Lipid Peroxidation Generates Body Odor Component trans-2-Nonenal Covalently Bound to Protein in Vivo
Lipid Peroxidation Generates Body Odor Component trans-2-Nonenal Covalently Bound to Protein in Vivo
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DOI：
10.1074/jbc.m109.068023
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发表时间：
2010-05-14
影响因子：
4.8
通讯作者：
Uchida, Koji
中科院分区：
文献类型：
--
作者：
Ishino, Kousuke;Wakita, Chika;Uchida, Koji
反式-2-壬烯醛是一种不饱和醛，在多不饱和脂肪酸的过氧化过程中内源性产生令人不愉快的油腻和青草味。2-壬烯醛通过醛优先与赖氨酸残基反应来共价修饰人血清白蛋白。修饰后的蛋白具有免疫原性，并从小鼠体内获得了与2-壬烯醛共价修饰的蛋白发生交叉反应的特异性单克隆抗体（mAb）27 Q4。为了验证体内蛋白结合的2-壬烯醛的存在，产生针对2-壬烯醛修饰的钥孔血蓝蛋白的mAb 27 Q4。发现一种新的2-壬烯醛-赖氨酸加合物，顺式-和反式-N-庚-3-[（庚-1-烯基）-4-己基-吡啶]赖氨酸（HHP-赖氨酸），构成抗体的表位。在暴露于次氮基三乙酸铁（一种诱导自由基介导的氧化组织损伤的铁螯合物）的大鼠肾脏中检测到mAb 27 Q4的免疫反应性物质。利用高效液相色谱在线电喷雾串联质谱，我们还建立了一个高灵敏度的检测方法的顺式和反式HHP-赖氨酸，并证实2-壬烯醛-赖氨酸加合物确实形成在脂质过氧化介导的蛋白质修饰过程中，在体外和体内。此外，我们研究了清道夫受体凝集素样氧化低密度脂蛋白受体-1在2-壬烯醛修饰的蛋白质的识别中的参与，并确定该受体识别HHP-赖氨酸加合物作为配体。
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-hexyl-pyridinium]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.