Fig. 3.

TRAF6 dimers with a single conjugate binding site are active. a Single-turnover assay using fluorescently labelled conjugate performed as in Fig. 2c. Mutant proteins that disrupted either the E2 interface (IL/DR) or ubiquitin binding (Arg^ZF1) were incubated with the conjugate alone, or following mixing (far right). Schematics indicate the site of each mutation on TRAF6. b The formation of diubiquitin from panel a was quantified. Experiments were performed in duplicate; error bars represent range of measurements. c Coomassie blue stained chain-building assay using TRAF6 RZ₃ mutants from panel a