Extended Data Fig. 6: S-Cyanylation of glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and glycerol-3-phosphate dehydrogenase (GPDH).
From: Regulation of mammalian cellular metabolism by endogenous cyanide production
(a) Schematic representation of chemical cleavage of polypeptide’s backbone occurring after S-cyanylation of target cysteine residues under alkaline conditions. Cleavage is then detected by SDS-PAGE followed by Coomassie-staining. Created with BioRender.com. (b) Human GAPDH contains three cysteine residues. S-cyanylation of GAPDH results in the generation of two visible bands that are consistent with one single cleavage reaction and one S-cyanylation site. (c) MS label-free quantification of S-cyanylation of C247 peptide. Peptide intensity is normalized to the total GAPDH in each sample. 0.55 µM GAPDH was treated with 10 µM KCN, 10 µM H2O2 or their combination (n=XX/group, technical replicates). (d) MS label-free quantification of GPDH treated with 10 µM KCN, 10 µM H2O2 or their combination (n = 5/group, biological replicates). ND – not detected. Data in c, d are expressed as the mean ± s.e.m. Data in c were analysed with a two-way ANOVA followed by Bonferroni’s multiple-comparisons test. **p < 0.01 indicates significant differences.
