Figure 7
Comparison of rP75-ΔECD-3CX and rP75-ΔECD-3CX-249C oligomerization rates. (A) The kinetics of rP75-ΔECD-3CX and rP75-ΔECD-3CX-249C dimerization based on SDS-page analysis (n = 3). k—the reaction rate constant calculated for irreversible second-order reaction. (B) The SDS-page analysis of purified proteins incorporated into DMPC:CHAPS (q = 1, LPR = 1,000:1) bicelles at the initial state and after 70 h of incubation without reducing agents (after dialysis). Full-length gels are presented in Supplementary Fig S9. (C) Comparison of rP75-ΔECD-3CX and rP75-ΔECD-3CX-249C oligomerization in the membrane of E. coli cells. Both strains were grown under the same conditions and the equivalent amount of cells (normalized to OD600) were harvested through 72 h after induction. After cell lysis, the membrane fraction was separated, solubilized in SDS containing buffer and analyzed by SDS-PAGE. 3CX rP75-ΔECD-3CX, 249C rP75-ΔECD-3CX-249C, M monomer, D dimer, T tetramer. The figure was prepared using the program Inkscape 0.92 (https://inkscape.org/).
