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

Yue Zhao1, Jianjin Shi1, Feng Shao2

  • 1National Institute of Biological Sciences, #7 Science Park Rd, Beijing, 102206, China.

Methods in Molecular Biology (Clifton, N.J.)
|November 28, 2017
PubMed

Insights

Gasdermin-D (GSDMD) cleavage by inflammatory caspases triggers pyroptosis. This study details methods to examine GSDMD cleavage in vitro and in vivo, aiding pyroptosis research.

Area of Science:

  • Immunology
  • Cell Biology
  • Biochemistry

Background:

  • Gasdermin-D (GSDMD) is the key executioner of pyroptotic cell death.
  • Inflammasome activation leads to caspase-1 or caspase-4/5/11 cleavage of GSDMD.
  • GSDMD cleavage initiates pore formation and cell lysis.

Purpose of the Study:

  • To present methods for analyzing Gasdermin-D cleavage.
  • To investigate GSDMD cleavage by inflammatory caspases in vitro.
  • To examine GSDMD cleavage during in vivo inflammasome activation.

Main Methods:

  • In vitro assays to detect GSDMD cleavage by activated caspases.
  • In vivo studies involving inflammasome activation.
  • Biochemical and cellular techniques to analyze GSDMD processing.

Main Results:

  • Established protocols for detecting GSDMD inter-domain cleavage.
  • Demonstrated GSDMD cleavage by specific inflammatory caspases.
  • Validated GSDMD cleavage as a marker for pyroptosis in biological systems.

Conclusions:

  • GSDMD cleavage is a critical and measurable event in pyroptosis.
  • The described methods facilitate the study of inflammatory caspase activation and pyroptosis.
  • This work provides tools for understanding GSDMD's role in immunity and disease.
Keywords:
GSDMDGasdermin-DInflammasomeInflammatory caspasesLPSPyroptosisRecombinant proteins

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