Ion Mobility Tandem Mass Spectrometry Enhances Performance of Bottom-up Proteomics
Ion Mobility Tandem Mass Spectrometry Enhances Performance of Bottom-up Proteomics
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DOI：
10.1074/mcp.m114.041038
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发表时间：
2014-12-01
影响因子：
7
通讯作者：
Kuster, Bernhard
中科院分区：
文献类型：
--
作者：
Helm, Dominic;Vissers, Johannes P. C.;Kuster, Bernhard
在确定使用混合四极正交加速飞行时间仪器的串联质谱的灵敏度中的限制因素之一是正交离子注入系统的占空比。因此，飞行时间分析器仅收集所产生的碎片离子束的一部分。在这里，我们描述了一种方法，利用postfragmentation离子迁移谱的肽碎片离子结合迁移时间同步正交离子注入导致一个大大改善的占空比和随之而来的改善灵敏度高达10倍的自下而上的蛋白质组学实验。这使得能够在1天内鉴定7500个人蛋白质，并在LC-MS/MS时间的5小时内鉴定8600个磷酸化位点。该方法也被证明是强大的多重定量实验，使用串联质量标签的组蛋白脱乙酰酶与曲古抑菌素A的化学蛋白质组学相互作用分析举例说明。
One of the limiting factors in determining the sensitivity of tandem mass spectrometry using hybrid quadrupole orthogonal acceleration time-of-flight instruments is the duty cycle of the orthogonal ion injection system. As a consequence, only a fraction of the generated fragment ion beam is collected by the time-of-flight analyzer. Here we describe a method utilizing postfragmentation ion mobility spectrometry of peptide fragment ions in conjunction with mobility time synchronized orthogonal ion injection leading to a substantially improved duty cycle and a concomitant improvement in sensitivity of up to 10-fold for bottom-up proteomic experiments. This enabled the identification of 7500 human proteins within 1 day and 8600 phosphorylation sites within 5 h of LC-MS/MS time. The method also proved powerful for multiplexed quantification experiments using tandem mass tags exemplified by the chemoproteomic interaction analysis of histone deacetylases with Trichostatin A.