Table 2.
ATP binding affinity of WT and mutant DnaA proteins: DnaA proteins were incubated with radioactive ATP and nucleotide binding was measured by filter retention assay (see Materials and Methods). Mean (±SD) values represents data from three independent set of experiments
| Amino acid change | ATP binding stoichiometry (fmoles ATP/fmoles protein) | K D (ATP binding) (μM) |
|---|---|---|
| WT | 0.30 ± 0.03 | 0.11 ± 0.03 |
| F76Y | 0.22 ± 0.03 | 0.14 ± 0.03 |
| F76A | 0.36 ± 0.07 | 0.12 ± 0.03 |
| H136Q | 0.19 ± 0.01 | 0.10 ± 0.02 |
| H136A | 0.08 ± 0.03 | 0.14 ± 0.04 |
| Q156E | 0.33 ± 0.05 | 0.10 ± 0.02 |
| Q156N | 0.04 ± 0.01 | 0.10 ± 0.02 |