Metabolic orchestration of NOD1 signaling by AMPK-mediated phosphorylation of ZDHHC5.

Innate immune responses must be coordinated with the cellular metabolic state. The intracellular pattern recognition receptor NOD1 detects bacterial peptidoglycans and initiates signaling from cellular membranes, a process regulated by ZDHHC5-mediated palmitoylation. Using biochemical analyses and immune-competent cell models, we show that AMP-activated protein kinase (AMPK) integrates metabolic cues into NOD1 signaling by regulating ZDHHC5. AMPK phosphorylates ZDHHC5 at Ser296 and Ser380, weakens its association with Golga7B, and promotes its displacement from the plasma membrane, thereby reducing NOD1 palmitoylation, membrane localization, and downstream signaling. Conversely, stimulation with the NOD1 agonist C12-iE-DAP suppresses AMPK activity, stabilizing ZDHHC5 at the membrane and sustaining NOD1 activation. These findings define a reciprocal regulatory circuit linking metabolic stress sensing and innate immune signaling, and illustrate how metabolic pathways govern the spatial control of pattern recognition receptors in innate immunity.