Structural characterization of N-glycopeptides by matrix-dependent selective fragmentation of MALDI-TOF/TOF tandem mass spectrometry
Structural characterization of N-glycopeptides by matrix-dependent selective fragmentation of MALDI-TOF/TOF tandem mass spectrometry
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DOI：
10.1021/ac049294n
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发表时间：
2004-10-15
影响因子：
7.4
通讯作者：
Nishimura, SI
中科院分区：
文献类型：
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作者：
Kurogochi, M;Nishimura, SI
迄今为止，基质辅助激光解吸/电离飞行时间（MALDI-TOF）质谱技术已被证明是一种方便、快速的蛋白质鉴定和表征方法。然而，对于复杂碳水化合物和糖肽的一般MALDI-TOF质谱分析仍然需要特别考虑电离和不稳定碳水化合物部分的断裂特征。在本研究中，我们将一种高效实用的方法称为MALDI-TOF/TOF质谱的基质依赖性选择性碎片化（MDSF）技术，该技术可以对低聚糖和n -糖肽进行高度敏感和可靠的碎片化。将MDSF技术应用于TOF/TOF质谱分析的结果表明，与传统的后源衰减相比，可以获得灵敏度高达170倍的产物离子峰。研究还表明，MDSF可以通过对不同电子激发态的单电荷前体离子的控制裂解来产生所需的结构信息。在TOF/TOF质谱中，MDSF观察到的理想产物离子峰有利于复杂低聚糖衍生物的结构表征，包括不稳定的Neu5Ac和Fuc残基以及n -糖肽。
The matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) MS technique described to date has proven to be a convenient and rapid method for identification and characterizations of proteins. However, the general MALDI-TOF MS analysis of complex carbohydrates and glycopeptides still entails special consideration of ionization and the fragmentation characteristics of labile carbohydrate moieties. In this study, an efficient and practical method we termed the matrix-dependent selective fragmentation (MDSF) technique of MALDI-TOF/TOF MS, which allows highly sensitive and reliable fragmentation of oligosaccharides and N-glycopeptides. Results from application of the MDSF technique to TOF/TOF MS analysis demonstrated that in comparison to the conventional postsource decay up to 170 times more sensitive product ion peaks could be obtained. It was also suggested that MDSF generates desired structural information based on the controlled cleavage of the singly charged precursor ion with different electronic excited states made by this method. Ideal product ion peaks observed by MDSF in TOF/TOF MS facilitated structural characterization of complex oligosaccharide derivatives including unstable Neu5Ac and Fuc residues and N-glycopeptides.