Electron Transfer Dissociation in Conjunction with Collision Activation To Investigate the Drosophila melanogaster Phosphoproteome
Electron Transfer Dissociation in Conjunction with Collision Activation To Investigate the Drosophila melanogaster Phosphoproteome
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DOI：
10.1021/pr800834e
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发表时间：
2009-06-01
影响因子：
4.4
通讯作者：
Aebersold, Ruedi
中科院分区：
文献类型：
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作者：
Domon, Bruno;Bodenmiller, Bernd;Aebersold, Ruedi
基于蛋白质的可逆磷酸化，更好地理解细胞如何调节和适应环境是当前分子和系统生物学研究的关键问题。在这项研究中，一个先进的质谱为基础的方法，利用电子转移解离（ETD）技术结合CID使用线性离子阱质谱仪进行描述。该技术被应用，第一次，从果蝇Kc167细胞系中分离的磷酸化肽的鉴定。我们表明，该方法是特别有用的大磷酸肽的表征，包括那些具有多个磷酸化位点，作为广泛的系列的C'和Z* 片段离子进行了观察。最后，我们应用了一个定向串联质谱工作流程，使用包含列表来增加识别肽的数量。
Better understanding how cells are regulated and adapt to their environment based on the reversible phosphorylation of proteins is a key question of current molecular and systems biology research. In this study, an advanced mass spectrometry based approach leveraging the electron transfer dissociation (ETD) technique in combination with CID using a linear ion trap mass spectrometer is described. The technique was applied, for the first time, to the identification of phosphorylated peptides isolated from the Drosophila melanogaster Kc167 cell line. We demonstrate that the method is particularly useful for the characterization of large phosphopeptides, including those with multiple phosphorylation sites, as extensive series of c' and z* fragment-ions were observed. Finally, we have applied a directed tandem mass spectrometric workflow using inclusion lists to increase the number of identified peptides.