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Nucleotide binding of chiDnaA
EcoDnaA (EcoWT), chiDnaA (ChiWT), and chiDnaA R285A (ChiRA) (1.9 pmol) were incubated at 0 °C for 15 min in the presence of various amounts (0–1 μm) of [α-32P]ATP or [3H]ADP, followed by filtration on a nitrocellulose membrane. Dissociation constant (Kd) and binding stoichiometry were determined using a Scatchard plot.
| DnaA |
Kd |
Stoichiometry |
||
|---|---|---|---|---|
| ATP | ADP | ATP | ADP | |
| nm | ||||
| EcoWT | 76 | 19 | 0.45 | 0.1 |
| ChiWT | 52 | 55 | 0.42 | 0.23 |
| ChiRA | 77 | 35 | 0.53 | 0.28 |
