Extended Data Fig. 6: Effects of detergent on the oligomeric state of the TOM complex.
From: Cryo-EM structure of the mitochondrial protein-import channel TOM complex at near-atomic resolution
a, Schematic diagram of the TOM complex purification procedure. Different detergent conditions (indicated by blue texts) were tested (specific conditions in b–e). b–e, Detailed SEC profiles of the purified TOM complex purified under different detergent conditions. “D” indicates the dimer peak, and “T” indicates the tetramer peak. Positions of the void peak (void) and peaks of molecular weight standards are indicated by arrowheads. TG, thyroglobulin (670 kDa); F, ferritin (440 kDa); ald, aldolase (156 kDa). Note that b–d is the same as in Fig. 4a–c, and b is the same experiment shown in Fig. 1a. f, SDS-PAGE analysis of peak fractions from the SEC purification shown in c. The peak positions are marked with “T” and “D”. The SDS gel was stained by Coomassie. g, Crude lysates prepared from cells overexpressing the TOM complex were solubilized with indicated detergent and subjected to BN-PAGE, followed by immunoblotting using an anti-Strep-tag antibody (detecting Tom40-Strep). A gradual decrease of mobility of the TOM complex accompanied by lowered detergent concentrations is likely due to an increased detergent micelle size. Source data for g are available with the paper online. The experiment in g was repeated twice with similar results.
