Fig. 4: KJE reverses misfolding and commits bound protein to fast folding.
a–d spFRET efficiency histograms measured during KJE-ATP assisted folding of FLucNC (a), FLucN (b), FLucNS (c) or FLucC (d). GuHCl-denatured FLuc was diluted to 50 pM in buffer containing 0.3 µM DnaK, 0.1 µM DnaJ, 5 mM ATP and 50 µM PBT, and the fE distribution of DnaK-bound FLuc was recorded, followed by initiation of folding with 0.5 µM GrpE. spFRET was recorded for 15 min either immediately (0 min) or after 60 min of folding. Representative measurements of three independent repeats are shown. e KJ-ATP converts compact, misfolded (MF) intermediates of FLuc to the DnaK-bound, expanded state. GuHCl-denatured FLucNC was diluted to 50 pM in buffer containing ATP and PBT, and the distance between N- and C-domains probed by spFRET as in (a). KJ-ATP were added after 1 h to generate DnaK-bound FLuc, followed by GrpE to initiate folding. Representative measurements of three independent repeats are shown. Spont., spontaneous folding. Source data are provided as a Source Data file.
