Inflammatory Caspases: Activation and Cleavage of Gasdermin-D In Vitro and During Pyroptosis

  • Yue Zhao
  • Jianjin Shi
  • Feng ShaoEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1714)


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.


Gasdermin-D GSDMD Pyroptosis Inflammatory caspases LPS Inflammasome Recombinant proteins 



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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© Springer Science+Business Media LLC 2018

Authors and Affiliations

  1. 1.National Institute of Biological SciencesBeijingChina

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