Chemoproteomics Maps Glycolytic Targetome in Cancer Cells

Hyperactivated glycolysis, favoring uncontrolled growth and metastasis by producing essential metabolic intermediates engaging bioenergetics and biosynthesis, is a metabolic hallmark of most cancers. Although sporadic information has revealed glycolytic metabolites also possess non-metabolic function as signaling molecules, it remains largely elusive how these metabolites interact and functionally regulate their binding targets. Here, we developed a Target Responsive Accessibility Profiling (TRAP) approach for mapping the glycolytic targetome in cancer cells. We identified 913 proteins and 2,487 interactions as target candidates for 10 metabolites involved in glycolysis. The elucidated targetome uncovers diverse regulatory modalities of glycolytic metabolites involving the direct perturbation of carbohydrate metabolism enzymes, intervention of transcriptional control, modulation of proteome-level acetylation and disruption of protein complex assemblies. The advantages gained by enhanced glycolysis in cancer cells through these distinct mechanisms are innovated by discovering lactate as a ligand for an orphan transcriptional regulator TRIM 28 that promotes p53 degradation, and by identifying pyruvate acting against a cell apoptosis inducer trichostatin A via attenuating protein acetylation. Lastly, the inhibition of glycolytic key enzymes led to identify an intrinsically active glycolytic intermediate, glyceraldehyde 3-phosphate, that elicits its cytotoxic effects by engaging with ENO1 and MTHFD1. Collectively, the mapped targetome constitutes a fertile source for understanding how glycolysis finely tunes cancer metabolism and signaling in favor of tumor growth, and fostering the exploitation of glycolytic targetome as promising nodes for cancer therapeutics development.