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 | |
Tao, Dingyin1,2; Qiao, Xiaoqiang1,2; Sun, Liangliang1,2; Hou, Chunyan1,2; Gao, Liang1,2; Zhang, Lihua1; Shan, Yichu1; Liang, Zhen1; Zhang, Yukui1 | |
刊名 | journal of proteome research |
2011-02-01 | |
卷号 | 10期号:2页码:732-738 |
关键词 | shotgun proteomics multidimensional HPLC integral membrane proteins serially coupled column ionic liquids |
ISSN号 | 待补充 |
通讯作者 | 张丽华 |
产权排序 | 1,1 |
中文摘要 | 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 |
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. |
学科主题 | 物理化学 |
WOS标题词 | science & technology ; life sciences & biomedicine |
类目[WOS] | biochemical research methods |
研究领域[WOS] | biochemistry & molecular biology |
关键词[WOS] | shotgun proteomic analysis ; tandem mass-spectrometry ; multidimensional separations ; parkinsons-disease ; chromatography ; technology ; exchange ; mudpit ; tissue ; lc |
收录类别 | SCI |
语种 | 英语 |
WOS记录号 | WOS:000286868500032 |
公开日期 | 2012-07-09 |
内容类型 | 期刊论文 |
源URL | [http://159.226.238.44/handle/321008/115317] |
专题 | 大连化学物理研究所_中国科学院大连化学物理研究所 |
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 |
推荐引用方式 GB/T 7714 | 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. |
APA | Tao, Dingyin.,Qiao, Xiaoqiang.,Sun, Liangliang.,Hou, Chunyan.,Gao, Liang.,...&Zhang, Yukui.(2011).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.journal of proteome research,10(2),732-738. |
MLA | Tao, Dingyin,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".journal of proteome research 10.2(2011):732-738. |
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