Gas-phase ion isomer analysis reveals the mechanism of peptide sequence scrambling.
Gas-phase ion isomer analysis reveals the mechanism of peptide sequence scrambling.
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DOI：
10.1021/ac401578p
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发表时间：
2014-03-18
影响因子：
7.4
通讯作者：
Li, Lingjun
中科院分区：
文献类型：
--
作者：
Jia, Chenxi;Wu, Zhe;Lietz, Christopher B.;Liang, Zhidan;Cui, Qiang;Li, Lingjun
在基于质谱的气相裂解分析期间肽序列扰乱导致肽和蛋白质的错误鉴定。因此，有必要开发一种有效的方法来探测气相碎片离子异构体相关的序列打乱和潜在的碎片机制，这将有助于生物信息学算法的蛋白质组学研究的发展。在这里，我们报告的第一次使用电子转移解离（ETD）产生的诊断碎片离子探测气相肽碎片离子异构体的成分。结合离子迁移谱（IMS）和甲醛标记，这种新的策略，使定性和定量分析的b型碎片离子异构体。ETD碎裂产生指示前体离子异构体组分的诊断碎片离子，并且随后的B离子异构体的IMS分析提供了它们的定量和结构信息。三个代表性的B离子（b9，b10，和b33从三个不同的肽）的异构体组分准确地通过这种方法进行了分析。b9离子异构体的IMS分析显示这些结构之间的动态转换。此外，分子动力学模拟预测的理论漂移时间值与实验测量值吻合得很好。我们的研究结果有力地支持了通过B离子环化的肽序列扰乱机制，并提供了第一个实验证据来支持从分子前体离子到环状B离子的转化（M→ c B）途径在能量（或动力学）上是较少有利的。
Peptide sequence scrambling during mass spectrometry-based gas-phase fragmentation analysis causes misidentification of peptides and proteins. Thus, there is a need to develop an efficient approach to probing the gas-phase fragment ion isomers related to sequence scrambling and the underlying fragmentation mechanism, which will facilitate the development of bioinformatics algorithm for proteomics research. Herein, we report on the first use of electron transfer dissociation (ETD)-produced diagnostic fragment ions to probe the components of gas-phase peptide fragment ion isomers. In combination with ion mobility spectrometry (IMS) and formaldehyde labeling, this novel strategy enables qualitative and quantitative analysis of b-type fragment ion isomers. ETD fragmentation produced diagnostic fragment ions indicative of the precursor ion isomer components, and subsequent IMS analysis of b ion isomers provided their quantitative and structural information. The isomer components of three representative b ions (b9, b10, and b33 from three different peptides) were accurately profiled by this method. IMS analysis of the b9 ion isomers exhibited dynamic conversion among these structures. Furthermore, molecular dynamics simulation predicted theoretical drift time values which were in good agreement with experimentally measured values. Our results strongly support the mechanism of peptide sequence scrambling via b ion cyclization, and provide the first experimental evidence to support that the conversion from molecular precursor ion to cyclic b ion (M→cb) pathway is less energetically (or kinetically) favored.
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影响因子：
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作者：
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通讯作者：
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发表时间：
2008-12-01
影响因子：
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作者：
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通讯作者：
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