High Concentration Trypsin Assisted Fast In-Gel Digestion for Phosphoproteome Analysis
文献类型：期刊论文
| 作者 | Liu Fang-Jie1,2; Ye Ming-Liang2; Pan Yan-Bo1,2; Zou Han-Fa2 |
| 刊名 | CHINESE JOURNAL OF ANALYTICAL CHEMISTRY
 |
| 出版日期 | 2015-10-01 |
| 卷号 | 43期号:10页码:1452-1458 |
| 关键词 | High Concentration Trypsin In-gel Digestion Phosphoproteome Analysis |
| ISSN号 | 0253-3820 |
| DOI | 10.1016/S1872-2040(15)60864-7 |
| 文献子类 | Article |
| 英文摘要 | Polyacrylamide gel electrophoresis (PAGE) is a powerful protein separation technology. Combined with mass spectrometry, it could identify thousands of proteins in proteomics analysis. However, the time-consuming procedure restricts its broad applications in proteomics study. In this work, it was found that high concentration of trypsin did not compromise subsequent phosphopeptide enrichment after in-gel digestion, but could promote the in-gel digestion. Hence, a new, fast and robust digestion method was established. Firstly, 50 mu g of HeLa cell protein was separated into 5 fractions by SDS-PAGE, and then the proteins were digested with high concentration trypsin for 30 min. Finally, the phosphopeptides were enriched by Ti-IMAC Tip. After liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis, about 2000 phosphorylation sites were identified in the experimental group, while less than 1500 phosphorylation sites were identified in control group. With the aid of high concentration of trypsin, in-gel digestion could be completed within only 30 min, and more phosphorylation site identifications and lower percentage of missed cleavages were acquired than those in control experiments. The experiment results demonstrated that high concentration of trypsin could not only accelerate the in-gel digestion, but also improve the phosphoproteome coverage. |
| WOS关键词 | ENZYMATIC DIGESTION ; MASS-SPECTROMETRY ; PROTEIN DIGESTION ; PROTEOMICS ; TECHNOLOGY |
| WOS研究方向 | Chemistry |
| 语种 | 英语 |
| WOS记录号 | WOS:000365738500002 |
| 出版者 | ELSEVIER SCIENCE INC |
| 源URL | [http://cas-ir.dicp.ac.cn/handle/321008/171606]  |
| 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 | Ye Ming-Liang |
| 作者单位 | 1.Chinese Acad Sci, Grad Sch, Beijing 100049, Peoples R China 2.Chinese Acad Sci, Dalian Inst Chem Phys, Key Lab Separat Sci Analyt Chem, Dalian 116023, Peoples R China |
| 推荐引用方式 GB/T 7714 | Liu Fang-Jie,Ye Ming-Liang,Pan Yan-Bo,et al. High Concentration Trypsin Assisted Fast In-Gel Digestion for Phosphoproteome Analysis[J]. CHINESE JOURNAL OF ANALYTICAL CHEMISTRY,2015,43(10):1452-1458. |
| APA | Liu Fang-Jie,Ye Ming-Liang,Pan Yan-Bo,&Zou Han-Fa.(2015).High Concentration Trypsin Assisted Fast In-Gel Digestion for Phosphoproteome Analysis.CHINESE JOURNAL OF ANALYTICAL CHEMISTRY,43(10),1452-1458. |
| MLA | Liu Fang-Jie,et al."High Concentration Trypsin Assisted Fast In-Gel Digestion for Phosphoproteome Analysis".CHINESE JOURNAL OF ANALYTICAL CHEMISTRY 43.10(2015):1452-1458. |
入库方式： OAI收割
来源：大连化学物理研究所
浏览0
下载0
收藏0
其他版本
除非特别说明，本系统中所有内容都受版权保护，并保留所有权利。