Quantitative Profiling of Protein S-Glutathionylation Reveals Redox-Dependent Regulation of Macrophage Function during Nanoparticle-Induced Oxidative Stress.
Quantitative Profiling of Protein S-Glutathionylation Reveals Redox-Dependent Regulation of Macrophage Function during Nanoparticle-Induced Oxidative Stress.
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DOI：
10.1021/acsnano.5b05524
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发表时间：
2016-01-26
期刊：
影响因子：
17.1
通讯作者：
Qian WJ
中科院分区：
文献类型：
--
作者：
Duan J;Kodali VK;Gaffrey MJ;Guo J;Chu RK;Camp DG;Smith RD;Thrall BD;Qian WJ
工程纳米颗粒（ENPs）越来越多地用于商业和医疗应用；因此，了解它们潜在的不利影响是一个重要的社会问题。在此，我们研究了蛋白质s -谷胱甘肽化（SSG）作为ENPs改变巨噬细胞先天免疫功能的潜在调节机制，使用定量氧化还原蛋白质组学方法对SSG修饰进行位点特异性测量。选择三种大批量生产的ENPs （SiO2、Fe3O4和CoO）作为代表，分别诱导低、中、高倾向刺激细胞活性氧（ROS）和破坏巨噬细胞功能。SSG修饰突出了一组广泛的氧化还原敏感蛋白和特定的Cys残基，它们与细胞氧化还原应激的总体水平和巨噬细胞吞噬功能的损害（CoO > Fe3O4比SiO2）密切相关。此外，我们的数据揭示了ENPs诱导中度和高水平ROS对SSG易感性的通路特异性差异。调控内质网应激反应的蛋白质翻译和蛋白质稳定性的途径以及参与吞噬的蛋白质在ENPs诱导亚细胞毒性氧化还原应激时对SSG最敏感。在较高水平的氧化还原应激下，SSG修饰模式显示出降低的特异性和更广泛的途径，包括经典应激反应和与凋亡机制相关的线粒体能量学（如糖酵解）。glutaredoxin（一种逆转SSG修饰的主要酶）的RNA沉默也证实了SSG在巨噬细胞先天免疫功能调节中的重要作用。我们的研究结果为作为ROS传感器的蛋白质信号和途径提供了独特的见解，这些信号和途径可能促进细胞适应enp，而不是与不可逆细胞结果相关的enp诱导的细胞内氧化应激靶标。
Engineered nanoparticles (ENPs) are increasingly utilized for commercial and medical applications; thus, understanding their potential adverse effects is an important societal issue. Herein, we investigated protein S-glutathionylation (SSG) as an underlying regulatory mechanism by which ENPs may alter macrophage innate immune functions, using a quantitative redox proteomics approach for site-specific measurement of SSG modifications. Three high-volume production ENPs (SiO2, Fe3O4, and CoO) were selected as representatives which induce low, moderate, and high propensity, respectively, to stimulate cellular reactive oxygen species (ROS) and disrupt macrophage function. The SSG modifications identified highlighted a broad set of redox sensitive proteins and specific Cys residues which correlated well with the overall level of cellular redox stress and impairment of macrophage phagocytic function (CoO > Fe3O4 ≫ SiO2). Moreover, our data revealed pathway-specific differences in susceptibility to SSG between ENPs which induce moderate versus high levels of ROS. Pathways regulating protein translation and protein stability indicative of ER stress responses and proteins involved in phagocytosis were among the most sensitive to SSG in response to ENPs that induce subcytoxic levels of redox stress. At higher levels of redox stress, the pattern of SSG modifications displayed reduced specificity and a broader set pathways involving classical stress responses and mitochondrial energetics (e.g., glycolysis) associated with apoptotic mechanisms. An important role for SSG in regulation of macrophage innate immune function was also confirmed by RNA silencing of glutaredoxin, a major enzyme which reverses SSG modifications. Our results provide unique insights into the protein signatures and pathways that serve as ROS sensors and may facilitate cellular adaption to ENPs, versus intracellular targets of ENP-induced oxidative stress that are linked to irreversible cell outcomes.