Table 2. Average rates of ATP hydrolysis determined by an NADH-coupled assay according to ref. 59.
| Enzyme | ATP hydrolysis rate (μmol l−1 min−1) | k (min−1) |
|---|---|---|
| WT ZRANB3 | 16.59±0.34 | 110.65±2.27 |
| ZRANB3-specific HNH insert deletion mutants | ||
| ZRANB3-Δ975–1013 | 5.76±0.03 | 38.41±0.23 |
| ZRANB3-Δ972–1010 | 4.83±0.90 | 32.24±6.00 |
| Basic residues HNH mutants | ||
| K984A | 20.93±2.01 | 139.56±13.45 |
| R987A | 6.34±0.40 | 42.31±2.70 |
| K988A | 18.83±1.94 | 125.55±12.95 |
| K998A | 15.12±0.48 | 100.85±3.22 |
| R1009A | 7.49±2.57 | 49.94±17.14 |
| K1046A, R1048A | 19.38±1.74 | 129.23±11.62 |
| HNH active site mutants | ||
| H1015A | 18.67±2.24 | 124.46±14.96 |
| D1020A | 12.94±0.67 | 86.32±4.49 |
| H1021A | 18.59±1.98 | 123.95±13.19 |
| N1036A | 14.05±2.46 | 93.71±16.43 |
| H1045A | 8.91±0.66 | 59.42±4.40 |
| PCNA binding mutants | ||
| PIP* | 20.71±0.86 | 138.10±5.76 |
| ΔAPIM | 20.24±0.70 | 134.95±5.22 |
| PIP* ΔAPIM | 20.40±4.40 | 136.02±29.39 |
| Cancer associated mutants | ||
| T66A | 0.45±0.03 | 3.02±0.21 |
| R169H | ND | ND |
| G401D | ND | ND |
| R947Q | 10.24±1.15 | 68.26±7.70 |
| R947* | 9.50±0.64 | 63.36±4.31 |
| D1020Y | 11.60±3.30 | 77.36±22.04 |
APIM, AlkB homologue 2 PCNA Interacting Motif; HNH, His-Asn-His; ND, not defined; PCNA, Proliferating Cell Nuclear Antigen; PIP, PCNA-Interacting Protein.
ATP hydrolysis rate was calculated using the molar absorption coefficient for NADH in water (6220 L mol−1 cm−1) and the light path of 0.424 cm.