Phosphoproteome analysis of drosophila metanogaster embryos
Phosphoproteome analysis of drosophila metanogaster embryos
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DOI：
10.1021/pr700696a
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发表时间：
2008-04-01
影响因子：
4.4
通讯作者：
Gygi, Steven P.
中科院分区：
文献类型：
--
作者：
Zhai, Bo;Villen, Judit;Gygi, Steven P.
蛋白质磷酸化是大多数细胞过程和发育中的一个关键调节事件。基于质谱学的蛋白质组学为大规模鉴定和表征磷酸化位点提供了一个框架。在这里，我们使用了成熟的磷酸肽浓缩和鉴定策略，包括强阳离子交换层析、固定化金属亲和层析和高精度质谱仪相结合的方法来研究果蝇胚胎发育过程中的磷酸化。总共从2702个蛋白质中发现了13720个不同的磷酸化位点，估计在多肽水平上的假发现率(FDR)为0.63%。由于数据集的大小，使用Motif-X算法提取了新的和已知的磷酸化基序，包括那些代表潜在有序磷酸化事件的基序。
Protein phosphorylation is a key regulatory event in most cellular processes and development. Mass spectrometry-based proteomics provides a framework for the large-scale identification and characterization of phosphorylation sites. Here, we used a well-established phosphopeptide enrichment and identification strategy including the combination of strong cation exchange chromatography, immobilized metal affinity chromatography, and high-accuracy mass spectrometry instrumentation to study phosphorylation in developing Drosophila embryos. In total, 13 720 different phosphorylation sites were discovered from 2702 proteins with an estimated false-discovery rate (FDR) of 0.63% at the peptide level. Because of the large size of the data set, both novel and known phosphorylation motifs were extracted using the Motif-X algorithm, including those representative of potential ordered phosphorylation events.