Summary
An amide-type adduct, hexanoyl-lysine (HEL) is generated from the reaction between n-6 fatty acid (FA)-derived lipid peroxide and lysine. Immunochemical and chemical methods can be used to detect the formation of HEL. For example, an ELISA kit using the monoclonal antibody to HEL is now commercially available. We recently identified propanoyl-lysine (propionyl-lysine, PRL) from the reaction of an n-3 FA and a lysine residue. The antibody to PRL has been prepared and characterized. Using these monoclonal antibodies, the localization of adducts in tissues has been confirmed. Moreover, both amide-type adducts, HEL and PRL, can be simultaneously measured using liquid chromatography mass spectrometry (LC/MS/MS) with isotope dilution methods. The LC/MS/MS analysis reveals the rigid amounts of the adducts in human urine. Both the chemical and immunochemical methods are useful for the estimation of amide-type adducts in vivo.
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References
Esterbauer, H., Schaur, R. J., and Zollner, H. (1991) Chemistry and biochemistry of 4-hydroxynonenal, malonaldehyde and related aldehydes. Free Rad. Biol. Med. 11, 81–128.
Kato, Y., Mori, Y., Makino, Y., Morimitsu, Y., Hiroi, S., Ishikawa, T., and Osawa, T. (1999) Formation of N(epsilon)-(hexanonyl)lysine in protein exposed to lipid hydroperoxide. A plausible marker for lipid hydroperoxide-derived protein modification. J. Biol. Chem. 274, 20406–20414.
Fukuchi, Y., Miura, Y., Nabeno, Y., Kato, Y., Osawa, T., and Naito, M. (2008) ImmunohistoÂchemical detection of oxidative stress biomarkers, dityrosine and N(epsilon)-(hexanoyl)lysine, and C-reactive protein in rabbit atherosclerotic lesions. J. Atheroscler. Thromb. 15, 185–192.
Kawai, Y., Kato, Y., Fujii, H., Makino, Y., Mori, Y., Naito, M., and Osawa, T. (2003) Immunochemical detection of a novel lysine adduct using an antibody to linoleic acid hydroperoxide-modified protein. J. Lipid Res. 44, 1124–1131.
Kawai, Y., Fujii, H., Kato, Y., Kodama, M., Naito, M., Uchida, K., and Osawa, T. (2004) Esterified lipid hydroperoxide-derived modification of protein: formation of a carboxyalkylamide-type lysine adduct in human atherosclerotic lesions. Biochem. Biophys. Res. Commun. 313, 271–276.
Kawai, Y., Fujii, H., Okada, M., Tsuchie, Y., Uchida, K., and Osawa, T. (2006) Formation of N(epsilon)-(succinyl)lysine in vivo: a novel marker for docosahexaenoic acid-derived protein modification. J. Lipid Res. 47, 1386–1398.
Kato, Y., Yoshida, A., Naito, M., Kawai, Y., Tsuji, K., Kitamura, M., Kitamoto, N., Osawa, T. (2004) Identification and quantification of N(epsilon)-(hexanoyl)lysine in human urine by liquid chromatography/tandem mass spectrometry. Free Radic. Biol. Med. 37, 1864–1874.
Kato, Y., Miyake, Y., Yamamoto, K., Shimomura, Y., Ochi, H., Mori, Y., and Osawa, T. (2000) Preparation of a monoclonal antibody to N(epsilon)-(hexanonyl)lysine: application to the evaluation of protective effects of flavonoid supplementation against exercise-induced oxidative stress in rat skeletal muscle. Biochem. Biophys. Res. Commun. 274, 389–393.
Kato, Y. and Osawa, T. (2002) Detection of lipid hydroperoxide-derived protein modification with polyclonal antibodies. In: Oxidative Stress Biomarkers and Antioxidant Protocols, (Donald Armstrong, ed.), Humana Press, pp. 37–44.
Hisaka, S., Kato, Y., Kitamoto, N., Yoshida, A., Kubushiro, Y., Naito, M., and Osawa, T. (2009) Chemical and Immunochemical identification of propanoyllysine derived from oxidized n-3 polyunsaturated fatty acid. Free Rad. Biol. Med. 46, 1463–1471.
Shimizu, K., Ogawa, F., Akiyama, Y., Muroi, E., Yoshizaki, A., Iwata, Y., Komura, K., Bae, S., and Sato, S. (2008) Increased serum levels of N(epsilon)-(hexanoyl)lysine, A new marker of oxidative stress, in systemic sclerosis. J. Rheumatol. 35, 2214–2219.
Minato, K., Gono, M., Yamaguchi, H., Kato, Y., and Osawa, T. (2005) Accumulation of N(epsilon)-(hexanoyl)lysine, an oxidative stress biomarker, in rice seeds during storage. Biosci. Biotechnol. Biochem. 69, 1806–1810.
Kageyama, Y., Takahashi, M., Nagafusa, T., Torikai, E., and Nagano, A. (2008) Etanerceptreduces the oxidative stress marker levels in patients with rheumatoid arthritis. Rheumatol. Int. 28, 245–251.
Garrity, J., Gardner, J. G., Hawse, W., Wolberger, C., Escalante-Semerena, J. C. (2007) N-Lysine propionylation controls the activity of propionyl-CoA synthetase. J. Biol. Chem. 282, 30239–30245.
Acknowledgments
We would like to thank Shinsuke Hisaka for his helpful technical assistance.
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Kato, Y., Osawa, T. (2009). Detection of a Lipid-Lysine Adduct Family with an Amide Bond as the Linkage: Novel Markers for Lipid-Derived Protein Modifications. In: Armstrong, D. (eds) Lipidomics. Methods in Molecular Biology™, vol 580. Humana Press. https://doi.org/10.1007/978-1-60761-325-1_7
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DOI: https://doi.org/10.1007/978-1-60761-325-1_7
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