CE-microreactor-CE-MS/MS for protein analysis
CE-microreactor-CE-MS/MS for protein analysis
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DOI：
10.1021/ac061638h
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发表时间：
2007-03-15
影响因子：
7.4
通讯作者：
Dovichi, Norman J.
中科院分区：
文献类型：
--
作者：
Schoenherr, Regine M.;Ye, Mingliang;Dovichi, Norman J.
我们提出了一种全自动自下而上的蛋白质表征方法的原理证明。蛋白质首先通过毛细管电泳分离。一个胃蛋白酶微反应器安装在毛细血管的远端。在反应器中形成的肽被转移到第二个毛细管，在那里它们被毛细管电泳分离并通过质谱分析表征。当一次消化产生的肽在第二根毛细管中分离时，下一个蛋白质部分在微反应器中进行消化。在第一维毛细管中的迁移时间是蛋白质的特征，而在第二维的迁移时间是肽的特征。二维分离的点容量为590。对细胞色素c和肌红蛋白的混合物进行MS/MS分析，细胞色素c的Mascot MOWSE得分为107，肌红蛋白的Mascot MOWSE得分为58。序列覆盖率分别为48%和22%。
We present a proof-of-principle for a fully automated bottom-up approach to protein characterization. Proteins are first separated by capillary electrophoresis. A pepsin microreactor is incorporated into the distal end of this capillary. Peptides formed in the reactor are transferred to a second capillary, where they are separated by capillary electrophoresis and characterized by mass spectrometry. While peptides generated from one digestion are being separated in the second capillary, the next protein fraction undergoes digestion in the microreactor. The migration time in the first dimension capillary is characteristic of the protein while migration time in the second dimension is characteristic of the peptide. Spot capacity for the two-dimensional separation is 590. A MS/MS analysis of a mixture of cytochrome c and myoglobin generated Mascot MOWSE scores of 107 for cytochrome c and 58 for myoglobin. The sequence coverages were 48% and 22%, respectively.