Fig. 1.

DnaK incorporates into the membrane of liposomes made of different lipids. Recombinant DnaK (1 μg) was incubated with POPS, POPG, POPE, or POPC liposomes (400 μg) in 50-mM Tris buffer pH 7.4 for 30 min at 25 °C. At the end of the incubation period, 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 LDS sample buffer, and liposome-incorporated proteins were separated by LDS-PAGE and analyzed by Western blotting using a monoclonal antibody against DnaK (SPA880, Enzo Life Sciences) and HRP-conjugated goat anti-mouse as secondary antibodies. Arrows indicate the location of monomeric (m) and dimeric (d) forms of DnaK, respectively