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Development of Online pH Gradient-Eluted Strong Cation Exchange Nanoelectrospray-Tandem Mass Spectrometry for Proteomic Analysis Facilitating Basic and Histidine-Containing Peptides Identification

文献类型：期刊论文

作者Xu, Jingjing2,3,4; Gao, Jing1,2,3; Yu, Chengli2,3,4; He, Han2,3; Yang, Yiming2; Figeys, Daniel1,5,6; Zhou, Hu1,2,3
刊名ANALYTICAL CHEMISTRY
出版日期2016-01-05
卷号88期号:1页码:583-591
ISSN号0003-2700
DOI10.1021/acs.analchem.5b04000
文献子类Article
英文摘要A novel one-dimensional online pH gradient-eluted strong cation exchange-nanoelectrospray ionization-tandem mass spectrometry (SCX-nano-ESI-MS/MS) method was developed for protein identification and tested with a mixture of six standard proteins, total lysate of HuH7 and N2a cells, as well as membrane fraction of N2a cells. This method utilized an online nanoflow SCX column in a nano-LC system coupled with a nanoelectrospray high-resolution mass spectrometer. Protein digests were separated on a nanoflow SCX column with a pH gradient and directly introduced into a mass spectrometer through nanoelectrospray ionization. More than five thousand unique peptides were identified in each 90 min LC-MS/MS run using 500 nanogram of protein digest either from total cell lysate or from membrane fraction. The unique peptide overlap between online strong cation exchange nano-ESI-MS/MS (SCXLC-MS/MS) and reverse phase nano-ESI-MS/MS (RPLC-MS/MS) is only <= 30%, which indicated these two methods were complementary to each other. The correlation coefficient of retention time and theoretical isoelectric point (pI) of identified peptides in SCXLC-MS/MS was higher than 0.4, which showed that peptides elution in SCXLC-MS/MS was dependent on their charge states. Furthermore, SCXLC-MS/MS showed identification capability for a higher proportion of basic peptides compared to the RPLC-MS/MS method, especially for histidine-containing peptides. Our SCXLC-MS/MS method is an excellent alternative method to the RPLC-MS/MS method for analysis of standard proteins, total cell and membrane proteomes.
WOS关键词LIQUID-CHROMATOGRAPHY ; PROTEIN IDENTIFICATION ; ANION-EXCHANGE ; RICH PEPTIDE ; SEPARATION ; QUANTIFICATION ; FRACTIONATION ; TRANSFECTION ; EXTRACTION ; GROWTH
资助项目National Natural Science Foundation of China[21375138] ; Ministry of Science and Technology of China[2013ZX09507001] ; Chinese Academy of Sciences[00000000] ; Innovation Project of Instrument and Equipment Function Development[YZ201542] ; Bureau of Goods, Chinese Academy of Sciences[00000000]
WOS研究方向Chemistry
语种英语
WOS记录号WOS:000367866100029
出版者AMER CHEMICAL SOC
源URL[http://119.78.100.183/handle/2S10ELR8/276181]  
专题分析化学研究室
通讯作者Figeys, Daniel
作者单位1.SIMMUOMICS Lab, Joint Res Lab Translat OMICS Shanghai Inst Mat Me, Beijing 100864, Peoples R China
2.Chinese Acad Sci, Shanghai Inst Mat Med, Dept Analyt Chem, Shanghai 201203, Peoples R China;
3.Chinese Acad Sci, Shanghai Inst Mat Med, CAS Key Lab Receptor Res, Shanghai 201203, Peoples R China;
4.Chinese Acad Sci, Univ Chinese Acad Sci, Beijing 100049, Peoples R China;
5.Univ Ottawa, Dept Biochem Microbiol & Immunol, Ottawa, ON K1H 8M5, Canada;
6.Univ Ottawa, Dept Chem & Biomol Sci, Ottawa, ON K1H 8M5, Canada;
推荐引用方式
GB/T 7714		 Xu, Jingjing,Gao, Jing,Yu, Chengli,et al. Development of Online pH Gradient-Eluted Strong Cation Exchange Nanoelectrospray-Tandem Mass Spectrometry for Proteomic Analysis Facilitating Basic and Histidine-Containing Peptides Identification[J]. ANALYTICAL CHEMISTRY,2016,88(1):583-591.
APA Xu, Jingjing.,Gao, Jing.,Yu, Chengli.,He, Han.,Yang, Yiming.,...&Zhou, Hu.(2016).Development of Online pH Gradient-Eluted Strong Cation Exchange Nanoelectrospray-Tandem Mass Spectrometry for Proteomic Analysis Facilitating Basic and Histidine-Containing Peptides Identification.ANALYTICAL CHEMISTRY,88(1),583-591.
MLA Xu, Jingjing,et al."Development of Online pH Gradient-Eluted Strong Cation Exchange Nanoelectrospray-Tandem Mass Spectrometry for Proteomic Analysis Facilitating Basic and Histidine-Containing Peptides Identification".ANALYTICAL CHEMISTRY 88.1(2016):583-591.

入库方式： OAI收割

来源：上海药物研究所

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