Fig. 1.

WW234 and L lock WWP1 in an inactive state. a Schematic of WWP1 domains showing an enzymatic activity summary derived from the autoubiquitination assay in b. b Autoubiquitination assay of Trx-tagged full-length (FL) WWP1 and various fragments. The weight markers belong to all separate gels. c GST pull-down assay of GST-tagged WW2L, WW2, L, LWW34, and WW34 with Trx-HECT. d WWP1 HECT autoubiquitination assay with different concentrations of Trx-WW2L, LWW34, Trx-WW2, Trx-WW34, and Trx-L. For the ubiquitination assay, the reactions were quenched after 15 min. Statistics for the enzymatic activities are shown below. Data are presented as the mean ± SD of triplicate experiments; ns, not significant, *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001 based on one-way analysis of variance (ANOVA) with Tukey’s multiple comparison test. Source data are provided as a Source Data file