source data of glycogen-G6PD phase separation

Glucose-6-phosphate (G6P) is a key metabolic molecule that regulates reactive oxygen species (ROS) homeostasis by initiating the pentose phosphate pathway (PPP) to generate NADPH that converts H2O2 to water by providing hydrogen. While both glucose phosphorylation and glycogenolysis result in G6P production, here we show that G6P derived from glycogenolysis, rather than glucose phosphorylation, flows to PPP for ROS clearance in CD8+ memory T (Tm) cells and inflammatory macrophages. This is because glycogenolysis-produced G1P allosterically promotes G6P dehydrogenase (G6PD) to bind to glycogen liquid-liquid phase separation (LLPS), thereby recruiting PPP enzymes to form compartmentalization for the rapid cascade reaction. Based on mechanistic elucidation, we demonstrated that G1P can act as an antitumor immunotherapeutic agent by modulating memory fitness and maintenance of tumor-reactive CD8+ T cells. These findings revealed a previously unknown function of glycogen metabolism, which is of paramount importance in the regulation of PPP and redox homeostasis in cells.