Binding of ADATMS13 and mutants to GST-VWF73 and multimeric vWF
| Constructs . | GST-VWF73 . | Multimeric VWF . | ||||
|---|---|---|---|---|---|---|
| BMax . | KD (×10−6M) . | BMax/KD (×103M−1) . | BMax . | KD (×10−6M) . | BMax/KD (×103M−1) . | |
| WT | 0.38 ± 0.01 | 28.4 ± 2.4 | 13.3 | 0.59 ± 0.02 | 12.8 ± 1.2 | 45.9 |
| del6aa | 0.46 ± 0.04 | 245.7 ± 45.7 | 1.9 | 0.44 ± 0.02 | 9.8 ± 1.6 | 45.2 |
| R659A | 0.39 ± 0.01 | 66.0 ± 26.3 | 5.9 | 0.38 ± 0.02 | 7.1 ± 1.3 | 53.8 |
| R660A | 0.40 ± 0.04 | 103.6 ± 26.3 | 3.9 | 0.57 ± 0.01 | 12.1 ± 1.6 | 47.2 |
| Y661A | 0.37 ± 0.01 | 35.5 ± 3.8 | 10.5 | 0.52 ± 0.03 | 12.3 ± 2.3 | 43.0 |
| G662A | 0.37 ± 0.01 | 25.2 ± 3.2 | 14.6 | 0.51 ± 0.02 | 9.7 ± 1.0 | 52.8 |
| E663A | 0.39 ± 0.01 | 32.3 ± 2.7 | 12.2 | 0.40 ± 0.02 | 10.6 ± 1.3 | 38.2 |
| E664A | 0.42 ± 0.02 | 47.6 ± 8.4 | 8.9 | 0.46 ± 0.03 | 10.2 ± 2.2 | 45.4 |
| Constructs . | GST-VWF73 . | Multimeric VWF . | ||||
|---|---|---|---|---|---|---|
| BMax . | KD (×10−6M) . | BMax/KD (×103M−1) . | BMax . | KD (×10−6M) . | BMax/KD (×103M−1) . | |
| WT | 0.38 ± 0.01 | 28.4 ± 2.4 | 13.3 | 0.59 ± 0.02 | 12.8 ± 1.2 | 45.9 |
| del6aa | 0.46 ± 0.04 | 245.7 ± 45.7 | 1.9 | 0.44 ± 0.02 | 9.8 ± 1.6 | 45.2 |
| R659A | 0.39 ± 0.01 | 66.0 ± 26.3 | 5.9 | 0.38 ± 0.02 | 7.1 ± 1.3 | 53.8 |
| R660A | 0.40 ± 0.04 | 103.6 ± 26.3 | 3.9 | 0.57 ± 0.01 | 12.1 ± 1.6 | 47.2 |
| Y661A | 0.37 ± 0.01 | 35.5 ± 3.8 | 10.5 | 0.52 ± 0.03 | 12.3 ± 2.3 | 43.0 |
| G662A | 0.37 ± 0.01 | 25.2 ± 3.2 | 14.6 | 0.51 ± 0.02 | 9.7 ± 1.0 | 52.8 |
| E663A | 0.39 ± 0.01 | 32.3 ± 2.7 | 12.2 | 0.40 ± 0.02 | 10.6 ± 1.3 | 38.2 |
| E664A | 0.42 ± 0.02 | 47.6 ± 8.4 | 8.9 | 0.46 ± 0.03 | 10.2 ± 2.2 | 45.4 |
BMax indicates the maximal binding; KD is the dissociation constant; and M is molar concentration. R, Y, G and E are single letter abbreviations of amino acids argine, tyrosine, glycine and glutamic acid.