BPS2026 – Exploring GSDMD-metal ion interactions

Gasdermin D (GSDMD) is a key regulator of pyroptosis, a programmed host defense mechanism that, when unregulated, leads to serious inflammatory pathologies. Upon infection, GSDMD is cleaved by caspases 1, 4, and 5 in humans to form pores in the cell membrane. This cleavage converts GSDMD from a full-length, cytoplasmic, auto-inhibited, monomeric state to an oligomerization-prone, membrane-soluble cytotoxic form. Such changes in structure, solubility properties, and oligomerization make finding GSDMD inhibitors particularly challenging. Here, we describe the screening of metal ion coordination complexes for GSDMD inhibition. We show concentration-dependent inhibition of GSDMD in biochemical pore formation assays and the use of site-directed mutagenesis to identify the metal ion binding sites.