Department Chemical and Pharmaceutical Biology, Groningen Research Institute of Pharmacy (GRIP), University of Groningen, Antonius Deusinglaan 1, 9713, AV, Groningen, The Netherlands.
Angew Chem Int Ed Engl. 2021 Sep 27;60(40):21875-21883. doi: 10.1002/anie.202106968. Epub 2021 Aug 31.
Lipoxygenase (LOX) activity provides oxidative lipid metabolites, which are involved in inflammatory disorders and tumorigenesis. Activity-based probes to detect the activity of LOX enzymes in their cellular context provide opportunities to explore LOX biology and LOX inhibition. Here, we developed Labelox B as a potent covalent LOX inhibitor for one-step activity-based labeling of proteins with LOX activity. Labelox B was used to establish an ELISA-based assay for affinity capture and antibody-based detection of specific LOX isoenzymes. Moreover, Labelox B enabled efficient activity-based labeling of endogenous LOXs in living cells. LOX proved to localize in the nucleus, which was rationalized by identification of a functional bromodomain-like consensus motif in 15-LOX-1. This indicates that 15-LOX-1 is not only involved in oxidative lipid metabolism, but also in chromatin binding, which suggests a potential role in chromatin modifications.
脂氧合酶 (LOX) 活性提供氧化脂质代谢物，这些代谢物参与炎症性疾病和肿瘤发生。基于活性的探针可在细胞环境中检测 LOX 酶的活性，为探索 LOX 生物学和 LOX 抑制提供了机会。在这里，我们开发了 Labelox B，作为一种有效的共价 LOX 抑制剂，用于具有 LOX 活性的蛋白质的一步活性标记。Labelox B 用于建立基于 ELISA 的测定法，用于亲和捕获和基于抗体的特异性 LOX 同工酶检测。此外，Labelox B 能够有效地对活细胞内的内源性 LOX 进行基于活性的标记。LOX 被证明定位于细胞核中，这通过在 15-LOX-1 中鉴定出一个功能性溴结构域样保守基序得到了合理化。这表明 15-LOX-1 不仅参与氧化脂质代谢，还参与染色质结合，这表明其在染色质修饰中可能具有潜在作用。
