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学科主题: 物理化学
题名: Prefractionation and separation by C8 stationary phase: Effective strategies for integral membrane proteins analysis
作者: Zhao, Qun1, 2;  Sun, Liangliang1, 2;  Liang, Yu1, 2;  Wu, Qi1, 2;  Yuan, Huiming1;  Liang, Zhen1;  Zhang, Lihua1;  Zhan, Yukui1
通讯作者: 张丽华
关键词: Reverse phase stationary phase ;  Hydrophobic peptides ;  Integral membrane protein ;  Prefractionation ;  mu RPLC-ESI-MS/MS
刊名: TALANTA
发表日期: 2012-01-15
DOI: 10.1016/j.talanta.2011.11.035
卷: 88, 页:567-572
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 ;  Physical Sciences
类目[WOS]: Chemistry, Analytical
研究领域[WOS]: Chemistry
英文摘要: Analysis of integral membrane proteins (IMPS) presents great challenges due to their hydrophobic nature. Recently, much attention has been paid to improve the solubilization of IMPs. However, besides that, the separation of hydrophobic peptides with high recovery is also a dominating factor, but with rare report. Here, the prefractionation of the digests by reverse phase trap column during desalting was presented to efficiently decrease the complexity of samples, with the identified hydrophobic peptides and IMPs increased by more than 43%. Furthermore, the effect of C18 and C8 stationary phases on the separation of membrane protein digests was studied. A total of 301 proteins (536 peptides) with C18 stationary phase and 398 proteins (703 peptides) with C8 stationary phase were identified by mu RPLC-ESI-MS/MS using an LCQ instrument in duplicate runs, with false discovery rate (FDR) less than 5% at peptide level. In addition, with C8 stationary phase, the number of identified hydrophobic peptides and IMPs was obviously improved by 29% and 20%, respectively, compared with that identified by C18 stationary phase, indicating that the polarity of stationary phase has evident effect on the analysis of membrane protein digests. All these results show that the prefractionation by reverse phase trap column during desalting and the separation by C8 stationary phases could facilitate the efficient identification of IMPs. (C) 2011 Elsevier B.V. All rights reserved.
关键词[WOS]: TANDEM MASS-SPECTROMETRY ;  SHOTGUN PROTEOMIC ANALYSIS ;  LIQUID-CHROMATOGRAPHY ;  ELEVATED-TEMPERATURE ;  LC-MS/MS ;  DIGESTION ;  ENRICHMENT ;  BRAIN ;  IDENTIFICATION ;  SOLUBILIZATION
语种: 英语
WOS记录号: WOS:000301159400079
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/118330
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, Key Lab Separat Sci Analyt Chem, Natl Chromatog R&A Ctr, Dalian 116023, Peoples R China
2.Chinese Acad Sci, Grad Sch, Beijing 100039, Peoples R China

Recommended Citation:
Zhao, Qun,Sun, Liangliang,Liang, Yu,et al. Prefractionation and separation by C8 stationary phase: Effective strategies for integral membrane proteins analysis[J]. TALANTA,2012,88:567-572.
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