Quantifying Caspase-1 Activity in Murine Macrophages

Methods Mol Biol. 2018;1725:163-176. doi: 10.1007/978-1-4939-7568-6_14.

Abstract

The caspase-1 protease is a core component of multiprotein inflammasome complexes, which play a critical role in regulating the secretion of mature, bioactive pro-inflammatory cytokines interleukin (IL)-1β and IL-18. The activity of caspase-1 is often measured indirectly, by monitoring cleavage of cellular caspase-1 substrates, processing of caspase-1 itself, or by quantifying cell death. Here we describe methods for eliciting caspase-1 activity in murine macrophages, via activation of the NLRP3, NAIP/NLRC4 or AIM2 inflammasomes. We then describe a simple fluorogenic assay for directly quantifying cellular caspase-1 activity.

Keywords: Biochemistry; Inflammasome; Inflammatory caspases; Protease.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis Regulatory Proteins / metabolism*
  • Bone Marrow / enzymology*
  • Bone Marrow / immunology
  • Bone Marrow / pathology
  • Caspase 1 / metabolism*
  • Cytokines / metabolism
  • Inflammasomes / immunology
  • Inflammasomes / metabolism*
  • Macrophages / enzymology*
  • Macrophages / immunology
  • Macrophages / pathology
  • Mice

Substances

  • Apoptosis Regulatory Proteins
  • Cytokines
  • Inflammasomes
  • Caspase 1