Chemoproteomic identification of a phosphohistidine acceptor: Insights into posttranslational regulation of glycolysis

Abstract

Protein phosphorylation is one of the most extensively studied posttranslational modifications (PTMs), controlling various biological phenomena. Among these modifications, phosphohistidine (pHis) is an underexplored form. With the development of pHis-specific antibodies and advances in phosphoproteomics, numerous pHis sites have been discovered. Some pHis acceptors, which bind to pHis proteins and get trans-phosphorylated, are involved in signaling and metabolic pathways, such as the bacterial two-component system (TCS) and the phosphoenolpyruvate:sugar transferase system (PTS). However, the presence and function of other pHis acceptors remain underexplored. Here, we report a chemoproteomic strategy using a stable pHis analog-based chemical probe to identify pHis acceptors. The putative pHis acceptors identified in this study include crucial metabolic enzymes. Further experiments indicate that histidine phosphorylation on a glycolytic enzyme can regulate glycolysis posttranslational.