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Fig. 4 | Microbial Cell Factories

Fig. 4

From: OhsR acts as an organic peroxide-sensing transcriptional activator using an S-mycothiolation mechanism in Corynebacterium glutamicum

Fig. 4

Reaction of OhsR-SOH with MSH but alkylation of OhsR with PEG-maleimide. a 20 μM reduced or OhsR-SOH was incubated with or without 200 μM MSH. Proteins were precipitated with TCA, treated with 5 mM PEG-maleimide, and evaluated on a CBB-stained 15% SDS-PAGE. b Proteins (50 μM) treated with 50 mM DTT were further incubated with or without 750 μM MSH, 250 μM CHP, or 250 μM CHP and 750 μM MSH, and samples were then separated by 15% nonreducing SDS-PAGE. c Analysis of sulfonic acid formed on OhsR by MALDI-TOF-TOF MS/MS. Bands (labeled (c)) excised from nonreducing gels in b were treated with trypsin and subjected to MS analysis. Only the relevant portion of each mass spectrum is shown

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