Heat-shock protein 27 (Hsp27) as a target of methylglyoxal in gastrointestinal cancer

BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR BASIS OF DISEASE

卷:1812

Article

Oya-Ito, Tomoko^1,2  Naito, Yuji¹  Takagi, Tomohisa¹  Handa, Osamu¹  Matsui, Hirofumi³  Yamada, Masaki⁴  Shima, Keisuke⁴  Yoshikawa, Toshikazu^1,2 

[1] Kyoto Prefectural Univ Med, Dept Mol Gastroenterol & Hepatol, Grad Sch Med Sci, Kyoto 6028566, Japan

[2] Kyoto Prefectural Univ Med, Dept Med Prote, Kyoto 6028566, Japan

[3] Univ Tsukuba, Div Gastroenterol, Grad Sch Comprehens Human Sci, Tsukuba, Ibaraki 3058575, Japan

[4] Shimadzu Co Ltd, Kyoto 6048511, Japan

关键词: Posttranslational modification;    Proteomics;    Heat-shock protein 27;    Methylglyoxal;    Apoptosis;    Cancer;   

DOI  :  10.1016/j.bbadis.2011.03.017

来源: Elsevier

【 摘 要 】

The molecular mechanisms underlying the posttranslational modification of proteins in gastrointestinal cancer are still unknown. Here, we investigated the role of methylglyoxal modifications in gastrointestinal tumors. Methylglyoxal is a reactive dicarbonyl compound produced from cellular glycolytic intermediates that reacts non-enzymatically with proteins. By using a monoclonal antibody to methylglyoxal-modified proteins, we found that murine heat-shock protein 25 and human heat-shock protein 27 were the major adducted proteins in rat gastric carcinoma mucosal cell line and human colon cancer cell line, respectively. Furthermore, we found that heat-shock protein 27 was modified by methylglyoxal in ascending colon and rectum of patients with cancer. However, methylglyoxal-modified heat-shock protein 25/heat-shock protein 27 was not detected in non cancerous cell lines or in normal subject. Matrix-associated laser desorption/ionization mass spectrometry/mass spectrometry analysis of peptide fragments identified Arg-75, Arg-79, Arg-89, Arg-94, Arg-127, Arg-136, Arg-140, Arg-188, and Lys-123 as methylglyoxal modification sites in heat-shock protein 27 and in phosphorylated heat-shock protein 27. The transfer of methylglyoxal-modified heat-shock protein 27 into rat intestinal epithelial cell line RIE was even more effective in preventing apoptotic cell death than that of native control heat-shock protein 27. Furthermore, methylglyoxal modification of heat-shock protein 27 protected the cells against both the hydrogen peroxide- and cytochrome c-mediated caspase activation, and the hydrogen peroxide-induced production of intracellular reactive oxygen species. The levels of lactate converted from methylglyoxal were increased in carcinoma mucosal cell lines. Our results suggest that posttranslational modification of heat-shock protein 27 by methylglyoxal may have important implications for epithelial cell injury in gastrointestinal cancer. (C) 2011 Elsevier B.V. All rights reserved.

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