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. Author manuscript; available in PMC: 2018 Jul 15.
Published in final edited form as: Biochem Pharmacol. 2017 Apr 9;136:86–98. doi: 10.1016/j.bcp.2017.03.025

Figure 8. H2S S-sulfhydrates LDHA.

Figure 8

MS/MS/LC was used to identify and quantity the post-translational modification of NaHS-induced S-sulfhydration (SSH) of LDHA. (A) Figure shows the area under the peak of SSH-Cys163 and the total digested protein without the SSH modification in both WT and C163A LDHA. (Total wild-type LDHA protein used in the assay was 1 μg; total C163A LDHA was increased to 2.5 μg in order to increase the detection of residual sulfhydrated cysteines.) Only minimal amount of sulfhydrated cysteines was detected in the C163A LDHA. (B) Figure represents the ratio of the area under the peak of SSH-Cys163 vs. the area under the peak of the digested peptide without S-sulfhydration modification in WT LDHA. (C) Extracted Ion Chromatogram (XIC) of WT LDHA (top) and C163A LDHA (bottom) treated with 10 μM NaHS. NaHS-treated WT LDHA produced a significant SSH-Cys163 peak at retention time (RT) 20.16 min, which was absent in NaHS-treated C163A LDHA.