ROS-dependent S-palmitoylation activates cleaved and intact gasdermin D PMID: 38599239

RQC and Longevity Regulation

2026-09-03

ROS-dependent S-palmitoylation activates cleaved and intact gasdermin D

Nature. 2024 Jun;630(8016):437-446. doi: 10.1038/s41586-024-07373-5.

Gang Du ^# ¹ ² , Liam B Healy ^# ³ ⁴ , Liron David ⁵ ⁶ ⁷ , Caitlin Walker ³ ⁴ ⁸ , Tarick J El-Baba ⁹ ¹⁰ , Corinne A Lutomski ⁹ ¹⁰ , Byoungsook Goh ¹¹ , Bowen Gu ⁴ ¹² , Xiong Pi ³ ⁴ , Pascal Devant ¹³ , Pietro Fontana ³ ⁴ , Ying Dong ³ ⁴ , Xiyu Ma ⁴ ¹² , Rui Miao ⁴ ¹² , Arumugam Balasubramanian ¹⁴ ¹⁵ , Robbins Puthenveetil ¹⁶ , Anirban Banerjee ¹⁶ , Hongbo R Luo ¹⁴ ¹⁵ , Jonathan C Kagan ¹³ , Sungwhan F Oh ¹¹ , Carol V Robinson ⁹ ¹⁰ , Judy Lieberman ⁴ ¹² , Hao Wu ¹⁷ ¹⁸

Affiliations

1. Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA, USA. [email protected].
2. Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, MA, USA. [email protected].
3. Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA, USA.
4. Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, MA, USA.
5. Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA, USA. [email protected].
6. Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, MA, USA. [email protected].
7. Seqirus, Cambridge, MA, USA. [email protected].
8. Department of Life Sciences, Ben-Gurion University of the Negev, Be'er Sheva, Israel.
9. Department of Chemistry, University of Oxford, Oxford, UK.
10. Kavli Institute for NanoScience Discovery, University of Oxford, Oxford, UK.
11. Center for Experimental Therapeutics and Reperfusion Injury, Department of Anesthesiology, Perioperative and Pain Medicine, Brigham and Women's Hospital, Boston, MA, USA.
12. Department of Pediatrics, Harvard Medical School, Boston, MA, USA.
13. Harvard Medical School and Division of Gastroenterology, Boston Children's Hospital, Boston, MA, USA.
14. Department of Pathology, Dana-Farber/Harvard Cancer Center, Harvard Medical School, Boston, MA, USA.
15. Department of Laboratory Medicine, Boston Children's Hospital, Boston, MA, USA.
16. Section on Structural and Chemical Biology, Neurosciences and Cellular and Structural Biology Division, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD, USA.
17. Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA, USA. [email protected].
18. Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, MA, USA. [email protected].
# Contributed equally.

Abstract

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 and forms a pore similar in structure to GSDMD-NT pores shown by cryogenic electron microscopy. ZDHHC5 and ZDHHC9 are the major palmitoyltransferases that mediate GSDMD palmitoylation, and their expression is upregulated by inflammasome activation and ROS. The other human gasdermins are also palmitoylated at their N termini. These data challenge the concept that cleavage is the only trigger for GSDMD activation. They suggest that reversible palmitoylation is a checkpoint for pore formation by both GSDMD-NT and intact GSDMD that functions as a general switch for the activation of this pore-forming family.