Combined phospho- and glycoproteome enrichment in nephrocalcinosis tissues of phytate-fed rats

Authors

  • TrangHuyen Tran,

    1. Lee Gil Ya Cancer and Diabetes Institute, Gachon University, Yeonsu-ku, Incheon, Republic of Korea
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    • These authors contributed equally to this work.

  • Jong-Moon Park,

    1. Lee Gil Ya Cancer and Diabetes Institute, Gachon University, Yeonsu-ku, Incheon, Republic of Korea
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    • These authors contributed equally to this work.

  • Ok-Hee Kim,

    1. Lee Gil Ya Cancer and Diabetes Institute, Gachon University, Yeonsu-ku, Incheon, Republic of Korea
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  • Bora Kim,

    1. Lee Gil Ya Cancer and Diabetes Institute, Gachon University, Yeonsu-ku, Incheon, Republic of Korea
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  • Do-young Choi,

    1. Department of Applied Chemistry, College of Applied Sciences, Kyung Hee University, Republic of Korea
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  • JeongHwa Lee,

    1. Department of Applied Chemistry, College of Applied Sciences, Kyung Hee University, Republic of Korea
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  • KwangPyo Kim,

    1. Department of Applied Chemistry, College of Applied Sciences, Kyung Hee University, Republic of Korea
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  • Byung-Chul Oh,

    Corresponding author
    1. Lee Gil Ya Cancer and Diabetes Institute, Gachon University, Yeonsu-ku, Incheon, Republic of Korea
    • Correspondence to: B.-C. Oh, Lee Gil Ya Cancer and Diabetes Institute, Gachon University, 7–45 Songdo-dong, Yeonsu-ku, Incheon 406–840, Republic of Korea.

      E-mail: bcoh@gachon.ac.kr

      Correspondence to: H. Lee, Gachon Institute of Pharmaceutical Sciences, College of Pharmacy, Gachon University, 7–45 Songdo-dong, Yeonsu-ku, Incheon 406–840, Republic of Korea.

      E-mail: hklee@gachon.ac.kr

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  • Hookeun Lee

    Corresponding author
    1. Lee Gil Ya Cancer and Diabetes Institute, Gachon University, Yeonsu-ku, Incheon, Republic of Korea
    2. College of Pharmacy, Gachon University, Yeonsu-ku, Incheon, Republic of Korea
    3. Gachon Medical Research Institute, Gil Medical Center, Incheon, Republic of Korea
    • Correspondence to: B.-C. Oh, Lee Gil Ya Cancer and Diabetes Institute, Gachon University, 7–45 Songdo-dong, Yeonsu-ku, Incheon 406–840, Republic of Korea.

      E-mail: bcoh@gachon.ac.kr

      Correspondence to: H. Lee, Gachon Institute of Pharmaceutical Sciences, College of Pharmacy, Gachon University, 7–45 Songdo-dong, Yeonsu-ku, Incheon 406–840, Republic of Korea.

      E-mail: hklee@gachon.ac.kr

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Abstract

RATIONALE

Protein post-translational modifications (PTMs) are directly involved in protein function and cellular activities. Among them, glycosylation and phosphorylation are particularly important modifications on proteins located at extracellular and intracellular domains, respectively. However, the combined detection using phospho- and glycoproteomics is limited mainly due to protocol differences.

METHODS

In this study, we developed a novel method for both phospho- and glycoproteome detection from a single sample batch, in which a titanium dioxide cartridge was used to capture the phosphoproteome, and the flow-through solution was processed for capturing N-linked glycopeptides using hydrazide resin.

RESULTS

By using 1 mg of protein from kidney tissue lysates from normal and diseased rats, we concurrently identified 437 glycosites/358 phosphosites and 468 glycosites/369 phosphosites in normal and disease kidneys, respectively, by liquid chromatography/tandem mass spectrometric analysis.

CONCLUSIONS

Compared with individual PTM analyses, the combined PTM analysis clearly provides more broad implications for PTMs related to the pathological status and discovery of biomarker candidates. Furthermore, the combined protocol thoroughly showed its advantages in enrichment efficiency and biological interpretation compared with current methods. Copyright © 2013 John Wiley & Sons, Ltd.

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