2010年8月
Chemical dephosphorylation for identification of multiply phosphorylated peptides and phosphorylation site determination
RAPID COMMUNICATIONS IN MASS SPECTROMETRY
- ,
- ,
- ,
- 巻
- 24
- 号
- 15
- 開始ページ
- 2277
- 終了ページ
- 2282
- 記述言語
- 英語
- 掲載種別
- 研究論文(学術雑誌)
- DOI
- 10.1002/rcm.4627
- 出版者・発行元
- WILEY
We have developed a novel strategy to improve the efficiency of identification of multiply phosphorylated peptides isolated by hydroxy acid modified metal oxide chromatography (HAMMOC). This strategy consists of alkali-induced chemical dephosphorylation (beta-elimination reaction) of phosphopeptides isolated by HAMMOC prior to analysis by liquid chromatography/mass spectrometry (LC/MS). This approach identified 1.9-fold more multiply phosphorylated peptides than the conventional approach without beta-elimination from a digested mixture of three standard phosphoproteins. In addition, the accuracy of phosphorylation site determination in synthetic phosphopeptides was significantly improved. Finally, we applied this approach to a cell lysate. By combining this dephosphorylation approach with the conventional approach, we successfully identified 1649 unique phosphopeptides, including 325 multiply phosphorylated phosphopeptides, from 200 mu g of cultured Arabidopsis cells. These results indicate that chemical dephosphorylation prior to LC/MS analysis increases the efficiency of identification of multiply phosphorylated peptides, as well as the accuracy of phosphorylation site determination. Copyright (C) 2010 John Wiley & Sons, Ltd.
- リンク情報
- ID情報
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- DOI : 10.1002/rcm.4627
- ISSN : 0951-4198
- eISSN : 1097-0231
- PubMed ID : 20623713
- Web of Science ID : WOS:000280495700012