Evidence that the ProPerDP method is inadequate for protein persulfidation detection due to lack of specificity
Evidence that the ProPerDP method is inadequate for protein persulfidation detection due to lack of specificity
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DOI：
10.1126/sciadv.abb6477
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发表时间：
2020-08-01
期刊：
影响因子：
13.6
通讯作者：
Liu, Huaiwei
中科院分区：
文献类型：
--
作者：
Fan, Kaili;Chen, Zhigang;Liu, Huaiwei
蛋白质过硫化（蛋白质-SSH）是近年来在真核和原核细胞中发现的一种先前未识别的修饰类型。尽管已经鉴定出一些过硫化蛋白质，但从蛋白质组水平分析蛋白质过硫化仍然是一个巨大的挑战。 ProPerDP是最近在Science Advances上报道的一种过硫化检测方法。作者声称，这种方法可以特异性检测细胞裂解物的过硫化蛋白，且假阳性率较小；因此，它可用于蛋白质过硫化的蛋白质组水平分析。然而，当使用该方法进行大肠杆菌细胞裂解物分析时，我们发现假阳性命中率>90%。我们对该方法进行了系统研究，发现碘乙酰基-PEG2-生物素标签错误标记是导致特异性低的原因。我们得出的结论是，ProPerDP 方法完全不足以进行过硫化分析。之前基于 ProPerDP 方法的研究结果需要重新研究。
Protein persulfidation (protein-SSH) is a previously unidentified type of modification found in both eukaryotic and prokaryotic cells in recent years. Although a few persulfidated proteins have been identified, analyzing protein persulfidation from a proteomic level is still a big challenge. ProPerDP is a persulfidation detection method recently reported in Science Advances. The authors claimed that this method could specifically detect persulfidated proteins of cell lysate with minor false-positive hits; hence, it could be used for proteomic-level analysis of protein persulfidation. However, when using this method for Escherichia coli cell lysate analysis, we found that the percentage of false-positive hit was >90%.We performed a systematic study on this method and discovered that iodoacetyl-PEG2-biotin tag mislabeling is the reason causing this low specificity. We concluded that the ProPerDP method is completely inadequate for persulfidation analysis. The previous findings based on the ProPerDP method need to be reinvestigated.