Endogenous formation of novel halogenated 2'-deoxycytidine. Hypohalous acid-mediated DNA modification at the site of inflammation.
A potential role of DNA damage by leukocyte-derived reactive species in carcinogenesis has been suggested. Leukocyte-derived peroxidases, such as myeloperoxidase and eosinophil peroxidase, use hydrogen peroxide and halides (Cl- and Br-) to generate hypohalous acids (HOCl and HOBr), halogenating intermediates. It has been suggested that these oxidants lead to the formation of halogenated products upon reaction with nucleobases. To verify the consequences of phagocyte-mediated DNA damage at the site of inflammation, we developed a novel monoclonal antibody (mAb2D3) that recognizes the hypohalous acid-modified DNA and found that the antibody most significantly recognized HOCl/HOBr-modified 2'-deoxycytidine residues. The immunoreactivity of HOCl-treated oligonucleotide was attenuated by excess methionine, suggesting that chloramine-like species may be the plausible epitopes of the antibody. On the basis of further characterization combined with mass spectrometric analysis, the epitopes of mAb2D3 were determined to be novel N4,5-dihalogenated 2'-deoxycytidine residues. The formation of the dihalogenated 2'-deoxycytidine in vivo was immunohistochemically demonstrated in the lung and liver nuclei of mice treated with lipopolysaccharides, an experimental inflammatory model. These results strongly suggest that phagocyte-derived oxidants, hypohalous acids, endogenously generate the halogenated DNA bases such as a novel dihalogenated 2'-deoxycytidine in vivo. Halogenation (chlorination and/or bromination) of DNA therefore may constitute one mechanism for oxidative DNA damage at the site of inflammation.
Kawai
Yoshichika
Y
Laboratory of Food and Biodynamics, Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya 464-8601, Japan. y-kawai@nutr.med.tokushima-u.ac.jp
Morinaga
Hiroshi
H
Kondo
Hajime
H
Miyoshi
Noriyuki
N
Nakamura
Yoshimasa
Y
Uchida
Koji
K
Osawa
Toshihiko
T
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry; D001959:Bromates / Q000494:pharmacology; D002700:Chloramines / Q000737:chemistry; D002713:Chlorine / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid; D002853:Chromatography, Liquid; D004247:DNA / Q000737:chemistry; D004249:DNA Damage; D003841:Deoxycytidine / Q000737:chemistry* / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000737:chemistry; D005260:Female; D018922:HL-60 Cells; D006801:Humans; D006997:Hypochlorous Acid / Q000494:pharmacology; D007150:Immunohistochemistry; D007249:Inflammation; D007962:Leukocytes / Q000378:metabolism; D008070:Lipopolysaccharides / Q000737:chemistry / Q000378:metabolism; D008099:Liver / Q000378:metabolism; D008168:Lung / Q000378:metabolism; D013058:Mass Spectrometry; D051379:Mice; D008810:Mice, Inbred C57BL; D008956:Models, Chemical; D009841:Oligonucleotides / Q000737:chemistry; D016877:Oxidants / Q000737:chemistry; D010100:Oxygen / Q000737:chemistry; D009195:Peroxidase / Q000737:chemistry; D010587:Phagocytosis; D017382:Reactive Oxygen Species; D012973:Sodium Hypochlorite / Q000494:pharmacology; D013997:Time Factors
D000911:Antibodies, Monoclonal; D001959:Bromates; D002700:Chloramines; D000939:Epitopes; D008070:Lipopolysaccharides; D009841:Oligonucleotides; D016877:Oxidants; D017382:Reactive Oxygen Species; D003841:Deoxycytidine; D002713:Chlorine; D006997:Hypochlorous Acid; D004247:DNA; D012973:Sodium Hypochlorite; D009195:Peroxidase; C027664:hypobromous acid; D010100:Oxygen