Online Alkaline-pH Reversed-Phase Nanoelectrospray-Tandem Mass Spectrometry Complements Traditional Phosphoproteomic Analysis via Influencing Charge State Distribution of Phosphopeptides
文献类型：期刊论文
| 作者 | Wang, Yuqiu6,7; Gao, Jing6; Xie, Wenfan5; Tang, Minchu2,6; Chen, Xin5; Chen, Lv4; Chen, Hongxu5; Yang, Zhicheng2,6; Gao, Qiang4; Liu, Yansheng1 |
| 刊名 | JOURNAL OF PROTEOME RESEARCH
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| 出版日期 | 2025-04-10 |
| 页码 | 11 |
| 关键词 | alkaline-pH-MS/MS phosphoproteomics chargestate mass spectrometry fractionation |
| ISSN号 | 1535-3893 |
| DOI | 10.1021/acs.jproteome.4c01091 |
| 英文摘要 | Phosphorylation (O-linked and N-linked) plays an important role in biological functions and cell signaling. Here, we employed a one-dimensional online alkaline-pH reversed-phase nanoelectrospray-tandem mass spectrometry (alkaline-pH-MS/MS) for the investigation of global phosphorylation. In this method, phosphopeptides were separated on a nanoflow C18 column with an alkaline-pH gradient and directly introduced to the mass spectrometer through nanoelectrospray ionization. Although the phosphosites and phosphopeptides identified by alkaline-pH-MS/MS were slightly lower than those of traditional online low-pH reversed-phase tandem MS (low-pH-MS/MS), these two methods were highly complementary to each other. This alkaline-pH-MS/MS may affect the actual polarity and CSD of phosphopeptides, consequently improving the identification of multiply phosphorylated peptides. Moreover, alkaline-pH-MS/MS was compatible with other peptide fractionation and phosphopeptide enrichment techniques, such as offline high-pH or low-pH reversed-phase liquid chromatography fractionation. The complementarity of alkaline-pH-MS/MS and low-pH-MS/MS was further demonstrated by the tandem mass tag (TMT)-based quantitative phosphoproteomic analysis of five pairs of hepatocellular carcinoma (HCC) tumors and normal adjacent tissues (NATs). Furthermore, unique information on significantly changed phosphosites was observed by alkaline-pH-MS/MS. This study provided an alternative and complementary tool for global analysis of both O- and N-phosphoproteome, which may be beneficial for the discovery of phosphoproteins with significant biological functions. |
| WOS关键词 | PHOSPHORYLATION ANALYSIS ; PROTEOGENOMIC CHARACTERIZATION ; HISTIDINE PHOSPHORYLATION ; IDENTIFICATION ; PEPTIDES ; RESOURCE ; GRADIENT |
| 资助项目 | National Key Research and Development Program[B2020-254(2)] ; National Key Research and Development Program[2020YFA0509000] ; National Key Research and Development Program[2022YFA1302902] ; Strategic Priority Research Program of the Chinese Academy of Sciences[XDB0830000] ; Shanghai Leading Talent Program of Eastern Talent Plan[LJ2023123] ; National Natural Science Foundation of China[22425703] ; National Natural Science Foundation of China[31800693] ; China Postdoctoral Science Foundation[GZB20230157] ; Shanghai Municipal Science and Technology Major Project ; Institutional Technology Service Center of Shanghai Institute of Materia Medica |
| WOS研究方向 | Biochemistry & Molecular Biology |
| 语种 | 英语 |
| WOS记录号 | WOS:001463385000001 |
| 出版者 | AMER CHEMICAL SOC |
| 源URL | [http://119.78.100.183/handle/2S10ELR8/316977]  |
| 专题 | 新药研究国家重点实验室 |
| 通讯作者 | Liu, Yansheng; Zhou, Hu |
| 作者单位 | 1.Yale Univ, Sch Med, Canc Biol Inst, West Haven, CT 06516 USA 2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China 3.Univ Chinese Acad Sci, Hangzhou Inst Adv Study, Sch Pharmaceut Sci & Technol, Hangzhou 310024, Peoples R China 4.Fudan Univ, Zhongshan Hosp, Liver Canc Inst, Minist Educ,Dept Liver Surg & Transplantat,Key Lab, Shanghai 200032, Peoples R China 5.Nanjing Univ Chinese Med, Sch Chinese Mat Med, Nanjing 210023, Peoples R China 6.Chinese Acad Sci, Shanghai Inst Mat Med, Dept Analyt Chem, State Key Lab Drug Res, Shanghai 201203, Peoples R China 7.Fudan Univ, Eye & ENT Hosp, Dept Otolaryngol, Shanghai 200031, Peoples R China |
| 推荐引用方式 GB/T 7714 | Wang, Yuqiu,Gao, Jing,Xie, Wenfan,et al. Online Alkaline-pH Reversed-Phase Nanoelectrospray-Tandem Mass Spectrometry Complements Traditional Phosphoproteomic Analysis via Influencing Charge State Distribution of Phosphopeptides[J]. JOURNAL OF PROTEOME RESEARCH,2025:11. |
| APA | Wang, Yuqiu.,Gao, Jing.,Xie, Wenfan.,Tang, Minchu.,Chen, Xin.,...&Zhou, Hu.(2025).Online Alkaline-pH Reversed-Phase Nanoelectrospray-Tandem Mass Spectrometry Complements Traditional Phosphoproteomic Analysis via Influencing Charge State Distribution of Phosphopeptides.JOURNAL OF PROTEOME RESEARCH,11. |
| MLA | Wang, Yuqiu,et al."Online Alkaline-pH Reversed-Phase Nanoelectrospray-Tandem Mass Spectrometry Complements Traditional Phosphoproteomic Analysis via Influencing Charge State Distribution of Phosphopeptides".JOURNAL OF PROTEOME RESEARCH (2025):11. |
入库方式： OAI收割
来源：上海药物研究所
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