Lipid peroxidation generates body odor component trans-2-nonenal covalently bound to protein in vivo.
trans-2-Nonenal is an unsaturated aldehyde with an unpleasant greasy and grassy odor endogenously generated during the peroxidation of polyunsaturated fatty acids. 2-Nonenal covalently modified human serum albumin through a reaction in which the aldehyde preferentially reacted with the lysine residues. Modified proteins were immunogenic, and a specific monoclonal antibody (mAb) 27Q4 that cross-reacted with the protein covalently modified with 2-nonenal was raised from mouse. To verify the presence of the protein-bound 2-nonenal in vivo, the mAb 27Q4 against the 2-nonenal-modified keyhole limpet hemocyanin was raised. It was found that a novel 2-nonenal-lysine adduct, cis- and trans-N(epsilon)-3-[(hept-1-enyl)-4-hexylpyridinium]lysine (HHP-lysine), constitutes an epitope of the antibody. The immunoreactive materials with mAb 27Q4 were detected in the kidney of rats exposed to ferric nitrilotriacetate, an iron chelate that induces free radical-mediated oxidative tissue damage. Using high performance liquid chromatography with on-line electrospray ionization tandem mass spectrometry, we also established a highly sensitive method for detection of the cis- and trans-HHP-lysine and confirmed that the 2-nonenal-lysine adducts were indeed formed during the lipid peroxidation-mediated modification of protein in vitro and in vivo. Furthermore, we examined the involvement of the scavenger receptor lectin-like oxidized low density lipoprotein receptor-1 in the recognition of 2-nonenal-modified proteins and established that the receptor recognized the HHP-lysine adducts as a ligand.
Ishino
Kousuke
K
Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya 464-8601.
Wakita
Chika
C
Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya 464-8601.
Shibata
Takahiro
T
Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya 464-8601.
Toyokuni
Shinya
S
Department of Pathology and Biological Responses, Graduate School of Medicine, Nagoya University, Nagoya 466-8550.
Machida
Sachiko
S
National Food Research Institute, 2-1-12 Kannondai, Tsukuba, Ibaraki 305-8642.
Matsuda
Shun
S
Research Center for Environmental Quality Management, Kyoto University, 1-2 Otsu, 520-0811, Japan.
Matsuda
Tomonari
T
Research Center for Environmental Quality Management, Kyoto University, 1-2 Otsu, 520-0811, Japan.
Uchida
Koji
K
Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya 464-8601. Electronic address: uchidak@agr.nagoya-u.ac.jp.
D000447:Aldehydes / Q000276:immunology / Q000378:metabolism*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D002851:Chromatography, High Pressure Liquid; D007150:Immunohistochemistry; D007668:Kidney / Q000187:drug effects / Q000378:metabolism; D015227:Lipid Peroxidation; D009682:Magnetic Resonance Spectroscopy; D008297:Male; D009812:Odorants; D011506:Proteins / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D021241:Spectrometry, Mass, Electrospray Ionization; D053719:Tandem Mass Spectrometry
D000447:Aldehydes; D000911:Antibodies, Monoclonal; D011506:Proteins; C013166:2-nonenal