Type-3 BRET, an improved competition-based bioluminescence resonance energy transfer assay.

ABSTRACT: We show that in conventional, competition-based bioluminescence resonance energy transfer (BRET) assays of membrane protein stoichiometry, the presence of competitors can alter tagged-protein density and artifactually reduce energy transfer efficiency. A well-characterized monomeric type I membrane protein, CD86, and two G protein-coupled receptors ?2AR and mCannR2, all of which behave as dimers in these conventional assays, exhibit monomeric behavior in an improved competition-based type-3 BRET assay designed to circumvent such artifacts.

SUBMITTER: Felce JH 

PROVIDER: S-EPMC4070276 | biostudies-other | 2014 Jun

REPOSITORIES: biostudies-other