Lipid peroxidation modification of protein generates Nε-(4- oxononanoyl)lysine as a pro-inflammatory ligand

Takahiro Shibata, Yuuki Shimozu, Chika Wakita, Noriyuki Shibata, Makio Kobayashi, Sachiko Machida, Rina Kato, Hiroyuki Itabe, Xiaochun Zhu, Lawrence M. Sayre, Koji Uchida

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

4-Oxo-2(E)-nonenal (ONE), a peroxidation product of ω-6 polyunsaturated fatty acids, covalently reacts with lysine residues to generate a 4-ketoamide-type ONE-lysine adduct, Nε-(4-oxononanoyl)lysine (ONL). Using an ONL-coupled protein as the immunogen, we raised the monoclonal antibody (mAb) 9K3 directed to the ONL and conclusively demonstrated that the ONL was produced during the oxidative modification of a low density lipoprotein (LDL) in vitro. In addition, we observed that the ONL was present in atherosclerotic lesions, in which an intense immunoreactivity was mainly localized in the vascular endothelial cells and macrophage- and vascular smooth muscle cell-derived foam cells. Using liquid chromatography with online electrospray ionization tandem mass spectrometry, we also established a highly sensitive method for quantification of the ONLand confirmed that theONLwas indeed formed during the lipid peroxidation-mediated modification of protein in vitro and in vivo. To evaluate the biological implications for ONL formation, we examined the recognition of ONL by the scavenger receptor lectin-like oxidized LDL receptor-1 (LOX-1). UsingCHOcells stably expressing LOX-1, we evaluated the ability of ONL to compete with the acetylated LDL and found that both the ONE-modified and ONL-coupled proteins inhibited the binding and uptake of the modified LDL. In addition, we demonstrated that the ONL-coupled protein was incorporated into differentiated THP-1 cells via LOX-1. Finally, we examined the effect of ONL on the expression of the inflammation-associated gene in THP-1 and observed that the ONL-coupled proteins significantly induced the expression of atherogenesis-related genes, such as the monocyte chemoattractant protein-1 and tumor necrosis factor-α, in a LOX-1-dependent manner. Thus, ONL was identified to be a potential endogenous ligand for LOX-1.

Original languageEnglish
Pages (from-to)19943-19957
Number of pages15
JournalJournal of Biological Chemistry
Volume286
Issue number22
DOIs
Publication statusPublished - Jun 3 2011
Externally publishedYes

Fingerprint

Lipid Peroxidation
Lysine
Ligands
Lipids
Oxidized LDL Receptors
Proteins
LDL Lipoproteins
Class E Scavenger Receptors
Genes
Scavenger Receptors
Electrospray ionization
Foam Cells
Electrospray Ionization Mass Spectrometry
Chemokine CCL2
Macrophages
Liquid chromatography
Endothelial cells
Tandem Mass Spectrometry
Unsaturated Fatty Acids
Vascular Smooth Muscle

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Shibata, T., Shimozu, Y., Wakita, C., Shibata, N., Kobayashi, M., Machida, S., ... Uchida, K. (2011). Lipid peroxidation modification of protein generates Nε-(4- oxononanoyl)lysine as a pro-inflammatory ligand. Journal of Biological Chemistry, 286(22), 19943-19957. https://doi.org/10.1074/jbc.M110.187047

Lipid peroxidation modification of protein generates Nε-(4- oxononanoyl)lysine as a pro-inflammatory ligand. / Shibata, Takahiro; Shimozu, Yuuki; Wakita, Chika; Shibata, Noriyuki; Kobayashi, Makio; Machida, Sachiko; Kato, Rina; Itabe, Hiroyuki; Zhu, Xiaochun; Sayre, Lawrence M.; Uchida, Koji.

In: Journal of Biological Chemistry, Vol. 286, No. 22, 03.06.2011, p. 19943-19957.

Research output: Contribution to journalArticle

Shibata, T, Shimozu, Y, Wakita, C, Shibata, N, Kobayashi, M, Machida, S, Kato, R, Itabe, H, Zhu, X, Sayre, LM & Uchida, K 2011, 'Lipid peroxidation modification of protein generates Nε-(4- oxononanoyl)lysine as a pro-inflammatory ligand', Journal of Biological Chemistry, vol. 286, no. 22, pp. 19943-19957. https://doi.org/10.1074/jbc.M110.187047
Shibata, Takahiro ; Shimozu, Yuuki ; Wakita, Chika ; Shibata, Noriyuki ; Kobayashi, Makio ; Machida, Sachiko ; Kato, Rina ; Itabe, Hiroyuki ; Zhu, Xiaochun ; Sayre, Lawrence M. ; Uchida, Koji. / Lipid peroxidation modification of protein generates Nε-(4- oxononanoyl)lysine as a pro-inflammatory ligand. In: Journal of Biological Chemistry. 2011 ; Vol. 286, No. 22. pp. 19943-19957.
@article{1b95d7472d1c47dc84eee681b48745c8,
title = "Lipid peroxidation modification of protein generates Nε-(4- oxononanoyl)lysine as a pro-inflammatory ligand",
abstract = "4-Oxo-2(E)-nonenal (ONE), a peroxidation product of ω-6 polyunsaturated fatty acids, covalently reacts with lysine residues to generate a 4-ketoamide-type ONE-lysine adduct, Nε-(4-oxononanoyl)lysine (ONL). Using an ONL-coupled protein as the immunogen, we raised the monoclonal antibody (mAb) 9K3 directed to the ONL and conclusively demonstrated that the ONL was produced during the oxidative modification of a low density lipoprotein (LDL) in vitro. In addition, we observed that the ONL was present in atherosclerotic lesions, in which an intense immunoreactivity was mainly localized in the vascular endothelial cells and macrophage- and vascular smooth muscle cell-derived foam cells. Using liquid chromatography with online electrospray ionization tandem mass spectrometry, we also established a highly sensitive method for quantification of the ONLand confirmed that theONLwas indeed formed during the lipid peroxidation-mediated modification of protein in vitro and in vivo. To evaluate the biological implications for ONL formation, we examined the recognition of ONL by the scavenger receptor lectin-like oxidized LDL receptor-1 (LOX-1). UsingCHOcells stably expressing LOX-1, we evaluated the ability of ONL to compete with the acetylated LDL and found that both the ONE-modified and ONL-coupled proteins inhibited the binding and uptake of the modified LDL. In addition, we demonstrated that the ONL-coupled protein was incorporated into differentiated THP-1 cells via LOX-1. Finally, we examined the effect of ONL on the expression of the inflammation-associated gene in THP-1 and observed that the ONL-coupled proteins significantly induced the expression of atherogenesis-related genes, such as the monocyte chemoattractant protein-1 and tumor necrosis factor-α, in a LOX-1-dependent manner. Thus, ONL was identified to be a potential endogenous ligand for LOX-1.",
author = "Takahiro Shibata and Yuuki Shimozu and Chika Wakita and Noriyuki Shibata and Makio Kobayashi and Sachiko Machida and Rina Kato and Hiroyuki Itabe and Xiaochun Zhu and Sayre, {Lawrence M.} and Koji Uchida",
year = "2011",
month = "6",
day = "3",
doi = "10.1074/jbc.M110.187047",
language = "English",
volume = "286",
pages = "19943--19957",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "22",

}

TY - JOUR

T1 - Lipid peroxidation modification of protein generates Nε-(4- oxononanoyl)lysine as a pro-inflammatory ligand

AU - Shibata, Takahiro

AU - Shimozu, Yuuki

AU - Wakita, Chika

AU - Shibata, Noriyuki

AU - Kobayashi, Makio

AU - Machida, Sachiko

AU - Kato, Rina

AU - Itabe, Hiroyuki

AU - Zhu, Xiaochun

AU - Sayre, Lawrence M.

AU - Uchida, Koji

PY - 2011/6/3

Y1 - 2011/6/3

N2 - 4-Oxo-2(E)-nonenal (ONE), a peroxidation product of ω-6 polyunsaturated fatty acids, covalently reacts with lysine residues to generate a 4-ketoamide-type ONE-lysine adduct, Nε-(4-oxononanoyl)lysine (ONL). Using an ONL-coupled protein as the immunogen, we raised the monoclonal antibody (mAb) 9K3 directed to the ONL and conclusively demonstrated that the ONL was produced during the oxidative modification of a low density lipoprotein (LDL) in vitro. In addition, we observed that the ONL was present in atherosclerotic lesions, in which an intense immunoreactivity was mainly localized in the vascular endothelial cells and macrophage- and vascular smooth muscle cell-derived foam cells. Using liquid chromatography with online electrospray ionization tandem mass spectrometry, we also established a highly sensitive method for quantification of the ONLand confirmed that theONLwas indeed formed during the lipid peroxidation-mediated modification of protein in vitro and in vivo. To evaluate the biological implications for ONL formation, we examined the recognition of ONL by the scavenger receptor lectin-like oxidized LDL receptor-1 (LOX-1). UsingCHOcells stably expressing LOX-1, we evaluated the ability of ONL to compete with the acetylated LDL and found that both the ONE-modified and ONL-coupled proteins inhibited the binding and uptake of the modified LDL. In addition, we demonstrated that the ONL-coupled protein was incorporated into differentiated THP-1 cells via LOX-1. Finally, we examined the effect of ONL on the expression of the inflammation-associated gene in THP-1 and observed that the ONL-coupled proteins significantly induced the expression of atherogenesis-related genes, such as the monocyte chemoattractant protein-1 and tumor necrosis factor-α, in a LOX-1-dependent manner. Thus, ONL was identified to be a potential endogenous ligand for LOX-1.

AB - 4-Oxo-2(E)-nonenal (ONE), a peroxidation product of ω-6 polyunsaturated fatty acids, covalently reacts with lysine residues to generate a 4-ketoamide-type ONE-lysine adduct, Nε-(4-oxononanoyl)lysine (ONL). Using an ONL-coupled protein as the immunogen, we raised the monoclonal antibody (mAb) 9K3 directed to the ONL and conclusively demonstrated that the ONL was produced during the oxidative modification of a low density lipoprotein (LDL) in vitro. In addition, we observed that the ONL was present in atherosclerotic lesions, in which an intense immunoreactivity was mainly localized in the vascular endothelial cells and macrophage- and vascular smooth muscle cell-derived foam cells. Using liquid chromatography with online electrospray ionization tandem mass spectrometry, we also established a highly sensitive method for quantification of the ONLand confirmed that theONLwas indeed formed during the lipid peroxidation-mediated modification of protein in vitro and in vivo. To evaluate the biological implications for ONL formation, we examined the recognition of ONL by the scavenger receptor lectin-like oxidized LDL receptor-1 (LOX-1). UsingCHOcells stably expressing LOX-1, we evaluated the ability of ONL to compete with the acetylated LDL and found that both the ONE-modified and ONL-coupled proteins inhibited the binding and uptake of the modified LDL. In addition, we demonstrated that the ONL-coupled protein was incorporated into differentiated THP-1 cells via LOX-1. Finally, we examined the effect of ONL on the expression of the inflammation-associated gene in THP-1 and observed that the ONL-coupled proteins significantly induced the expression of atherogenesis-related genes, such as the monocyte chemoattractant protein-1 and tumor necrosis factor-α, in a LOX-1-dependent manner. Thus, ONL was identified to be a potential endogenous ligand for LOX-1.

UR - http://www.scopus.com/inward/record.url?scp=79957601907&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=79957601907&partnerID=8YFLogxK

U2 - 10.1074/jbc.M110.187047

DO - 10.1074/jbc.M110.187047

M3 - Article

C2 - 21471194

AN - SCOPUS:79957601907

VL - 286

SP - 19943

EP - 19957

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 22

ER -