A novel approach for identification and characterization of glycoproteins using a hybrid linear ion trap/FT-ICR mass spectrometer
A novel approach for identification and characterization of glycoproteins using a hybrid linear ion trap/FT-ICR mass spectrometer
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DOI：
10.1016/j.jasms.2005.10.008
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发表时间：
2006-02-01
影响因子：
3.2
通讯作者：
Mulholland, JJ
中科院分区：
文献类型：
--
作者：
Peterman, SM;Mulholland, JJ
结合源碰撞诱导解离（CID）和串联质谱采集在伪MS 3实验中使用线性离子阱的结果在一个高度选择性和敏感的方法来识别糖肽洗脱蛋白质消化。增加的灵敏度部分归因于源CID的非选择性性质，其允许同时活化所有电荷状态和共洗脱糖型，从而产生更大的质荷比（m/z）204和/或366氧鎓离子的离子丰度。与单独的源CID不同，伪MS 3方法增加了选择性，同时通过消除线性离子阱中的氧鎓离子的串联质谱采集期间的化学噪声来提高灵敏度。在混合线性离子阱/傅立叶变换-离子回旋共振（FT-ICR）中进行实验使得随后的高分辨率/高质量准确度全扫描质谱（MS）和线性离子阱中的MS/MS的并行采集能够在伪MS 3扫描之后的2秒内直接完成，以在一个实验扫描循环中整理糖肽的鉴定和表征。使用组合的假MS 3、高分辨率MS和数据依赖性MS/MS事件对牛胎球蛋白消化物进行分析，结果鉴定出4种N-连接和2种O-连接的糖肽，分析前未对糖部分进行酶裂解或释放唾液酸。此外，在一次分析运行中鉴定了超过95%的总蛋白质序列。
Combining source collision-induced dissociation (CID) and tandem mass spectral acquisition in a pseudo-MS3 experiment using a linear ion trap results in a highly selective and sensitive approach to identifying glycopeptide elution from a protein digest. The increased sensitivity is partially attributed to the nonselective nature of source CID, which allows simultaneous activation of all charge states and coeluting glycoforms generating greater ion abundance for the mass-to-charge (m/z) 204 and/or 366 oxonium ions. Unlike source CID alone, a pseudo-MS3 approach adds selectivity while improving sensitivity by eliminating chemical noise during the tandem mass spectral acquisition of the oxonium ions in the linear ion trap. Performing the experiments in the hybrid linear ion trap/Fourier transform-ion cyclotron resonance (FT-ICR) enables subsequent high-resolution/high-mass accuracy full-scan mass spectra (MS) and parallel acquisition of MS/MS in the linear ion trap to be completed in 2 s directly following the pseudo-MS3 scan to collate identification and characterization of glycopeptides in one experimental scan cycle. Analysis of bovine fetuin digest using the combined pseudo-MS3, high-resolution MS, and data-dependent MS/MS events resulted in identification of four N-linked and two O-linked glycopeptides without enzymatic cleavage of the sugar moiety or release of the sialic acids before analysis. In addition, over 95% of the total protein sequence was identified in one analytical run.