Table 1 Steady-state kinetic parameters for nucleotide incorporation by exonuclease mutant saPolX-H435A on 1 nucleotide gapped DNA substrates.
Templating nucleotide | Incoming nucleotide | kcat (min−1) | KM (μM) | kcat/KM (μM−1 min−1) | fincorporation |
|---|---|---|---|---|---|
dA | dTTP | 5.82 × 10−2 ± 5.3 × 10−3 | 6.38 ± 0.63 | 9.14 × 10−3 | 1 |
dT | dATP | 8.07 × 10−1 ± 5.3 × 10−2 | 1.32 ± 0.16 | 0.616 | 1 |
dG | dCTP | 7.68 × 10−1 ± 5.9 × 10−2 | 3.76 ± 0.39 | 0.205 | 1 |
dC | dGTP | 7.68 × 10−1 ± 9.9 × 10−2 | 1.87 ± 0.27 | 0.410 | 1 |
dT | dGTP | 1.37 × 10−2 ± 1.5 × 10−3 | 47.00 ± 7.72 | 2.95 × 10−4 | 4.8 × 10−4 |
dA | rUTP | ND | ND | ND | – |
dT | rATP | ND | ND | ND | – |
dG | rCTP | 2.1 × 10–2 ± 3.3 × 10–3 | 137.63 ± 9.40 | 1.52 × 10–4 | 7.4 × 10–4 |
dC | rGTP | 1.17 × 10–2 ± 1.9 × 10–3 | 246.50 ± 42.11 | 4.74 × 10–5 | 1.15 × 10–4 |
dG | rGTP | ND | ND | ND | - |
dA | 8oxodGTP | 1.04 × 10–2 ± 1.5 × 10–3 | 57.35 ± 10.39 | 1.83 × 10–4 | 0.02 |
dC | 8oxodGTP | ND | ND | ND | - |
dT | 2-OH-dATP | 3.76 × 10–2 ± 5.0 × 10–3 | 4.46 ± 0.70 | 8.55 × 10–3 | 0.014 |
dG | 2-OH-dATP | ND | ND | ND | - |
dC | 2-OH-dATP | ND | ND | ND | - |
