Fig. 3

Attenuating effect of p.P243L on E3 activity in CHIP. a Impaired E3 activity of the STUB1-p.P243L mutant. The WT or p.P243L mutant of Myc-STUB1 was transiently expressed in HEK293T cells. Then, the anti-Myc immunoprecipitates were subjected to in vitro ubiquitination assay, and samples were immunoblotted with the indicated antibodies. b Crystal structure of the mouse CHIP U box dimer in complex with E2 (Ubc13 and Uev1a) (PDB: 2c2v). One protomer in the CHIP dimer is colored cyan and the other blue. Ubc13 is colored magenta. P243 in the human CHIP (P244 in mouse STUB1) is shown in orange. c Close-up view around the β-sheet of the CHIP U box (PDB: 2c2v). The β-sheet is shown as sticks. The backbone hydrogen bonds in the β-sheet are indicated by dashed lines. Side chains other than proline on the β-sheet are omitted. The residue numbers indicated are those of human CHIP