Fig. 2: Effects of point mutations on TRAP1 stability.
a T600 and its surrounding residues are visualized on the 3D structure of the TRAP1 homodimer. b Residues surrounding T600 in both protomer A (light blue) and B (dark yellow) of TRAP1. Numbering is relative to zTrap1 sequence as in PDB code 4IPE. To convert to hTrap1 numbering 15 should be subtracted. In the zoomed views the structure is rotated and centered on the mutated position. c Change in DF score to mutated positions in protomer A or B for each residue in going from WT to T600P (indicated by red circle) projected onto the protein 3D structure (P600A, P600B). Color code for DF scores: blue areas (positive values) correspond to lower mechanical coordination in Trap1 mutants with respect to the wild-type protein, whereas orange ones (negative values) indicate a higher coordination in the former. Gray/white areas are those unaffected by the mutation. d Western blot assessing TRAP1 protein levels in sMPNST cells re-expressing the human WT or T600P TRAP1 forms after knocking-out endogenous TRAP1. Where indicated, cells were treated with the proteasomal inhibitor MG132 10 μM for 6 h. Data are reported as average ± SEM of 3 independent experiments with a two-tail unpaired t-test. e Western blot assessing TRAP1 protein levels in sMPNST cells re-expressing the indicated mutant forms of TRAP1 after knocking-out endogenous TRAP1. Where indicated, cells were treated with 10 μM of MG132 for 6 h. Data are reported as average ± SEM of 3 independent experiments.
