Top-down proteomics with a quadrupole time-of-flight mass spectrometer and collision-induced dissociation
Top-down proteomics with a quadrupole time-of-flight mass spectrometer and collision-induced dissociation
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DOI：
10.1002/rcm.3925
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发表时间：
2009-03-01
影响因子：
2
通讯作者：
Damonte, Gianluca
中科院分区：
文献类型：
--
作者：
Armirotti, Andrea;Benatti, Umberto;Damonte, Gianluca
通过对仪器参数的轻微修改，我们证明了在四极杆飞行时间（qTOF）仪器上使用碰撞诱导解离（CID）串联质谱法可以获得令人满意的自上而下数据。蛋白质鉴定是通过N和c端序列标签和BLAST数据库搜索实现的。多重带电片段离子（主要是y型和b型）的精确质量测量补充了有限的裂解位点集，并提供了高度的序列覆盖率（90-100%）。翻译后的修改问题也可以解决。这种方法可能会帮助那些无法负担高分辨率仪器的质谱（MS）核心设施，从而在全球质谱社区扩大自上而下的蛋白质分析的好处。版权所有John Wiley & Sons， Ltd. 2009
With slight modifications of the instrumental parameters, we demonstrate that satisfactory top-down data can be obtained with collision-induced dissociation (CID) tandem mass spectrometry on a quadrupole time-of-flight (qTOF) instrument not originally designed for this purpose. Protein identification is achieved with both N- and C-terminal sequence tags and BLAST database searches. The accurate mass measurement of multiply charged fragment ions (mostly y and b-type) supplements the limited set of cleavage sites and provides a high degree of sequence coverage (90-100%). Post-translational modification issues can be addressed too. This approach might help those mass spectrometry (MS) core facilities that are not able to afford very high-resolution instruments, thus expanding the benefits of top-down protein analysis over the worldwide MS community. Copyright (C) 2009 John Wiley & Sons, Ltd.