Table 1.
Reconstitution of protein using SDS.
Table 2.
Reconstitution of protein using Triton X-100 removed with Bio-Beads.
Table 3.
Reconstitution of protein using DM.
Figure 1.
Circular dichroism spectra of MBP-cytochrome b6 fusion protein in different detergent solutions.
Table 4.
Predicted per cent secondary structure content of the MBP-b6 in detergent solutions (Triton X-100, DM or SDS) from CD analysis.
Figure 2.
Solubilization of liposomes (black circles) by SDS (A), Triton X-100 (B) and DM (C) monitored by light scattering.
The onset of solubilization concentration and the total lysis concentration are marked with dashed lines. As a control, a buffer without detergent (empty circles) was titrated into liposomes.
Figure 3.
Zeta potential of liposomes during the reconstitution process for SDS removed by dialysis (A) or with Bio-Beads (B); Triton X-100 removed with Bio-Beads (C) and DM removed by dialysis (D) or with Bio-Beads (E).
Plotted are the mean values of n = 5 independent experiments with error bars depicting standard deviations. Asterisks denote significant differences between control and reconstitution experiments. LUV – initial liposome solutions (the same initial batch was measured at this point, which later was divided in two and processed in parallel for control and reconstitution experiments); Post. mix. – proteoliposomes in post-reconstitution mixture (directly after detergent removal); Proteolip. – final proteoliposomes (after centrifugation and washing). Control experiments underwent the same routine only the detergent solution without protein was added to them.
Figure 4.
Orientation of MBP-apocytochrome b6 in proteoliposomes reconstituted with dodecyl maltoside removed with Bio-Beads was analyzed by Western blotting with anti-MBP antibodies.
Purified, concentrated MBP-apocytochrome b6 in DM solution (1); DM solution of MBP-apocytochrome b6 added for reconstitution (2); untreated proteoliposomes (3); proteoliposomes treated with proteinase K (4). Lanes 3 and 4 contain the same amount of proteoliposomes.