Fig. 3.

The incorporation of DnaK into POPS liposomes is not affected by the presence of nucleotides. Recombinant DnaK (1 μg) was incubated with POPS liposomes (400 μg) in 50-mM Tris buffer pH 7.4 for 30 min at 25 °C in the absence or presence of MgCl₂ (1 mM), ATP + MgCl₂ (1 mM each), ADP + MgCl₂ (1 mM each), GTP + MgCl₂ (1 mM each), and GDP + MgCl₂ (1 mM each). The liposomes were centrifuged at 100,000×g for 40 min at 4 °C. Pellets were resuspended in Na₂CO₃ buffer (pH 11.5) and centrifuged again. Liposome pellets were then solubilized in LDL sample buffer, and liposome-incorporated proteins were separated by LDS-PAGE and detected by Coomassie Brilliant Blue staining