Immunochemical method to detect proteins that undergo selective modification by 1,2-naphthoquinone derived from naphthalene through metabolic activation
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- Miura Takashi
- Doctoral Program in Life System Medical Sciences, Graduate School of Comprehensive Human Sciences, University of Tsukuba
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- Kumagai Yoshito
- Doctoral Program in Life System Medical Sciences, Graduate School of Comprehensive Human Sciences, University of Tsukuba
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抄録
Naphthalene undergoes biotransformation by a variety of enzymes to yield 1,2-naphthoquinone (1,2-NQ), a reactive metabolite that binds covalently to proteins. Because this covalent modification is thought to account for naphthalene toxicity, a procedure to detect 1,2-NQ bound to macromolecules is required. In this study, we prepared a polyclonal antibody against 1,2-NQ and examined the specificities of the antibody for various aromatic structures and for the regiochemistry of the quinone functionality. Western blot analysis revealed that the antibody prepared against 1,2-NQ recognized the naphthalene moiety with the ortho-dicarbonyl group, but not with the para-dicarbonyl group; in addition, little cross-reactivity of ortho-quinones with different numbers of aromatic rings (n = 1, 3, 4, 5, 6) was seen. Dot blot and Western blot analyses with the polyclonal antibody enabled quantitative determination of the formation of protein-bound 1,2-NQ during the metabolic activation of naphthalene. The present method can be expected to applicable for the identification of the molecular targets of 1,2-NQ derived from naphthalene in cells and tissues.
収録刊行物
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- The Journal of Toxicological Sciences
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The Journal of Toxicological Sciences 35 (6), 843-852, 2010
一般社団法人 日本毒性学会
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詳細情報 詳細情報について
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- CRID
- 1390001204900770176
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- NII論文ID
- 10026963043
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- NII書誌ID
- AN00002808
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- ISSN
- 18803989
- 03881350
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- NDL書誌ID
- 10931777
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- 本文言語コード
- en
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- データソース種別
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- JaLC
- NDL
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