Fig. 4: Impact of nb.C11 on NEDD4-2 HECT domain catalytic activity.
From: Ion channel inhibition by targeted recruitment of NEDD4-2 with divalent nanobodies
a Time course of WW4-NEDD4-2 HECT catalytic activity assessed by monitoring formation of polyubiquitin chains in an in vitro ubiquitination assay probed with anti-ubiquitin antibody. Blot is representative of 3 independent experiments with similar results. b Time course of HECT domain in vitro catalytic activity for either purified WW4-HECT4-2 (black circles, n = 3) or full-length NEDD4-2 (red circles, n = 2). c Time course of WW4-HECT4-2 in vitro autoubiquitination visualized using anti-WW4-HECT4-2 antibody. Blot is representative of 3 independent experiments with similar results. d Impact of purified nb.C11 on capacity of WW4-HECT4-2 to catalyze formation of polyubiquitin chains in vitro. e Population data showing impact of nb.C11 on WW4-HECT4-2 in vitro catalytic activity (n = 6). *p < 0.0001 compared to WW4-HECT4-2 control by one-way ANOVA and Tukey’s multiple comparisons test. f Impact of purified nb.C11 on capacity of full-length NEDD4-2 to catalyze formation of polyubiquitin chains in vitro. g Population data showing impact of nb.C11 on full-length NEDD4-2 in vitro catalytic activity (n = 4). *p < 0.0001 compared to full-length NEDD4-2 control by one-way ANOVA and Tukey’s multiple comparisons test.
