Fig. 6
An ordered Cdc34 CTDprox extension is involved in Ub discharge. a Left, crystal structure of hCdc34BΔdist conjugated to Ub (gold) at active site Cys to Lys mutation in presence of CC0651 inhibitor, shown as in Fig. 5c but rotated 90°. Top right, zoomed in view of Cdc34 CTDprox trajectory over the Ubc core with contacting residues shown as sticks. Bottom right, zoomed in view of CTDprox as in Fig. 5c. b Left, cartoon representation of hCdc34BΔdist-Ub as in a but rotated ~90° with CC0651 shown as gray sticks and isopeptide bond indicated. Right, zoomed in view of Cdc34 CTDprox, Ubc core, and Ub(t) interfaces with contacting residues shown as sticks. c K48-diUb assays for the indicated mutants with (top) and without (middle) minimal E3 ligase. Data are represented by mean ± SD with individual replicates shown as gray circles. Biochemical assays have four independent replicates, unless indicated by an asterisk when there are three replicates. Representative replicates are labeled above. Bottom, single turnover K48-diUb assay with indicated Ub mutants (n = 4). Data are shown as above with representative image labeled above with (−) lanes to show initial thioester formation and ( + ) to indicate subsequent addition of E3 ligase and excess UbA for single Ub discharge. Source data are provided as a Source Data file
