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

Research output: Contribution to journalArticle

10 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

Fingerprint

Phosphopeptides
Liquid chromatography
Mass spectrometry
Peptides
Phosphorylation
Phosphoproteins
Caseins
Muscle
Cells

ASJC Scopus subject areas

  • Analytical Chemistry
  • Spectroscopy
  • Organic Chemistry

Cite this

Sequential Fe3O4/TiO2 enrichment for phosphopeptide analysis by liquid chromatography/tandem mass spectrometry. / Choi, Sunkyu; Kim, Jaeyoon; Cho, Kun; Park, Gunwook; Yoon, Jong Hyuk; Park, Sehoon; Yoo, Jong Shin; Ryu, Sung Ho; Kim, Young Hwan; Kim, Jeongkwon.

In: Rapid Communications in Mass Spectrometry, Vol. 24, No. 10, 30.05.2010, p. 1467-1474.

Research output: Contribution to journalArticle

Choi, Sunkyu ; Kim, Jaeyoon ; Cho, Kun ; Park, Gunwook ; Yoon, Jong Hyuk ; Park, Sehoon ; Yoo, Jong Shin ; Ryu, Sung Ho ; Kim, Young Hwan ; Kim, Jeongkwon. / Sequential Fe3O4/TiO2 enrichment for phosphopeptide analysis by liquid chromatography/tandem mass spectrometry. In: Rapid Communications in Mass Spectrometry. 2010 ; Vol. 24, No. 10. pp. 1467-1474.
@article{720fe0955a9147468f4168c1e66f4d9a,
title = "Sequential Fe3O4/TiO2 enrichment for phosphopeptide analysis by liquid chromatography/tandem mass spectrometry",
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.",
author = "Sunkyu Choi and Jaeyoon Kim and Kun Cho and Gunwook Park and Yoon, {Jong Hyuk} and Sehoon Park and Yoo, {Jong Shin} and Ryu, {Sung Ho} and Kim, {Young Hwan} and Jeongkwon Kim",
year = "2010",
month = "5",
day = "30",
doi = "10.1002/rcm.4541",
language = "English",
volume = "24",
pages = "1467--1474",
journal = "Rapid Communications in Mass Spectrometry",
issn = "0951-4198",
publisher = "John Wiley and Sons Ltd",
number = "10",

}

TY - JOUR

T1 - Sequential Fe3O4/TiO2 enrichment for phosphopeptide analysis by liquid chromatography/tandem mass spectrometry

AU - Choi, Sunkyu

AU - Kim, Jaeyoon

AU - Cho, Kun

AU - Park, Gunwook

AU - Yoon, Jong Hyuk

AU - Park, Sehoon

AU - Yoo, Jong Shin

AU - Ryu, Sung Ho

AU - Kim, Young Hwan

AU - Kim, Jeongkwon

PY - 2010/5/30

Y1 - 2010/5/30

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=77951884001&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=77951884001&partnerID=8YFLogxK

U2 - 10.1002/rcm.4541

DO - 10.1002/rcm.4541

M3 - Article

C2 - 20411586

AN - SCOPUS:77951884001

VL - 24

SP - 1467

EP - 1474

JO - Rapid Communications in Mass Spectrometry

JF - Rapid Communications in Mass Spectrometry

SN - 0951-4198

IS - 10

ER -