期刊
JOURNAL OF SEPARATION SCIENCE
卷 33, 期 13, 页码 1879-1887出版社
WILEY-V C H VERLAG GMBH
DOI: 10.1002/jssc.200900718
关键词
Fractionating; Human liver; Phosphoproteome; Strong anion-exchange; Trap column
资金
- National Natural Science Foundation of China [20675081, 20735004]
- China State Key Basic Research Program [2007CB914104]
- China High Technology Research Program [2006AA02A309]
- CAS [KJCX2.YW.HO9]
- DICP
It is one of the key issues to develop powerful fractionating method to increase the identification of the low-abundance phosphopeptides. In this study, a semi-online 2-D LC separation strategy based on three-step fractionation of the enriched peptides on strong anion-exchange trap column was developed. It was demonstrated that the sensitivity and phosphoproteome coverage obtained by this fractionating method with strong anion-exchange trap column is much higher than those by the conventional methods based on C18 trap column. In addition, when the same amount of sample was loaded, the number of identified phosphopeptides had increased 108%. Combination of this three-step fractionation method with RPLC-MS/MS analysis by 300 min RP-gradient separation was applied to phosphoproteome analysis of human liver proteins, and 853 unique phosphopeptides was positively identified from 500 mu g tryptic digest of human liver proteins. After three cycles' consecutive analyses, 1554 unique phosphopeptides and 1566 phosphorylated sites were totally identified from 735 phosphorylated proteins at a false discovery rate of < 1% in about 54h of analysis time.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据