Fig. 3
Redox response of OhsR detected by nonreducing SDS-PAGE. a 50 μM proteins treated with 50 mM DTT were further incubated with or without 150 μM H2O2, 75 μM HClO, 100 μM CHP, 200 μM t-BHP, 500 μM MSH, 150 μM CHP and 500 μM MSH, or 300 μM t-BHP and 500 μM MSH, and samples were then separated by 15% non-reducing SDS-PAGE (Left panel) and reducing SDS-PAGE (Right panel). b–d Analysis of S-mycothiolation and formed on OhsR by MALDI-TOF-TOF MS/MS. Bands (labeled (b–d)) excised from non-reducing gels in a were treated with trypsin and subjected to MS analysis. Only the relevant portion of each mass spectrum is shown. e S-mycothiolation of OhsR was monitored by biotin switch assay