Comprehensive Profiling of the Rice Ubiquitome Reveals the Significance of Lysine Ubiquitination in Young Leaves
Comprehensive Profiling of the Rice Ubiquitome Reveals the Significance of Lysine Ubiquitination in Young Leaves
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DOI：
10.1021/pr5009724
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发表时间：
2015-05-01
影响因子：
4.4
通讯作者：
Wang, Guo-Liang
中科院分区：
文献类型：
--
作者：
Xie, Xin;Kang, Houxiang;Wang, Guo-Liang
蛋白质泛素化是一种重要的翻译后修饰，调节真核生物的发育、凋亡、对环境提示的反应和其他过程。虽然已经在水稻中发现了几种泛素化蛋白，但由于目前分析方法的局限性，大规模的仪式泛素组分析还没有报道。在这里，我们报道了第一个水稻泛素组，通过高灵敏的免疫亲和纯化和高分辨率的LC-MS/MS鉴定，我们在水稻叶细胞的464个蛋白质中鉴定了861个含二甘氨酸-赖氨酸的多肽。对泛素组的生物信息学分析确定了泛素化蛋白的多种细胞功能和不同的亚细胞定位，还揭示了水稻中可能的7个泛素化基序。与结合和催化活性相关的蛋白质被预测为赖氨酸泛素化的优先靶点。蛋白质相互作用网络和KEGG分析表明，水稻中广泛的信号和代谢途径受到蛋白质泛素化的调控。我们的结果证明了显著改进的方法在植物蛋白质组泛素化检测中的有效性。水稻叶片中464个泛素化蛋白的鉴定为分析这些泛素化相关蛋白的生理功能奠定了基础。
Protein ubiquitination is a major post-translational modification that regulates development, apoptosis; responses to environmental cues, and other processes in eukaryotes. Although several ubiquitinated proteins have been identified in rice, large-scale profiling of the rite ubiquitome has not been reported because of limitations in the current analytical methods. Here, we report the first rice ubiquitome, determined by combining highly sensitive immune affinity purification and high resolution LC-MS/MS. We identified 861 di-Gly-Lys-containing peptides in 464 proteins in rice leaf cells. Bioinformatic analyses of the ubiquitome identified a variety of cellular functions and diverse subcellular localizations for the ubiquitinated proteins, and also revealed seven putative ublquitination motifs in rice. Proteins related to binding and catalytic activity were predicted to be the preferential targets of lysine ubiquitination. A protein interaction network and KEGG analysis indicated that a wide range of signaling and metabolic pathways are modulated by protein ubiquitination in rice. Our results demonstrate the usefulness of the significantly improved method for assaying proteome-wide ubiquitination in plants. The identification of the 464 ubiquitinated proteins in rice leaves provides a foundation for the analysis of the physiological roles of these ubiquitination-related proteins.