Extended Data Fig. 9: Comparison of E2-Ub(T)-E3HECT in states 1, 2 and 7 (this study) with published structures of E3sHECT and structure-function analysis of the interface between N and C-lobes in E3HECT.
From: Structural basis for transthiolation intermediates in the ubiquitin pathway
a, Comparison of states 1, 2 and 7 of E2-Ub(T)-E3HECT showing similar orientation of the N and C-lobes of E3HECT and distinct positioning and conformation of Ub(T) compared to crystal structures of E2~Ub(T):E3HECT complex (Protein Data Bank (PDB) ID: 3JW0) and E3HECT~Ub(T) (PDB ID: 4BBN). b, Magnified view of the interface between N and C-lobes in the structures shown in (a). c, Histograms derived by quantification of the in-gel fluorescence of the gels presented in (Supplementary Fig. 4). Bars represent mean ± s.d of n = 3 replicates. Statistical differences between wild-type and mutants were determined by two-tailed unpaired t-test: ***P < 0.001. d, Histograms derived by quantification of the in-gel fluorescence of the gels presented (Supplementary Fig. 4). EQ/EQ indicate E451Q/E455Q mutation in E3. Bars represent mean ± s.d of n = 3 replicates. Data were analyzed by two-sided one-way ANOVA with Tukey’s test: ***P < 0.001, ns, not significant. e, Orthogonal views of surfaces for models for E2 and E3 for states 1 through 7 with ubiquitin shown as in cartoon representation next to similar depictions of PDB ID: 3JW0 and 4BBN to illustrate the rotation and translation of ubiquitin as it transits between states 1–7 to its product conformation in 3JW0 and 4BBN. Cartoon of ubiquitin shown at the bottom aligned at its C-terminus in respective complexes to provide another view of Ub(T) transitioning through states 1–7 compared to Ub(T) in PDB ID: 3JW0 and 4BBN.
