論文

査読有り
2016年12月

High-Confidence Sequencing of Phosphopeptides by Electron Transfer Dissociation Mass Spectrometry Using Dinuclear Zinc(II) Complex

ANALYTICAL CHEMISTRY
  • Daiki Asakawa
  • ,
  • Issey Osaka

88
24
開始ページ
12393
終了ページ
12402
記述言語
英語
掲載種別
研究論文(学術雑誌)
DOI
10.1021/acs.analchem.6b03645
出版者・発行元
AMER CHEMICAL SOC

Phosphorylation is the most abundant protein modification, and tandem mass spectrometry (MS2) with electron transfer dissociation (ETD) has proven to be a promising method for phosphoproteomic applications owing to its ability to determine phosphorylation sites on proteins.. However, low precursor charge states hinder the ability to obtain useful information through peptide sequencing by ETD, and the presence of acidic phosphate groups contributes to a low charge state of peptide ions. In the present report, we used a dinudear zinc complex, (Zn2L)(3+) (L = alkoxide form of 1,3-bis[bis(pyridin-2-ylmethyl)amino]propan-2-ol) for electrospray ionization (ESI), followed by ETD-MS2 analysis. Since (Zn2L)(3+) selectively bound to phosphopeptide with addition of a positive charge per phosphate group, the use of (Zn2L)(3+) for ESI improved the ionization yield of phosphopeptides in phosphoprotein digest. Additionally, an increase in the charge state of phosphopeptides were observed by addition of (Zn2L)(3+), facilitating phosphopeptide sequencing by ETD-MS2. Since the binding between (Zn2L)(3+) and the phosphate group was retained during the ETD process, a comparison between the ETD mass spectra obtained using two dinudear zinc complex derivatives containing different zinc isotopes, namely ((Zn2L)-Zn-64)(3+) and ((Zn2V)-Zn-68)(3+); provided information about the number of phosphate groups in each fragment ion, allowing the phosphorylation site to be unambiguously determined. The details of the fragmentation processes of the (Zn2L)(3+)-phosphopeptide complex were investigated using a density functional theory calculation. As in the case of protonated peptides, ETD induced peptide backbone dissociation in the (Zn2L)(3+) phosphopeptide complex proceeded through an aminoketyl radical intermediate.

リンク情報
DOI
https://doi.org/10.1021/acs.analchem.6b03645
Web of Science
https://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=JSTA_CEL&SrcApp=J_Gate_JST&DestLinkType=FullRecord&KeyUT=WOS:000390621000059&DestApp=WOS_CPL
ID情報
  • DOI : 10.1021/acs.analchem.6b03645
  • ISSN : 0003-2700
  • eISSN : 1520-6882
  • Web of Science ID : WOS:000390621000059

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