Fig. 7: C1P binds to the DGR domain of KEAP1 to promote the dissociation of NRF2. | Experimental & Molecular Medicine

Fig. 7: C1P binds to the DGR domain of KEAP1 to promote the dissociation of NRF2.

From: Ceramide kinase-mediated C1P metabolism attenuates acute liver injury by inhibiting the interaction between KEAP1 and NRF2

Fig. 7

a C1P-coated beads were used to pull down the proteins that directly bind to C1P in AML12 cells with H2O2-induced oxidative damage. The KEAP1 and NRF2 levels in the pulled-down proteins were measured via western blotting. b Ceramide-coated beads were used to pull down the proteins that directly bind to ceramide in AML12 cells with H2O2-induced oxidative damage. The KEAP1 and NRF2 levels in the pulled-down proteins were measured via western blotting. c The binding mode and binding site of the C1P and KEAP1 proteins (PDBID: 7C60) were analyzed. AutoDock Vina was used for docking. d The domain structure of the mouse KEAP1 protein is shown. Only the deletion of the DGR domain inhibited the binding of C1P to KEAP1. e FLAG-tagged full-length KEAP1 and domain deletion mutants of KEAP1 were overexpressed in HEK-293T cells and pulled down with C1P-coated beads. In the pull-down assay of domain deletion mutants of KEAP1 with C1P-coated beads, the input proteins were analyzed. f The domain deletion mutants of KEAP1 that can bind to C1P were pulled down and analyzed by western blotting.

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