Bottom-Up Two-Dimensional Electron-Capture Dissociation Mass Spectrometry of Calmodulin
Bottom-Up Two-Dimensional Electron-Capture Dissociation Mass Spectrometry of Calmodulin
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DOI：
10.1007/s13361-017-1812-y
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发表时间：
2018-01-01
影响因子：
3.2
通讯作者：
O'Connor, Peter B.
中科院分区：
文献类型：
--
作者：
Floris, Federico;van Agthoven, Maria A.;O'Connor, Peter B.
二维质谱法（2D MS）是一种串联质谱法技术，其通过在碎片化之前调制ICR池中的离子回旋频率，允许混合物中的所有前体的数据独立碎片化，而无需预先分离。它作为一种分析技术的力量已被证明，特别是蛋白质组学。最近，1D和2D MS之间的比较研究已经进行了使用红外多光子解离（IRMPD）钙调蛋白（CaM），突出了自上而下（TDP）和自下而上的蛋白质组学（BUP）的技术的能力。这项工作的目标是扩大这项研究的钙调素使用电子捕获解离（ECD）2D MS作为一个单一的互补BUP实验，以提高切割覆盖率的蛋白质分析。通过将BUP 2D ECD MS的结果添加到CaM的2D IRMPD MS分析中，总切割覆盖率从近似40%增加到近似68%。
Two-dimensional mass spectrometry (2D MS) is a tandem mass spectrometry technique that allows data-independent fragmentation of all precursors in a mixture without previous isolation, through modulation of the ion cyclotron frequency in the ICR-cell prior to fragmentation. Its power as an analytical technique has been proven particularly for proteomics. Recently, a comparison study between 1D and 2D MS has been performed using infrared multiphoton dissociation (IRMPD) on calmodulin (CaM), highlighting the capabilities of the technique in both top-down (TDP) and bottom-up proteomics (BUP). The goal of this work is to expand this study on CaM using electron-capture dissociation (ECD) 2D MS as a single complementary BUP experiment in order to enhance the cleavage coverage of the protein under analysis. By adding the results of the BUP 2D ECD MS to the 2D IRMPD MS analysis of CaM, the total cleavage coverage increased from similar to 40% to similar to 68%.