Fig. 7: ClpP β-hairpin mutants are affected in stimulating ClpC ATPase and unfolding activities.
From: Structure of the central Staphylococcus aureus AAA+ protease MecA/ClpC/ClpP
a ATPase activities of MecA/ClpC complexes were determined in the absence and presence of ClpP wild type (WT) or β-hairpin mutants. b Unfolding of MecA-mEOS3.2 by ClpC was monitored in the absence or presence of ClpP WT and indicated β-hairpin mutants. Initial MecA-mEOS3.2 fluorescence was set as 100% and MecA-mEOS3.2 unfolding rates were determined. c ATPase activities of ΔN-ClpC-R443A were determined in the presence of ClpP WT or mutants as indicated. d Unfolding of Kaede-SsrA by ΔN-ClpC-R443A was monitored in the absence or presence of ClpP WT and β-hairpin mutants. Initial Kaede-SsrA fluorescence was set as 100% and unfolding rates of ΔN-ClpC-R443A were determined. e ATPase activities of ClpC WT (+MecA), N659A and alloW (E620A/K621A) and alloR (E692A/K694A) mutants were determined in the absence and presence of ClpP. The activity of ClpC WT (+MecA) was set to 100%. f Unfolding rates of MecA-mEOS3.2 by ClpC WT, N659A and alloW (E620A/K621A) and alloR (E692A/K694A) mutants were determined in the absence and presence of ClpP. Unfolding activity of ClpC WT was set at 100%. Error bars represent standard deviations (n = 3) (a–f).
