Comment on "Evidence that the ProPerDP method is inadequate for protein persulfidation detection due to lack of specificity".
Comment on "Evidence that the ProPerDP method is inadequate for protein persulfidation detection due to lack of specificity".
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DOI：
10.1126/sciadv.abe7006
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发表时间：
2021-04
期刊：
影响因子：
13.6
通讯作者：
Nagy P
中科院分区：
文献类型：
--
作者：
Dóka É;Arnér ESJ;Schmidt EE;Dick TP;van der Vliet A;Yang J;Szatmári R;Ditrói T;Wallace JL;Cirino G;Olson K;Motohashi H;Fukuto JM;Pluth MD;Feelisch M;Akaike T;Wink DA;Ignarro LJ;Nagy P
Fan等人的文章不准确地指出ProPerDP不足以检测蛋白质过硫化物。Fan等人最近的报告声称ProPerDP方法不足以检测蛋白质过硫化。在仔细评估他们的工作后，我们得出结论，Fan等人提出的主张没有得到他们的数据支持，而是建立在方法论的缺陷上。应理解，ProPerDP方法产生含半胱氨酸和不含半胱氨酸的肽的混合物。相反，Fan等人认为，检测到不含半胱氨酸的肽表明在非半胱氨酸残基上发生了非特异性烷基化。然而，如果是真的，那么这些肽将不会通过还原释放，因此不会在报告的工作流程中作为产物出现。此外，作者使用大肠杆菌突变体对ProPerDP进行的生物学评估是基于未经其他方法证实的假设。我们得出结论，Fan等人没有严格评估该方法，ProPerDP仍然是分析蛋白质过/多硫化的可靠方法。
The article by Fan et al. inaccurately states that ProPerDP is inadequate for protein persulfide detection. The recent report by Fan et al. 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 et al. 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 et al. 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 Escherichia coli mutants was based on assumptions that have not been confirmed by other methods. We conclude that Fan et al. did not rigorously assess the method and that ProPerDP remains a reliable approach for analyses of protein per/polysulfidation.
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