Profiling Glutathionylome in CD38-Mediated Epithelial–Mesenchymal Transition
Profiling Glutathionylome in CD38-Mediated Epithelial–Mesenchymal Transition
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分析 CD38 介导的上皮-间质转化中的谷胱甘肽组
DOI：
10.1021/acs.jproteome.1c00893
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发表时间：
2022
影响因子：
4.4
通讯作者：
Haiteng Deng
中科院分区：
文献类型：
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作者：
Yingying Ma;Songbiao Zhu;Meiqi Yi;Wenhao Zhang;Yuanyuan Xue;Xiaohui Liu;Haiteng Deng
蛋白质S-谷胱甘肽化是一种重要的翻译后修饰，调节各种细胞过程。然而，谷胱甘肽在上皮间质转化（EMT），胚胎发育，伤口愈合，癌的进展和转移的关键细胞过程中的变化，尚未得到充分的表征。我们以前的研究表明，CD 38过表达降低了细胞的烟酰胺腺嘌呤二核苷酸（NAD+）水平，并导致细胞发生EMT。在本研究中，我们设计了一个表达谷胱甘肽合成酶（GS）突变体的细胞系统，该突变体催化叠氮基丙氨酸形成谷胱甘肽类似物，以描述CD 38过表达细胞中谷胱甘肽组的变化。我们鉴定了1298种谷胱甘肽化蛋白，并揭示了谷胱甘肽化水平改变的蛋白质参与EMT相关通路，包括上皮粘附连接、肌动蛋白细胞骨架和整合素信号传导。此外，谷胱甘肽水平的15-羟前列腺素脱氢酶（15-PGDH）的CD 38过表达细胞增加。我们进一步证明了15-PGDH中Cys 63残基的谷胱甘肽化导致酶活性降低，该酶活性可以通过增加前列腺素E2（PGE 2）来促进EMT。综上所述，这些结果表明，可点击谷胱甘肽是用于谷胱甘肽组分析的有效探针，并且Cys 63上的15-PGDH的谷胱甘肽化抑制其促进EMT的酶活性。
Protein S-glutathionylation is an important posttranslational modification that regulates various cellular processes. However, changes in glutathionylome in epithelial–mesenchymal transition (EMT), a crucial cellular process for embryonic development, wound healing, and carcinoma progression and metastasis, have not been fully characterized. Our previous study revealed that CD38 overexpression decreased cellular nicotinamide adenine dinucleotide (NAD+) levels and caused cells to undergo EMT. In the present study, we engineered a cell system in which the glutathione synthetase (GS) mutant was expressed that catalyzed the formation of a glutathione analogue from azido-alanine to profile changes of glutathionylome in CD38-overexpressing cells. We identified 1298 glutathionylated proteins and revealed that proteins with changed glutathionylation levels involved in EMT associated pathways including epithelial adherens junction, actin cytoskeleton, and integrin signaling. Moreover, the glutathionylation level of 15-hydroxyprostaglandin dehydrogenase (15-PGDH) was increased in CD38-overexpressing cells. We further demonstrated that glutathionylation of Cys63 residue in 15-PGDH led to decreased enzymatic activity that could promote EMT by increasing prostaglandin E2(PGE2). Taken together, these results indicate that the clickable glutathione is an effective probe for glutathionylome profiling, and glutathionylation of 15-PGDH on Cys63 inhibits its enzymatic activity to promote EMT.