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Highly specific enrichment of phosphopeptides by zirconium dioxide nanoparticles for phosphoproteome analysis
Author(s) -
Zhou Houjiang,
Tian Ruijun,
Ye Mingliang,
Xu Songyun,
Feng Shun,
Pan Chensong,
Jiang Xiaogang,
Li Xin,
Zou Hanfa
Publication year - 2007
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200600718
Subject(s) - zirconium dioxide , nanoparticle , chemistry , zirconium , nanotechnology , materials science , inorganic chemistry , composite material
Large‐scale characterization of phosphoproteins requires highly specific methods for the purification of phosphopeptides because of the low abundance of phosphoproteins and substoichiometry of phosphorylation. A phosphopeptide enrichment method using ZrO 2 nanoparticles is presented. The high specificity of this approach was demonstrated by the isolation of phosphopeptides from the digests of model phosphoproteins. The strong affinity of ZrO 2 nanoparticles to phosphopeptides enables the specific enrichment of phosphopeptides from a complex peptide mixture in which the abundance of phosphopeptides is two orders of magnitude lower than that of nonphosphopeptides. Superior selectivity of ZrO 2 nanoparticles for the enrichment of phosphorylated peptides than that of conventional immobilized metal affinity chromatography was observed. Femtomole phosphopeptides from digestion products could be enriched by ZrO 2 nanoparticles and can be well detected by MALDI mass spectrometric analysis. ZrO 2 nanoparticles were further applied to selectively isolate phosphopeptides from the tryptic digestion of mouse liver lysate for phosphoproteome analysis by nanoliter LC MS/MS (nano‐LC‐MS/MS) and MS/MS/MS. A total of 248 defining phosphorylation sites and 140 phosphorylated peptides were identified by manual validation using a series of rigid criteria.