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.
Similar content being viewed by others
References
Bergsbaken T, Fink SLand Cookson BT (2009) Pyroptosis: host cell death and inflammation. Nat Rev Microbiol 7:99–109
Miao EA, Leaf IA, Treuting PM et al (2010) Caspase-1-induced pyroptosis is an innate immune effector mechanism against intracellular bacteria. Nat Immunol 11:1136–1142
Kepp O, Galluzzi L, Zitvogel L et al (2010) Pyroptosis - a cell death modality of its kind? Eur J Immunol 40:627–630
Shi J, Zhao Y, Wang K et al (2015) Cleavage of GSDMD by inflammatory caspases determines pyroptotic cell death. Nature 526:660–665
Kayagaki N, Stowe IB, Lee BL et al (2015) Caspase-11 cleaves gasdermin D for non-canonical inflammasome signalling. Nature 526:666–671
Ding J, Wang K, Liu W et al (2016) Pore-forming activity and structural autoinhibition of the gasdermin family. Nature 535:111–116
Aglietti RA, Estevez A, Gupta A et al (2016) GsdmD p30 elicited by caspase-11 during pyroptosis forms pores in membranes. Proc Natl Acad Sci U S A 113:7858–7863
Liu X, Zhang Z, Ruan J et al (2016) Inflammasome-activated gasdermin D causes pyroptosis by forming membrane pores. Nature 535:153–158
Sborgi L, Ruhl S, Mulvihill E et al (2016) GSDMD membrane pore formation constitutes the mechanism of pyroptotic cell death. EMBO J 35:1766–1778
Shi J, Zhao Y, Wang Y et al (2014) Inflammatory caspases are innate immune receptors for intracellular LPS. Nature 514:187–192
Garcia-Calvo M, PetersonEP RDM et al (1999) Purification and catalytic properties of human caspase family members. Cell Death Differ 6:362–369
Zhao Y, Yang J, Shi J et al (2011) The NLRC4 inflammasome receptors for bacterial flagellin and type III secretion apparatus. Nature 477:596–600
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.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2018 Springer Science+Business Media LLC
About this protocol
Cite this protocol
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
Download citation
DOI: https://doi.org/10.1007/978-1-4939-7519-8_9
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7518-1
Online ISBN: 978-1-4939-7519-8
eBook Packages: Springer Protocols