Fig. 4: Folding landscapes of translocated protein substrates.
From: Transmembrane protein rotaxanes reveal kinetic traps in the refolding of translocated substrates
a Bivariate histograms representing the dwell times in levels 2′ and 3′ for oligo(dC)30-V5-oligo(dA)30 threaded C terminus-first. Data were acquired with three different pores. Each panel corresponds to the indicated refolding time (n: number of events). When level 2′ or 3′ was not observed, a dwell time of 0.2 ms, the temporal detection limit, was assigned. The “Native” panel records values obtained from Trx V5 that had never been unfolded. The “All” panel displays the data from all the other panels in a single plot to provide clearer visualization of three populations. b Kinetic model giving the lowest AIC and BIC values with the kinetic constants derived from the fit. c Time-course evolution of population A (red circles), population B (yellow circles), native Trx V5 (purple circles) and unfolded Trx V5 (blue circles). The solid lines represent the best fits to the model with the lowest AIC and BIC values. d Three destabilizing mutations were introduced into Trx V5 to form Trx V2. A rotaxane was formed, and the refolding was examined as in ‘a’. When level 2′ or 3′ was not observed, a dwell time of 0.2 ms was assigned. e Kinetic model giving the lowest AIC and BIC values with the kinetic constants derived from the fit. f Time-course evolution of population A (red circles), native Trx V2 (purple circles) and unfolded Trx V2 (blue circles). The solid lines represent the best fits.
