Online Alkaline-pH Reverse Phase Nanoelectrospray-Tandem Mass Spectrometry Complements Conventional Low-pH Method for Global Proteomic Analysis
文献类型：期刊论文
| 作者 | Gao, Jing4; Wang, Yuqiu3,4; Zhang, Shu2; Yang, Zhicheng4; Zhu, Hongwen4; Luo, Xuanyang1,4; Lu, Dayun1,4; Gao, Qiang2; Zhou, Hu1,4
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| 刊名 | JOURNAL OF PROTEOME RESEARCH
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| 出版日期 | 2023-05-05 |
| 卷号 | 22期号:5页码:1446-1454 |
| 关键词 | online alkaline-pH reverse phase mass spectrometry proteomics amino acid variant |
| DOI | 10.1021/acs.jproteome.2c00765 |
| 文献子类 | Article |
| 英文摘要 | The global proteome analysis was limited by the identification of peptides with low abundance or specific physiochemical properties. Here, a one-dimensional online alkaline-pH reverse phase nanoelectrospray-tandem mass spectrometry (alkaline-pH-MS/MS) method was developed and optimized for global proteomic analysis. In this method, peptides were separated on a nanoflow C18 column with an alkaline-pH mobile phase (pH = 8.0) and directly injected into the mass spectrometer. The unique peptides overlapped between alkaline-pH-MS/MS and conventional online low-pH reverse phase nanoelectrospray-tandem mass spectrometry (low-pH-MS/MS) were as low as 45%, strongly indicating that these two methods were complementary to each other. In addition, alkaline-pH-MS/MS showed identification capacity for a higher proportion of peptides with negative grand average of hydropathy (GRAVY) or high isoelectric point (pI). Compared to low-pH-MS/MS, alkaline-pH-MS/MS enabled enrichment preference toward histidine-, lysine-, methionine-, and proline-containing peptides. The complementarity of alkaline-pH-MS/MS and low-pH-MS/MS was further demonstrated for the analysis of tryptic digests from 15 intrahepatic cholangiocarcinoma (iCCA) cell lines. The alternating 60 min alkaline-pH-MS/MS plus 60 min low-pH-MS/MS method outperformed the conventional 120 min low-pH-MS/MS method in both the identification of amino acid variants and protein groups. Therefore, we established the alkaline-pH-MS/MS method as a simple, competitive, alternative method to low-pH-MS/MS for global proteomic analysis. |
| WOS关键词 | FRACTION CONCATENATION ; LIQUID-CHROMATOGRAPHY ; GRADIENT ; SENSITIVITY ; RESOLUTION ; PEPTIDES ; DRAFT ; HPLC |
| WOS研究方向 | Biochemistry & Molecular Biology |
| 语种 | 英语 |
| WOS记录号 | WOS:000963396600001 |
| 出版者 | AMER CHEMICAL SOC |
| 源URL | [http://119.78.100.183/handle/2S10ELR8/309702]  |
| 专题 | 新药研究国家重点实验室 |
| 通讯作者 | Zhou, Hu |
| 作者单位 | 1.Univ Chinese Acad Sci, Beijing 100049, Peoples R China 2.Fudan Univ, Zhongshan Hosp, Liver Canc Inst, Dept Liver Surg & Transplantat,Key Lab Carcinogene, Shanghai 200032, Peoples R China; 3.East China Univ Sci & Technol, Sch Bioengn, Shanghai 200237, Peoples R China; 4.Chinese Acad Sci, Shanghai Inst Mat Media, Analyt Res Ctr Organ & Biol Mol, CAS Key Lab Receptor Res, Shanghai 201203, Peoples R China; |
| 推荐引用方式 GB/T 7714 | Gao, Jing,Wang, Yuqiu,Zhang, Shu,et al. Online Alkaline-pH Reverse Phase Nanoelectrospray-Tandem Mass Spectrometry Complements Conventional Low-pH Method for Global Proteomic Analysis[J]. JOURNAL OF PROTEOME RESEARCH,2023,22(5):1446-1454. |
| APA | Gao, Jing.,Wang, Yuqiu.,Zhang, Shu.,Yang, Zhicheng.,Zhu, Hongwen.,...&Zhou, Hu.(2023).Online Alkaline-pH Reverse Phase Nanoelectrospray-Tandem Mass Spectrometry Complements Conventional Low-pH Method for Global Proteomic Analysis.JOURNAL OF PROTEOME RESEARCH,22(5),1446-1454. |
| MLA | Gao, Jing,et al."Online Alkaline-pH Reverse Phase Nanoelectrospray-Tandem Mass Spectrometry Complements Conventional Low-pH Method for Global Proteomic Analysis".JOURNAL OF PROTEOME RESEARCH 22.5(2023):1446-1454. |
入库方式： OAI收割
来源：上海药物研究所
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