TABLE 2.
| Mutants | Maximal cleavage activity (μM min−1) | Half-maximal activation concn (μM) | |
|---|---|---|---|
| The residues in peptide binding pockets | AlgW | 165.4 ± 4.46 | 2.02 ± 0.21 |
| AlgW-W281A | 10.32 ± 1.51 | 1.51 ± 0.72 | |
| AlgW-L282A | 13.24 ± 1.59 | 2.76 ± 1.19 | |
| AlgW-V284A | 63.13 ± 3.82 | 3.56 ± 0.73 | |
| AlgW-E285A | 57.5 ± 4.9 | 3.84 ± 1.09 | |
| AlgW-K287A | 49.42 ± 3.59 | 6.53 ± 1.38 | |
| AlgW-M342A | NA | NA | |
| AlgW-R374A | 5.45 ± 0.68 | ND | |
| The residues involved in domain-domain interactions | AlgW-D149A | 8.16 ± 0.85 | ND |
| AlgW-E151A | 34.93 ± 1.87 | 2.99 ± 0.56 | |
| AlgW-Y212A | 39.46 ± 6.17 | 2.12 ± 0.62 | |
| AlgW-E266A | 60.51 ± 5.07 | 1.08 ± 0.5 | |
| AlgW-R279A | 26.87 ± 3.21 | 2.19 ± 0.99 | |
| AlgW-R347A | 63.5 ± 8.42 | 1.85 ± 0.99 |
a
Apparent activation parameters were determined using the formula of Michaelis-Menten [Y = Vmax · X/(Km + X)]; the parameters listed are the means of three independent determinations.
b
NA, no detectable activity; ND, not determined.