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. 2021 Sep 3;7(36):eabg4674. doi: 10.1126/sciadv.abg4674

Fig. 2. Probing pore-loop conformational dynamics using smFRET.

Fig. 2.

(A) Disaggregation activity of ClpB WT and PL1 mutants. Results show that all nonlabeled pore-loop mutants (NL) were active, but only the mutant S236C maintained activity after labeling (L). Error bars were calculated from two experiments. (B) FRET-efficiency histograms of the PL1 variant without substrate (light red) and in the presence of 20 μM κ-casein (dark red). (C) Donor-acceptor fluorescence cross-correlation functions of the PL1 variant; light red, without substrate; dark red, with substrate. (D) Relative change of the low FRET-efficiency population in histograms (fig. S3A) as a function of κ-casein concentration. (E) FRET-efficiency histograms of the PL2 variant without substrate (light brown) and in the presence of 20 μM κ-casein (dark brown). (F) FRET-efficiency histograms of the PL3 variant without substrate (light orange) and in the presence of 20 μM κ-casein (dark orange).