Improved Recovery and Identification of Membrane Proteins from Rat Hepatic Cells using a Centrifugal Proteomic Reactor
文献类型：期刊论文
| 作者 | Zhou, Hu1,3,5 ; Wang, Fangjun1,4,5; Wang, Yuwei1,5; Ning, Zhibin1,5; Hou, Weimin1,2,5; Wright, Theodore G.1,5; Sundaram, Meenakshi1,5; Zhong, Shumei1,5; Yao, Zemin1,5; Figeys, Daniel1,2,5
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| 刊名 | MOLECULAR & CELLULAR PROTEOMICS
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| 出版日期 | 2011-10 |
| 卷号 | 10期号:10 |
| ISSN号 | 1535-9476 |
| DOI | 10.1074/mcp.O111.008425 |
| 文献子类 | Article |
| 英文摘要 | Despite their importance in many biological processes, membrane proteins are underrepresented in proteomic analysis because of their poor solubility (hydrophobicity) and often low abundance. We describe a novel approach for the identification of plasma membrane proteins and intracellular microsomal proteins that combines membrane fractionation, a centrifugal proteomic reactor for streamlined protein extraction, protein digestion and fractionation by centrifugation, and high performance liquid chromatography-electrospray ionization-tandem MS. The performance of this approach was illustrated for the study of the proteome of ER and Golgi microsomal membranes in rat hepatic cells. The centrifugal proteomic reactor identified 945 plasma membrane proteins and 955 microsomal membrane proteins, of which 63 and 47% were predicted as bona fide membrane proteins, respectively. Among these proteins, >800 proteins were undetectable by the conventional in-gel digestion approach. The majority of the membrane proteins only identified by the centrifugal proteomic reactor were proteins with >= 2 transmembrane segments or proteins with high molecular mass (e.g. > 150 kDa) and hydrophobicity. The improved proteomic reactor allowed the detection of a group of endocytic and/or signaling receptor proteins on the plasma membrane, as well as apolipoproteins and glycerolipid synthesis enzymes that play a role in the assembly and secretion of apolipoprotein B100-containing very low density lipo-proteins. Thus, the centrifugal proteomic reactor offers a new analytical tool for structure and function studies of membrane proteins involved in lipid and lipoprotein metabolism. Molecular & Cellular Proteomics 10: 10.1074/mcp.O111.008425, 1-11, 2011. |
| WOS关键词 | LOW-DENSITY LIPOPROTEINS ; TRIGLYCERIDE TRANSFER PROTEIN ; B-CONTAINING LIPOPROTEINS ; TANDEM MASS-SPECTROMETRY ; LIVER PLASMA-MEMBRANE ; APOLIPOPROTEIN-B ; MCA-RH7777 CELLS ; LC-MS/MS ; APO-B ; SECRETION |
| 资助项目 | NSERC[00000000] ; J.-Louis Levesque Foundation[00000000] ; China-Canada Health Research Initiative[CCI 92211] ; Canadian Institute for Health Research[NMD 15486] |
| WOS研究方向 | Biochemistry & Molecular Biology |
| 语种 | 英语 |
| WOS记录号 | WOS:000295773800023 |
| 出版者 | AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC |
| 源URL | [http://119.78.100.183/handle/2S10ELR8/278390]  |
| 专题 | 分析化学研究室 |
| 通讯作者 | Figeys, Daniel |
| 作者单位 | 1.Univ Ottawa, Ottawa Inst Syst Biol, Ottawa, ON K1H 8M5, Canada; 2.Univ Ottawa, Dept Chem, Fac Sci, Ottawa, ON K1N 6N5, Canada; 3.Chinese Acad Sci, Shanghai Inst Mat Med, Shanghai 201203, Peoples R China; 4.Chinese Acad Sci, Dalian Inst Chem Phys, Key Lab Separat Sci Analyt Chem, Natl Chromatog Res & Anal Ctr, Dalian 116023, Peoples R China 5.Univ Ottawa, Dept Biochem Microbiol & Immunol, Fac Med, Ottawa, ON K1H 8M5, Canada; |
| 推荐引用方式 GB/T 7714 | Zhou, Hu,Wang, Fangjun,Wang, Yuwei,et al. Improved Recovery and Identification of Membrane Proteins from Rat Hepatic Cells using a Centrifugal Proteomic Reactor[J]. MOLECULAR & CELLULAR PROTEOMICS,2011,10(10). |
| APA | Zhou, Hu.,Wang, Fangjun.,Wang, Yuwei.,Ning, Zhibin.,Hou, Weimin.,...&Figeys, Daniel.(2011).Improved Recovery and Identification of Membrane Proteins from Rat Hepatic Cells using a Centrifugal Proteomic Reactor.MOLECULAR & CELLULAR PROTEOMICS,10(10). |
| MLA | Zhou, Hu,et al."Improved Recovery and Identification of Membrane Proteins from Rat Hepatic Cells using a Centrifugal Proteomic Reactor".MOLECULAR & CELLULAR PROTEOMICS 10.10(2011). |
入库方式： OAI收割
来源：上海药物研究所
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