Polyclonal Antibodies to a Fluorescent 4-Hydroxy-2-nonenal (HNE)-Derived Lysine-Lysine Cross-Link: Characterization and Application to HNE-Treated Protein and in Vitro Oxidized Low-Density Lipoprotein

详细信息    查看全文

• 作者：Guozhang Xu ; Yahua Liu ; and Lawrence M. Sayre
• 刊名：Chemical Research in Toxicology
• 出版年：2000
• 出版时间：May 2000
• 年：2000
• 卷：13
• 期：5
• 页码：406 - 413
• 全文大小：240K
• 年卷期：v.13,no.5(May 2000)
• ISSN：1520-5010

文摘

Oxidative modification of low-density lipoprotein (LDL) is thought to play a key role in theetiology of atherosclerosis. Oxidized LDL that accumulates in atherosclerotic plaques is knownto exhibit a characteristic fluorescence with excitation and emission near 360 and 430 nm,respectively. (E)-4-Hydroxy-2-nonenal (HNE), formed during LDL oxidation, is capable ofmodifying LDL to generate the same fluorescent signature. The HNE-derived fluorophore wasshown by us to possess a 2-hydroxy-2-pentyl-1,2-dihydropyrrol-3-one iminium (HPDPI)structure. We herein report the synthesis of the HPDPI-derived lysine-lysine cross-link neededas a standard reference for HPLC quantitation of the cross-link in protein hydrolysates. Themain focus of the current work, however, is the design and development of two polyclonalantibodies against the HPDPI epitope. Utilizing these antibodies, levels of the HPDPI epitopewere estimated in HNE-treated bovine serum albumin and in copper-oxidized LDL by anenzyme-linked immunosorbent assay. Our results are consistent with the premise that thefluorescent HPDPI cross-link is a key contributor to the fluorescence exhibited by atheroscleroticlesions.