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Changes in electrostatics of the redox-regulated molecular switch of the HMGB1 A-box domain as a function of the oxidation status of C23 and C45, revealed through Gd-based measurements of ϕENS potentials. The measurement for the reduced state was conducted in the presence of 5 mM DTT to maintain the thiol forms of C23 and C45. In the oxidized state, C23 and C45 form a disulfide bond. The bar graph shows changes in ϕENS potentials upon formation of the C23–C45 disulfide bond.
