Identification of Protein Targets of 12/15-Lipoxygenase-Derived Lipid Electrophiles in Mouse Peritoneal Macrophages Using Omega-Alkynyl Fatty Acid
Identification of Protein Targets of 12/15-Lipoxygenase-Derived Lipid Electrophiles in Mouse Peritoneal Macrophages Using Omega-Alkynyl Fatty Acid
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DOI：
10.1021/acschembio.7b01092
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发表时间：
2018-04-01
影响因子：
4
通讯作者：
Arita, Makoto
中科院分区：
文献类型：
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作者：
Isobe, Yosuke;Kawashima, Yusuke;Arita, Makoto
12/15-脂氧合酶（12/15-LOX）以位置特异性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物，包括脂质衍生的亲电子试剂（LDE）。腹膜巨噬细胞是小鼠中 12/15-LOX 表达最高的部位。然而，该酶在巨噬细胞调节中的作用尚不完全清楚。在这里，我们描述了一种化学蛋白质组学方法来识别酶促生成的 LDE 的靶标。通过用 omega-炔基花生四烯酸 (aAA) 处理小鼠腹腔巨噬细胞，我们鉴定了一系列由 12/15-LOX 催化反应产生的 LDE 加合的蛋白质。通路分析揭示了参与能量代谢的蛋白质显着富集，并发现糖酵解通量和线粒体呼吸受到 12/15-LOX 表达的显着影响。我们的发现利用化学蛋白质组学使用 aAA 来识别酶促生成的 LDE 的细胞内靶标。
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 utility of chemoproteomics using aAA for identifying intracellular targets of enzymatically generated LDE.