Fig 3.

Analysis of MerA glutathionylation with biotinylated glutathione (BioGSSG). Prereduced MerA was incubated for 1 h in the presence of BioGSSG (2 mM) with or without prior incubation with both 2-iodoacetamide (IAM) (100 mM) and N-ethylmaleimide (NEM) (20 mM). The reversibility of the glutathionylation of MerA was assessed by treatment with dithiothreitol (DTT) (50 mM). Proteins were resolved by nonreducing SDS-PAGE and transferred onto nitrocellulose for Western blotting with antibiotin antibodies. Coomassie staining of the gel shows equal loading in each lane. The results of two independent tests are shown.