Figure 5.

ER-specific disulfide rearrangements occur in a partially ER-exposed disintegrin intermediate. A–C, non-reducing SDS-PAGE shows either total translation product (V5) or ER-specific species (NE) for the 146 intermediate of the disintegrin domain when translated in reducing (Red), redox-balanced (Bal), and oxidizing (Ox) lysate (A); when translated in oxidizing lysate with G6Pi added after translation and samples taken at specific time points after G6Pi treatment (B, lanes 4–6); and when translated in oxidizing lysate to which G6Pi was added after translation in the presence or absence of auranofin (Aur) (C, lanes 4–7). Control samples for B and C were translated in reducing, redox-balanced, and oxidizing lysates (lanes 1–3 in both cases) for gel mobility comparison. The experiments in A and B were repeated three times and those in C twice, with representative data shown. Symbols indicate reduced preprotein (asterisks), reduced mature protein (circles), and oxidized mature protein (inverted triangles).