Comment on "Evidence that the ProPerDP method is inadequate for protein persulfidation detection due to lack of specificity".

Abstract

The recent report by Fan <i>et al</i> alleged that the ProPerDP method is inadequate for the detection of protein persulfidation. Upon careful evaluation of their work, we conclude that the claim made by Fan <i>et al</i> is not supported by their data, rather founded in methodological shortcomings. It is understood that the ProPerDP method generates a mixture of cysteine-containing and non-cysteine-containing peptides. Instead, Fan <i>et al</i> suggested that the detection of non-cysteine-containing peptides indicates nonspecific alkylation at noncysteine residues. However, if true, then such peptides would not be released by reduction and therefore not appear as products in the reported workflow. Moreover, the authors' biological assessment of ProPerDP using <i>Escherichia coli</i> mutants was based on assumptions that have not been confirmed by other methods. We conclude that Fan <i>et al</i> did not rigorously assess the method and that ProPerDP remains a reliable approach for analyses of protein per/polysulfidation.