Evaluation of Front-End Higher Energy Collision-Induced Dissociation on a Benchtop Dual-Pressure Linear Ion Trap Mass Spectrometer for Shotgun Proteomics
Evaluation of Front-End Higher Energy Collision-Induced Dissociation on a Benchtop Dual-Pressure Linear Ion Trap Mass Spectrometer for Shotgun Proteomics
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DOI：
10.1021/ac203210a
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发表时间：
2012-02-07
影响因子：
7.4
通讯作者：
MacCoss, Michael J.
中科院分区：
文献类型：
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作者：
Bereman, Michael S.;Canterbury, Jesse D.;MacCoss, Michael J.
我们报告了前端高能碰撞诱导解离（fHCD）的台式双压线性离子阱上的实施。采用软件和硬件修改，详细描述如下，以允许分离的离子在仪器的中间多极（q 00）中经历与环境气体分子的碰撞。比较fHCD和共振激发碰撞诱导解离（RE-CID）在注射时间，扫描总数，效率，质量测量精度（MMA），独特的肽鉴定和光谱质量的不稳定的修饰肽的性能的结果。fHCD的效率约为RE-CID的23%，并且根据搜索算法，使用Mascot或Sequest搜索算法，fHCD从可溶性全细胞裂解物（秀丽隐杆线虫）中识别的肽分别比RE-CID多6.6%或少15%（q < 0.01）。fHCD为磷酸化和糖基化（O-GlcNAc）肽的分析提供了明显的优势，因为使用fHCD的光谱的平均互相关分数（XCorr）在统计学上大于（p < 0.05）使用RE-CID收集的光谱。
We report the implementation of front-end higher energy collision-induced dissociation (fHCD) on a benchtop dual-pressure linear ion trap. Software and hardware modifications were employed, described in detail vide-infra, to allow isolated ions to undergo collisions with ambient gas molecules in an intermediate multipole (q00) of the instrument. Results comparing the performance of fHCD and resonance excitation collision-induced dissociation (RE-CID) in terms of injection time, total number of scans, efficiency, mass measurement accuracy (MMA), unique peptide identifications, and spectral quality of labile modified peptides are presented. fHCD is approximately 23% as efficient as RE-CID, and depending on the search algorithm, it identifies 6.6% more or 15% less peptides (q < 0.01) from a soluble whole-cell lysate (Caenorhabditis elegans) than RE-CID using Mascot or Sequest search algorithms, respectively. fHCD offers a clear advantage for the analysis of phosphorylated and glycosylated (O-GlcNAc) peptides as the average cross-correlation score (XCorr) for spectra using fHCD was statistically greater (p < 0.05) than for spectra collected using RE-CID.