Fig. 4: Cryo-EM structure of the human DDB1-DDA1-DET1-COP1-Ube2e2-c-Jun complex.
From: Cryo-EM structure of the human COP1-DET1 ubiquitin ligase complex
A Coomassie blue-stained SDS-PAGE analysis of the purified DDB1-DDA1-DET1-COP1-Ube2e2-c-Jun complex. MW, molecular weight. The experiment was repeated at least three times independently with similar results. Source data are provided as a Source Data file. B Gel filtration (Superose 6 Increase 10/300 GL) profile of the purified DDD-COP1 (black) and DDD-Ube2e2-COP1-c-Jun complex (red). C, D The negative stain micrograph and 2D class averages of gel filtration fractions collected from the DDD-COP1 (C) and the DDD-Ube2e2-COP1-c-Jun complex (D). Scale bar, 100 nm. E The chimera model of the human DDD-Ube2e2-COP1-c-Jun complex. The subunits are colored as follows: DDB1, orange; DDA1, purple; DET1, thistle; COP1, dark sea green; Ube2e2, cornflower blue. F Circular plot of cross-linked DDD-Ube2e2-COP1-c-Jun subunits identified by XL-MS. Purple lines indicate intra-links, whereas green lines indicate inter-links. Date in F are processed once. G Sequence alignment of reported COP1 substrate and adaptor. The conserved VP motif is shown in red. Close-up views of the interface between COP1WD40: Trib1(PDB 5IGQ) and COP1WD40:c-Jun (model). H Pull-down experiment of MBP-c-Jun in cells transfected with wild-type or mutated GST-COP1. The eluents were analyzed by SDS-PAGE and western blotting. Data in H are representative of three independent experiments. Source data are provided as a Source Data file.
