Department of Chemistry , Wayne State University , Detroit , Michigan 48202 , United States.
J Proteome Res. 2019 Apr 5;18(4):1806-1818. doi: 10.1021/acs.jproteome.8b00986. Epub 2019 Mar 11.
Reactive oxygen species (ROS) are important signaling molecules, but their overproduction is associated with many cardiovascular diseases, including cardiomyopathy. ROS induce various oxidative modifications, among which glutathionylation is one of the significant protein oxidations that occur under oxidative stress. Despite previous efforts, direct and site-specific identification of glutathionylated proteins in cardiomyocytes has been limited. In this report, we used a clickable glutathione approach in a HL-1 mouse cardiomyocyte cell line under exposure to hydrogen peroxide, finding 1763 glutathionylated peptides with specific Cys modification sites, which include many muscle-specific proteins. Bioinformatic and cluster analyses found 125 glutathionylated proteins, whose mutations or dysfunctions are associated with cardiomyopathy, many of which include sarcomeric structural and contractile proteins, chaperone, and other signaling or regulatory proteins. We further provide functional implication of glutathionylation for several identified proteins, including CSRP3/MLP and complex I, II, and III, by analyzing glutathionylated sites in their structures. Our report establishes a chemoselective method for direct identification of glutathionylated proteins and provides potential target proteins whose glutathionylation may contribute to muscle diseases.
活性氧（ROS）是重要的信号分子，但它们的过度产生与许多心血管疾病有关，包括心肌病。ROS 诱导各种氧化修饰，其中谷胱甘肽化是氧化应激下发生的重要蛋白质氧化之一。尽管之前已经做了很多努力，但在心肌细胞中直接和特异性地鉴定谷胱甘肽化蛋白仍然受到限制。在本报告中，我们在暴露于过氧化氢的 HL-1 小鼠心肌细胞系中使用了一种可点击的谷胱甘肽方法，发现了 1763 个具有特定 Cys 修饰位点的谷胱甘肽化肽，其中包括许多肌肉特异性蛋白。生物信息学和聚类分析发现了 125 个谷胱甘肽化蛋白，其突变或功能障碍与心肌病有关，其中许多包括肌节结构和收缩蛋白、伴侣蛋白以及其他信号或调节蛋白。我们通过分析其结构中的谷胱甘肽化位点，进一步提供了几种已鉴定蛋白（包括 CSRP3/MLP 和复合物 I、II 和 III）的谷胱甘肽化的功能意义。我们的报告建立了一种用于直接鉴定谷胱甘肽化蛋白的化学选择性方法，并提供了可能导致肌肉疾病的谷胱甘肽化潜在靶蛋白。
