Sequential Fe3O4/TiO2 enrichment for phosphopeptide analysis by liquid chromatography/tandem mass spectrometry

Sunkyu Choi, Jaeyoon Kim, Kun Cho, Gunwook Park, Jong Hyuk Yoon, Sehoon Park, Jong Shin Yoo, Sung Ho Ryu, Young Hwan Kim, Jeongkwon Kim

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11 Citations (Scopus)

Abstract

Protein phosphorylation regulates a wide range of cellular functions and is associated with signaling pathways in cells. Various strategies for enrichment of phosphoproteins or phosphopeptides have been developed. Here, we developed a novel sequential phosphopeptide enrichment method, using magnetic iron oxide (Fe3O4) and titanium dioxide (TiO2) particles, to detect mono- and multi-phosphorylated peptides. In the first step, phosphopeptides were captured on Fe3O4 particles. In a subsequent step, any residual phosphopeptides were captured on TiO2 particles. The particles were eluted and rinsed to yield phosphopeptide-enriched fractions that were combined and analyzed using liquid chromatography/tandem mass spectrometry (LC/MS/MS). The validity of this sequential Fe3O4/TiO2 enrichment strategy was demonstrated by the successful enrichment of bovine a-casein phosphopeptides. We then applied the sequential Fe3O4/TiO2 enrichment method to the analysis of phosphopeptides in L6 muscle cell lysates and successfully identified mono- and multi-phosphorylated peptides.

Original languageEnglish
Pages (from-to)1467-1474
Number of pages8
JournalRapid Communications in Mass Spectrometry
Volume24
Issue number10
DOIs
Publication statusPublished - 30 May 2010
Externally publishedYes

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ASJC Scopus subject areas

  • Analytical Chemistry
  • Spectroscopy
  • Organic Chemistry

Cite this

Choi, S., Kim, J., Cho, K., Park, G., Yoon, J. H., Park, S., Yoo, J. S., Ryu, S. H., Kim, Y. H., & Kim, J. (2010). Sequential Fe3O4/TiO2 enrichment for phosphopeptide analysis by liquid chromatography/tandem mass spectrometry. Rapid Communications in Mass Spectrometry, 24(10), 1467-1474. https://doi.org/10.1002/rcm.4541