Method to Measure Ubiquitination of NLRs

Methods Mol Biol. 2023:2696:105-114. doi: 10.1007/978-1-0716-3350-2_7.

Abstract

Posttranslational modifications are crucial in determining the functions of proteins in the cell. Modification of the NLRP3 inflammasome by the ubiquitin system has recently emerged as a new level of regulation of the inflammasome complex. Here we describe a method to detect poly-ubiquitination of NRLP3 using two different approaches: (i) detection with a ubiquitin antibody or (ii) using TUBEs (Tandem Ubiquitin Binding entities). This approach can be used to detect ubiquitination of other NLRs or other proteins.

Keywords: Immunoprecipitation; Inflammasome; NLRP3; TUBE; Ubiquitination.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Inflammasomes* / metabolism
  • NLR Family, Pyrin Domain-Containing 3 Protein* / metabolism
  • Protein Processing, Post-Translational
  • Ubiquitin / metabolism
  • Ubiquitination

Substances

  • Inflammasomes
  • NLR Family, Pyrin Domain-Containing 3 Protein
  • Ubiquitin