Detection of N.EPSILON.-(hexanoyl)lysine in the tropomyosin 1 protein in N-methyl-N'-nitro-N-nitrosoguanidine-induced rat gastric cancer cells

• Okada Hitomi

  Molecular Gastroenterology and Hepatology, Kyoto Prefectural University of Medicine

• Naito Yuji

  Molecular Gastroenterology and Hepatology, Kyoto Prefectural University of Medicine

• Takagi Tomohisa

  Molecular Gastroenterology and Hepatology, Kyoto Prefectural University of Medicine

• Takaoka Megumi

  Molecular Gastroenterology and Hepatology, Kyoto Prefectural University of Medicine

• Oya-Ito Tomoko

  Molecular Gastroenterology and Hepatology, Kyoto Prefectural University of Medicine

• Fukumoto Kohei

  Molecular Gastroenterology and Hepatology, Kyoto Prefectural University of Medicine

• Uchiyama Kazuhiko

  Molecular Gastroenterology and Hepatology, Kyoto Prefectural University of Medicine

• Handa Osamu

  Molecular Gastroenterology and Hepatology, Kyoto Prefectural University of Medicine

• Kokura Satoshi

  Molecular Gastroenterology and Hepatology, Kyoto Prefectural University of Medicine

• Nagano Yumiko

  Department of Gastroenterology, Institute of Clinical Medicine, University of Tsukuba

• Matsui Hirofumi

  Department of Gastroenterology, Institute of Clinical Medicine, University of Tsukuba

• Kato Yoji

  School of Human Science and Environment, University of Hyogo

• Osawa Toshihiko

  Department of Nutritional Science, Aichi Gakuin University

• Yoshikawa Toshikazu

  Molecular Gastroenterology and Hepatology, Kyoto Prefectural University of Medicine

Bibliographic Information

Other Title

• Detection of Nε-(hexanoyl)lysine in the tropomyosin 1 protein in N-methyl-N'-nitro-N-nitrosoguanidine-induced rat gastric cancer cells

Abstract

N^ε-(Hexanoyl)lysine, formed by the reaction of lysine with n-6 lipid hydroperoxide, is a lipid peroxidation marker during the initial stage of oxidative stress. The aim of the present study is to indentify N^ε-(hexanoyl)lysine-modified proteins in neoplastic transformed gastric mucosal cells by N-methyl-N’-nitro-N-nitrosoguanidine, and to compare the levels of these proteins between gastric mucosal cells and normal gastric cells. Much greater fluorescence of 2-[6-(4'-hydroxy)phenoxyl-3H-xanthen-3-on-9-yl]benzoic acid, an index of the intracellular levels of reactive oxygen species, was observed for gastric mucosal cells compared to normal gastric cells. N^ε-(Hexanoyl)lysine-modified proteins were detected by SDS-PAGE or two-dimensional electrophoresis and Western blotting using anti-N^ε-(hexanoyl)lysine polyclonal antibody, and a protein band of between 30–40 kDa was clearly increased in gastric mucosal cells compared to normal gastric cells. Two N^ε-(hexanoyl)lysine-modified protein spots in gastric mucosal cells were identified as the tropomyosin 1 protein by mass spectrometry using a MASCOT search. The existence of N^ε-(hexanoyl)lysine modification in tropomyosin 1 was confirmed by Western blotting of SDS-PAGE-separated or two-dimensional electrophoresis-separated proteins as well as by the immunoprecipitation with anti-tropomyosin 1 antibody. These data indicate that N^ε-(hexanoyl)lysine modification of tropomyosin 1 may be related to neoplastic transformation by N-methyl-N'-nitro-N-nitrosoguanidine in gastric epithelial cells.<br>

Journal

• Journal of Clinical Biochemistry and Nutrition

  Journal of Clinical Biochemistry and Nutrition 50 (1), 47-52, 2011

  SOCIETY FOR FREE RADICAL RESEARCH JAPAN

Keywords

• lipid peroxide
• gastric cancer
• hexanoyl-lysine
• tropomyosin 1

Details

• CRID: 1390282679649595648
• NII Article ID: 130004466669
• DOI: 10.3164/jcbn.11-39
• ISSN: 18805086
• ISSN: 09120009
• Text Lang: en
• Data Source:
  • JaLC
  • Crossref
  • CiNii Articles
  • KAKEN
• Abstract License Flag: Disallowed