Figure 3.

E. coli DnaK binds to ScOrc4p and disassembles oligomers. (a) SDS/PAGE of soluble lysates and Ni²⁺ affinity chromatography peak fractions for proteins in Fig. 2a and RepA. The arrow points to the 70-kDa copurifying band. (Top) Western blot with anti-DnaK polyclonal antiserum. (b) (Top) FPLC gel filtration analysis of purified DnaK-ScOrc4p (ΔC367) complexes (lane L) after incubation with ATP, at 4°C or 37°C. Elution positions of standards (Fig. 2b) and unbound ATP/ADP are marked on the absorption profiles. (Bottom) SDS/PAGE and densitometric ratios for significant fractions.