Dóka Éva, Arnér Elias S J, Schmidt Edward E, Dick Tobias P, van der Vliet Albert, Yang Jing, Szatmári Réka, Ditrói Tamás, Wallace John L, Cirino Giuseppe, Olson Kenneth, Motohashi Hozumi, Fukuto Jon M, Pluth Michael D, Feelisch Martin, Akaike Takaaki, Wink David A, Ignarro Louis J, Nagy Péter
Department of Molecular Immunology and Toxicology, National Institute of Oncology, 1122 Budapest, Hungary.
Department of Selenoprotein Research, National Institute of Oncology, 1122 Budapest, Hungary.
Sci Adv. 2021 Apr 21;7(17). doi: 10.1126/sciadv.abe7006. Print 2021 Apr.
The recent report by Fan 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 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 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 mutants was based on assumptions that have not been confirmed by other methods. We conclude that Fan did not rigorously assess the method and that ProPerDP remains a reliable approach for analyses of protein per/polysulfidation.
范最近的报告称，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，范提出的主张没有得到他们数据的支持，而是基于方法上的缺陷。据了解，ProPerDP方法会产生含半胱氨酸和不含半胱氨酸的肽混合物。相反，范认为检测不含半胱氨酸的肽表明在非半胱氨酸残基处发生了非特异性烷基化。然而，如果真是这样，那么这些肽不会通过还原释放出来，因此不会在所报道的流程中作为产物出现。此外，作者使用突变体对ProPerDP进行的生物学评估是基于尚未被其他方法证实的假设。我们得出结论，范没有严格评估该方法，而ProPerDP仍然是分析蛋白质过硫化/多硫化的可靠方法。
