Fig. 5.

The incorporation of DnaK and Hsp70 into POPS liposomes results in different states of oligomerization. Recombinant DnaK (1 μg) or recombinant human Hsp70 (1 μg) were incubated with POPS liposomes (400 μg) in 50-mM Tris buffer pH 7.4 for 30 min at 25 °C. The liposomes were centrifuged at 100,000×g for 40 min at 4 °C. Pellets were resuspended in Tris buffer (pH 7.4), Na₂CO₃ buffer (pH 11.5), or glycine buffer (pH 2) and centrifuged again. Liposome pellets were then solubilized in LDS sample buffer, and liposome-incorporated proteins were separated by LDS-PAGE and detected by Coomassie Brilliant Blue staining. Arrows indicate the location of monomeric (m), dimeric (d) and oligomeric (o) forms of human Hsp70 and bacterial DnaK, respectively