Overview of the ADAMTS13 mutants constructed containing single or multiple amino acid substitutions. Cloning strategy and mutations affecting the particular domain(s) are denoted.
Cloning strategy . | Amino acid exchange in wt ADAMTS13 . | Domain(s) affected . |
|---|---|---|
| Overlapping primer PCR | ||
| W7 | R7W | Signalpeptide (S) |
| X44 | Q44X | Propeptide (P) |
| E448 | Q448E | Cysteine-rich (Cys) |
| Inverse PCR | ||
| A618 | P618A | Spacer (Sp) |
| V732 | A732V | Tsp1-2 |
| W1336 | R1336W | Cub2 |
| A-V | P618A-A732V | Sp/Tsp1-2 |
| Switching restriction fragments | ||
| W1-A | R7W-P618A | P/Sp |
| W1-V | R7W-A732V | P/Tsp1-2 |
| E-A | Q448E-P618A | Cys/Sp |
| V-W2 | A732V-R1336W | Tsp1-2/Cub2 |
| E-A-V | Q448E-P618A-A732V | Cys/Sp/Tsp1-2 |
| W1-A-V | R7W-P618A-A732V | S/Sp/Tsp1-2 |
| A-V-W2 | P618A-A732V-R1336W | Sp/Tsp1-2/Cub2 |
| W1-E-A | R7W-Q448E-P618A | P/Cys/Sp |
| W1-E-W2 | R7W-Q448E-R1336W | P/Cys/Cub2 |
| W1-E-A-V | R7W-Q448E-P618A-A732V | S/Cys/Sp/Tsp1-2 |
| W1-A-V-W2 | R7W-P618A-A732V-R1336W | S/Sp/Tsp1-2/Cub2 |
| W1-E-A-V-W2 | R7W-Q448E-P618A-A732V-R1336W | S/Cys/Sp/Tsp1-2/Cub2 |
Cloning strategy . | Amino acid exchange in wt ADAMTS13 . | Domain(s) affected . |
|---|---|---|
| Overlapping primer PCR | ||
| W7 | R7W | Signalpeptide (S) |
| X44 | Q44X | Propeptide (P) |
| E448 | Q448E | Cysteine-rich (Cys) |
| Inverse PCR | ||
| A618 | P618A | Spacer (Sp) |
| V732 | A732V | Tsp1-2 |
| W1336 | R1336W | Cub2 |
| A-V | P618A-A732V | Sp/Tsp1-2 |
| Switching restriction fragments | ||
| W1-A | R7W-P618A | P/Sp |
| W1-V | R7W-A732V | P/Tsp1-2 |
| E-A | Q448E-P618A | Cys/Sp |
| V-W2 | A732V-R1336W | Tsp1-2/Cub2 |
| E-A-V | Q448E-P618A-A732V | Cys/Sp/Tsp1-2 |
| W1-A-V | R7W-P618A-A732V | S/Sp/Tsp1-2 |
| A-V-W2 | P618A-A732V-R1336W | Sp/Tsp1-2/Cub2 |
| W1-E-A | R7W-Q448E-P618A | P/Cys/Sp |
| W1-E-W2 | R7W-Q448E-R1336W | P/Cys/Cub2 |
| W1-E-A-V | R7W-Q448E-P618A-A732V | S/Cys/Sp/Tsp1-2 |
| W1-A-V-W2 | R7W-P618A-A732V-R1336W | S/Sp/Tsp1-2/Cub2 |
| W1-E-A-V-W2 | R7W-Q448E-P618A-A732V-R1336W | S/Cys/Sp/Tsp1-2/Cub2 |