Table I. Cleavage of GST-VWF73-H and plasma VWF by ADAMTS13 and various mutants.
The signs +++, ++, and +/− indicate >50%, 10–50%, and <10% activity, respectively, compared with normal human plasma. S, indicates the predicted Tyr–Met bond was cleaved; NS, indicates the bond other than the Tyr–Met bond was cleaved. The domain structures of full-length ADAMTS13 (FL) and the mutants (from M to delCUB) are illustrated in detail in Fig. 1. The cleavage of GST-VWF73-H or plasma VWF was determined by 12% or 5% Tris-HCl SDS-gel and Western blotting with anti-GST IgG or anti-VWF IgG as described under “Experimental Procedures.”
| Constructs | Cleavage of VWF73
|
Cleavage of VWF,16 h | |
|---|---|---|---|
| 3 h | 16–24 h | ||
| M | − | + NS | − |
| MD | +/− | + S | − |
| MT | − | + S | − |
| MS | − | + S | − |
| MDT | + | ++ S and NS | − |
| MCS | +/− | ++ S | − |
| MTCS | +/− | ++ S | − |
| MDTC | ++ | ++ S | − |
| MDTCS | +++ | ++ S | ++ S |
| delCUB | +++ | ++ S | ++ S |
| FL | +++ | ++ S | ++ S |