Identification of a Bromodomain‐like Region in 15‐Lipoxygenase‐1 Explains Its Nuclear Localization

• Published in: Angewandte Chemie International Edition 2021-09, Vol.60 (40), p.21875-21883
• Main Authors: Chen, Deng, Xiao, Zhangping, Guo, Hao, Gogishvili, Dea, Setroikromo, Rita, Wouden, Petra E., Dekker, Frank J.
• Format: Article
• Language: English
• Subjects: activity-based probes; Antibodies; Arachidonate 15-Lipoxygenase - metabolism; bromodomains; Chromatin; Enzyme Inhibitors - chemistry; Enzyme Inhibitors - pharmacology; Enzyme-linked immunosorbent assay; Humans; Inflammatory diseases; inhibitors; Isoenzymes; Labeling; Lipid metabolism; Lipids; Lipoxygenase; lipoxygenases; Liquid oxygen; Localization; Metabolites; Molecular Conformation; Oxidative metabolism; oxidoreductases; Tumorigenesis
• ISSN: 1433-7851; 1521-3773; 1521-3773
• DOI: 10.1002/anie.202106968

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. Activity‐based labeling of lipoxygenase (LOX) activity using the novel one‐step labeling probe Labelox B enables detection of isoenzyme inhibitory selectivity in an ELISA‐based assay and visualization of the localization of LOX activity in living cells. The observed nuclear localization was rationalized by identification of a bromodomain‐like region in the 15‐LOX‐1 isoenzyme.