Table 1.
BRCA2 domains | ssDNA (nM) | p53N (μM) | p53 TAD2WT (μM) | p53 TAD2Mut1 (μM) | p53 TAD2Mut2 (μM) |
OB1 | 10,000 ± 1,000 | n.q.* | n.q.* | n.q.* | n.q.* |
OB2 | 150 ± 7 | 5.2 ± 0.3 | 6.0 ± 0.5 | 50 ± 4 | n.q.* |
OB3 | 110 ± 6 | 2.5 ± 0.2 | 3.2 ± 0.4 | 43 ± 3 | n.q.* |
OB2 + OB3 | 40 ± 5 | 3.0 ± 0.4 | 4.0 ± 0.3 | 39 ± 5 | 120 ± 10 |
ssDNA (dT)15, p53 TAD (1–93), p53TAD2 WT (35–57), p53TAD2 Mut1 (35–57): L43A/L45A/I50A/W53A/F54A and p53TAD2 Mut2 (35–57): D41K/D42K/D48K/D49K/E51K were used. Experiments were carried out in a buffer containing 25 mM Hepes, 150 mM NaCl, 3 mM DTT, pH 7.4 at 20 °C. The dissociation constant was obtained by fitting to a cooperative binding model and gave a Hill constant of ∼2.2.
*n.q.: not quantified (> 250 μM).