Extended Data Fig. 8: The overall shape of SdeA and the function of SdeA CTD.

a, Superimposition of various Ub structures (PDB codes: 5M93 (orange), 1UBQ (pink), 5CRA (chain C, cyan), 3ZLZ (chain B, yellow) and 4BOZ (chain B, grey)) onto the SdeA mART–Ub–NADH structure with R42 residues of all the Ub molecules shown in stick representation. SdeA mART and Ub from the SdeA mART–Ub–NADH complex are shown in green and magenta, respectively. b, In vitro GST pull-down assays to detect the interactions of SdeA with IcmS or its complexes. GST-fused SdeA protein was incubated with IcmS, the IcmS–IcmW complex, the IcmS–IcmW–DotLc (residues 656–783 of DotL) ternary complex or the IcmS–IcmW–DotLc–LvgA quaternary complex. The protein samples bound to glutathione resins were washed three times and analysed by SDS–PAGE and Coomassie blue staining. IcmS/W represents IcmS + IcmW. The band marked with an asterisk represents the degraded GST tag. Similar results were obtained in three independent experiments. Uncropped blots and gel images are shown in Supplementary Fig. 1. c, Experimental PDDFs (pair distance distribution function) for SdeA(231–1190), SdeA(1–1499), SdeA(1–1190) and SdeA(1092–1496). d, Overlay of the experimental scattering profiles (exp) from the four samples in the SAXS analysis with the back-calculated scattering profile of the crystal structure of SdeA(231–1190) (cal). e, Fitting the crystal structure of SdeA(231–1190) into the SAXS envelope of SdeA(231–1190). Two perpendicular views are shown. f, Superimposition of the SAXS envelopes of SdeA(231–1190) (coloured as in e) and SdeA(1–1190) (light magenta) with the crystal structure of SdeA(231–1190) fitted. g, SAXS envelopes of SdeA(1092–1496). h, Superimposition of the SAXS envelopes of SdeA(1–1190) (light magenta), SdeA(1092–1496) (cyan) and SdeA(1–1499) (wheat) with the crystal structure of SdeA(231–1190) fitted.