Comparative glycoproteomics based on lectins affinity capture of N-linked glycoproteins from human Chang liver cells and MHCC97-H cells

Authors

  • Zhibin Xu,

    1. Glycoconjugate Key Laboratory, Ministry of Health, Department of Biochemistry and Molecular Biology, Fudan University, Shanghai, P. R. China
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  • Xinwen Zhou,

    1. Department of Chemistry, Fudan University, Shanghai, P. R. China
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  • Haojie Lu,

    1. Department of Chemistry, Fudan University, Shanghai, P. R. China
    2. Institutes for Biomedical Sciences, Fudan University, Shanghai, P. R. China
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  • Nan Wu,

    1. Glycoconjugate Key Laboratory, Ministry of Health, Department of Biochemistry and Molecular Biology, Fudan University, Shanghai, P. R. China
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  • Hongbo Zhao,

    1. Glycoconjugate Key Laboratory, Ministry of Health, Department of Biochemistry and Molecular Biology, Fudan University, Shanghai, P. R. China
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  • Lineng Zhang,

    1. Glycoconjugate Key Laboratory, Ministry of Health, Department of Biochemistry and Molecular Biology, Fudan University, Shanghai, P. R. China
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  • Wen Zhang,

    1. Glycoconjugate Key Laboratory, Ministry of Health, Department of Biochemistry and Molecular Biology, Fudan University, Shanghai, P. R. China
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  • Yu Long Liang,

    1. Glycoconjugate Key Laboratory, Ministry of Health, Department of Biochemistry and Molecular Biology, Fudan University, Shanghai, P. R. China
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  • Liying Wang,

    1. Glycoconjugate Key Laboratory, Ministry of Health, Department of Biochemistry and Molecular Biology, Fudan University, Shanghai, P. R. China
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  • Yinkun Liu,

    1. Institutes for Biomedical Sciences, Fudan University, Shanghai, P. R. China
    2. Liver Cancer institute Zhong Shan Hospital, Fudan University, Shanghai, P. R. China
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  • Pengyuan Yang,

    1. Department of Chemistry, Fudan University, Shanghai, P. R. China
    2. Institutes for Biomedical Sciences, Fudan University, Shanghai, P. R. China
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  • Xiliang Zha Dr.

    Corresponding author
    1. Glycoconjugate Key Laboratory, Ministry of Health, Department of Biochemistry and Molecular Biology, Fudan University, Shanghai, P. R. China
    • Department of Biochemistry and Molecular Biology, Shanghai Medical College, Fudan University, 138 Yi Xue Yuan Road, Shanghai 200032, China Fax: +86-21-6417-9832
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Abstract

We present here an effective technique for the large-scale separation and identification of N-linked glycoproteins from Chang liver cells, the human normal liver cells. To enrich N-linked glycoproteins from the whole cells, a procedure containing the lysis of human liver cells, the solubilization of total proteins, lectin affinity chromatography including Concanavalin A and wheat germ agglutinin was established. Furthermore, captured N-linked glycoproteins were separated by 2-DE, and identified by MS and database searching. Finally, we found 63 N-glycoproteins in Chang liver cells. In addition, using the above method, we identified 7 remarkably up-regulated glycoproteins from MHCC97-H cells, highly metastatic liver cancer cells, compared to Chang liver cells. These up-regulated glycoproteins were associated with liver cancer and might be used as biomarkers for tumor diagnosis. Results showed that we established a high-throughput proteomic analysis for separating N-linked glycoproteins from human liver cells. This strategy greatly improved the glycoprotein analysis method associated with proteome-wide glycosylation changes related to liver cancer. Our work was part of the HUPO Human Liver Proteome Project (HLPP) studies and was supported by CHINA HUPO.

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