Fig. 2
From: A biosensor-based framework to measure latent proteostasis capacity
FRET reports on barnase foldedness and aggregation. a Bait protein barnase structure is shown (PDB ID 1A2P) with the location of destabilizing mutations used to tune K f (and thus ΔG F ). b Urea denaturation curves, assayed by FRET, are shown of barnase constructs expressed in mammalian lysate fitted to a two-state unfolding model (one representative replicate per mutant of n = 3). c A confocal micrograph image of representative cells expressing a destabilized barnase variant (I25A, I96G) in HEK293T cells. Scale bar=10 µm. The wild-type barnase variant does not form visible aggregates. The middle graph shows fluorescence spectra (excitation 405 nm) for cells with only diffuse biosensor vs. cells with visible aggregates. The right graph shows a proxy measure for FRET of these cells (means ± SEM)
