Abstract
Gasdermin D is an essential mediator of pyroptosis. Under resting conditions gasdermin D is inactive in the cytosol. Following inflammasome activation, gasdermin D undergoes processing and oligomerization to create membrane pores and induce pyroptosis to release mature IL-1 and IL-18. Biochemical methods for the analysis of gasdermin D activation states are important for assessing gasdermin D function. Here we describe the biochemical methods for assessing gasdermin D processing and oligomerization and its inactivation using small molecule inhibitors.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Kayagaki N, Stowe IB, Lee BL, O’Rourke K, Anderson K, Warming S, Cuellar T, Haley B, Roose-Girma M, Phung QT, Liu PS, Lill JR, Li H, Wu J, Kummerfeld S, Zhang J, Lee WP, Snipas SJ, Salvesen GS, Morris LX, Fitzgerald L, Zhang Y, Bertram EM, Goodnow CC, Dixit VM (2015) Caspase-11 cleaves gasdermin D for non-canonical inflammasome signalling. Nature 526:666. https://doi.org/10.1038/nature15541. https://www.nature.com/articles/nature15541#supplementary-information
Liu X, Zhang Z, Ruan J, Pan Y, Magupalli VG, Wu H, Lieberman J (2016) Inflammasome-activated gasdermin D causes pyroptosis by forming membrane pores. Nature 535:153. https://doi.org/10.1038/nature18629. https://www.nature.com/articles/nature18629#supplementary-information
Shi J, Zhao Y, Wang K, Shi X, Wang Y, Huang H, Zhuang Y, Cai T, Wang F, Shao F (2015) Cleavage of GSDMD by inflammatory caspases determines pyroptotic cell death. Nature 526(7575):660–665. https://doi.org/10.1038/nature15514
Hu JJ, Liu X, Xia S, Zhang Z, Zhang Y, Zhao J, Ruan J, Luo X, Lou X, Bai Y, Wang J, Hollingsworth LR, Magupalli VG, Zhao L, Luo HR, Kim J, Lieberman J, Wu H (2020) FDA-approved disulfiram inhibits pyroptosis by blocking gasdermin D pore formation. Nat Immunol 21(7):736–745. https://doi.org/10.1038/s41590-020-0669-6
Humphries F, Shmuel-Galia L, Ketelut-Carneiro N, Li S, Wang B, Nemmara VV, Wilson R, Jiang Z, Khalighinejad F, Muneeruddin K, Shaffer SA, Dutta R, Ionete C, Pesiridis S, Yang S, Thompson PR, Fitzgerald KA (2020) Succination inactivates gasdermin D and blocks pyroptosis. Science:eabb9818. https://doi.org/10.1126/science.abb9818
Humphries F, Shmuel-Galia L, Ketelut-Carneiro N, Li S, Wang B, Nemmara VV, Wilson R, Jiang Z, Khalighinejad F, Muneeruddin K, Shaffer SA, Dutta R, Ionete C, Pesiridis S, Yang S, Thompson PR, Fitzgerald KA (2020) Succination inactivates gasdermin D and blocks pyroptosis. Science 369(6511):1633–1637. https://doi.org/10.1126/science.abb9818
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Humphries, F. (2023). Biochemical Methods for Assessing Gasdermin D Inactivation in Macrophages. In: Fink, S.L. (eds) Pyroptosis. Methods in Molecular Biology, vol 2641. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3040-2_10
Download citation
DOI: https://doi.org/10.1007/978-1-0716-3040-2_10
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-3039-6
Online ISBN: 978-1-0716-3040-2
eBook Packages: Springer Protocols