An effective method for de novo peptide sequencing based on phosphorylation strategy and mass spectrometry.
An effective method for de novo peptide sequencing based on phosphorylation strategy and mass spectrometry.
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DOI：
10.1016/j.talanta.2010.12.035
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发表时间：
2011-05
期刊：
影响因子：
6.1
通讯作者：
Dongmei Zhang;Hongxia Liu;Shusheng Zhang;Xiaolan Chen;Shangfu Li;Cunlong Zhang;Xiangming Hu;Kaishun Bi;Xiaohui Chen;Yuyang Jiang
中科院分区：
文献类型：
--
作者：
Dongmei Zhang;Hongxia Liu;Shusheng Zhang;Xiaolan Chen;Shangfu Li;Cunlong Zhang;Xiangming Hu;Kaishun Bi;Xiaohui Chen;Yuyang Jiang
本文提出了一种基于磷酸化策略和UPLC-MS /MS的有效多肽测序方法。将模型肽溶液与磷酸化溶液混合进行磷酸化反应。利用UPLC-MS /MS对优化后的斜坡碰撞能量模式下磷酸化肽进行分析和表征。结果表明，这种磷酸化方式显著增强了修饰肽光谱中a1和b系列离子的信号强度。它还可以有效地区分肽中的谷氨酰胺(Q)和赖氨酸(K)残基。通过分析胰蛋白酶消化的牛血清白蛋白，验证了该方法的可行性。数据表明，该方法可为蛋白质组学研究中的从头肽测序提供有用的工具。
An effective method for peptide sequencing based on phosphorylation strategy and UPLC–MS/MS is proposed in this report. A phosphorylation reaction was carried out by mixing model peptide solution with phosphorylation solution. UPLC–MS/MS was used to analyze and characterize the phosphorylated peptides in the optimized ramp collision energy mode. The results illustrated that this phosphorylation approach significantly strengthened the signal intensity of both a1and b series ions of the spectra of the modified peptides. It also can be used to effectively distinguish glutamine (Q) and lysine (K) residues in peptides. The feasibility of this approach was validated by analyzing the trypsin-digested BSA. Data suggested that this proposed method could be a useful tool for the de novo peptide sequencing in proteome research.