The 12/15-lipoxygenase (12/15-LOX) enzyme introduces peroxyl groups, in a position-specific manner, into polyunsaturated fatty acids to form various kinds of bioactive lipid metabolites, including lipid-derived electrophiles (LDE). The resident peritoneal macrophage is the site of highest 12/15-LOX expression in the mouse. However, the role of the enzyme in the regulation of resident macrophages is not fully understood. Here, we describe a chemoproteomic method to identify the targets of enzymatically generated LDE. By treating mouse peritoneal macrophages with omega-alkynyl arachidonic acid (aAA), we identified a series of proteins adducted by LDE generated through a 12/15-LOX catalyzed reaction. Pathway analysis revealed a dramatic enrichment of proteins involved in energy metabolism and found that glycolytic flux and mitochondrial respiration were significantly affected by the expression of 12/15-LOX. Our findings thus highlight the utility of chemoproteomics using aAA for identifying intracellular targets of enzymatically generated LDE.

中文翻译：

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使用ω-炔基脂肪酸在小鼠腹膜巨噬细胞中鉴定12 / 15-脂氧合酶衍生的脂质亲电子蛋白的蛋白质靶标

12 / 15-脂加氧酶（12 / 15-LOX）酶以位置特定的方式将过氧基引入多不饱和脂肪酸中，形成各种生物活性脂质代谢产物，包括脂质衍生的亲电试剂（LDE）。常驻腹膜巨噬细胞是小鼠中最高12 / 15-LOX表达的位点。然而，该酶在常驻巨噬细胞调节中的作用尚未完全了解。在这里，我们描述了一种化学旋转方法来识别酶促生成的LDE的目标。通过用ω-炔基花生四烯酸（aAA）处理小鼠腹膜巨噬细胞，我们鉴定了一系列由LDE加成的蛋白质，这些LDE通过12 / 15-LOX催化反应生成。途径分析揭示了参与能量代谢的蛋白质的大量富集，并发现糖酵解通量和线粒体呼吸受12 / 15-LOX的表达显着影响。因此，我们的发现突出了使用aAA的化学蛋白质组学在鉴定酶促生成的LDE的细胞内靶标方面的实用性。