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学科主题: 物理化学
题名: Development of a Highly Efficient 2-D System with a Serially Coupled Long Column and Its Application in Identification of Rat Brain Integral Membrane Proteins with Ionic Liquids-Assisted Solubilization and Digestion
作者: Tao, Dingyin1, 2;  Qiao, Xiaoqiang1, 2;  Sun, Liangliang1, 2;  Hou, Chunyan1, 2;  Gao, Liang1, 2;  Zhang, Lihua1;  Shan, Yichu1;  Liang, Zhen1;  Zhang, Yukui1
通讯作者: 张丽华
关键词: shotgun proteomics ;  multidimensional HPLC ;  integral membrane proteins ;  serially coupled column ;  ionic liquids
刊名: JOURNAL OF PROTEOME RESEARCH
发表日期: 2011-02-01
DOI: 10.1021/pr100893j
卷: 10, 期:2, 页:732-738
收录类别: SCI
文章类型: Article
部门归属: 18
项目归属: 1810
产权排名: 1,1
WOS标题词: Science & Technology ;  Life Sciences & Biomedicine
类目[WOS]: Biochemical Research Methods
摘要: Development of a Highly Efficient 2-D System with a Serially Coupled Long Column and Its Application in Identification of Rat Brain Integral Membrane Proteins with Ionic Liquids-Assisted Solubilization and Digestion
研究领域[WOS]: Biochemistry & Molecular Biology
英文摘要: Two dimensional high performance liquid chromatography-electrospray ionization tandem mass spectrometry (2D-HPLC-ESI-MS/MS) is one of the most powerful techniques for high resolution, efficiency, and throughput separation and identification of proteomes. For a bottom-up strategy-based proteome analysis, usually multistep salt elution was needed in the first dimension separation by SCX, to simplify the peptides for the further second dimensional separation by RPLC. Here, by using a 30 cm-long serially coupled long column (SCLC) in the second dimension, we reduced the salt steps of SCX from 13 to 5 to shorten the total analysis time. Compared to the commonly applied 2D-HPLC with over 10-step salt elution in SCX and microRPLC with a short column (SC), named as SC-2D, the peak capacity of 2D-HPLC with a SCLC column, named as SCLC-2D, was increased 3.3-folds while the analysis time was increased by only 1.17-folds. Therefore, the time-based protein identification efficiency was similar to 55 protein groups/h, nearly 2-fold of that for SC-2D (similar to 28 protein groups/h). With the further combination of assisted solubilization by ionic liquids and SCLC-2D, 608 integral membrane proteins (IMPs) (27.66% of the total 2198 proteins, FDR < 1%) were identified from rat brain, more than those obtaind by the traditional urea method (252 unique IMPs, occupying 17.03% of total 1480 proteins). All of these results demonstrate the promise of the developed technique for large-scale proteome analysis.
关键词[WOS]: SHOTGUN PROTEOMIC ANALYSIS ;  TANDEM MASS-SPECTROMETRY ;  MULTIDIMENSIONAL SEPARATIONS ;  PARKINSONS-DISEASE ;  CHROMATOGRAPHY ;  TECHNOLOGY ;  EXCHANGE ;  MUDPIT ;  TISSUE ;  LC
语种: 英语
WOS记录号: WOS:000286868500032
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/115317
Appears in Collections:中国科学院大连化学物理研究所_期刊论文

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作者单位: 1.Chinese Acad Sci, Dalian Inst Chem Phys, Natl Chromatog R&A Ctr, Key Lab Separat Sci Analyt Chem, Dalian 116023, Peoples R China
2.Chinese Acad Sci, Grad Sch, Beijing 100039, Peoples R China

Recommended Citation:
Tao, Dingyin,Qiao, Xiaoqiang,Sun, Liangliang,et al. Development of a Highly Efficient 2-D System with a Serially Coupled Long Column and Its Application in Identification of Rat Brain Integral Membrane Proteins with Ionic Liquids-Assisted Solubilization and Digestion[J]. JOURNAL OF PROTEOME RESEARCH,2011,10(2):732-738.
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