Characterization of glycopeptides by combining collision-induced dissociation and electron-transfer dissociation mass spectrometry data
Characterization of glycopeptides by combining collision-induced dissociation and electron-transfer dissociation mass spectrometry data
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DOI：
10.1002/rcm.3850
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发表时间：
2009-01-01
影响因子：
2
通讯作者：
Novotny, Milos V.
中科院分区：
文献类型：
--
作者：
Alley, William R., Jr.;Mechref, Yehia;Novotny, Milos V.
糖肽的结构表征不容易通过碰撞诱导解离（CID）获得，这是由于聚糖部分的广泛断裂和肽骨架的最小断裂。在这项研究中，我们利用了电子转移解离（ETD）作为肽片段化的补充方法的潜力。模型糖蛋白，包括核糖核酸酶B，胎球蛋白，辣根过氧化物酶，和触珠蛋白，在这里使用。在ETD中，自由基阴离子将电子转移到肽骨架并诱导N-C α键的裂解。聚糖部分保留在肽骨架上，基本上不受ETD过程的影响。因此，ETD不仅允许鉴定糖肽的氨基酸序列，还允许明确分配其糖基化位点。当从两种裂解技术获得的数据相结合时，可以全面表征整个糖肽。这是通过能够在LC/MS/MS分析期间在CID和ETD之间动态交替的质谱仪实现的。这里用几种胰蛋白酶糖肽证明了这一点。版权所有（C）2008约翰威利父子有限公司
Structural characterization of a glycopeptide is not easily attained through collision-induced dissociation (CID), due to the extensive fragmentation of glycan moieties and minimal fragmentation of peptide backbones. In this study, we have exploited the potential of electron-transfer dissociation (ETD) as a complementary approach for peptide fragmentation. Model glycoproteins, including ribonuclease B, fetuin, horseradish peroxidase, and haptoglobin, were used here. In ETD, radical anions transfer an electron to the peptide backbone and induce cleavage of the N-C alpha bond. The glycan moiety is retained oil the peptide backbone, being largely unaffected by the ETD process. Accordingly, ETD allows not only the identification of the amino acid sequence of a glycopeptide, but also the unambiguous assignment of its glycosylation site. When data acquired from both fragmentation techniques are combined, it is possible to characterize comprehensively the entire glycopeptide. This is being achieved with a mass spectrometer capable of alternating between CID and ETD on-the-fly during an LC/MS/MS analysis. This is demonstrated here with several tryptic glycopeptides. Copyright (C) 2008 John Wiley & Sons, Ltd.