Abstract
Gasdermin-D (also known as GSDMD), the newly identified executioner of pyroptotic cell death, is cleaved by activated caspase-1 downstream of canonical inflammasome activation or caspase-4, 5, and 11 upon their ligation and activation by cytosolic LPS. Upon a single cleavage between the two domains in Gasdermin-D, the N-terminal domain binds to membrane lipids and lyses cells by forming pores of an inner diameter of 10–14 nm within the membrane. The inter-domain cleavage of Gasdermin-D is a reliable marker for the activation of inflammatory caspases and cell pyroptosis. Here, we describe the methods for examining Gasdermin-D cleavage by activated inflammatory caspases in vitro and upon inflammasome activation in vivo.
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Acknowledgments
The work was supported by the grants from the China Ministry of Science and Technology and China National Science Foundation, the Chinese Academy of Sciences, Howard Hughes Medical Institute, and Beijing municipal government to F. S.
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Zhao, Y., Shi, J., Shao, F. (2018). Inflammatory Caspases: Activation and Cleavage of Gasdermin-D In Vitro and During Pyroptosis. In: De Nardo, D., De Nardo, C. (eds) Innate Immune Activation. Methods in Molecular Biology, vol 1714. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7519-8_9
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DOI: https://doi.org/10.1007/978-1-4939-7519-8_9
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