Abstract
Gasdermins (GSDMs) are pore-forming proteins that mediate pyroptosis and contribute to inflammatory and cancer-related processes. Although GSDME shares structural similarity with other gasdermins, its activation and pore assembly mechanisms remain incompletely defined. Here we determine high-resolution cryo-electron microscopy structures of 27- and 28-fold human GSDME pores at 3.64 Å and 3.58 Å resolution. The structures reveal conserved structural architecture together with distinct features, including an extended transmembrane β-barrel and a comparatively compact membrane-engagement geometry. Structure-guided mutagenesis identifies lipid-binding and oligomerization interfaces required for pore formation. We further demonstrate that caspase-3 activates GSDME through direct recognition of a DMPD tetrapeptide motif within the interdomain linker, independently of the GSDME C-terminal domain. Following proteolytic activation, S-palmitoylation of GSDME N-terminal domain enhances pore-forming efficiency, with Cys180 serving as the primary functional site. Together, these findings establish a coordinated structural and regulatory framework in which proteolytic licensing and lipid modification sequentially control GSDME pore formation.
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Data availability
The atomic coordinates of the 27- and 28-fold GSDME pores have been deposited in the Protein Data Bank (PDB) under accession numbers 9PE0 and 9PDU, respectively. The associated cryo-EM density maps have been deposited in the Electron Microscopy Data Bank (EMDB) under accession numbers EMD-71549 and EMD-71544, respectively. All other data are available from the corresponding author upon request. Several structural coordinates in the PDB database were used in this study, which can be located by accession numbers 6CB8, 5B5R, 6N9O, 6N9N, 6VFE, 7V8H, and 3CVR. The source data are provided as a Source Data file. Source data are provided with this paper.
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Acknowledgements
This work was supported by UConn Health Start-up fund and US National Institutes of Health grants R01AI158435 to J.R.
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J.R. conceived the study. E.T. expressed and purified the proteins. E.T. and J.R. reconstituted the GSDME pores. C.W. froze the cryo-EM grids. J.W. and D.N. screened and collected the cryo-EM datasets. J.R. and D.N. processed the cryo-EM datasets. J.R. built the atomic model. J.R., T.T., and E.T. analyzed the structures and performed biochemical experiments. T.T., C.W., E.T., and X.W. performed cellular experiments. J.R. and V.R. supervised cellular experiments and analyzed the data. All authors organized and analyzed data. J.R. wrote the paper with input from all authors.
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Teran, E., Tian, T., Wang, C. et al. Structural basis and regulation of GSDME pore formation. Nat Commun (2026). https://doi.org/10.1038/s41467-026-70643-5
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DOI: https://doi.org/10.1038/s41467-026-70643-5
