Subunit Interface Residues F129 and H294 of Human RAD51 Are Essential for Recombinase Function
Figure 2
Mutation of HsRAD51(F129) and HsRAD51(H294) residues affect ATP turnover.
(A) Purification of wild type and HsRAD51 substitution mutant proteins. Protein (1 µg) analyzed by 12% SDS-PAGE. (B) Steady-state ATPase activity with ssDNA in the presence of 150 mM KCl. (C) ATPγS binding by wild type and HsRAD51 substitution mutant proteins in the presence of ssDNA and 150 mM KCl. (D) ADP binding by wild type and HsRAD51 substitution mutant proteins in the presence of ssDNA and 150 mM KCl. (E) ATP turnover (kcat) with ssDNA and dsDNA in the presence and absence of KCl (K+). kcat values were calculated by Michaelis-Menten analysis. Error bars indicate standard deviation from at least three independent experiments.