Kwon Oh Kwang, Kim Sun Ju, Lee You-Mie, Lee Young-Hoon, Bae Young-Seuk, Kim Jin Young, Peng Xiaojun, Cheng Zhongyi, Zhao Yingming, Lee Sangkyu
College of Pharmacy, Research Institute of Pharmaceutical Sciences, Kyungpook National University, Daegu, South Korea.
School of Life Sciences, KNU Creative BioResearch Group (BK21 plus program), Kyungpook National University, Daegu, Korea.
Proteomics. 2016 Jan;16(1):136-49. doi: 10.1002/pmic.201500017. Epub 2015 Dec 7.
The zebrafish (Danio rerio) is a popular animal model used for studies on vertebrate development and organogenesis. Recent research has shown a similarity of approximately 70% between the human and zebrafish genomes and about 84% of human disease-causing genes have common ancestry with that of the zebrafish genes. Zebrafish embryos have a number of desirable features, including transparency, a large size, and rapid embryogenesis. Protein phosphorylation is a well-known PTM, which can carry out various biological functions. Recent MS developments have enabled the study of global phosphorylation patterns by using MS-based proteomics coupled with titanium dioxide phosphopeptide enrichment. In the present study, we identified 3500 nonredundant phosphorylation sites on 2166 phosphoproteins and quantified 1564 phosphoproteins in developing embryos of zebrafish. The distribution of Ser/Thr/Tyr phosphorylation sites in zebrafish embryos was found to be 88.9, 10.2, and 0.9%, respectively. A potential kinase motif was predicted using Motif-X analysis, for 80% of the identified phosphorylation sites, with the proline-directed motif appearing most frequently, and 35 phosphorylation sites having the pSF motif. In addition, we created six phosphoprotein clusters based on their dynamic pattern during the development of zebrafish embryos. Here, we report the largest dataset of phosphoproteins in zebrafish embryos and our results can be used for further studies on phosphorylation sites or phosphoprotein dynamics in zebrafish embryos.
斑马鱼（Danio rerio）是一种常用于脊椎动物发育和器官发生研究的热门动物模型。最近的研究表明，人类和斑马鱼基因组之间的相似度约为70%，约84%的人类致病基因与斑马鱼基因有共同的祖先。斑马鱼胚胎具有许多理想的特征，包括透明度高、体型大以及胚胎发育迅速。蛋白质磷酸化是一种众所周知的翻译后修饰（PTM），它可以执行各种生物学功能。最近质谱技术的发展使得通过基于质谱的蛋白质组学结合二氧化钛磷酸肽富集来研究全局磷酸化模式成为可能。在本研究中，我们在斑马鱼发育胚胎中鉴定出2166个磷酸化蛋白质上的3500个非冗余磷酸化位点，并对1564个磷酸化蛋白质进行了定量。发现斑马鱼胚胎中丝氨酸/苏氨酸/酪氨酸磷酸化位点的分布分别为88.9%、10.2%和0.9%。使用Motif-X分析预测了80%已鉴定磷酸化位点的潜在激酶基序，脯氨酸导向基序出现频率最高，有35个磷酸化位点具有pSF基序。此外，我们根据斑马鱼胚胎发育过程中的动态模式创建了六个磷酸化蛋白质簇。在此，我们报告了斑马鱼胚胎中最大的磷酸化蛋白质数据集，我们的结果可用于进一步研究斑马鱼胚胎中的磷酸化位点或磷酸化蛋白质动态。
