Comprehensive comparison of collision induced dissociation and electron transfer dissociation.
Comprehensive comparison of collision induced dissociation and electron transfer dissociation.
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DOI：
10.1021/ac8007785
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发表时间：
2008-07-01
影响因子：
7.4
通讯作者：
Pandey, Akhilesh
中科院分区：
文献类型：
--
作者：
Molina, Henrik;Matthiesen, Rune;Kandasamy, Kumaran;Pandey, Akhilesh
电子转移解离（ETD）是最近引入的质谱技术，已被证明是一个很好的工具，用于阐明不稳定的翻译后修饰，如磷酸化和丝氨酸和苏氨酸残基的O-GlcNAc酰化。然而，与已经研究了几十年的碰撞诱导解离（CID）不同，基于ETD的碎片化的复杂性尚未得到牢固的建立或系统的解决。在该分析中，我们系统地比较了大多数不含此类不稳定修饰的肽的CID和ETD片段化模式。使用标准的48种蛋白质混合物，我们能够测量实验的假阳性率，并且还评估了大量肽以详细比较CID和ETD片段化模式。对来自标准蛋白质和复杂蛋白质样品的约19000种肽的分析显示：（i）CID比ETD多鉴定50%的肽;（ii）ETD导致氨基酸序列覆盖率比CID增加约20%;以及（iii）CID和ETD片段化的组合将平均胰蛋白酶肽的序列覆盖率增加至92%。有趣的是，我们的分析显示，近60%的ETD鉴定的肽带有两个正电荷，这与普遍接受的情况形成鲜明对比。我们还提出了一种新的策略，用于自动验证肽分配的基础上识别的肽连续CID和ETD片段在交替模式。
Electron transfer dissociation (ETD) is a recently introduced mass spectrometric technique which has proven to be an excellent tool for the elucidation of labile post-translational modifications such as phosphorylation and O-GlcNAcylation of serine and threonine residues. However, unlike collision induced dissociation (CID), which has been studied for decades, the intricacies of ETD-based fragmentation have not yet been firmly established or systematically addressed. In this analysis, we have systematically compared the CID and ETD fragmentation patterns for the large majority of the peptides that do not contain such labile modifications. Using a standard 48 protein mix, we were able to measure false-positive rates for the experiments and also assess a large number of peptides for a detailed comparison of CID and ETD fragmentation pattern. Analysis of ∼19 000 peptides derived from both standard proteins and complex protein samples revealed that (i) CID identified 50% more peptides than ETD; (ii) ETD resulted in ∼20% increase in amino acid sequence coverage over CID; and (iii) combining CID and ETD fragmentation increased the sequence coverage for an average tryptic peptide to 92%. Interestingly, our analysis revealed that nearly 60% of all ETD-identified peptides carried two positive charges, which is in sharp contrast to what has been generally accepted. We also present a novel strategy for automatic validation of peptide assignments based on identification of a peptide by consecutive CID and ETD fragmentation in an alternating mode.
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影响因子：
15
作者：
Breuker, K;Oh, HB;McLafferty, FW
通讯作者：
McLafferty, FW
DOI：
10.1073/pnas.0607084104
发表时间：
2007-02-13
影响因子：
11.1
作者：
Chi, An;Huttenhower, Curtis;Hunt, Donald F.
通讯作者：
Hunt, Donald F.
影响因子：
16.6
作者：
Savitski, Mikhail M.;Kjeldsen, Frank;Zubarev, Roman A.
通讯作者：
Zubarev, Roman A.
DOI：
10.1073/pnas.0405549101
发表时间：
2004-09-14
影响因子：
11.1
作者：
Olsen, JV;Mann, M
通讯作者：
Mann, M
DOI：
10.1016/j.jasms.2005.08.019
发表时间：
2006-01-01
影响因子：
3.2
作者：
O'Connor, PB;Cournoyer, JJ;McLuckey, SA
通讯作者：
McLuckey, SA