Fig. 2: AP Profiling resolves folding at codon resolution. | Nature Communications

Fig. 2: AP Profiling resolves folding at codon resolution.

From: Arrest Peptide Profiling resolves co-translational folding pathways and chaperone interactions in vivo

Fig. 2

a A truncation library of the G-domain from EF-G (GEF-G) is inserted into the AP Profiling construct. After library expression, cells are sorted into gates by their log10(GFP/mCherry) ratio. Deep sequencing of individual gates yields the distribution of each clone in the library. b AP scores (top panel) for wild-type GEF-G (red) and a destabilized variant (F/A mutant, gray). AP scores are mostly similar, except for the region around ncodons = 330, where signals for the wild-type sequence are higher than for the mutant, indicating folding into a stable tertiary structure. The folding score (bottom panel), calculated as the difference between wild-type and F/A mutant scores, reveals the position of tertiary folding around ncodons = 330. c Single-molecule optical tweezers experiments with ribosome-nascent chain complexes show the unfolding signature of wild-type GEF-G, whereas the F/A mutant lacks a clear unfolding transition, indicating that it is not stably structured. Source data are provided as a Source Data file.

Back to article page