The initial phases of neuronal differentiation are key to neuronal function. A particularly informative model to study these initial phases are retinoic acid-stimulated SH-SY5Y cells. Although these progressions are associated with redox-sensitive processes, it is largely undefined how the cellular proteome underpins redox dynamics and the management of reactive oxygen species. Here, we map the global cysteine-based redox landscape of SH-SY5Y cells using quantitative redox proteomics. We find evidence that redox alterations occurred early in differentiation and affect the expression of neuronal marker proteins and the extension of neurites. The spatiotemporal analysis of reactive oxygen species suggests a NOX2-dependent peak in cytoplasmic superoxide anions/hydrogen peroxide generation 2 h after retinoic acid stimulation. At the same time point, 241 out of 275 proteins with an altered cysteine redox state are reversibly oxidized in response to retinoic acid. Our analyses pinpoint redox alterations of proteins involved in the retinoic acid homeostasis and cytoskeletal dynamics.

中文翻译：

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支持视黄酸诱导的神经元分化的半胱氨酸氧化的蛋白质组学景观。

神经元分化的初始阶段是神经元功能的关键。研究这些初始阶段的一个特别有用的模型是视黄酸刺激的 SH-SY5Y 细胞。尽管这些进展与氧化还原敏感过程有关，但细胞蛋白质组如何支持氧化还原动力学和活性氧的管理在很大程度上是不确定的。在这里，我们使用定量氧化还原蛋白质组学绘制了 SH-SY5Y 细胞的全球基于半胱氨​​酸的氧化还原景观。我们发现有证据表明氧化还原改变在分化早期发生并影响神经元标记蛋白的表达和神经突的延伸。活性氧的时空分析表明，在视黄酸刺激后 2 小时，细胞质超氧阴离子/过氧化氢生成中出现 NOX2 依赖性峰。在同一时间点，在 275 种半胱氨酸氧化还原状态改变的蛋白质中，有 241 种被视黄酸可逆地氧化。我们的分析确定了参与视黄酸稳态和细胞骨架动力学的蛋白质的氧化还原变化。