Characterization of top-down ETD in a travelling-wave ion guide
Characterization of top-down ETD in a travelling-wave ion guide
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DOI：
10.1016/j.ymeth.2015.05.019
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发表时间：
2015-11-01
期刊：
影响因子：
4.8
通讯作者：
Sobott, Frank
中科院分区：
文献类型：
--
作者：
Lermyte, Frederik;Verschueren, Tim;Sobott, Frank
自上而下的测序方法对于蛋白质表征变得越来越相关，特别是电子捕获（ECD）和电子转移解离（ETD），其允许以最小的副反应进行广泛的主链切割。获得序列特异性片段的同时保持高阶结构的方面以及氘标记在HID交换中的位置的能力已经吸引了结构蛋白质组学领域的科学家的兴趣。最近，ETD也已结合离子迁移率的商业可用的四极/飞行时间仪器，这种实施铺平了道路，以新颖的结构研究和调查的ETD过程本身。在目前的工作中，我们研究了使用ETD的标准肽和蛋白质的片段化，并提供了一个详细的描述的参数控制的反应的时间和效率的效果。我们还强调了如何与电子转移后的离子迁移率分离的组合提供了扩展的分析优势，如分配片段的前体的特定的电荷减少状态。(C)2015 Elsevier Inc. All rights reserved.
Top-down sequencing methods are becoming increasingly relevant for protein characterization, in particular electron capture (ECD) and electron transfer dissociation (ETD) which allow for extensive backbone cleavage with minimal side reactions. The ability to obtain sequence-specific fragments while maintaining aspects of the higher-order structure, as well as the position of deuterium labels in HID exchange, has attracted interest from scientists in the field of structural proteomics. Recently, ETD has also been combined with ion mobility on commercially available quadrupole/time-of-flight instruments, and this implementation paves the way to novel structural studies and investigation of the ETD process itself. In the current work, we investigate the use of ETD for fragmentation of standard peptides and proteins and provide a detailed description of the effect of the parameters controlling the time and efficiency of the reaction. We also highlight how the combination with ion mobility separation after electron transfer provides extended analytical benefits, such as assignment of fragments to a specific charge-reduced state of the precursor. (C) 2015 Elsevier Inc. All rights reserved.