Chemoproteomic Mapping of Glycolytic Targetome in Cancer Cells

Abstract Hyperactivated glycolysis is a metabolic hallmark of most cancer cells. Although sporadic information has revealed that glycolytic metabolites possess non-metabolic functions as signaling molecules, it remains largely elusive how these metabolites interact with and functionally regulate their binding targets. Here we introduce a Target Responsive Accessibility Profiling (TRAP) approach that measures ligand binding-induced accessibility changes for target identification through globally labeling reactive proteinaceous lysines. With TRAP, we mapped 913 target candidates and 2,487 interactions for 10 major glycolytic metabolites in a model cancer cell line. The wide targetome depicted by TRAP unveils diverse regulatory modalities of glycolytic metabolites involving direct perturbation of carbohydrate metabolism enzymes, intervention of orphan transcriptional proteins’ interactions, and modulation of targetome-level acetylation. These results deepen our understanding of how glycolysis orchestrates signaling pathways in cancer cells in support of their survival and inspire the exploitation of the glycolytic targetome for cancer therapy development.