TABLE 2.
Properties of CPV resolvase metal cofactors
ND, not determined due to weak binding; n = binding sites per resolvase dimer.
| Mg2+ | Mn2+ | |
|---|---|---|
| Wild-type resolvasea | ||
| Kd (μm) | ND | 1190 ± 30 |
| ΔH (kcal/mol) | ND | −6.14 ± 0.05 |
| Resolvase E59A·HJ (A site binding) | ||
| Kd (μm) | 277 ± 14 | 24.7 ± 0.8 |
| ΔH (kcal/mol) | 3.93 ± 0.08 | 2.76 ± 0.02 |
| Nb | 2 | 2.02 ± 0.01 |
| Resolvase D131A·HJ (B site binding) | ||
| Kd (μm) | 392 ± 16 | 5.75 ± 0.21 |
| ΔH (kcal/mol) | 4.52 ± 0.08 | 8.11 ± 0.04 |
| Nb | 2 | 1.75 ± 0.10 |
| Resolvase D130N/E59A·HJ (A site binding) a | ||
| Kd (μm) | ND | 151 ± 24 |
| ΔH (kcal/mol) | ND | 2.26 ± 0.12 |
| Bulged duplex substrate cleavage | ||
| Km (μm) | 750 ± 330 | 50 ± 17 |
a Stoichiometry was fixed at n = 2 (see “Experimental Procedures”).
b Stoichiometry was fixed at n = 2 when [resolvase·HJ]/Kd < 4].