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Targeted Proteo-Glycomics Analysis of Sialyl Lewis X Antigen Expressing Glycoproteins Secreted by Human Hepatoma Cell Line

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Abstract

Sialyl Lewis X (SLEX) antigen, Neu5Acα2–3Galβ1–4 (Fucα1–3) GlcNAc-R, plays important roles in cell-to-cell interaction: for example, the E- and P-selectin-mediated influx of SLEX expressing leukocytes into inflamed areas. A human hepatocellular carcinoma cell line, HepG2 cells, was highly expressed SLEX on secreted glycoproteins and cell surface, in contrast with HuH-7 cells. We identified SLEX expressing glycoproteins in HepG2 cultured medium by two-dimensional polyacrylamide gel electrophoresis, followed by in gel digestion and peptide mass fingerprint using matrixassisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOFMS), including transferrin, α1-antitrypsin, α2-HS glycoprotein and β2-glycoprotein. We analyzed N-glycans of these glycoproteins by MALDI-TOFMS in combination with exoglycosidase digestion; our results indicate increases in poly-fucosylated and high-branched N-glycans. High α1,3-fucosylation in glycoproteins would be caused by increased expression of α1,3-fucosyltransferase activities in HepG2 cells.

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Authors and Affiliations

  1. Department of Clinical Chemistry, School of Pharmaceutical Sciences, Toho University, Miyama 2-2-1, Funabashi, Chiba, 274-8510, Japan

    Koji Higai, Kumiko Shibukawa & Kojiro Matsumoto

  2. Department of Disease Analysis, School of Pharmaceutical Sciences, Toho University, Miyama 2-2-1, Funabashi, Chiba, 274-8510, Japan

    Satoshi Muto

Authors
  1. Koji Higai
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  2. Kumiko Shibukawa
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  3. Satoshi Muto
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  4. Kojiro Matsumoto
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Correspondence to Koji Higai.

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Higai, K., Shibukawa, K., Muto, S. et al. Targeted Proteo-Glycomics Analysis of Sialyl Lewis X Antigen Expressing Glycoproteins Secreted by Human Hepatoma Cell Line. ANAL. SCI. 19, 85–92 (2003). https://doi.org/10.2116/analsci.19.85

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  • DOI: https://doi.org/10.2116/analsci.19.85

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