Combination of online enzyme digestion with stable isotope labeling for high-throughput quantitative proteome analysis
文献类型：期刊论文
| 作者 | Wang, Fangjun2; Wei, Xiaoluan2; Zhou, Hu1,3 ; Liu, Jing2; Figeys, Daniel3; Zou, Hanfa2
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| 刊名 | PROTEOMICS
 |
| 出版日期 | 2012-11 |
| 卷号 | 12期号:21页码:3129-3137 |
| 关键词 | Miscleavage Online enzyme digestion Quantitative proteome analysis Stable isotope labeling Technology |
| ISSN号 | 1615-9853 |
| DOI | 10.1002/pmic.201200162 |
| 文献子类 | Article |
| 英文摘要 | Various enzyme reactors and online enzyme digestion strategies have been developed in recent years. These reactors greatly enhanced the detection sensitivity and proteome coverage in qualitative proteomics. However, these devices have higher rates of miscleavage in protein digestion. Therefore, we investigated the effect of online enzyme digestion on the quantification accuracy of quantitative proteomics using chemical or metabolic isotope labeling approaches. The incomplete digestion would introduce some unexpected variations in comparative quantification when the samples are digested and then chemically isotope labeled in different aliquots. Even when identical protein aliquots are processed on these devices using post-digestion chemical isotope labeling and the CVs of the ratios controlled to less than 50% in replicate analyses, about 10% of the quantified proteins have a ratio greater than two-fold, whereas in theory the ratio is 1:1. Interestingly, the incomplete digestion with enzyme reactor is not a problem when metabolic isotope labeling samples were processed because the proteins are isotopically labeled in vivo prior to their simultaneous digestion within the reactor. Our results also demonstrated that both high quantification accuracy and high proteome coverage can be achieved in comparative proteome quantification using online enzyme digestion even when a limited amount of metabolic isotope labeling samples is used (1683 proteins comparatively quantified from 105 Hela cells). |
| WOS关键词 | TANDEM MASS-SPECTROMETRY ; PHASE LIQUID-CHROMATOGRAPHY ; TRYPSIN REACTOR ; COUPLED ONLINE ; CELL-CULTURE ; AMINO-ACIDS ; IDENTIFICATION ; PROTEOLYSIS ; CAPILLARY ; SILAC |
| 资助项目 | Creative Research Group Project by NSFC[21021004] ; China State Key Basic Research Program Grant[2012CB910601] ; China State Key Basic Research Program Grant[2012CB-910101] ; Analytical Method Innovation Program of MOST[2009IM031800] ; Analytical Method Innovation Program of MOST[2010IM030500] ; "Hundred Talent Young Scientist Program" by DICP[00000000] ; Natural Sciences and Engineering Research Council of Canada (NSERC)[00000000] |
| WOS研究方向 | Biochemistry & Molecular Biology |
| 语种 | 英语 |
| WOS记录号 | WOS:000310564100002 |
| 出版者 | WILEY-BLACKWELL |
| 源URL | [http://119.78.100.183/handle/2S10ELR8/277893]  |
| 专题 | 分析化学研究室 |
| 通讯作者 | Zou, Hanfa |
| 作者单位 | 1.Chinese Acad Sci, Shanghai Inst Mat Med, Shanghai 200031, Peoples R China 2.Chinese Acad Sci, Dalian Inst Chem Phys, CAS Key Lab Separat Sci Analyt Chem, Natl Chromatog Res & Anal Ctr, Dalian 116023, Peoples R China; 3.Univ Ottawa, Ottawa Inst Syst Biol, Ottawa, ON, Canada; |
| 推荐引用方式 GB/T 7714 | Wang, Fangjun,Wei, Xiaoluan,Zhou, Hu,et al. Combination of online enzyme digestion with stable isotope labeling for high-throughput quantitative proteome analysis[J]. PROTEOMICS,2012,12(21):3129-3137. |
| APA | Wang, Fangjun,Wei, Xiaoluan,Zhou, Hu,Liu, Jing,Figeys, Daniel,&Zou, Hanfa.(2012).Combination of online enzyme digestion with stable isotope labeling for high-throughput quantitative proteome analysis.PROTEOMICS,12(21),3129-3137. |
| MLA | Wang, Fangjun,et al."Combination of online enzyme digestion with stable isotope labeling for high-throughput quantitative proteome analysis".PROTEOMICS 12.21(2012):3129-3137. |
入库方式： OAI收割
来源：上海药物研究所
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