biostudies-literatureDu GNICHD NIH HHSNCCIH NIH HHSNIDDK NIH HHSNIAID NIH HHSNHLBI NIH HHSNCI NIH HHS437-446https://www.ebi.ac.uk/biostudies/studies/S-EPMC11283288biostudies-literatureUnknown630(8016)Gasdermin D (GSDMD) is the common effector for cytokine secretion and pyroptosis downstream of inflammasome activation and was previously shown to form large transmembrane pores after cleavage by inflammatory caspases to generate the GSDMD N-terminal domain (GSDMD-NT)^1-10. Here we report that GSDMD Cys191 is S-palmitoylated and that palmitoylation is required for pore formation. S-palmitoylation, which does not affect GSDMD cleavage, is augmented by mitochondria-generated reactive oxygen species (ROS). Cleavage-deficient GSDMD (D275A) is also palmitoylated after inflammasome stimulation or treatment with ROS activators and causes pyroptosis, although less efficiently than palmitoylated GSDMD-NT. Palmitoylated, but not unpalmitoylated, full-length GSDMD induces liposome leakage aNatureROS-dependent S-palmitoylation activates cleaved and intact gasdermin D.PMC11283288R01 AI124491R01 AI182369R37 AI116550R01 AI139914P01 HL158688R01 CA240955P30 DK034854DP1 HD087988R01 AT010268R01 AI116550R01 AI167993El-Baba TJDong YLutomski CAMiao RMa XDevant PRobinson CVBanerjee AWu HLuo HRGu BPuthenveetil ROh SFPi XBalasubramanian AFontana PDu GWalker CKagan JCHealy LBDavid LGoh BLieberman JfalseROS-dependent S-palmitoylation activates cleaved and intact gasdermin D.Gasdermin D (GSDMD) is the common effector for cytokine secretion and pyroptosis downstream of inflammasome activation and was previously shown to form large transmembrane pores after cleavage by inflammatory caspases to generate the GSDMD N-terminal domain (GSDMD-NT)^1-10. Here we report that GSDMD Cys191 is S-palmitoylated and that palmitoylation is required for pore formation. S-palmitoylation, which does not affect GSDMD cleavage, is augmented by mitochondria-generated reactive oxygen species (ROS). Cleavage-deficient GSDMD (D275A) is also palmitoylated after inflammasome stimulation or treatment with ROS activators and causes pyroptosis, although less efficiently than palmitoylated GSDMD-NT. Palmitoylated, but not unpalmitoylated, full-length GSDMD induces liposome leakage a2024-01-01T00:00:00Z2024 Jun2026-06-03T05:28:30.79Z2025-04-06T22:24:32.778ZS-EPMC112832883859923910.1038/s41586-024-07373-5