Molecular mechanism of J-domain-triggered ATP hydrolysis by Hsp70 chaperones. Kityk et al.
Figure S2: Immunoblot analysis of the expression of wild type and mutant dnaK. The ∆dnaK52 strain (BB1553) carrying a lacIq plasmid was transformed with plasmids encoding mutant and wild-type dnaK under the control of an IPTG-inducible promoter. Cells were grown under the conditions used in complementation assays (50 µM IPTG, 40 °C), extract prepared, separated by SDS-PAGE and DnaK detected by immunoblotting using a DnaK-specific polyclonal antiserum. Lanes from left to right: pre-stained molecular weight marker, extract of cells expressing dnaKwt, vector control, dnaK-Q378A, dnaK-T420A, dnaK-D477A, empty lane.