| 纳升级电喷雾离子源-四极杆-飞行时间质谱中磷酸化肽的离子抑制作用 |
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| 引用本文： | 邹瑶,姜武辉,邹丽娟,李秀玲,梁鑫淼. 纳升级电喷雾离子源-四极杆-飞行时间质谱中磷酸化肽的离子抑制作用[J]. 色谱, 2013, 31(4): 367-371. DOI: 10.3724/SP.J.1123.2012.11024 |
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| 作者姓名： | 邹瑶 姜武辉 邹丽娟 李秀玲 梁鑫淼 |
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| 作者单位： | 1. 大连医科大学附属第二医院, 辽宁 大连 116023; 2. 中国科学院大连化学物理研究所, 中国科学院分离分析化学重点实验室, 辽宁 大连 116023; 3. 辽宁宇洁环保咨询有限公司, 辽宁 沈阳 110000 |
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| 基金项目： | “十二五”科技支撑计划项目(2012BAD33B03);国家自然科学基金项目(81171486) |
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| 摘 要： | 目前磷酸化蛋白质组学研究中的主要技术是蛋白质酶解产生的磷酸化肽的质谱检测。但是实际样品中的磷酸化肽(特别是多磷酸化肽)很难被检测到。其原因普遍认为是由于质谱检测时,非磷酸化肽抑制磷酸化肽。但也有认为非磷酸化肽对磷酸化肽没有抑制作用。另外磷酸化肽之间是否存在离子抑制作用还没有报道。本文采用相同氨基酸序列的标准磷酸化肽和非磷酸化肽,将其单独和混合进行质谱检测,通过对比混合前后磷酸化肽的信号强度,证明了非磷酸化肽对磷酸化肽有离子抑制作用；单磷酸化肽对二磷酸化肽有一定的抑制作用,但不太显著;单磷酸化肽对三磷酸化肽、二磷酸化肽对三磷酸化肽均有显著的离子抑制作用。该研究为今后单磷酸化肽和多磷酸化肽的分段富集和检测提供了有力的证明。
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| 关 键 词： | 离子化抑制 磷酸化肽 质谱 |
| 收稿时间： | 2012-11-28 |
Ionization suppression effect of phosphopeptides in nano-electrospray ionization quadrupole time-of-flight mass spectrometry |
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| ZOU Yao,JIANG Wuhui,ZOU Lijuan,LI Xiuling,LIANG Xinmiao. Ionization suppression effect of phosphopeptides in nano-electrospray ionization quadrupole time-of-flight mass spectrometry[J]. Chinese journal of chromatography, 2013, 31(4): 367-371. DOI: 10.3724/SP.J.1123.2012.11024 |
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| Authors: | ZOU Yao JIANG Wuhui ZOU Lijuan LI Xiuling LIANG Xinmiao |
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| Affiliation: | 1. The Second Affiliated Hospital of Dalian Medical University, Dalian 116023, China; 2. Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China; 3. Liaoning Yujie Environmental Consulting Limited Company, Shenyang 110000, China |
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| Abstract: | Protein phosphorylation is an important post-translational modification (PTM), and involves in many cell activities and disease processes. Nowadays, mass spectrometry (MS) is a powerful tool in peptide-based proteomics. But phosphopeptides, especially multiply phosphorylated peptides, are difficult to be detected by MS. One opinion is that the presence of unphosphorylated peptides lead to the ionization suppression of phosphopeptides. But contrary opinion exists. Moreover, the suppression effect caused by different types of phosphopeptides hasn’t been studied yet. In this study, a set of synthetic standard phosphopeptides (singly, doubly and triply phosphorylated peptides) with the same amino acid sequence, and the unphosphorylated peptides were injected into mass spectrometer with or without mixing. Through the ratios of signal intensities of the phosphopeptides before and after mixing, we confirmed that the signals of phosphopeptides were suppressed by those of the unphosphorylated peptides. In addition, the signals of the doubly phosphorylated peptides were slightly suppressed by those of the singly phosphorylated peptides with the same amino acid sequence. The signals of the triply phosphorylated peptides were notably suppressed by the singly and doubly phosphorylated peptides with the same amino acid sequence. Therefore, the singly and multiply phosphorylated peptides should be fractionated and enriched before mass spectrometry analysis. |
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| Keywords: | phosphopeptides mass spectrometry (MS) ionization suppression |
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