Measuring NLR Oligomerization III: Detection of NLRP3 and NLRC4 Complex by Bioluminescence Resonance Energy Transfer

Methods Mol Biol. 2023:2696:93-103. doi: 10.1007/978-1-0716-3350-2_6.

Abstract

Bioluminescent resonance energy transfer (BRET) is a natural phenomenon resulting from a non-radiative energy transfer between a bioluminescent donor (Renilla luciferase) and a fluorescent protein acceptor. BRET signal is dependent on the distance and the orientation between the donor and the acceptor and could be used to study protein-protein interactions and conformational changes within proteins at real-time in living cells. This protocol describes the use of BRET technique to study NLRP3 oligomerization in living cells before and during NLRP3 inflammasome activation.

Keywords: BRET; Bioluminescence; NLRC4; NLRP3; Sensor.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bioluminescence Resonance Energy Transfer Techniques / methods
  • Energy Transfer
  • Luciferases, Renilla / genetics
  • Luminescent Measurements / methods
  • NLR Family, Pyrin Domain-Containing 3 Protein*
  • Proteins*

Substances

  • NLR Family, Pyrin Domain-Containing 3 Protein
  • Proteins
  • Luciferases, Renilla