Photoaffinity labelling-based chemoproteomic strategy identifies PEBP1 as the target of ethyl gallate against macrophage activation.
Wei YuMin LiaoYang ChenRui XueXiao-Meng ShiDan LiuFang-Fang ZhuoHui TangZhi-Yuan LuPeng-Fei TuBo HanXin JiaKe-Wu ZengPublished in: Chemical communications (Cambridge, England) (2023)
Ulcerative colitis (UC) is an inflammatory disease of the colon with an unmet need for therapeutic targets. Ethyl gallate (EG) is a natural small molecule for UC treatment, but its cellular target is unknown. By labelling EG with a diazirine photocrosslinker and a click chemistry handle, we identified phosphatidyl-ethanolamine binding protein1 (PEBP1) as a direct cellular target of EG by forming hydrogen bonds with Asp70 and Tyr120. In particular, hydrogen/deuterium exchange mass spectrometry indicated that EG induced the sequence (residues 141-153) embedding to inhibit S153 phosphorylation of PEBP1. Additionally, the EG-mediated sequence (residues 108-122) exposure significantly enhanced PEBP1-Raf-1 interaction to block the downstream NF-κB inflammatory pathway in macrophages. Moreover, PEBP1 siRNA substantially reversed the EG-dependent down-regulation of the phosphorylation of IKKβ, IκBα and NF-κB, demonstrating that the NF-κB signal functioned as an essential anti-inflammation mechanism of PEBP1. Collectively, we revealed PEBP1 as a previously undescribed cellular target in macrophages for UC therapy and identified a new allosteric site for PEBP1 biology study using EG as a chemical probe.
Keyphrases
- oxidative stress
- small molecule
- signaling pathway
- mass spectrometry
- lps induced
- binding protein
- pi k akt
- nuclear factor
- ulcerative colitis
- diabetic rats
- adipose tissue
- genome wide
- stem cells
- liquid chromatography
- bone marrow
- high glucose
- quantum dots
- endothelial cells
- gene expression
- single cell
- drug delivery
- protein protein
- protein kinase
- amino acid
- stress induced
- gas chromatography
- capillary electrophoresis
- living cells
- drug discovery