Modified sites and functional consequences of 4-oxo-2-nonenal adducts in HDL that are elevated in familial hypercholesterolemia
Modified sites and functional consequences of 4-oxo-2-nonenal adducts in HDL that are elevated in familial hypercholesterolemia
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DOI：
10.1074/jbc.ra119.009424
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发表时间：
2019-12-13
影响因子：
4.8
通讯作者：
Davies, Sean S.
中科院分区：
文献类型：
--
作者：
May-Zhang, Linda S.;Yermalitsky, Valery;Davies, Sean S.
脂质醛4-氧代-2-壬烯醛（ONE）是一种高反应性蛋白质交联剂，由n-6多不饱和脂肪酸的过氧化反应产生，与4-羟基壬烯醛（HNE）一起生成。脂质过氧化产物介导的高密度脂蛋白（HDL）中蛋白质的交联导致HDL功能障碍，并有助于动脉粥样硬化的形成。虽然HNE是相对较好的研究，在动脉粥样硬化和修改HDL的ONE的作用是未知的。在此，我们发现家族性高胆固醇血症（FH）患者的HDL（54.6 +/- 33.8 pmol/mg）中的单酮酰胺（赖氨酸）加合物显著高于健康对照组（15.3 +/- 5.6 pmol/mg）。ONE交联的载脂蛋白A-I（apoA-I）在HDL上，浓度为> 3 mol ONE/10 mol apoA-I（0.3 eq.），其比HNE低100倍，但与有效的蛋白质交联剂异丁香苷相当。ONE修饰的HDL部分抑制HDL对脂多糖（LPS）诱导的小鼠巨噬细胞中肿瘤坏死因子a（Tnfa）和白细胞介素-1 β（IL-1 β）基因表达的保护能力。在3当量下，ONE显著降低apoA-I与HDL的交换，从46.5%降至18.4%（P < 0.001）。令人惊讶的是，HDL或apoA-I的一种修饰并不改变巨噬细胞胆固醇流出能力。LC-MS/MS分析显示，apoA-I中的Lys-12、Lys-23、Lys-96和Lys-226被一个酮酰胺加合物修饰。与其他二羰基清除剂相比，戊基吡哆胺（PPM）最有效地阻断了ONE诱导的HDL中的蛋白质交联，并在体外炎症模型中预防了HDL功能障碍。我们的研究结果表明，ONE-HDL加合物导致HDL功能障碍，并在患有严重高胆固醇血症的FH患者中升高。
The lipid aldehyde 4-oxo-2-nonenal (ONE) is a highly reactive protein cross-linker derived from peroxidation of n-6 polyunsaturated fatty acids and generated together with 4-hydroxynonenal (HNE). Lipid peroxidation productmediated cross-linking of proteins in high-density lipoprotein (HDL) causes HDL dysfunction and contributes to atherogenesis. Although HNE is relatively well studied, the role of ONE in atherosclerosis and in modifying HDL is unknown. Here, we found that individuals with familial hypercholesterolemia (FH) had significantly higher ONE-ketoamide (lysine) adducts in HDL (54.6 +/- 33.8 pmol/mg) than healthy controls (15.3 +/- 5.6 pmol/mg). ONE crosslinked apolipoprotein A-I (apoA-I) on HDL at a concentration of > 3 mol ONE per 10 mol apoA-I (0.3 eq.), which was 100-fold lower than HNE, but comparable to the potent protein crosslinker isolevuglandin. ONE-modified HDL partially inhibited HDL's ability to protect against lipopolysaccharide (LPS)-induced tumor necrosis factor a (Tnfa) and interleukin-1 beta (Il-1 beta) gene expression in murine macrophages. At 3 eq., ONE dramatically decreased apoA-I exchange from HDL, from similar to 46.5% to similar to 18.4% (P < 0.001). Surprisingly, ONE modification of HDL or apoA-I did not alter macrophage cholesterol efflux capacity. LC-MS/MS analysis revealed that Lys-12, Lys-23, Lys-96, and Lys-226 in apoA-I are modified by ONE ketoamide adducts. Compared with other dicarbonyl scavengers, pentyl-pyridoxamine (PPM) most efficaciously blocked ONE-induced protein cross-linking in HDL and also prevented HDL dysfunction in an in vitro model of inflammation. Our findings show that ONE-HDL adducts cause HDL dysfunction and are elevated in individuals with FH who have severe hypercholesterolemia.