Dissolving capability difference based sequential extraction: A versatile tool for in-depth membrane proteome analysis
文献类型：期刊论文
| 作者 | Fang, Fei1,2; Zhang, Yukui2; Zhao, Qun2; Li, Xiao2; Liang, Zhen2; Zhang, Lihua2 |
| 刊名 | ANALYTICA CHIMICA ACTA
 |
| 出版日期 | 2016-11-16 |
| 卷号 | 945页码:39-46 |
| 关键词 | Sequential Extraction Sample Preparation Membrane Proteome Profiling Missing Proteins |
| ISSN号 | 0003-2670 |
| DOI | 10.1016/j.aca.2016.09.032 |
| 文献子类 | Article |
| 英文摘要 | Profiling membrane proteins would facilitate revealing disease mechanism and discovering new drug targets as they play essential roles in cellular signaling, substrate transport, and cell adhesion. However, the analysis of membrane proteins still remains a challenge due to their high hydrophobicity, as well as the suppression effect of high abundant soluble proteins. In this work, to achieve a membrane proteome profiling, a sample preparation strategy based on sequential extraction at the protein level assisted by a range of extraction reagents with different dissolving capabilities, followed by nano-RPLC-ESI-MS/MS analysis was developed and applied for HeLa cell line analysis. It was found that with progressively harsher extraction reagents (i.e., 2 M NaCl, 4 M urea, 0.1 M Na2CO3, and 10% 1-dodecyl-3- methylimidazolium chloride (C12ImCl) performed, much more high hydrophobic proteins and low abundant proteins were identified. With our developed strategy, 5553 of the identified proteins (4419 gene products) were annotated to be membrane proteins and 2573 proteins (2183 gene products) have at least one transmembrane domain, to our best knowledge, which is the most comprehensive membrane proteome dataset for HeLa cell line. Notably, 110 of the identified membrane proteins were discovered in the "missing proteins" list referred to those in the neXtProt database. All above results indicated that our strategy has great potential to tackle the difficult but relevant task of identifying and profiling membrane proteins. (C) 2016 Elsevier B.V. All rights reserved. |
| WOS关键词 | CANCER-CELL-LINE ; SAMPLE PREPARATION ; DRUG TARGETS ; PROTEINS ; IDENTIFICATION ; SOLUBILIZATION ; EXPRESSION ; SEPARATION |
| WOS研究方向 | Chemistry |
| 语种 | 英语 |
| WOS记录号 | WOS:000388107500005 |
| 出版者 | ELSEVIER SCIENCE BV |
| 源URL | [http://cas-ir.dicp.ac.cn/handle/321008/169756]  |
| 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 |
| 通讯作者 | Zhang, Lihua |
| 作者单位 | 1.Chinese Acad Sci, Grad Sch, Beijing 100039, Peoples R China 2.Chinese Acad Sci, Natl Chromatog Res & Anal Ctr, Dalian Inst Chem Phys, Key Lab Separat Sci Analyt Chem, 457 Zhongshan Rd, Dalian 116023, Peoples R China |
| 推荐引用方式 GB/T 7714 | Fang, Fei,Zhang, Yukui,Zhao, Qun,et al. Dissolving capability difference based sequential extraction: A versatile tool for in-depth membrane proteome analysis[J]. ANALYTICA CHIMICA ACTA,2016,945:39-46. |
| APA | Fang, Fei,Zhang, Yukui,Zhao, Qun,Li, Xiao,Liang, Zhen,&Zhang, Lihua.(2016).Dissolving capability difference based sequential extraction: A versatile tool for in-depth membrane proteome analysis.ANALYTICA CHIMICA ACTA,945,39-46. |
| MLA | Fang, Fei,et al."Dissolving capability difference based sequential extraction: A versatile tool for in-depth membrane proteome analysis".ANALYTICA CHIMICA ACTA 945(2016):39-46. |
入库方式： OAI收割
来源：大连化学物理研究所
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