Extended Data Fig. 9: Validation of ENO1 as the binding target of G3P.
From: Chemoproteomic mapping of the glycolytic targetome in cancer cells
(a) Summarized Rtreated/control values of the TRPs in ENO1 assigned for G3P using the multiplexed-TRAP data. Of note, the letter before K denotes the Type A/B/C of TRPs. Data represent the mean ± SEM (n=3 biologically independent samples) and P values were determined using an unpaired two-tailed Student’s t-test. (b) Thermal shift assay validating the engagement of G3P (500 μM) with ENO1 using HCT116 cell lysates. Experiments were repeated (n=3 biologically independent samples) with one representative sample shown. (c) Thermal shift assay showing G3P-mediated stabilization of ENO1 using HCT116 cell lysates. Experiments were repeated (n=2 biologically independent samples) with one representative sample shown. (d) Volcano plot of the TRPs of G3P (500 μM) assigned by TRAP analysis of the human recombinant ENO1 (n=3 biologically independent samples). TRPs (RG3P/control >1.5 or <0.67, p <0.05 by unpaired two-sided Student’s t-test) were determined and highlighted in red. (e) SPR analysis showing weakened affinity of G3P to the ENO1 carrying K330E point mutation (Mutant-ENO1). The experiment was conducted once.
