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Western blot analysis of maleimide-labeled and purified single-cysteine mutants. Western blots were decorated with streptavidin coupled to alkaline phosphatase to detect biotinylated proteins (A, C, and E) or antibodies against the His-tag to compare the amounts of protein (B, D, and F). Single-cysteine mutants were labeled with biotin maleimide with (+) or without (−) prior incubation with AMdiS, as indicated, in whole cells (A and B) and inside-out membrane vesicles (C–F).
