutants
| Protein name | Mutation(s) | Domain(s) affected | Charge change | log binding affinity (pK) |
|---|---|---|---|---|
| Annexin V-128 | None (wild-type) | None | 0 | 30.8 |
| Affinity mutants | ||||
| Annexin V-131 | D303N | 4 | 1 | 22.4 |
| Annexin V-138 | E72Q | 1 | 1 | 22.4 |
| Annexin V-136 | D144N, E228Q | 2, 3 | 2 | 13.9 |
| Annexin V-139 | E72Q, D303N | 1, 4 | 2 | 15.8 |
| Annexin V-137 | D144N, E228Q, D303N | 2, 3, 4 | 3 | 38%* |
| Annexin V-143 | E72Q, D144N, D303N | 1, 2, 4 | 3 | 38%* |
| Annexin V-145 | E72Q, D144N, E228Q, D303N | 1, 2, 3, 4 | 4 | 18%* |
| Charge mutants | ||||
| Annexin V-177 | R18E | 1 | −2 | 30.9 |
| Annexin V-178 | R6Q | 1 | −1 | 32.1 |
| Annexin V-158 | E234Q | 3 | 1 | 31.3 |
| Annexin V-175 | E78Q, E234Q | 1, 3 | 2 | 28.8 |
↵* Percentage of pK value for wild-type protein.
Mutations present are given in single-letter amino acid code. RBC-binding affinity was determined by calcium titration as described (20). Binding affinity of annexin V-137, -143, and -145 was determined by phospholipid vesicle-binding assay (6) because binding affinity is too weak to measure by RBC-binding assay.