DYn-2 Based Identification of Arabidopsis Sulfenomes
DYn-2 Based Identification of Arabidopsis Sulfenomes
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DOI：
10.1074/mcp.m114.046896
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发表时间：
2015-05-01
影响因子：
7
通讯作者：
Messens, Joris
中科院分区：
文献类型：
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作者：
Akter, Salma;Huang, Jingjing;Messens, Joris
鉴定应激细胞的亚砜化状态正在成为检测关键活性氧信号蛋白的一种战略方法。在这里，我们优化了一种利用基于二聚酮的DYn-2探针在过氧化氢（H2O2）胁迫下植物细胞中捕获半胱氨酸磺酸的体内方法。我们证明DYn-2特异性地以H2O2剂量和时间依赖的方式检测亚砜化事件。通过质谱分析，我们发现拟南芥细胞经过H2O2处理后，有226个磺化蛋白存在于细胞质中（123）；质体(68);线粒体(14);核(10);内质网，高尔基体和质膜(7)和过氧化物酶体(4)。其中，123个磺化蛋白在植物中进行半胱氨酸氧化翻译后修饰之前从未报道过。总而言之，通过这种DYn-2方法，我们已经确定了新的亚砜化蛋白，并对拟南芥亚砜体的位置有了初步的了解。
Identifying the sulfenylation state of stressed cells is emerging as a strategic approach for the detection of key reactive oxygen species signaling proteins. Here, we optimized an in vivo trapping method for cysteine sulfenic acids in hydrogen peroxide (H2O2) stressed plant cells using a dimedone based DYn-2 probe. We demonstrated that DYn-2 specifically detects sulfenylation events in an H2O2 dose- and time-dependent way. With mass spectrometry, we identified 226 sulfenylated proteins after H2O2 treatment of Arabidopsis cells, residing in the cytoplasm (123); plastid (68); mitochondria (14); nucleus (10); endoplasmic reticulum, Golgi and plasma membrane (7) and peroxisomes (4). Of these, 123 sulfenylated proteins have never been reported before to undergo cysteine oxidative post-translational modifications in plants. All in all, with this DYn-2 approach, we have identified new sulfenylated proteins, and gave a first glance on the locations of the sulfenomes of Arabidopsis thaliana.