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. Author manuscript; available in PMC: 2023 Nov 19.
Published in final edited form as: J Thromb Haemost. 2023 Apr 8;21(8):2137–2150. doi: 10.1016/j.jtha.2023.03.034

FIGURE 4.

FIGURE 4

Sulfenylation by H2O2 promotes PDI disulfide formation. (A) Differential alkylation to detect PDI disulfide formation. The free cysteine thiols were labeled and blocked with maleimide-Cy3. Cysteine disulfides and other reversible oxidative modifications, including sulfenylation, were reduced with TCEP, and the newly available free thiols were labeled with maleimide-Cy5. (B) H2O2 promoted disulfide formation on PDI compared to buffer-treated conditions. (C) H2O2-mediated disulfide formation on PDI was prevented by arsenite. (D) Redox state of the PDI a (C53-C56) and a′ (C397-C400) active-site cysteines determined by differential cysteine alkylation and mass spectrometry. p-values were determined by unpaired Student’s t-test in (B) and one-way ANOVA with Tukey’s post hoc analysis in (C). Data represented as mean ± SEM.