Figure 6
Isolation, ubiquitination activity and structural characterisation of the Hsp90:p53-TMGST:CHIP complex. (a) p53-TMGST ubiquitination assay mediated by Hsp90 and CHIP. For the western blot, an anti-p53 antibody was used. (b) Formation of the Hsp90:p53-TMGST:CHIP complex. Top, size-exclusion profile of the complex (continuous line) compared with the profiles for Hsp90 (dotted line), p53-TMGST (broken lines), and the Hsp90:CHIP complex (grey broken line). Bottom, the selected fractions were analysed by SDS-PAGE and stained with Coomassie Blue. (c) Three orthogonal views of the Hsp90:p53-TMGST:CHIP complex 3D reconstruction. (d) The same views with docking of Hsp90, p53-TMGST (p53DBD and GST), and CHIP. The DTSSP and DSSO crosslinks depicted in the images are described in Supplementary Fig. 7c. Only crosslinks between residues located in regions with known atomic structure are depicted in the figure. Not all the crosslinks are depicted in every view. Bar, 100 Å for (c,d).
