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Conformational Difference in HMGB 1 Proteins of Human Neutrophils and Lymphocytes Revealed by Epitope Mapping of a Monoclonal Antibody

  • Ito Ichiaki
    Department of Biological Science and Technology, Science University of Tokyo
  • Mitsuoka Nobuyoshi
    Department of Biological Science and Technology, Science University of Tokyo
  • Sobajima Junko
    Department of Rheumatology and Clinical Immunology, Kyoto University Graduate School of Medicine
  • Uesugi Hiroko
    Saiseikai Noe Hospital
  • Ozaki Shoichi
    Division of Rheumatology and Allergy, Department of Internal Medicine, St. Marianna University School of Medicine
  • Ohya Kazuhiko
    Department of Product Development, Medical and Biological Laboratories Company, Ltd.
  • Yoshida Michiteru
    Department of Biological Science and Technology, Science University of Tokyo

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Abstract

HMGB 1 and HMGB 2 are abundant nonhistone chromosomal proteins in eukaryotic organisms. Their respective primary sequences are highly conserved. Our previous studies showed that these proteins are novel autoantigens of anti-neutrophil cytoplasmic antibodies in sera from patients with ulcerative colitis (UC), rheumatic disease and autoimmune hepatitis (AIH). In the present paper, we showed that antiHMGB 1 and HMGB 2 antibodies in sera of patients with UC do not recognize HMGB 1 in neutrophils while they recognize the protein in lymphocytes. Anti-HMGB 2 mono-clonal antibody FBH 7, recognizing HMGB 1 in lymphocytes, showed a similar profile to the antibodies in the patients' sera. In order to elucidate the difference in immuno-reactivity to HMGB 1 between neutrophils and lymphocytes, we mapped the epitope for FBH 7 by means of several methods. The results showed that FBH 7 recognizes the intact conformation composed of 52-56 residues of HMGB 1 in lymphocytes. This suggested that HMGB 1 in neutrophils is conformationally changed in the epitope or the peripheral structure of the epitope from the protein in lymphocytes. The apparent conformational change of HMGB 1 between neutrophils and lymphocytes will be important for understanding the functional difference of HMGBl in these cells.

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