Rapidly alternating transmission mode electron-transfer dissociation and collisional activation for the characterization of polypeptide ions
Rapidly alternating transmission mode electron-transfer dissociation and collisional activation for the characterization of polypeptide ions
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DOI：
10.1021/ac7022734
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发表时间：
2008-05-01
影响因子：
7.4
通讯作者：
McLuckey, Scott A.
中科院分区：
文献类型：
--
作者：
Han, Hongling;Xia, Yu;McLuckey, Scott A.
在四极杆/飞行时间串联质谱仪（QqTOF）的高压碰撞池（Q2）中，快速连续地进行阳离子传输/电子转移试剂阴离子存储模式电子转移离子/离子反应和束型碰撞激活多肽离子，电子转移试剂阴离子在其中积累。电子转移离解（ETD）和碰撞诱导离解（CID）实验的占空比都比离子捕获方法有所提高，因为ETD实验没有离散的离子存储和反应步骤，CID实验没有离散的离子存储步骤和频率调谐。对于这种技术，由于通过TOF分析仪进行质量分析，也获得了中等高的分辨率和质量精度。这种相对简单的方法已经用三带电荷的胰蛋白酶肽、三带电荷的胰蛋白酶磷酸肽和三带电荷的胰蛋白酶n链糖肽证明了。对于色氨酸，由于这两种解离方法提供的互补信息，序列鉴定比单一方法更确定。由于ETD和CID分离方法获得的互补信息，确定了肽的肽序列和翻译后修饰（PTM）位点。这种结合ETD和CID的方法对于表征糖肽特别有用，因为ETD产生关于肽序列和糖基化位点位置的信息，而CID提供关于聚糖结构的信息。
Cation transmission/electron-transfer reagent anion storage mode electron-transfer ion/ion reactions and beam-type collisional activation of the polypeptide ions are performed in rapid succession in the high-pressure collision cell (Q2) of a quadrupole/time-of-flight tandem mass spectrometer (QqTOF), where the electron-transfer reagent anions are accumulated. Duty cycles for both electron-transfer dissociation (ETD) and collision-induced dissociation (CID) experiments are improved relative to ion trapping approaches since there are no discrete ion storage and reaction steps for ETD experiments and no discrete ion storage step and frequency tuning for CID experiments. For this technique, moderately high resolution and mass accuracy are also obtained due to mass analysis via the TOF analyzer. This relatively simple approach has been demonstrated with a triply charged tryptic peptide, a triply charged tryptic phosphopeptide, and a triply charged tryptic N-linked glycopeptide. For the tryptic peptide, the sequence is identified with more certainty than would be available from a single method alone due to the complementary information provided by these two dissociation methods. Because of the complementary information derived from both ETD and CID dissociation methods, peptide sequence and post-translationAl modification (PTM) sites for the phosphopeptide are identified. This combined ETD and CID approach is particularly useful for characterizing glycopeptides because ETD generates information about both peptide sequence and locations of the glycosylation sites, whereas CID provides information about the glycan structure.