Extended Data Fig. 6: PGDLV motif of TRIM28 interacts with C-terminal of CtBP.
From: Reciprocal stabilization of CtBP and TRIM28 represses autophagy to promote metastasis
(a) ITC titration and fitting curve measurement of interaction between TRIM28-C term and CtBP1 (left) or CtBP2 (right). (b) Schematic diagram showing CtBP2 protein major domains and the truncation mutants. (c) Gluc PCA assay of CtBP2 truncation mutants interaction with TRIM28 in HEK-293T, each dot represents the luminescence intensity per 100 μl reaction system (n = 5). (d) Structure prediction of interaction between TRIM28-C-terminal and CtBP2 (333-445) by Alphafold3 (AlphaFold Server - Google DeepMind). This structural image was generated by the PyMOL Molecular Graphics System (Version 3.0 Schrödinger, LLC). TRIM28-C-terminal is shown in cyan, PGDLV motif in magenta, CtBP2 (333-445) in green. The interaction details are enlarged and shown in right figure, yellow dash line means hydrogen bound. (e–i) Gluc PCA assay of CtBP2 and CtBP2 truncation mutants (1-445, 1-127, 127-333, 333-445) (e) and CtBP2 truncation mutants (1-333, 127-445) (f) interaction, each dot represents the luminescence intensity per 100 μl reaction system (n = 5), CtBP2 truncation mutants (333-358) (g) and (333-445) (h) interaction with TRIM28 or TRIM28 mutants (n = 5), and CtBP1 or CtBP2 interaction with CtBP2 dimerization defect mutants (i) in HEK-293T cells (n = 4). Data are shown as the mean with s.d. (c,e-i); Statistical analysis was performed using a two-tailed, unpaired Student’s t-test (c,e-i); n.s. P > 0.05, *P < 0.05, **P < 0.01 and ***P < 0.001.
