Abstract
The gasdermin family represents a type of membrane pore-forming proteins. The gasdermin family is extensively characterized as the executioner of pyroptotic cell death in mammals; recent studies suggest that gasdermin-like pore-forming proteins are also present in bacteria and fungi. In humans, gasdermin D (GSDMD) is activated through inter-domain cleavage by caspase-1 in the canonical inflammasome pathway and cytosolic LPS-activated caspase-4 or caspase-5. The cleavage disrupts the autoinhibition of GSDMD and liberates the N-terminal gasdermin-N domain that binds to membrane lipids and forms pores of an inner diameter of ~18 nm on the membrane, responsible for cell pyroptosis. Here, we describe the methods of determining the phospholipid-binding and pore-forming activity of gasdermins in a robust in vitro system. We also introduce a method of specifically detecting the caspase-cleaved form of GSDMD in pyroptotic cells.
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Acknowledgments
The work was supported by grants from the Ministry of Science and Technology of China, China National Natural Science Foundation of China, Chinese Academy of Sciences, and Chinese Academy of Medical Sciences to F.S.
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Wang, K., Ding, J., Shao, F. (2023). Determination of Gasdermin Pores. In: Pelegrín, P., Di Virgilio, F. (eds) NLR Proteins. Methods in Molecular Biology, vol 2696. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3350-2_11
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DOI: https://doi.org/10.1007/978-1-0716-3350-2_11
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