POMGNT1 Is Glycosylated by Mucin-Type <i>O</i>-Glycans

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Author(s)

    • Xin Xin Xin Xin
    • Molecular Glycobiology, Research Team for Mechanism of Aging, Tokyo Metropolitan Geriatric Hospital and Institute of Gerontology|Laboratory of Cancer Biology and Molecular Immunology, Graduate School of Pharmaceutical Sciences, The University of Tokyo
    • Akasaka-Manya Keiko Shinohara Yasuro
    • Laboratory of Medical and Functional Glycomics, Graduate School of Advanced Life Science, and Frontier Research Center for Post-Genome Science and Technology, Hokkaido University
    • Endo Tamao
    • Molecular Glycobiology, Research Team for Mechanism of Aging, Tokyo Metropolitan Geriatric Hospital and Institute of Gerontology
    • Akasaka-Manya Keiko
    • Molecular Glycobiology, Research Team for Mechanism of Aging, Tokyo Metropolitan Geriatric Hospital and Institute of Gerontology
    • Manya Hiroshi
    • Molecular Glycobiology, Research Team for Mechanism of Aging, Tokyo Metropolitan Geriatric Hospital and Institute of Gerontology
    • Furukawa Jun-ichi
    • Laboratory of Medical and Functional Glycomics, Graduate School of Advanced Life Science, and Frontier Research Center for Post-Genome Science and Technology, Hokkaido University
    • Kuwahara Naoyuki
    • Structural Biology Research Center, Photon Factory, Institute of Materials Structure Science, High Energy Accelerator Research Organization (KEK)
    • Okada Kazue
    • Laboratory of Medical and Functional Glycomics, Graduate School of Advanced Life Science, and Frontier Research Center for Post-Genome Science and Technology, Hokkaido University
    • Tsumoto Hiroki
    • Proteome Research, Research Team for Mechanism of Aging, Tokyo Metropolitan Geriatric Hospital and Institute of Gerontology
    • Higashi Nobuaki
    • Laboratory of Cancer Biology and Molecular Immunology, Graduate School of Pharmaceutical Sciences, The University of Tokyo
    • Kato Ryuichi
    • Structural Biology Research Center, Photon Factory, Institute of Materials Structure Science, High Energy Accelerator Research Organization (KEK)

Abstract

Protein <i>O</i>-linked mannose β1,2-<i>N</i>-acetylglucosaminyltransferase 1 (POMGNT1) is a Golgi glycosyltransferase that catalyzes the formation of the <i>N</i>-acetylglucosamine (GlcNAc) β1→2Man linkage of <i>O</i>-mannosyl glycan. POMGNT1 is not modified by <i>N</i>-glycans because there are no potential <i>N</i>-glycosylation sites; however, it is not clear whether POMGNT1 is modified by <i>O</i>-glycans. To determine whether POMGNT1 is <i>O</i>-glycosylated, we prepared recombinant human POMGNT1 from HEK293T cells. The recombinant POMGNT1 was recognized by <i>Sambucus sieboldiana</i> lectin (SSA), and sialidase digestion of POMGNT1 decreased SSA reactivity and enhanced the reactivity of <i>Arachis hypogaea</i> lectin (PNA). These results suggest that POMGNT1 is modified by a sialylated core-1 <i>O</i>-glycan. Next, we analyzed the structures of the <i>O</i>-glycans on POMGNT1 by β-elimination and pyrazolone-labeling methods in combination with mass spectrometry. We identified several mucin-type <i>O</i>-glycans containing (NeuAc)<sub>1</sub>(Hex)<sub>1</sub>(HexNAc)<sub>1</sub>, (NeuAc)<sub>2</sub>(Hex)<sub>1</sub>(HexNAc)<sub>1</sub>, and (NeuAc)<sub>2</sub>(Hex)<sub>2</sub>(HexNAc)<sub>2</sub>. To examine whether the <i>O</i>-glycans affect the functions and properties of POMGNT1, we compared glycosylated and non-glycosylated forms of recombinant sPOMGNT1 for their activity and surface hydrophobicity using the hydrophobic probe 1-anilino-8-naphthalene sulfonate (ANS). POMGNT1 activity and surface hydrophobicity were not affected by the presence or absence of <i>O</i>-glycans.

Journal

  • Biological and Pharmaceutical Bulletin

    Biological and Pharmaceutical Bulletin 38(9), 1389-1394, 2015

    The Pharmaceutical Society of Japan

Codes

  • NII Article ID (NAID)
    130005096979
  • NII NACSIS-CAT ID (NCID)
    AA10885497
  • Text Lang
    ENG
  • ISSN
    0918-6158
  • NDL Article ID
    026699387
  • NDL Call No.
    Z53-V41
  • Data Source
    NDL  J-STAGE 
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